Program and Abstract Book

Functional and healthy food ingredients generated through state-of-the-art biotechnology

Program and Abstract Book

https://food4health.uni-hohenheim.de

2 3

Distinguished scientists and guests, ladies and gentlemen,

We warmly welcome each and every one of you to the Chinese-German Symposium on “Functional and healthy food ingredients generated through state-of-the-art biotechnology”. Food science is one of the key disciplines in developments regarding feeding the world. In the light of growing populations, longer life expectancies, as well as consumer demands for safe and healthy food the development of novel and natural functional foods has become the fastest growing sector in food industry. Current topics in this field are for instance the nutritional needs of an aging population or the requirements of adapted diets (e.g. because of adverse, allergic food effects or other health im- pairments). Customized food ingredients and formu- lations can be instrumental to ensure balanced and healthy nutrition. Thus, one of the key research aspects is the production of functional food ingre-dients through state-of-the-art biotechnological processes that opens new doors to healthy food rather than remaining in traditional food processing technologies only.Starting in 2014, Jiangnan University in Wuxi and University of Hohenheim in Stuttgart have built up a long-term and in-depth cooperation platform in the field of food science and biotechnology. Since then, a series of joint activities has been implemen-ted, e.g. visits and exchanges of researchers and students through all academic levels, studying ab-road at the partner university, joint research, and an academic conference on food processing in Wuxi in September 2015.

Based on these activities all partners involved have developed mutual trust and a shared scientific vision. To take the cooperation to the next level, collaboration will be intensified and at the same time extended to other Chinese and German experts in the field of functional food processing. Concerted effort is not only key but of utmost importance to boost higher education and research in that field.This symposium is a first step towards this goal and we would like to thank everyone contributing to the inspiring scientific program. Outstanding resear-chers from both countries will present their work and knowledge, discuss cutting-edge research questions as well as future challenges for the bio-technological production of functional and healthy food ingredients. The symposium will cover basic questions and novel developments in biocatalysis, fermentation, downstream processing and analysis in the area of food science, as well as new enginee-ring and processing approaches. The exchange of knowledge about all these disciplines and corre-sponding technologies shall lead to a Chinese-German research agenda that focuses on new stra-tegies for tailored functional food ingredients, as well as their quality, safety, and authenticity con-trol. The symposium shall serve as an incubator to foster bi-national research partnerships. The parti-cipation of young scientists will pave the way for the next generation of experts, shall nurture fruitful discussions and the development of new ideas. We are looking forward to an open and constructive exchange that will strengthen and extend research cooperation between the participants of this sym-posium in particular, and between China and Germany strategically.

This Symposium has been kindly supported by the

Sino-German Center for Research Promotion

Welcome to the CDZ SymposiumFunctional and healthy food ingredients generated through state-of-the-art biotechnology12 - 14 September 2018, University of Hohenheim, Stuttgart

Prof. Dr. Bo JiangJiangnan University

Prof. Dr. Lutz FischerUniversity of Hohenheim

4 5

Scientific Board MembersProf. Dr. rer. nat. Lutz FischerUniversity of HohenheimVice-Dean of the Faculty of Natural Sciences, Institute of Food Science and Biotechnology, Chair of the Department of Biotechnology and Enzyme Science

Prof. Dr. Bo JiangJiangnan UniversityDirector, International Office.Professor, State Key Laboratory of Food Science and Technology

1800 Lihu Avenue, Wuxi, Jiangsu 214122, ChinaPhone: +86-510-85328307 | Fax: +86-510-85913622 [email protected]

Garbenstr. 25, D-70593 StuttgartPhone: +49 (0)711 459 23018 | Fax.: +49 (0)711 459 [email protected]

Prof. Dr. Wanmeng MuJiangnan UniversityDeputy Director, State Key Laboratory of Food Science and Technology

Junior-Prof. Dr. Yanyan ZhangUniversity of HohenheimInstitute of Food Science and Biotechnology, Chair of the Department Flavor Chemistry

1800 Lihu Avenue, Wuxi, Jiangsu 214122, ChinaPhone/Fax: [email protected]

Fruwirthstr. 12, Verfügungsgebäude 221, 70599 StuttgartPhone: +49 711 45924871 | Fax:+49 711 [email protected]

Founded in 1818 after devastating famines, the Uni versity of Hohenheim is not only engaged in intensive basic research but has traditionally also been committed to developing innovative solutions for some of society‘s pressing problems. To do so, the University of Hohenheim engages in a combi-nation of scientific disciplines that is unique among German universities.

Today, the University of Hohenheim is the leading University in agricultural research and food scien-ces, as well as strong and unparalleled in natural, social, business, economic, and communication sciences. The combination makes it possible to find solutions for many global challenges.

For more information, visit www.uni-hohenheim.de

The Hohenheim Research Center for Health Sciences provides a dynamic platform for researchers, lectur-ers, young scientists and students dedicated to life science and societal health topics and promotes high-level research across several disciplines in accordance with the modified „One Health“ concept by• joining expertise, e.g. in biology, immunology,

health care and medicine, agriculture and food sciences, economics and social sciences

• building bridges between bench scientists, clini-cal investigators, health researchers, business and public stakeholders

• strengthening national and international re-search networks for exchange and productive partnerships

• obtaining funds for integrated research projects focusing on major scientific and societal topics, including e.g. growth, development, demogra-phic change, lifestyle, nutrition, aging as well as their social and economic impact.

For more information about the Research Center, visit www.health.uni-hohenheim.de

University of Hohenheim

Research Center for Health Sciences

The availability of high-quality and safe foodstuffs is a tremendous challenge for societies in the 21th century. Consequently, research and teaching in the discipline of food science focus on the optimal use and processing of raw materials from agricultural production into foodstuffs with high standards of quality and safety on the basis of modern technological developments. The natural scientific and engineering aspects of the trans-formation of plant- and animal-based raw materials into consumable, high-quality foodstuffs, dietary supplements, as well as functional active and value-added compounds are taught at the Institute of Food Science and Biotechnology. The value-added food chain is considered as a whole and the complex interdependencies between ingredients,

processing and functionality of foodstuffs is inten- sely studied across all departments. Ten professors and about 100 researchers with backgrounds in the engineering and natural scien-tif ic aspects of food science offer a vast scope and extraordinary quality in research and teaching. Modern laboratories equipped with the latest analytics and technical centers with pilot plants connect research with practical application. The annual Lebensmittelwissenschaftliche und Bio-technologische Kolloquium organised by the Insti-tute as well as national and international conferen-ces organised by individual departments provide an excellent stage for exchange between teaching and practical applications.

Institute of Food Science and Biotechnology

6 7

Hohenheim Palace, 70599 Stuttgart, Germany How to reach the Venue ‘Palace Hohenheim‘

Public transportation

From Stuttgart Airport

From the arrival terminal’s building, turn to the right to reach the bus station in approximately 3 minutes.

Please follow the sign with the bus. Take bus 122 (direction „Esslingen (N) ZOB“) to the stop „Plieningen

Post“. There, on the other side of the street change to one of the busses 70 or 73 until „Plieningen Garbe“.

Travel time: 25 minutes. Walk from there towards Hohenheim Palace (red dotted line, appr. 15 - 20 min)

Alternative route:

Take the S2 or S3 in Terminal 1, on Level 1 and get off at Vaihingen. There, catch the tram U3 to Plienin-

gen. Travel time: 30 to 45 minutes.

From Stuttgart Railway Station

From the main station at Stuttgart take the tram U7 (Ostfi ldern) to the station „Ruhbank“. Then change to

bus 70 (Plieningen) until the stop „Plieningen Garbe“. Travel time: 30 minutes.

Alternative route:

Take the tram U5 (Leinfelden Bf), U6 (Fasanenhof) or U12 (Dürrlewang) to „Möhringen Bahnhof“, and

change to tram U3 to the fi nal stop „Plieningen Universität Hohenheim“.

Travel time: 32 minutes. Walk from there towards Hohenheim Palace (red dotted line, appr. 15 - 20 min).

Useful numbers and links

Taxi: +49 711 9484409 | +49 171 5411841

Öff entlicher Nahverkehr | Stuttgarter Verkehrsverbundhttp://www.vvs.de/home-vvs/

Please follow the sign with the bus. Take bus 122 (direction „Esslingen (N) ZOB“) to the stop „Plieningen

Post“. There, on the other side of the street change to one of the busses 70 or 73 until „Plieningen Garbe“.

Travel time: 32 minutes. Walk from there towards Hohenheim Palace (red dotted line, appr. 15 - 20 min).

8 9

Table of Contents

Thank you to our Supporters .....................................................................................................................................2

Welcome Address ........................................................................................................................................................3

Scientific Board Members ..........................................................................................................................................4

Hosting Institution: University of Hohenheim / Hohenheim Research Center for Health Sciences 5

Venue .............................................................................................................................................................................6

How to reach the Venue SHMT ...................................................................................................................................7

Table of Contents .........................................................................................................................................................9

Program .......................................................................................................................................................................10

Abstracts – Oral Presentations ................................................................................................................................14

O1 – O35 Abstracts ....................................................................................................................................................16

Abstracts – Posters ...................................................................................................................................................52

P1 – P14 Abstracts .....................................................................................................................................................54

List of Speakers .........................................................................................................................................................68

Authors & Co-Authors Index ....................................................................................................................................70

Notes ...........................................................................................................................................................................72

10 11

ProgramWEDNESDAY, 12 SEPTEMBER 2018 (DAY 1)

REGISTRATION AND WELCOME

08:30 Registration09:00 Welcome Address

Rolf SCHMID, Steinbeis Asia Technology Consulting09:10 Welcome and Introduction to the Symposium

Lutz FISCHER, University of Hohenheim and Bo JIANG, Jiangnan University

SESSION 1: Chairs: Mingyong XIE and Herbert SCHMIDT09:30 Key Technology of Probiotic Fermented Fruit and Vegetable as well as Its Application in

Industry Mingyong XIE

09:50 Production of Vitamin B12 During Tofu Fermentation Bernward BISPING

10:10 Research and Industrialization Progress on Aqueous Extraction of Edible Oil in ChinaRuijin YANG

10:30 Coffee Break / Poster Exhibition

11:00 Exopolysaccharide Production by Food-Grade Lactic and Acetic Acid BacteriaFrank JAKOB

11:20 The Microbial Diversity and Community Dynamics of Chinese Traditional Fermented VegetablesTao XIONG

11:40 Simultaneous Production of Microbial Lipids and Gluconic Acid with Newly Isolated Oleaginous Yeast StrainsKatrin OCHSENREITHER

12:00 Lunch Break

SESSION 2: Chairs: Bo JIANG and Jörg HINRICHS

13:00 Modification of Functional Properties of Egg White Protein by Dietery Glycerol Monolaurate Supplementation Fengqin FENG

13:20 Production of Human Milk Oligosaccharides by E. coliGeorg SPRENGER

13:40 Recent Approaches for Improving Monascus Pigment Production Li NI

14:00 Enhanced Biocatalytic Conversion Strategies of Renewables to Tailor Made Glycolipid Biosurfactants in Uncommon Reaction MediaJens RUDAT

14:20 Stability of High Protein Food Matrix Peng ZHOU


15:10 Process Engineering of Enzymatic Processes in the Hydrolysis and Fractionation of Functional PeptidesUlrich KULOZIK

15:30 Strategy for Biosynthesis of Low-Calorie SugarsBo JIANG

15:50 Production of Prebiotic Sugars Through Enzymatic Lactose ConversionLutz FISCHER

16:10 Anti-Diabetic Mechanism of Dietary Polysaccharides Based on Their Gastrointestinal FunctionsShaoping NIE

16:30 Continuous Production of Low-Calorie Sweeteners Using a New Recombinant Fructosyl- transferase

Peter CZERMAK

17:00 Welcome Reception and Scientific Evaluation of Hohenheimer Spirits at the Department of Yeast Genetics and Fermentation Technology | Host: Ralf KOELLING-PATERNOGA

18:00 Welcome Dinner at the Restaurant Garbe, Plieningen

THURSDAY, 13 SEPTEMBER 2018 (DAY 2)

SESSION 3: Chairs: Wanmeng MU and Lutz FISCHER

09:00

09:20

09:40

10:00

Recent Progress on Ketose 3-Epimerase and Its Application in Enzymatic Production of D-AlluloseWanmeng MU Continuous Enzymatic Production of Technofunctional Casein HydrolysatesTimo STRESSLERThe Strategies to Improve the Expression of Recombinant Protein in Bacillus subtilisXin YANModern Biooxidation by Heme-Thiolate PeroxidasesJan KIEBIST

12 13


10:40 Characterization of Gelatin/Zein Nanofibers Fabricated by ElectrospinningHui ZHANG

11:00 Enzyme-Based Strategies for Structuring Foods with Improved Functionality Jochen WEISS

11:20 Degradation of Leaf Cell Wall for Native ComponentsChen ZHANG

11:40 Flavor and Enzymatic Impacts in the Processing of Yeast Fermented BeveragesKai BÜCHNER

12:00 Lunch Break

SESSION 4: Chairs: Li NI and Jochen WEISS

13:00

13:20

13:40

14:00

14:20

14:40

Senseoproteomics – Effect-Directed Analysis of Taste-Active Peptides in FoodKarin SEBALDArabinoxylan Alleviates Acute Colitis by Altering Colon Symptoms and Host Immune Response Jielun HUVirulence Mechanisms of Enterohemorrhagic Escherichia coli – The Role of Phage-Encoded O-Acetyl EsterasesHerbert SCHMIDT The Reciprocal Interactions between Tea Phenolics and Human Gut MicrobiotaZhibin LIUBacteriophages in Food Products and Their Influence on the Human Intestinal Microbiota Joerg HINRICHS

Coffee Break / Poster Exhibition

SESSION 5: Chairs: Fengqin FENG and Yanyan ZHANG

15:10 Effect-Directed Analysis of Functional Food IngredientsGertrud MORLOCK

15:30 Analyses of the Chemical and Oenological Factors Influencing the Taste Characteristics of Wines Applying Time Related Sensory MethodsUlrich FISCHER

15:50 Comprehensive Strategies for Food Authentication Markus FISCHER

16:10 Small-Group Workshops: Discussions on emerging topics

18:00 Dinner at Ristorante Pulcinella (Speakers only)

FRIDAY, 14 SEPTEMBER 2018 (DAY 3)

SESSION 5 cont.:Chairs: Peng ZHOU and Georg SPRENGER

09:00 Fungal Enzymes for Safer Food Ralf G. BERGER

09:20 Bioflavor Production with Basidiomycetes: Potential in Developing Novel Nonalcoholic BeveragesYanyan ZHANG

09:40 Innovative Products from Fungal CulturesHolger ZORN


10:30 Panel Discussion: Research Gaps and Trends

11:00 Small-Group Workshops: Towards a Joint Research Agenda

12:30 Lunch Break

13:30 Small-Group Workshops: Towards a Joint Research Agenda continued

15:00 Coffee Break

15:30 Wrap-up and Presentation of a Draft Research Agenda/ Next StepsLutz FISCHER, University of Hohenheim and Wanmeng MU, Jiangnan University

15:50 Closing RemarksHai SUN, Baden-Wuerttemberg International

CONTACTS

Scientific Directors:Prof. Dr. Lutz FISCHER and Jun.-Prof. Dr. Yanyan ZHANGInstitute of Food Science and Biotechnology, Hohemheim UniversityProf. Dr. Bo JIANG and Prof. Dr. Wanmeng MUState Key Laboratory of Food Science and Technology, Jiangnan University

Organization | Head Office of the Hohenheim Research Center for Health Sciences, University of HohenheimPhone: +49 (0)711 459 24615, [email protected]

For Press and Media | Florian Klebs and Dorothea ElsnerDepartment of Media and Marketing, University of Hohenheim, +49 (0)711 459 23289, [email protected]

14 15

ABSTRACTS

OralPresentations

16 17

O1 Key Technology of Probiotic Fermented Fruit and Vegetable as well as Its Application in Industry

QIANQIAN GUAN, TAO XIONG, SHAOPING NIE, AND MINGYONG XIEState Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, Jiangxi, 330047, ChinaCorresponding author: [email protected]

Chronic diseases, such as cardiovascular di-sease, cancer, diabetes, and hypertension, are seriously threatening the health of people. The disorder of intestinal flora and the insufficient intake of dietary fiber are two important causes of the occurrence of chronic diseases. Intake of probiotics and prebiotics may help to improve the structure of intestinal flora. Probiotic products such as probiotic fermented yogurt, probiotic capsules, probiotic powder etc., are currently popular all over the world, while there are few probiotic fermented fruit and vegetable products on the market. We introduced for the first time the fermentation technology into fruit and vegetable processing and developed an innovative techno-logical system to solve the problems in fruit and vegetable fermentation industry such as lacking bacterial strains specific for fruit and vegetable fermentation, less developed technology of bac-

terial agent preparation, and limited sorts of fruit and vegetable fermentation products. This system comprises upstream, midstream, and downstream of the industrial chain, which has broken through the technology bottleneck of high-density cul-ture and industrial agent preparation. By this technology, a series of probiotic fermented fruit, and vegetable products have been developed, promoting the establishment of entire new industry of fruit and vegetable fermentation. On this basis, the safety and functionality of these probiotics and their fermented fruit and vegetable products were studied. Due to the combination of probiotics and prebiotics, fermented fruit and vegetable products have the functionality of improving intestinal health, relieving chronic disease. Thus, the products are of great market potential and will bring the revolutionary influence on probiotics as well as fruit and vegetable processing industry.

O2 Co-fermentation of Tofu and Plant Material for Enrichment of Vitamin B12 by Propionibacterium freudenreichii ssp. freudenreichii DSM 20271 andLactobacillus reuteri DSM 20016

BERNWARD BISPINGUniversity of Hamburg, Hamburg School of Food Science, Division of Food Microbiology and Biotechnology, Ohnhorststr. 18, 22609 Hamburg, Germany, Phone: +49-40-42816 642, Fax: +49-40-42816 644Corresponding author: [email protected]

Session 1 Session 1

Vegan and vegetarian foodstuffs do not contain vitamin B12, as vitamin B12 is always only produced by Procaryotes (Bacteria and Archaea). As the re-commended daily intake amounts to 3 µg of vitamin B12 per day, people with such forms of nutrition have to supplement vitamin B12.The aim of this study is to unlock a new plant based source of cobalt to increase vitamin B12 production during fermentation of tofu. Therefore tofu was co-fermented with plant material in a submerged system to enrich it with vitamin B12.69 edible plants with a reported high cobalt content were screened by ICP-MS for their ability to be used as a cobalt source. 47 samples revealed higher cobalt concentrations than the used tofu itself. 40 of those were co-fermented together with tofu by Propionibacterium freudenreichii ssp. freudenreichii DSM 20271 and Lactobacillus reuteri DSM 20016 in a glucose fed fed-batch process with the supplementation of betaine and riboflavin. The 11 plant materials that resulted in the highest vitamin B12 concentrations after fermentation, were

extracted by hot water and this liquid was used in different fermentation experiments. For the finally selected Cicer arietinum L. seeds an optimization experiment of the extraction was performed by varying the swelling time, the breaking of chickpeas, and the cooking temperature. No regression model could be fitted to the optimization data for the re-sults of vitamin B12 that scattered between 110 ng/mL and 172 ng/mL in the fermentation broth. For the yield of extraction a regression model could be fitted with a cross-validated R2 = 0.944. The bioavailability of cobalt is plant dependent and cannot be reduced to its concentration. The highest concentration of vitamin B12 achieved by tofu fermentation with a chickpea extract amoun-ted to 0.172 µg/mL in the fermentation broth. That corresponds to a consumption of 17 mL of fermentation broth to meet the RDI of 3 µg. Further-more the prediction of yield of extract during the extraction process is possible for chickpeas so that economic and environmental considerations can be taken into account.

18 1918

O3 Research and Industrialization Progress on Aqueous Extraction of Edible Oil in China

RUIJIN YANG1, WENBIN ZHANG2, QIYAN ZHAO3, PENGFEI LI4, AND XIN HAN4

1 State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, Jiangsu2 Collaborative innovation center of food safety and quality control in Jiangsu Province, Jiangnan University, Wuxi 214122, Jiangsu 3 School of Food Science and Technology, Jiangnan University, Wuxi 214122, Jiangsu4 Jiangsu JunQi Biotechnology co.,LTD., Nantong 226600, Jiangsu Corresponding author: [email protected]

Traditional approach for extracting oil from oil-seeds are mechanical pressing and solvent ex-traction, which both have ineradicable weak-nesses. Mechanical pressing is less efficient while solvent extraction has inherent disadvantages of poor quality of protein in oil cake (meal), and the hexane used has been classified as a neurotoxin and a volatile organic compound. Hence, aque-ous extraction process (AEP) has gained much interests and regarded as an alternative method. The recent research achievements and the pro-gress in industrialization of the process in China was summarized.Three derivative processes developed by AEP, direct aqueous extraction process, enzyme asis-

ted aqueous extraction process and ethanol as-sisted aqueous extraction process, and different approaches for sufficient utilization of by-pro-ducts including protein, polysaccharide and fiber, were introduced. At present, the oil yields of AEP has achieved breakthrough. The free oil yields of peanut, camellia seed, rapeseed, sunflower seed, corn germ, peony seed and flaxseed have totally reached over 92% at the laboratory scale. Among these, AEP of peanut oil and camellia seed oil have realized industrialization in China, gaining free oil more than 92% and 91% respec-tively. With the development of technology and equipment, this process will be more mature in the near future.

19

O4 Exopolysaccharide Production by Food-Grade Lactic and Acetic Acid Bacteria

FRANK JAKOB AND RUDI F. VOGELTechnical University of Munich, Chair for Technical Microbiology, Gregor-Mendel-Straße 4, 85354 Freising Corresponding author: [email protected]

Lactic (LAB) and acetic (AAB) acid bacteria are traditionally used for the manufacture of fermented foods. In addition to their main function as acid producers during food fermentations, many food-grade LAB and AAB produce considerable amounts of high-molecular weight exopolysaccharides (EPS), which are secreted into the environment. Due to their water-binding (hydrocolloid) properties, such EPS improve the structure, stability or shelf-life of foods and often exhibit health-promoting properties as dietary fibers or prebiotics. In situ EPS production by LAB such as Streptococcus thermophilus and Lactobacillus bulgaricus is especially exploited in the dairy industry for the manufacture of yoghurt. However, our recent researches revealed diverse no-vel LAB and

AAB derived EPS as functional hydrocolloids for improvement of pastries, beverages and meat/plant products. A key finding of our recent stu-dies is that specific EPS-types can be directively modified regarding their macromolecular pro-perties (molecular weight, particle shapes, abundance/type of branches and concomitant rheological properties) by proper and knowledge-based adjustment of the fermentation conditions, which opens multiple novel possibilities for (food) biotechnological exploitation of EPS produced by LAB and AAB.

Here, a brief overview about our current research progress on EPS biosynthesis, structures and func-tionalities by / of food-grade LAB and AAB is given.

Session 1 Session 1

20 2120

O5 The Microbial Diversity and Community Dynamics of Chinese Traditional Fermented Vegetables

TAO XIONG, QIANQIAN GUAN, CHANGGEN LIU, AND YANGSHENG XIAOState Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, Jiangxi, 330047, ChinaCorresponding author: [email protected]

Fermentation is one of the oldest food processing and preservation methods, there are various kinds of traditional fermented food in China, especially the traditional fermented vegetable products. In almost every region of China, there are traditional fermented vegetable products with regional characteristics, such as pickle in Sichuan, sauerkraut in Northeast China, serofluid dish in Northwest China, Fuling Zhacai in Chongqing, Sour Bamboo Shoots in Guangdong and Guangxi areas, etc. These traditional fermented vegetable products are abundant of lactic acid bacteria (LAB), and the distribution of LAB taxa is highly variable from one sample to another. We have collected more than 2,000 samples of traditional fermented vegetables from various regions of China and have fully explored and standardized the traditional techniques and microbial resources. We have so far preserved

more than 6,000 strains of lactic acid bacteria and established the first dedicated strain library with independent intellectual property rights for fruit and vegetable fermentation in China. In addi-tion, we fully analyzed the microbial diversity and community dynamics in traditional fermented vegetables, illustrated the metabolic mechanism of multi-strain synergistic fermentation and revealed the relationship between functional micro-organism and product flavor in Sichuan pickle preliminarily. This research is of profound signi-ficance. Firstly, it effectively promoted the in- heritance of traditional fermented vegetable pro-duction techniques that have been circulating in various ethnic regions of China for thousands of years; Secondly, It is of great significance to carry forward Chinese traditional food culture; Finally, It has laid a solid theoretical foundation for the long-term development of Chinese probiotics industry.

21

Session 1 Session 1

O6 Simultanous Production of Microbial Lipids and Gluconic Acid with NewlyIsolated Oleaginous Yeast Strains

QIAN X.1, JIANG M.2, AND OCHSENREITHER, K.11 Karlsruhe Institute of Technology (KIT), Institute of Process Engineering in Life Sciences, Section II: Technical Biology, Karlsruhe, Germany

2 Nanjing Tech University, College of Biotechnology and Pharmaceutical Engineering, Nanjing, China Corresponding author: [email protected]

Single cell oils (SCOs) are intracellular storage lipids comprising of triacyglycerols (TAGs). They are produced by oleaginous microorganisms which are able to accumulate between 20% and up to 80 % lipid per dry biomass. SCOs are chemically equi-valent to plant oils and can be used for nutrition, oleochemistry and the production of fuels. Especially polyunsaturated fatty acids of the ω-3 and ω-6 class (e.g., ω-linolenic acid, linoleic acid) are essential for maintaining biofunctions in mammalians like humans and are essential ingredient of infant nutrition. The newly identified SCO producing yeast species Cryptococcus pod-zolicus and Trichosporon porosum were isolated from beat bog samples. During fermentation under SCO producing conditions gluconic acid was formed extracellular by both isolates in considerable amounts (Schulze et al. 2014) competing for substrate availability. Beside SCO production, gluconic acid is a second value-added product, as gluconic acid and its derivates find wide application in the food and pharmaceutical industries.

Due to the simultaneous formation of both products and the resulting competition for carbon source, yields and concentrations of both products are lo-wered. The objectives of this study are therefore to identify parameters which inhibit the synthesis of one product in order to optimize the production of the second product. Furthermore, it was deter-mined if the identified regulation mechanisms are similar for both species. By using a complex nitrogen source, e.g. peptone, instead of ammonium-salts, single cell oil production is favored and gluconic acid production is de-layed. Using this approach production phases of both products can be clearly separated. Further-more, high glucose concentrations and high aeration rates have an impact on gluconic acid pro-duction. Therefore, carbon flow can be directed to one of the products in order to optimize yield and final product concentration by changing process parameters and medium composition.

22 2322

O7 Modification of Functional Properties of Egg White Protein by Dietery Glycerol Monolaurate Supplementation

FENGQIN FENGDepartment of Food Science and Nutrition, Zhejiang University, Hangzhou, Zhejiang 310058, ChinaCorresponding author: [email protected]

Feed additives are widely used in poultry indus-try to maintain and improve the animal health and poultry production. Understanding the interac-tions between feed additives and egg white func-tional properties may offer a novel insight into the effect of feed additives on laying hens. Glycerol monolaurate (GML) is a kind of widely used in feed/food industry as an effective anti-bacterial emulsifier. In this work, the effects of three feed supplementation dose of GML (150, 300, 450 mg/kg) on the functional properties of egg white protein (EWP) were investigated. The hardness of the egg white protein gel was signifi-cantly improved by medium (300 mg/kg) and high (450 mg/kg) dose supplementation of GML. The

foaming capacity and foaming stability were both increased with the increasing dose supplementa-tion of GML. The emulsifying activity and stability were maintained and the creaming process was slowed down in the supplementation groups. The heat denaturation temperature and rheological property of EWP were also modified.Overall, there is a significant dose-effect re-lationship between GML concentration in feed and EWP functional properties. These results demonstrated that GML supplementation has a potential to modify functional properties of egg white protein, which broadened the application range of GML and provides a new perspective to evaluate the efficacy of feed additives.

23

O8 Production of Human Milk Oligosaccharides by E. coli

FLORIAN BAUMGÄRTNER, GEORG A. SPRENGER, AND CHRISTOPH ALBERMANNInstitute of Microbiology, University of Stuttgart, Allmandring 31, D-70569 Stuttgart, GermanyCorresponding author: [email protected]

Topic: Beneficial effects of human milk oligosac-charides (HMOs) on infants’ well-being have raised attention for HMOs as potential nutritional additives for infant formula [1a, 7]. However, chemical or in vitro enzymatic syntheses are laborious or costly [1a,b]. Objective of this work was the construction of plasmid-free Escherichia coli strains capable of synthesizing HMOs using recombinant glyco-syltransferases in combination with intracellularly generated nucleotide-activated sugars (GDP-L-fucose, UDP-activated sugars) to allow further research on these compounds.Results and Conclusions: Strain construction was based on the E. coli K-12 strains (JM109 or LJ110) using a site-specific λ-red recombineering technique for chromosomal integration of heterologous genes in combination with a screening on differential agar plates [2]. Strain evaluations and HMO syntheses were conducted in shake-flask cultivations and fed-batch fermentations, allowing quantification and improvement of intracellular precursor molecule levels via HPLC. Plasmid-free E. coli strains (with up to 6 consecutive integrationsin one strain) were constructed for the synthesis ofdifferent HMOs. Different copy-numbers of genes

enabled leveling of gene expression in order to raise HMO productivity. Chromosomal stability of the recombinant genes allowed syntheses in fed-batch fermentations without the need for antibiotics as selection markers. Product titers of up to 20 g/L (2´-fucosyllactose) with full consumption of lactose were reached [3,4]. Next, we could demonstrate the combination of specific bacterial glycosyltransferases (LgtA, WbgO) to-gether with enhanced intracellular synthesis of UDP-nucleotide-activated sugars for the efficient preparative synthesis of oligosaccharide core-structures such as lacto-N-tetraose (LNT) with ti-ters of >12 g/l (with similar values of the precursor, LNT II, as major by-product) from galactose and lactose in fed-batch cultivations [4,5]. Subsequent combination of these syntheses of core-structures with chromosomally integrated genes for fucosyl-transferases and enhanced intracellular supply of GDP-L-fucose resulted in fucosylated HMOs up to penta- and hexasaccharides such as LNF I and LNDFH II [6]. Thus, chromosomal integration turned out to be a powerful tool for synthetic microbiology, allowing multiple rapid and site-specific insertions and construction of genetically stable strains.

Session 2 Session 2

References:[1] a) L. Bode, Glycobiology 22 (2012), p. 1147–1162; b) X. Chen, Adv. Carbohyd. Chem. Biochem. 72 (2015) , p.113-190.[2] C. Albermann, N. Trachtmann, G. A. Sprenger, Biotechnol. J. 5 (2010), p. 32–38.[3] F. Baumgärtner, L. Seitz, G. A. Sprenger, C. Albermann, Microb. Cell Fact. 12 (2013), 40.[4] F. Baumgärtner, G. A. Sprenger, C. Albermann, Enzyme Microb. Technol. 75-76 (2015), p. 37–43.[5] F. Baumgärtner, J. Conrad, G. A. Sprenger, C. Albermann, Chembiochem 15 (2014), p. 1896–1900.[6] F. Baumgärtner, L. Jurzitza, J. Conrad, U. Beifuss, G. A. Sprenger, C. Albermann, Bioorg. Med. Chem. 23 (2015), p. 6799–6806.[7] G.A. Sprenger, F. Baumgärtner, C. Albermann, J. Biotechnol. 258 (2017) p. 79-91.

24 2524

O9 Recent Approaches for Improving Monascus Pigment Production

XIAOPING ZHU, LILING CHEN, ZHIBIN LIU, WEN ZHANG, CHEN ZHANG, AND LI NIInstitute of Food Science and Technology, Fuzhou University, Fuzhou, Fujian, 350108, ChinaCorresponding author: [email protected]

Monascus spp., an important microbial resour-ce, has been used in the production of certain fermented foods in East Asia, particularly China and Japan. Monascus pigment is one of the ma-jor secondary metabolites. Its components are complex and usually divided into three catego-ries according to the distribution of its maximum absorption wavelength: red pigment, orange pig-ment, and yellow pigment. At present, there are more than 50 kinds of monascus pigment compo-nents found, but only 16 types of structures have been determined, including Monascorubramine, Rubropunctamine, Monascorubrin, Rubropuncta-tin, Ankaflavin, Monascin etc. Recently, many studies showed that monascus pigment has spe-cial physiological activities, such as antibacterial activity, blood lipid reduction, anti-cancer, an-ti-inflammation and so on, which imply that it has a big potential as a functional food additive or in-gredient. Therefore, how to improve and regulate the pigment production has attracted increasing attention.

The major approaches to improving the monascus pigment production include the excellent strain screening by using mutagenesis techno-logy or genetic engineering technology, culture medium, and fermentation conditions optimizing. In recent years, the effect of some exogenous additives such as surfactants, metal ions, vege-table oils, antibiotics, and antioxidants on the monascus fermentation has been investigated. In present study, we focused on the regulation of monascus liquid fermentation by nine kinds of antioxidants including glutathione, SOD, vitamin E, lipoic acid, tea polyphenols, quercetin, rutin, ellagic acid, and shannephenol. Among which, tea polyphenols had the best regulatory effect. The regulation mechanism of tea polyphenols to the synthesis of monascus pigment from three aspects, including the expression of key regula-tory genes, oxidative stress, and transcriptome were further explored.

25

Session 2 Session 2

O10 Enhanced Biocatalytic Conversion Strategies of Renewables to Tailor Made Glycolipid Biosurfactants in Uncommon Reaction Media

JENS RUDAT, ANDRÉ DELAVAULT, JENS GRÜNINGER, REBECCA HOLLENBACH, SASCHA SIEBENHALLER, AND CHRISTOPH SYLDATKKarlsruhe Institute of Technology (KIT), Institute of Engineering in Life Sciences, Section II: Technical Biology, Karlsruhe, Germany Corresponding author: [email protected]

We encounter surface-active molecules such as surfactants every day, for example in cleaning agents or body care products. They are also widely used for food, cosmetic and pharmaceutical applications. Of particular interest for industrial applications are glycolipids. Due to the increased environmental awareness of consumers and legal requirements, products with reduced carbon footprints and complete biodegradability are in high demand. This requires the introduction of new compounds and production processes based on renewable raw materials. Nevertheless, their production on a commercial scale has only been realized in very few cases. Up to now, they are only found in niche applications due to their distinctly higher prices compared to traditional surfactants. Common rea-sons for their rare use in industry are expensive substrates, limited product concentrations and yields – often caused by substrate and/or product inhibitions – and the formation of product mixtures instead of pure compounds, which cause high costs for downstream processing.Glycolipids can also be obtained by chemical or enzymatic synthesis from renewable materials, the latter method using regio-, stereo-, and substrate specificity of enzymes as advantages. As biocata-lysts hydrolytic enzymes are used, which have the capability under conditions of reduced water acti-vity in uncommon reaction media or at very high

substrate concentrations, to reverse the normal hy-drolytic reaction and to synthesize ester-, peptide- or even glycosylbonds. The enzymatic synthesis of glycolipids using lipases or esterases has already become a common technique, but it is still limited to the use of a small number of lipases and production of non-ionic sugar esters. Furthermore, the resul-ting products are often only stable in a narrow pH range. Here, the successful formation of more sta-ble ether bonds or glycosidic bonds by the reverse reaction catalyzed e.g. by glycosidases, etherases would be of high interest. However, this is still a challenge, mainly because only a small number of enzymes is commercially available and these en-zymes show only low stability in organic solvents.

Here we present results of our investigations on the development of different strategies for the en-zymatic synthesis of surface-active glycolipids with new structures. In particular, the results of different enzymatic syntheses of sugarlipids, alditollipids and alkylglycosides in a deep eutectic solvent sys-tem are presented. We will discuss the different methods and aspects of enzymatic glycolipid pro-duction from an academic and industrial point of view and will try to propose strategies to overcome some of the still existing limitations mentioned above in future.

26 2726

O11 Stability of High Protein Food Matrix

PENG ZHOUState Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, ChinaCorresponding author: [email protected]

Dairy proteins have been widely used to develop various food products, such as bakery foods, in-fant formula and foods for sports nutrition, many of those foods are in solid and semi-solid states and are often thought to be relatively stable. However, the high protein containing foods or food ingredients in such state was often found losing their quality during storage. Physical changes were often observed, including tex-ture hardening, decreasing in solubility, structu-ral collapse, particle agglomeration, molecular migration/crystallization, and phase separation, as well as chemical reactions such as protein aggregation and Maillard reaction. Factors, such as moisture content, the presence of certain ion,

the state of proteins, pH, and the environmental temperature and relative humidity, have signifi-cant effects on the protein stability in food matrix, resulting in changes of quality of such foods. This presentation focuses on two major deterioration, the loss of stability for dairy protein ingredients and the textural hardening of intermediate moisture foods containing dairy proteins. The overall goal is to identify the molecular mechanism and controlling factors for such deterioration, which are expected to lead to new strategies for controlling the protein stability during long-term storage thereby providing such dairy protein ingredients or foods better quality and longer shelf life.

27

O12 Process Engineering of Enzymatic Processes in the Hydrolysis and Fractionation of Functional Peptides

ULRICH KULOZIKChair for Food and Bioprocess Engineering, TUM School of Life Sciences Weihenstephan, Technical University of Munich, Weihenstephaner Berg 1, D-85354 FreisingCorresponding author: [email protected]

Session 2 Session 2

The production of food grade functional peptides can be realised by enzymatic hydrolysis. Despite of the application of specific enzymes, enzymatic hydrolysis of food proteins represents until today a complex and hardly manageable system resulting in a mixture of numerous different peptides.Production processes, such as substrate pre-treat-ment by the thermal denaturation of proteins and controlled hydrolysis by the regulation of environ-mental conditions, increase the specific release of particular bioactive peptides. To economically implement large-scale enzymatic processes, the immobilized enzyme represents an alternative approach due to the possibility of re-using the enzyme and producing enzyme-free hydrolysates. Following enzymatic process,

peptides with different functionalities must be separated and enriched to achieve high-yield products. Membrane separation methods, such as ultrafiltration and nanofiltration, are commonly applied in current food industry due to the feasible cost. To increase the selectivity of membrane filtration,electromembrane separation technologies are sug-gested, which use electric charge and molecular mass to affect peptide separation in one-step. In addition, and with high selectivity, membrane adsorption chromatographic techniques are attrac-tive due to the largely improved fractionation effi-ciency by increasing process capacity and flow rates compared with conventional bead-based chromatography.

ReferencesLeeb, E., Gotz, A., Letzel, T., Cheison, S. C., & Kulozik, U. (2015). Influence of denaturation and aggregation of beta-lactoglobulin on its tryptic hydrolysis and the release of functional peptides. Food Chemistry, 187, 545-554.Leeb, E., Holder, A., Letzel, T., Cheison, S. C., Kulozik, U., & Hinrichs, J. (2014). Fractionation of dairy based functional peptides using ion-exchange membrane adsorption chromatography and cross-flow electro membrane filtration. International Dairy Jour-nal, 38(2), 116-123.Mao, Y., & Kulozik, U. (2018). Selective hydrolysis of whey proteins using a flow-through monolithic reactor with large pore size and immobilised trypsin. International Dairy Journal, 85, 96-104.

28 2928

O13 Strategy for Biosynthesis of Low-Calorie Sugars

BO JIANGState Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, ChinaCorresponding author: [email protected]

Low-calorie sugars, used as sucrose substitutes, provide consumers with a sweet taste with rela-tively low calories. Many low-calorie sugars are non-nutritive sugars because they are not meta-bolized and pass through the body unchanged. Except for the low calories, they have various other biological functions and enormous poten-tials for applications in pharmaceutical, cos-metics, food, and flavor industries. Currently, D-tagatose, D-psicose, D-allose, xylitol, epilac-tose, and other rare sugars that exist in naturein very small quantities have been studied exten-sively. The traditional extraction method from thenatural products or the traditional fermentationcan no longer satisfy the amount of needs anymore, and the enzymatic synthesis greatly lowers

the price of the sugars and extends the applica-tions of the low-calorie sugars. Biological production of these low-calorie sugars depends on various enzymes including aldo-keto isomerases (EC 5.3.1), carbohydrate epimerases (EC 5.1.3), and oxidoreductases (EC 1.1). Molecular techniques including cloning and expression of proteins of interest, site-directed mutagenesis, homology modeling, molecular docking, and molecular dynamics simulation are applied for the bio-production of low-calorie sugars. Cloning of genes from various bacteria and molecular modification provide us with more efficient engineering bacteria that lower the production cost for the low-calorie sugars.

29

O14 Production of Prebiotic Sugars Through Enzymatic Lactose Conversion

LUTZ FISCHERUniversity of Hohenheim, Institute of Food Science and Biotechnology, Department of Biotechnology and Enzyme ScienceCorresponding author: [email protected]

Prebiotic sugars are indigestible food ingredients by humans and benefically affect the host by selec-tively stimulating the growth and activity of probiotic bacteria species in the colon. This supports the ba-lance in the intestinal ecosystem of the human host and improves health (Kovács et al. 2014, Adv. Bio-chem. Eng. Biotechnol. 143: 257). Only two groups of sugars accomplish these criteria today, when following the recommendations by the European Food Safety Authority (EFSA). These are Galac-tooligosaccharides (GOS) and Fructosides (FOS), including lactulose and inulin. However, several other sugars such as Xyloseoligo-saccharides (XOS) or epilactose are discussed as potential prebiotics.Since lactose is globally generated in millions of metric tons it is a promising substrate for further creation of valuable ingredients using biocatalysis. In former studies we showed the suitability of par-ticular microbial β-galactosidases in the production of galactooligosaccharides (GOS) and lactulose, respectively. The enzyme screening, kinetic pro-

perties and technical suitability for production pro-cesses will be discussed briefly.Recently, some thermostable cellobiose-2-epimerases (CE) showed a surprising catalytic side reaction, that is the intramolecular isomerisation of glucose to fructose when lactose (D-Gal-β1,4-D-Glc) is the substrate. This side reaction of CEwas used in our group for the in situ productionof lactulose (D-Gal-β1,4-D-Fru) in milk. On theother hand, the common epimerisation reactionof CE is the intramolecular conversion of glucoseto mannose in cellobiose or lactose, respectively.Here, lactose was enzymatically epimerised to socalled epilactose (D-Gal-β1,4-D-Man), a potentialnew prebiotic, in a gram-scale production process.All enzymes used were of microbial origin, eitherfound by conventional screening methods, in themetagenome or in protein data bases (in silico). Thediscovery of novel enzmes by combination of me-tagenome and in silico screening wil be discussedfinally.

Session 2 Session 2

30 3130

O15 Anti-Diabetic Mechanism of Dietary Polysaccharides Based on Their Gastrointestinal Functions

SHAOPING NIE, JIELUN HU, HAISHAN LI, QIXING NIE, QIQIONG LI, QINGYING FANG, AND MINGYONG XIEState Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, Jiangxi, 330047, ChinaCorresponding author: [email protected]

Diabetes mellitus (DM) is an impaired carbohy-drate, fat and protein metabolic syndrome in-duced by insufficient insulin secretion or decrea-sed tissue sensitivity to insulin. DM is attracting global attention, which has severely affected the quality of life of human and is related to several severe complications. It has been realized that the optimal selection of food and dietary factors play key roles in pre-venting and reducing the risk of lifelong Type 2 diabetes mellitus (T2DM). In terms of food and dietary factors, the polysaccharide shows pro-tective effect on T2DM. Dietary polysaccharides were mainly from natural sources, namely plants, fungi, algae, and etc. They were resistant to hu-man digestion and absorption in human small intestine with complete or partial fermentation in the large bowel and have shown anti-diabe-tic ability. In this presentation, we aimed to pro-

vide information on anti-diabetic mechanism of the dietary polysaccharides, based on the whole gastrointestinal process, which was a new angle of view for understanding the anti-diabetic me-chanism of dietary polysaccharides. The dietary polysaccharides could attenuate diabetes by the mechanisms of gastrointestinal viscosity, gastro-intestinal satiety, large bowel fermentation, and gastrointestinal anti-inflammation effects. Thus, it could slow down gastric emptying, reduce fat and carbohydrate digestion, inhibit α-amylase and α-glucosidase activities, lower glucose ab-sorption efficacy and postprandial glycaemia, raise satiety, alter microbiota and short-chain fat-ty acid production, and attenuate inflammation in diabetes. Further researches could take efforts on the mechanisms of the polysaccharide action through host-microbiota interactions targeting diabetes.

31

O16 Continuous Production of Low-Calorie Sweeteners Using a New Recombinant Fructosyltransferase

JAN PHILIPP BURGHARDT1,2, AMAD UR REHMAN1, MEHRDAD EBRAHIMI1, DOREEN GERLACH3, AND PETER CZERMAK1,2,3,4

1 Institute of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, Giessen, Germany

2 Faculty of Biology and Chemistry, Justus Liebig University, Giessen/Germany3 Department of Bioresources, Fraunhofer Institute for Molecular Biology and Applied Ecology (IME), Giessen4 Department of Chemical Engineering, Kansas State University, Manhattan (Kansas), USA Corresponding author: [email protected]

References[1] Ur Rehman A, Z Kovacs, M Ebrahimi, H Quitmann, P Czermak (2016): Enzymatic production of fructooligosaccharides from

inexpensive and abundant substrates using a membrane reactor system, Separation Science and Technology, 51. pp. 1537-1545[2] Spohner S C, P Czermak (2016): Heterologous expression of Aspergillus terreus fructosyltransferase in Kluyveromyces lactis,

New Biotechnology, 33. pp. 473-479

Oligosaccharides, e.g. fructooligosaccharides (FOS) are an emerging prebiotic constituent in healthy nutrition. FOS are mainly of interest for indigestibility by human digestive system making them essentially calorie free. Also, FOS have a sweetness level of 30-60% as high as that of sucrose depending upon the chain. FOS are used for masking the after taste of other alternative sweeteners. Furthermore, they are termed to enhance the growth of bifidobacteria in human digestive system and have positive impact on general health.For the production of FOS pure sucrose is used most of the time as substrate. Molasses a rather in-expensive viscous by-product of sugar processing and still rich in sugar could be an alternative source of FOS production. Currently, molasses is used as a substrate for certain biological conversions and transformations and has only a limited application in the nutritional industry so far. In this study an en-zyme membrane reactor (EMR) was used for pro-cessing of the substrate [1]. In this study the FOS are produced with a commercial available enzyme

(Pectinex Ultra SP-L from Aspergillus aculeatus, Novo-zyme A/S, Denmark) and with a new recom-binant fructosyltransferase. This new enzyme was recombinantly expressed and produced extracellu-larly in the yeast Kluyveromyces lactis GG799 [2].During enzyme reaction glucose accumulates as a by-product and inhibits at higher concentration the FOS production. Therefore, enzyme membrane reactor (EMR) system was used for continuous product removal to minimize inhibitory effects of glucose and to supply a continuous fed of sucrose. Diluted molasses were used as substrate in comparison with pure sucrose in these enzymatic reactions. An ultrafiltration separation of used enzyme ensured a continuous production of FOS. In the present study 89% and 69% conversion were achieved with 63 % and 47% of FOS, 24 % and 21% glucose and 3% and 2% of fructose from molasses and pure sucrose. The EMR system for the production of FOS from inexpensive molasses was proven to be an efficient process fit for industrial application.

Session 2 Session 2

32 3332

O17 Recent Progress on Ketose 3-Epimerase and Its Application in Enzymatic Production of D-Allulose

WANMENG MU, WENLI ZHANG, AND BO JIANGState Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, ChinaCorresponding author: [email protected]

D-allulose, also known as D-psicose, an epimerof D-fructose, is a rare monosaccharide thatexists in extremely small quantities in nature. Itis an ideal substitute for sucrose, because it has70% of the sweetness of sucrose and ultra-lowthe energy. In addition, it has received sustainedattention because of its unique physiologicalfunctions and potential health benefits. However,it is scarce in nature, and difficult to chemicallysynthesize. Thus, biological production of D- al-lulose is attractive to researchers.It has been demonstrated that ketose 3-epi-merase plays an irreplaceable role in the bio-conversion of D-fructose to D-allulose. In ourrecent works, eight D-allulose-producing ketose3-epimerases were identified from Rhodobactersphaeroides SK011, Clostridium cellulolyticumH10, Clostridium scindens 35704, Desmosporasp. 8437, Clostridium sp. BNL1100, Clostridiumbolteae ATCC BAA-613, Dorea sp. CAG317, andTreponema primitia ZAS-1. Molecular modifica-

tion by site-directed mutagenesis was performed to improve the thermostability and catalytic acti-vity of the D-allulose-producing enzyme from C. bolteae.Most of previous works focused on the ketose 3-epimerase expression using E. coli as a hostcell. However, the safety problems in the usageof genetically engineered bacteria and antibioticsresistance have become global issues. The se-lection of safe hosts and construction of enginee-ring strains without antibiotic resistance genesare the development direction of future research.In the present study, the recombinant Bacillussubtilis expressing ketose 3-epimerase free ofantibiotic resistance genes was constructed. Theapproaches included: (1) replicative plasmidswith an auxotrophic marker; (2) genomic integra-tion with tandem repeat genes; (3) spore surfacedisplay. All these three approaches finally suc-ceeded, showing great potentials for the industri-al production of D-allulose.

33

O18 Continuous Enzymatic Production of Technofunctional Casein Hydrolysates

TIMO STRESSLER, JACOB EWERT, AND LUTZ FISCHERUniversity of Hohenheim, Institute of Food Science and Biotechnology, Department of Biotechnology and Enzyme ScienceCorresponding author: [email protected]

Milk proteins are being used in the food indus-try due to their nutritional value and technofunc-tionality. The technofunctionality of proteins in-cludes the water-holding capacity, oil- and fat- binding properties, gelling properties and the ability to stabilize oil–water (emulsion) or gas–water (foam) interphases. In addition to the na-tive food proteins used commonly, modified pro-teins can also be applied. These modifications can improve the technofunctional properties, such as the attachment onto the interphase. An example for these modifications is the enzy-matic protein hydrolysis. During the hydrolysis, proteins are cleaved into peptides of different sizes and free amino acids. Possible consequen-ces of a hydrolysis are: a decreased molecular weight, an increased number of ionizable groups, an exposure of hydrophobic groups from the in-ner protein molecule, an increased solubility, a decreased viscosity and, consequently, changes regarding the environmental interactions at the interphase. A benefit of the enzymatic hydrolysis is that it provides a more uniform product, can be performed under milder conditions and, thus,

avoid the conditions required for chemical treat-ments. The specificity of the peptidase used for hydrolysis and the resulting degree of hydrolysis of the protein hydrolysate are the major parame-ters for the obtained technofunctional proper-ties. The protein hydrolysis can be performed in discontinuous batch processes or in continuous systems using immobilized enzymes (soluble or insoluble). The enzymatic hydrolyses of proteins in batch processes has several disadvantages compared to continuous systems. Exemplarily, batch hydrolyses are more expensive because the enzymes cannot be reused.

In recent studies, we investigate the techno-functional properties of caseinate hydrolysates produced in a batch processes, using an enzyme-membrane reactor and a novel developed biocatalyst-filter reactor (BFR). The BFR displayed an easy-to-build and easy-to-apply process for a continuous process development. Thereby, the BFR seems especially useful for applications where the substrate or product has a high viscosity or poor fluidity.

Session 3 Session 3

34 3534

O19 The Strategies to Improve the Expression of Recombinant Protein inBacillus subtilis

XIN YAN, CHAOYANG ZHOU, BIN YE, AND LEIZHEN ZHAOKey Laboratory of Agricultural Environmental Microbiology, Ministry of Agriculture, College of Life Sciences, Nanjing Agricultural University, Nanjing, Jiangsu, 210095, ChinaCorresponding author: [email protected]

Due to its excellent fermentation properties, high product yields and GRAS (generally regarded as safe) status, Gram-positive bacterium Bacillus subtilis has been developed as an attractive host in the production of industrial enzyme. However, generally, proteins derived from a Bacillus sp. are expressed much better (in the grams/l range), but others are expressed very poorly (in the mg/l ran-ge). Therefore, how to improve the expression le-vel of recombinant protein in B. subtilis is of great interest. The secretion of recombinant protein in B. subtilis involves several key steps including

transcription, translation, secretion and folding. In this presentation, I will introduce several stra-tegies employed in our lab to optimize the tran-scription and translation of target genes in B. subtilis. The transcription level was enhanced through engineering the promoter region and in-creasing the copy number of the expression cas-sette. The translation efficiency was improved by optimizing the translation initiation region and genetic codon. The results show that these stra-tegies are effective in improving the expression of recombinant protein in Bacillus subtilis.

35

O20 Modern Biooxidation by Heme-Thiolate Peroxidases

JAN KIEBIST1, MARTIN HOFRICHTER2, AND KATRIN SCHEIBNER1

1 Institute of Biotechnology, BTU Cottbus-Senftenberg, Senftenberg, Germany 2 International Institute Zittau, TU Dresden, Zittau, Germany Corresponding author: [email protected]

References:1 Hofrichter M. et al.: Fungal unspecific peroxygenases: heme-thiolate proteins that combine peroxidase and cytochrome P450 properties. Adv. Exp. Med. Biol. (2015), 851:341-368.

2 Barková K. et al.: Regioselective hydroxylation of diverse flavonoids by an aromatic peroxygenase. Tetrahedron (2011), 67:4874-4878.3 Aranda C. et al.: Selective synthesis of resveratrol analogue 4,4’-dihydroxy-trans-stilbene and stilbenoids modification by fungal peroxygenases. Catal. Sci. Technol. (2018), 8:2394-2401.4 Babot E. et al.: Regioselective hydroxylation in the production of 25-hydroxyvitamin D by Coprinopsis cinerea peroxygenase. Chem CatChem (2015), 7:283-290.

In the last decades, the selective enzymatic oxi-dation of inert or poorly activated carbons has driven an increased interest for organic chemistry. Unspecific peroxygenases (UPOs, EC 1.11.2.1), found in most fungal organisms, are secreted glycosylated enzymes that constitute a distinct superfamily of heme-thiolate proteins. UPOs ca-talyze efficiently and selectively the transfer of peroxide-borne oxygen into various organic mo-lecules including less-activated hydrocarbons. The substrate spectrum of UPOs as well as the portfolio of catalyzed reactions resemble those of cytochrome P450 monooxygenases and clas-sical heme peroxidases1. Among these reactions are hydroxylations, epoxidations, dealkylations, oxygenations of heteroatoms and halides as well as one-electron oxidations. Substance classes that can be oxyfunctionalized comprise n-al-kanes, cycloalkanes, aromatics, heterocycles, terpenes and steroids. Altogether more than 400 compounds have been found to serve as sub-strates for UPOs.Among these oxyfunctionalized organic mole-cules several bioactive compound were synthe-

sized or modified, especially polyphenols. Poly-phenols represent a group of antioxidants found naturally in fruits and vegetables and known for their anti-inflammatory, antimicrobial and an-tiviral activities and therefore their potential to reduce chronic diseases. UPOs are capable to selectively oxyfunctionalize diverse flavonoids including flavones, isoflavones, flavonones and flavonols2 and also non-flavonoid polyphenolics like stilbenes. For instance, trans-stilbene could be converted to 4,4’-dihydroxy-trans-stilben and pinosylvin to resveratrol; both have emerged as fascinating compounds because of their wide spectrum of biological effects3. In addition to po-lyphenols, 25-hydroxylated derivatives of Vita-min D2 and D3 were synthesized by UPOs and exhibit positive effects on diseases, such as hy-perglycemia, chronic kidney disease and for an-ephric patients4. These and more examples proved peroxy-genases as very useful tools for the synthesis or modification of functional and healthy food ingre-dients playing important roles in the prevention of life-style related diseases.

Session 3 Session 3

36 3736

O21 Characterization of Gelatin/Zein Nanofibers Fabricated by Electrospinning

HUI ZHANG Department of Food Science and Nutrition, Zhejiang University, Hangzhou, Zhejiang 310058, ChinaCorresponding author: [email protected]

Electrospinning is a novel fabrication technique to produce nanofibers from polymers under a strong electrostatic field. Based on the food safety concerns in industry, it is necessary to produce nanofibers that are solely or mainly composed of food biopolymers. Gelatin is a commonly used food biopolymer that has been electrospun to form hydrophilic fibers, while zein is a class of alcohol-soluble prolamine proteins to generate hydrophobic fibers. In this study, we hypothesize that zein is uniformly dispersed in the gelatin network by electrospinning to improve the water resistance of nanofibers.30% (w/v) proteins at different weight ratios were dissolved in 80% acetic aqueous solution, and electrospun to fabricate nanofibers. The field emission scanning electron microscopy obser-vations showed that the diameter of gelatin/zein nanofibers was significantly increased with the

increasing gelatin ratio. The Fourier transform infrared spectroscopy and differential scanning calorimetry measurements indicated that gelatin and zein strongly interacted via hydrogen bon-ding, and this homogeneous mixing resulted in the crystalline structure of the gelatin/zein nano-fibers at a 1:1 ratio, in agreement with the wa-ter contact angle results showing a hydrophobic surface at 118°. However, the gelatin/zein nano-fibers at higher zein ratios (1:2 and 1:3) gave rise to a much higher elongation at break of 87.9% and 69.0%, indicating good deformability and fle-xibility. The confocal laser scanning microscopy observations suggested that in contrast to the pure gelatin or zein fibers, the gelatin/zein nano-fibers maintained the 3D porous structures after immersed in water or ethanol for 24 h, leading to the improved solvent resistance.

37

O22 Enzyme-Based Strategies for Structuring Foods with Improved Functionality

JOCHEN WEISSUniversity of Hohenheim, Institute of Food Science and Biotechnology, Department of Food Physics and Meat ScienceCorresponding author: [email protected]

The creation of stable food structures is key to the manufacture of existing and novel foods. Since foods are most commonly multiphase dispersions, their properties and stabilities are largely influenced by the nature of their interfaces. A deliberate modification of surface active food molecules such as proteins, polar lipids, or saponins that are adsorbed at food system inter-faces using a combination of physicochemical techniques and enzymes has recently been pro-ven to be very promising to engineer new food structures. In this presentation, we will highlight results from two Ph.D. thesis (one completed, and one in progress), in which we used this approach to manufacture two food dispersion systems with enhanced technofunctional properties.

Cross-linking enzymes such as laccase were used to promote the formation of intra- and inter-molecular bonds between biopolymers adsorbed or deposited at the interface of oil-in-water emul-sions to improve their stability and functionality. In particular, stability to freeze-thaw cycling, and to the exposure of salt at various pH was found to increase due to biopolymers forming a more dense and rigid interface at the oil-water inter-face. The susceptibility to Ostwald ripening was also decreased, which allows emulsion to be made from highly soluble lipids such as for ex-ample essential oils. At high droplet concentra-tions, droplets were networked to form so called

emulsions gels with elastic properties. A critical oil droplet concentration was identified where the crosslinking mechanism transitioned from intra- to interdroplet crosslinking. This was attributed to an increased contact time and a decreased sepa-ration distance of droplets in highly concentrated emulsions.

More recently, transglutaminase was used to transform emulsified organogels to yield animal fat replacement systems. Emulsions were hot homogenized using a mixture of solid and liquid fats, quench cooled to induce crystallization of the lipid organogelator, and then crosslinked with transglutaminase. The system had both elastic as well as melting properties to mimic pork back fat. Comminution experiments showed that inclusion in raw meat batters is possibly yielding structures characteristic of pure animal-derived raw fer-mented sausages. Current studies are elaborating the influence of solid – liquid fat content and ratio, protein content, and enzyme type and activity to modulate the thermal and mechanical properties of the system for future uses in vegan products.

In future studies, we intend to investigate the use of combinations of crosslinking and hydrolyzing enzymes to create novel structures, with the idea of sequencing the treatments and mixing treated systems in order to molecularly tailor the structures for novel food applications.

Session 3 Session 3

38 3938

O23 Degradation of Leaf Cell Wall for Native Components

CHEN ZHANG, ZHIBIN LIU, AND LI NIInstitute of Food Science and Technology, Fuzhou University, Fuzhou, Fujian 350108, ChinaCorresponding author: [email protected]

Leaves were considered as potential food and medicines resources. However, the production and application of their components are limited due to the rigidity of cell walls. To develop biore-finery technology targeting on cell wall for native components is a key to overcome the limitation.This study was first using alkaline extraction with different conditions to investigate degrada-tion sequence of cell wall components, and then developing enzymatic methods to degrade cell wall for native protein using green tea leaves as model material. It was found that alkaline protein extraction was not facilitated by increased solu-bility or hydrolysis of protein, but positively cor-related to leaf tissue disruption. HG pectin, RGII pectin, polyphenols, and organic acids can be extracted before protein. Protein extraction can then be followed by the extraction of cellulose and hemi-cellulose. RGI pectin and lignin yield were both linearly correlated to protein yield,

which indicated that they are likely to be the key limitation to leaf protein extraction. Based on the above findings, enzymes namely Viscozyme, pectinase, arabanase, galactanase, cellulase, and hemicellulase were tested. Arabi-nanase, galactanase, and hemi-cellulase have no or minor influence on the hydrolysis of side chains of Rhamnogalacturonan I pectin (arabinan and galactanan) and hemi-cellulose when they were used individually. Cellulase and pectinase not only hydrolyzed cellulose and pectin, but also released the other components. Although cel-lulose is considered to be the skeleton of plant cell walls, its full extraction did not lead to the solubilization of all components in plant cells. Results indicate that our enzyme-aided extrac-tions are functioning on the primary plant cell wall, which mainly consists of pectin and cellulose that are cross-linked. To obtain more components, a method for lignin degradation is needed.

39

O24 Flavor and Enzymatic Impacts in the Processing of Yeast Fermented Beverages

KAI BÜCHNER & THOMAS BECKERChair of Brewing and Beverage Technology, TUM School of Life Sciences Weihenstephan, Technical University of Munich, Weihenstephaner Steig 20, 85354 FreisingCorresponding author: [email protected]

Yeast is the primary microorganism for fermented beverages such as beer, wine and sake. However, existing individual strains will not completely fulfill future demands for an efficient and high-quality fermentation. The biochemical forma-tion of yeast-derived sensory-active metabolites like higher alcohols and esters via expression of endogenous enzymes determines the different characteristics of aroma and taste in fermented beverages. In a yeast fermentation process, a large number of environmental factors act on the production of volatile aroma compounds. Factors like substrate composition in fermentation media as well as process parameters influencing these flavor-active metabolites have already been de-scribed. These factors can act on the expressi-on of yeast genes coding for enzymes involved in aroma metabolism, resulting in concentration differences in esters and higher alcohols import-ant for flavor. In subsequent experiments, the effect of amino acid supply on the transcription level of genes involved in the biosynthesis of aro-ma-relevant higher alcohols and esters was in-vestigated. Amino acids were chosen according to the previous multivariate data analysis. The-se results show that the supply of single amino

acids can determine the final concentrations of some important aroma-contributing metabolites by significant up- or downregulation of genes in-volved in the biosynthesis of higher alcohols, en-coding transaminases (BAT1 and BAT2), decar-boxylases (PDC1, PDC5, PDC6 and ARO10), and alcohol dehydrogenase (ADH1). The regulation of genes involved in the biosynthesis of esters such as fatty acid synthases (FAS1 and FAS2), alcohol acetyltransferases (ATF1 and ATF2), acyl-CoeA:ethanol O-acyltransferases (ETH1 and EEB1) shows also significant up- und down-regulation under given fermentation conditions. In the following research, self-cloning strains of the industrial brewing yeast Saccharomyces pa-storianus TUM 34/70 were constructed to over-express the gene ATF1 under the control of tem-perature inducible promoters (PSSA3, PHSP104 and PUBI4). After temperature shift (comparable to beer maturation phase), the concentrations of ethyl acetate and isoamyl acetate were signifi-cantly increased in comparison to the wild type. Therefore, self-cloning yeast is an innovative biotechnological tool to control the flavor of the final beverage (beer, wine, sake) via expression of yeast enzymes.

Session 3 Session 3

40 4140

O25 Sensoproteomics – Effect-Directed Analysis of Taste-Active Peptides inFood

KARIN SEBALD, ANDREAS DUNKEL, AND THOMAS HOFMANNChair of Food Chemistry and Molecular Sensory Science, Technische Universität München; Lise-Meitner-Str. 34, 85354 Freising, GermanyCorresponding author: [email protected]

In fermented foods, such as dairy products, co-coa or soy sauce, sensory active peptides have been shown to have an impact on the complex taste profile modulating bitter and salt taste in-tensity or kokumi sensation. In the last decades, these taste-active peptides have been identified by using a laborious fractionation approach. Targeted and untargeted proteomics combined with molecular sensory science tools enables a straightforward identification of taste-active peptides in complex food peptide mixtures. To identify bitter tasting peptides in an alternatively produced cream cheese, LC-MS-based approaches using QQQ-MS and ToF-MS in

combination with computationally predicted and empirically refined fragmentation parameters as well as data independent acquisition strategies were developed. By applying defined selection criteria, candidate marker peptides were selected and their presence finally verified using synthetic references. Further, the taste qualities and human taste thresholds were determined by sensory analy-sis. Finally, due to quantitative studies of these tar-get analytes in both cream cheese manufactu-ring procedures, differences in technological processes can be uncovered.

41

O26 Arabinoxylan Alleviates Acute Colitis by Altering Colon Symptoms andHost Immune Response

JIELUN HU, SHAOPING NIE, QIAO DING, JUNYI YIN, DANFEI HUANG, AND MINGYONG XIEState Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, Jiangxi, 330047, ChinaCorresponding author: [email protected]

Inflammatory bowel disease (IBD), an idiopathic inflammatory disease in the gastrointestinal tract, is one of the claimed diseases that caused by dysbiosis and affects a significant global human population. Nowadays, several treatments have been used to cure IBD; however, traditional treatments often have side effects. Therefore, new treatments which are based on natural products and have little side-effects are in urgent need. Polysaccharides, as natural products, were re-ported to have beneficial effects on treatment of IBD, especially acute colitis. Arabinoxylan is one representative kind of polysaccharide. Recent-ly, we have isolated a pure and homogeneous

arabinoxylan from the seeds of Plantago asiati-ca L. It was given orally to mice before, during and after dextran sodium sulfate (DSS)-induced acute colitis. Treatments with different doses of arabinoxylan could reduce the weight loss in-duced by DSS. Administrations of arabinoxylan also resulted in an obvious reduction in colitis related symptoms in colon tissues. In addition, arabinoxylan intake could attenuate colitis-as-sociated gene expression and production of cytokines in colon tissues. Our study highlights extraordinary potential of prebiotics in colonic in-flammation and can be adapted to the study of other inflammations.

Session 4 Session 4

42 4342

O27 Virulence mechanisms of Enterohemorrhagic Escherichia coli – The Roleof Phage-Encoded O-Acetyl Esterases

HERBERT SCHMIDT1, NADJA SAILE1, STEFANIE FEUERBAUM1, GOTTFRIED POHLENTZ2, ANDJOHANNES MÜTHING2

1 Department of Food Microbiology and Hygiene, Institute of Food Science and Biotechnology, University of Hohenheim2 Institute for Hygiene, University of Münster, Germany Corresponding author: [email protected]

Enterohemorrhagic Escherichia coli (EHEC) are the causative agents of hemorrhagic colitis (HC) and the hemolytic uremic syndrome (HUS). EHEC are mainly transmitted to humans by the ingestion of raw or undercooked food. The major pathoge-nicity factor of EHEC strains is the production of one or more Shiga toxins (Stx). Stx are encoded in the genome of lambdoid prophages. Besides these Stx-phages, further non-Stx-encoding lambdoid prophages are present in varying num-bers in the chromosome of EHEC strains. The foodborne EHEC O157:H7 strain EDL933 and the E. coli O104:H4 outbreak strain LB226692 harbor numerous lambdoid prophages, some of them encoding Stx. The stx genes are located in distinct positions close to the antiterminator Q in the late transcribed region of the respective pro-phages. Upon induction of the prophages, stx is cotranscribed together with the late transcribed phage genes.We could show that in many EHEC strains, a large open reading frame is located in 3’-direction close to the stx genes. This large open reading frame is homologous to the chromosomal nanS gene, the latter of which is present in most E. coli strains. NanS is an esterase, which is able to cleave an acetyl residue from 5-N-acetyl-9-O -acetyl neuraminic acid (Neu5,9Ac2), a sialic

acid derivative occurring in intestinal mucus. The remaining Neu5Ac can then be used as a carbon source. Bioinformatic analysis has shown that in the chromosome of different EHEC strains large and varying numbers of nanS homologs are present, which we have designated nanS-p. We could verify that recombinant gene products of these genes function as O -acetyl esterases. Growth experiments with E. coli O157:H7 strain EDL933 demonstrated that, by using these enzymes, the strains were capable to grow well on Neu5,9Ac2 as a carbon source, but mutants with deletion of all seven nanS-p alleles did not. E. coli O104:H4 strain LB226692 harbors fivenanS-p genes and we could show that it canoutcompete commensal E. coli when Neu5,9Ac2

was used as carbon source. Moreover, analysis ofbovine maxillary gland mucin (BSM) degradationby recombinant NanS-p enzymes using NanoESIMS demonstrated cleavage of up to three acetylresidues from sialic acids.We hypothesize that multiple prophage-locatednanS-p genes represent a mobile gene pool,improving substrate utilization in the intestinalmucus and therefore growth and maintenanceof the infecting bacterial population in the largeintestine.

43

O28 The Reciprocal Interactions between Tea Phenolics and Human GutMicrobiota

ZHIBIN LIU1,2,3, MARIEKE ELISABETH BRUINS3, LI NI2, AND JEAN-PAUL VINCKEN1

1 Laboratory of Food Chemistry, Wageningen University, P.O. Box 17, 6700 AA Wageningen, The Netherlands2 Institute of Food Science and Technology, Fuzhou University, Fuzhou 350108, China3 Food and Biobased Research, Wageningen University, PO Box 17, 6700 AA Wageningen, The Netherlands Corresponding authors: [email protected]; [email protected]

Tea, originating from the leaves of Camellia si-nensis, is the most widely consumed beverage in the world. A considerable number of epide-miological and human intervention studies has demonstrated the importance of regular tea con-sumption in human health. The pharmaceutical benefits are generally attributed to the phenolic compounds in tea. However, the low bioavailabi-lity of phenolics has long been recognized, which does not seem in accordance with their in vivo bioactivities.The interaction of tea phenolics and gut microbiota has long been overlooked and might provide clues to understand the health-beneficial effects of tea phenolics. In this study, phenolics from green tea, oolong tea and black tea were used to intervene the development of obesity of mice. After the intervention, gut microbiota changes were also investigated. It was revealed that besides the profound anti-obesity effect of tea phenolics, they also substantially increased the diversity of gut microbiota and altered the structure.

Linear discriminant analysis effect size algorithm identified 30 key phylotypes in response to high-fat diet and tea, including Alistipes, Rikenella, Lachnospiraceae, Akkermansia, Bacteroides, Allobaculum, Parabacteroides, etc. Spearman’s correlation analysis indicated that these key phylotypes might have a close association with the obesity related indexes of the host.In addition to the in vivo study, the degradation pathway of (‒)-epigallocatechin-3-O -gallate, a typical tea phenolics, in mimic colonic environ-ment were also studied. It was revealed that va-lerolactones and a series of phenolic acids were the predominant metabolites of (‒)-epigalloca-techin-3-O -gallate. These microbial metabolites are readily absorbed in the large intestine and might exert health benefits to the host, rather than their original forms.Summarizing, the reciprocal interactions bet-ween tea phenolics and gut microbiota might eventually exert some health benefits to the host.

Session 4 Session 4

44 4544

O29 Bacteriophages in Food Products and Their Influence on the Human Intestinal Microbiota

MEIKE SAMTLEBE1, NATALIA WAGNER2, HORST NEVE2, CHARLES M.A.P. FRANZ2, AND JÖRG HINRICHS1

1 Department of Soft Matter Science and Dairy Technology, Institute of Food Science and Biotechnology, Universität Hohenheim, Stuttgart, Germany 2 Department of Microbiology and Biotechnology, Max Rubner-Institut, Kiel, Germany

Corresponding author: [email protected]

Potential applications of bacteriophages for therapeutic use are nowadays receiving increasing attention, as phages reveal valuable properties: Phages are i) ubiquitous in the environment, ii) can be easily isolated and propagated and iii) have narrow host ranges. Therefore, they may be suitable means to control and minimize undesirable bacteria. Phages have previously been used to control bacterial diseases, and more recently the application of phages in food processing to reduce pathogens and spoilage bacteria has been approved. The human gut contains about 1015 individual phage particles, and the gut phage populations of individuals vary significantly in terms of phage biodiversity and phage titers. Various studies have proposed a correlation between the gut phageome, human health and diseases. Nevertheless, the targeted application of phages in the human gastrointestinal tract faces numerous challenges, i.e., their limited host ranges, bacterial resistances to phages, manufacturing issues, delivery systems and sensitivity to gastrointestinal conditions.

Our hypothesis is that phages that are specifical-ly integrated into food matrixes may shape and modulate the microbiota associated with the hu-man gastrointestinal tract. Hence, a preliminary research study aimed at encapsulating phages to improve their viability under gastrointestinal acid conditions and, hence, delivering them to the in-testine in active form. Lactococcus lactis phages were selected as a simple model system. The effects of different encapsulation techniques, gastrointestinal pH and enzymes were investi-gated in in vitro experiments simulating human digestive conditions. The data obtained indicated that - in comparison to free phages-encapsulated phages are not inactivated during their transit of the stomach. Furthermore, under simulated inte-stine conditions, an effective release of phages from the capsules was achieved. In a subsequent study, the stability of encapsulated and non-en-capsulated phages were analyzed in a dynamic in vitro gastrointestinal model simulating condi-tions of the human upper gastrointestinal tract. First results of this study will be presented and discussed.

45

O30 Effect-Directed Analysis of Functional Food Ingredients

GERTRUD E. MORLOCKChair of Food Science, Institute of Nutritional Science, and Interdisciplinary Research Center (IFZ), Justus Liebig University Giessen, Heinrich-Buff-Ring 26-32, 35392 Giessen, GermanyCorresponding author: [email protected]

On the one hand, the separation of thousands of compounds in a complex extract is thrilling, but may be still unsatisfactorily due to coelu-tions. Hence, the question arises where to stop in high-sophisticated separation science? Which technical effort is economically justifiable in routine? On the other hand, the separation itself does not imply an effect-directed answer to questions such as “Which compounds out of the thousands are effective?” In contrast to high-sophisticated, comprehensive separation science, a simple, streamlined metho-dology is presented that is able to answer these effect-directed questions without the need for a perfect separation [1, 2].The first part of the streamlined methodology is an effect-directed screening of up to 22 raw ex-tracts in parallel. It is a non-targeted bioprofiling (HPTLC-UV/Vis/FLD-bioassay). Depending on the bioassay or enzymatic assay selected, for example, antibiotics, estrogens, androgens and

inhibitors of acetylcholinesterase, α-/ß-gluco-sidase, α-amylase, xanthine oxidase or tyrosi-nase are discovered in complex samples [3-5]. Examples of the HPTLC-UV/Vis/FLD-(bio)assay methodology are given for functional food ingre-dients, taking 3 to 20 min per sample for the dis-covery of bioactive components. The second part is a highly targeted characterization of the effective compounds discovered via the hyphenation to structure elucidating techniques [6]. Information on effec-tive compounds in a complex sample and their sum formulae can be obtained from a single chromatographic run. HPTLC may serve as a survey on effect-directed components in complex samples. Benefits result from the side-by-side sample comparison, matrix-tolerance, avoidance of carry over and of discrimination as well as the always fresh adsorbent, the comparatively low-tech workflow and the multifold evaluation of the separated sample saved on the plate.

References: Samtlebe, Meike; Denis, Sylvain; Chalancon, Sandrine; Atamer, Zeynep; Wagner, Natalia; Neve, Horst et al. (2018): Bacterio-phages as modulator for the human gut microbiota: Release from dairy food systems and survival in a dynamic human gastroin-testinal model. In: LWT - Food Science and Technology 91, S. 235–241. DOI: 10.1016/j.lwt.2018.01.033.

References: [1] Morlock, G.E.: Bioassays and further effect-directed detections in chromatography, in Worsfold P.J., Poole, C., Townshend,A., Miro, M. (Eds.): Reference Module in Encyclopedia of Analytical Science Third Edition, Elsevier Science, Amsterdam, in print[2] G.E. Morlock, ACS Symposium Series 1185 (2013) 101-121.[3] G.E. Morlock, I. Klingelhöfer, Anal. Chem. 86 (2014) 8289–8295.[4] I. Klingelhöfer, G.E. Morlock, Anal. Chem. 87 (2015) 11098−11104.[5] M. Jamshidi-Aidji, G.E. Morlock, Anal. Chem. 88 (2016) 10979−10986.[6] I. Yüce, G.E. Morlock, J. Chromatogr. A 1469 (2016) 120-127.

Session 4 Session 4

46 4746

O31 Analyses of the Chemical and Oenological Factors Influencing the Taste Characteristics of Wines Applying Time Related Sensory Methods

ULRICH FISCHERInstitute for Viticulture and Oenology, Dienstleistungszentrum Ländlicher Raum (DLR) – RheinpfalzBreitenweg 71, D-67435 Neustadt an der Weinstrasse, GermanyCorresponding author: [email protected]

The sensory evaluation of wine is a non-targeted analytical method, which is able to bridge the gap between the chemical composition of wine and flavor perception of connoisseurs and consumers. Traditional assessments such as dif-ference tests or descriptive analysis provide only a snapshot of the dynamic process during consumption of food and beverages. Time re-lated methods such as time intensity or temporal dominance of sensations however, quantify the perceived gustatory, orthonasal olfactory and trigeminal modalities along a time axis. This presentation will highlight the benefits of these time related methods in context of wine con-stituents responsible for the bitter taste, the effect of partial dealcoholization of wine, skin contact during grape processing and aging of red wine in different oak barrels.Time intensity records over time the change in intensity of one sensory modality alone. This is useful to study in detail temporal properties of bitterness or astringency of phenolic compounds present in wine or the impact of human physiology such as salivary flow. Applying this technique

it became evident that wine consumer with low salivary flow rates perceive the same phenolic stimulus by far more bitter and astringent as well as for a longer time period. However, recording only one sensory property alone poses the risk that other perceived modalities may interfere with the evaluated attribute. To overcome this dumping effect, temporal dominance of sensations provides an array of six to ten oral-ly perceived attributes of which the one is dyna-mically selected, which is perceived as the most dominant. Partial dealcoholization of wine clearly diminished bitter and alcoholic aftertaste in white and red wines and enhanced fruity and sour perception. Extending skin contact however did not reinforce bitterness perception, but lowered sourness. In contrast, diminishing grape solids in the grape juice prior fermentation lead to a significant decline of bitter aftertaste in white wines.In conclusion, assessment of temporal properties of wine is essential to evaluate winemaking and quality and yields new insights, which are not revealed by traditional static sensory approaches.

O32 Comprehensive Strategies for Food Authentication

MARKUS FISCHERHamburg School of Food Science, Universität Hamburg, Germanywww.hsfs.org, www.food-profiling.orgCorresponding author: [email protected]

Food fraud, defined by Spink and Moyer in 2011, as the “deliberate and intentional substitution, ad-dition, tampering, or misrepresentation of food, food ingredients, or food packing” or “false or misleading statements about a product for eco-nomic gain” is a growing challenge these days, but not really new and can be tracked down till to the ancient world. However, together with increa-singly complex commodity flows, triggered by the rising globalization of markets, cases of falsifying food products have risen at the same time, as recently shown by the worldwide OPSON opera-tions of Europol and Interpol. In this scope, fal-sified food valued at app. 230 million Euro was seized during OPSON VI in 2016/17, which until then had been the greatest case in the food area. Food fraud is usually profit-motivated and affects all types of products and all regions of the world. The most frequently counterfeited products are oils especially olive oil, milk, honey and spices. Economically motivated adulteration (EMA) is powered by the fact that the consumer is willing to spend more money for extraordinary traits. In this course, deliberate and intentional false de-claration of the geographical origin, misleading statements about the production process (orga-nic/conventional production), and false or mislea-ding statements regarding the biological identity (species, varieties) are the main issues.

Food can be unambiguously described using a sufficient number of valid and stable bi-omarkers (sequences, molecules or elements/isotopes). Depending on the question, the entirety of the DNA (genome), the proteins (proteome), the me-

tabolic products (metabo-lome) and the element or isotopic profiles (isotopolome) are analyzed. The technologies used (genomics, proteomics, metabolomics and isotopolomics) are derived from the above-mentioned terms. Since the re-sulting hypothesis-free and highly resolved mo-lecular finger-prints are unambiguous, it is pos-sible to distinguish between a reference and an unknown sample. However, in this approach comparatively large amounts of data are obtained, which must be reduced by software-based methods to the key components with the greatest variance between different sample populations. The differences (marker groups) identified in this way can subse-quently be determined quantitatively by directed, so-called targeted analyzes. More particularly, due to the holistic approach, i.e. by the crosslin-ked and multi-dimensional detection of all re-levant molecular and atomic levels, a unique, systemwide and forgeryproof image of a sample is obtained and on this basis its individuality is clearly defined.To give the consumer peace of mind, more re-liable analytical strategies and solutions are required to address the parameters mentioned above. This process must also attend the fact that many of our “modern-day fraudsters” also have a scientific background and a sound wor-king knowledge of the methods used within cor-porate quality control or regulatory surveillance.

The talk is going to give an overview about state-of-the art technical approaches to cover the ori-ginality of food.

Session 5 Session 5

References[1] Sokolowski, M., Fischer, U. (2012): Evaluation of bitterness in white wine applying descriptive analysis, time-intensity analy-

sis, and temporal dominance of sensations analysis. Anal Chim Acta.. 732, 46-52.[2] Sokolowski, M., Rosenberger, A., Fischer, U.(2015) Sensory impact of skin contact on white wines characterized by

descriptive analysis, time–intensity analysis and temporal dominance of sensations analysis, Food Qual. Pref., 39, 285-297.

48 4948

O33 Fungal Enzymes for Safer Food

RALF G. BERGERInstitute of Food Chemistry, Gottfried Wilhelm Leibniz University Hannover, Callinstraße 5, D-30167 Hannover, Germany Corresponding author: [email protected]

Traditional food processes, such as sulphuring, curing or smoking often inevitably involve the formation of unwanted side-products, such as nitrosamines, polycyclic aromatic hydrocarbons or aromatic amines. Enzymes are the perfect tools to selectively intervene into the multi-component chemistry of foods. Contaminants or risk compounds may be reduced, and beneficial compounds, such as flavours, may be fortified. The problem is to obtain the right enzyme in sufficient quality and quantity. In the past decade, it was found that higher fungi, especially Basidiomycota, are a rich source of enzymes with unique properties. Recent examples are:

■ The preparation of savoury protein hydroly-sates using peptidase cocktails instead ofhydrochloric acid,

■ The creation of better digestible coffee byenzymatic degradation of chlorogenic acids,

■ The replacement of gelatine by firm gels fromcross-linked wheat bran extract,

■ The conversion of a side-stream from winemaking to a natural antimicrobial,

■ The enzymatic improvement of doughstructure without baking agents,

■ The generation of natural volatile flavours,such as nootkatone,

■ The replacement of traditional smoking by acold smoke flavour,

■ The reduction of sodium chloride by thegeneration of salt-taste enhancing peptidesusing cleavage specific peptidases.

Savings of energy, water, and chemicals often go along with the new bioeconomic processes. At the same time, high-volume side-streams of industrial food processing, such as brans, peels, molasses find new economic uses. A problem to be solved is the often low yield of heterologously expressed basidiomycetous enzymes.

49

O34 Bioflavor Production with Basidiomycetes: Potential in Developing NovelNonalcoholic Beverages

YANYAN ZHANG1, MARCO ALEXANDER FRAATZ2, AND HOLGER ZORN2

1 Department of Flavor Chemistry, Institute of Food Science and Biotechnology, University of Hohenheim, Fruwirthstr. 12, Stuttgart 70599, Germany2 Institute of Food Chemistry and Food Biotechnology, Justus Liebig University, Heinrich-Buff-Ring 17, Giessen 35392, Germany


Basidiomycetes represent the highest developed class of fungi. They are able to synthesize phar-macological relevant secondary metabolites, na-tural flavor compounds, and highly sought after enzymes. Different from microorganisms used in traditional fermented beverages, such as yeasts, lactic acid bacteria, Acetobacter species, and lower fungi, basidiomycetes possess the unique capability of producing a wide range of natural flavors in submerged and solid-state cultures. Driven by consumers’ demand for food delivering hedonic flavors, edible basidiomycetes produ-cing pleasant bioflavor compounds are ideal can-didates to develop novel fermentation systems for producing nonalcoholic fermented beverages.Utilization of the diversity of basidiomycetes novel fermentation systems were established based on the different substracts such as un-hopped wort. The optimal combinations with their correspon-ding fermentation times were screened by senso-

ry evaluation. Afterwards, aroma compounds of the beverages with pleasant flavors were isola-ted by liquid-liquid extraction (LLE) and by headspace solid phase microextraction (HS-SPME). The key odor-active compounds were analyzed by aroma extract dilution analysis (AEDA) using a gas chromatography system equipped with a tan-dem mass spectrometry detector and an olfacto-ry detection port (GC-MS/MS-O). For HS-SPME, a revised AEDA method was developed by se-quentially increasing the GC inlet split ratio (aro-ma dilution analysis, ADA). The key odorants of the beverages were analyzed in kinetic studies, quantified, and verified by aroma reconstitution. The biosynthetic pathway of the most important odorant of a beverage was elucidated by means of isotopic labeling experiments. To prepare for the market introduction of the beverages, com-prehensive safety assessments and nutritionally relevant parameters were performed.

References: Nieter A., Kelle S., Linke D., Berger R. G.: A p-coumaroyl esterase from Rhizoctonia solani with a pronounced chlorogenic acid esterase activity, New Biotechnology 2017, 37, 153–161. D. Linke, S. J. L. Riemer, S. Schimanski, A. Nieter, U. Krings, R.G. Berger: Cold generation of smoke flavor by the first phenol aciddecarboxylase from a filamentous ascomycete - Isaria farinosa, Fungal Biol. doi.org/10.1016/j.funbio.2017.05.006.K. Schulz; A. Nieter; A.-K. Scheu; J. L. Copa-Patiño; L. Popper; R. G. Berger: A type D ferulic acid esterase from Streptomyceswerraensis affects the volume of wheat dough pastries. Appl. Microbiol. Biotechnol. 2018, 102(3), 1269-1279.Th. Detering, D. Linke, S. Gounel, N. Mano, R. G. Berger: Laccase-catalysed cleavage of malvidin-3O-galactoside to 2,6-dime-thoxy-1,4-benzoquinone and a coumarin galactoside, Mycol. Prog., doi.org/10.1007/s11557-018-1380-y.M. Siebert, R. G. Berger, A. Nieter: Enzymatic mitigation of 5-O-chlorogenic acid for an improved digestibility of coffee, FoodChem. 2018, 258, 124-128.R. Bel-Rhlid, R. G. Berger, I. Blank: Bio-Mediated Generation of Food Flavors – Towards Sustainable Flavor Production Inspired byNature, Trends Food Sci. Technol. 2018, 78, 134-143.

Session 5 Session 5

50 5150

O35 Innovative Products from Fungal Cultures

HOLGER ZORNInstitute of Food Chemistry and Food Biotechnology, Justus Liebig University, Heinrich-Buff-Ring 17, Giessen 35392, GermanyCorresponding author: [email protected]

Based on their unique biochemical potential, their non-toxicity, and their aerobic growth, edi-ble mushrooms represent ideal candidates for the development of food and food ingredients. Submerged cultures of basidiomycetes may e.g. be employed for the secretion of food enzymes and the generation of natural flavours. Apart from that, options for upcycling of various agricultural side streams to valuable proteins will be discus-sed.

To replace pregastric esterases derived from the tongue of goats in the production of cheese, numerous basidiomycetes were screened for the secretion of lipases with the desired cata-lytic properties. A novel esterase derived from Flammulina velutipes was biochemically characterized and its encoding cDNA was cloned. Application tests revealed highly similar characteristics to those of the traditional goat enzymes.

A number of methyl branched aldehydes impart interesting flavor impressions, and especially 12-methyltridecanal is a highly sought after fla-voring compound. To produce natural branched

aldehydes, various fungi were screened for the formation of iso and anteiso fatty acids. Suitable fungi were successfully grown in submerged cultures. After lipase-catalyzed hydrolysis, the released fatty acids were converted to the corres-ponding methyl branched aldehydes by means of an α-dioxygenase. Their flavour impressions and threshold values were determined by GC-MS-olfactometry.

Basidiomycetes were grown submerged using side streams of the food industry as the sole carbon source, and the fungal mycelia were evaluated for their nutritional properties. An animal study was performed using Zucker rats. Rats that were fed with 5% P. sajor-caju showed significantly lower levels of liver triacylglycerols and cholesterol when compared to rats that were fed the control diet. Histopathological examinations confirmed these results. A microarray analysis of the liver revealed significantly higher mRNA expression levels of some genes of the β-oxidation and the lipid secretion pathways while reporting lower mRNA expression levels of some genes of the cholesterol synthesis pathway.

51

Session 5

52 53

ABSTRACTS

POSTER

54 5554

P1 Investigating a Dietary Supplement Administered for Histamine Intolerance

LUCAS KETTNER, INES SEITL, TIMO STRESSLER, AND LUTZ FISCHERInstitute of Food Science and Biotechnology, Department of Biotechnology and Enzyme Science, University of HohenheimCorresponding author: [email protected]

An emerging subarea within food intolerances is the sensitivity of the human body towards the biogenic amine histamine. The ingestion of small amounts of exogenous histamine can lead to adverse physiological reactions like diarrhea, headaches or itching in some susceptible in-dividuals. This intolerance derives from a dise-quilibrium between the amount of ingested histamine and the capacities of the histamine degrading enzyme diamine oxidase (DAO) in the human intestine. If DAO is not available in sufficient amounts, histamine accumulates in the human bloodstream and causes different allergy-like reactions. Individuals that suffer from this sensitivity can attain relief mainly by being abstinent from histamine containing foods. As a gleam of hope for the affected individuals, a dietary supplement that contains the histamine degrading DAO enzyme from a pig kidney extract is commercially available and can be taken before histamine containing meals. Our aim was to investigate this dietary supplement regarding its

capability to degrade histamine in a buffered test system with histamine concentrations that can be found in typical histamine containing foods (150 ppm). Since the dietary supplement has to release the enzyme in the human intestine, the capsule preparation must be able to withstand the acidic environment of the human stomach, which was investigated utilizing simulated gastric fluid. Within 5 hours, the applied amount of histamine was reduced merely by 12 %. As a comparison, a partially purified diamine oxidase extract from pig kidney achieved a histamine reduction of 24 % at the same time. Our histamine bioconversion experiments showed that for entire complementation of the native DAO in histamine sensitive humans, the DAO amount in the cap-sules might not be sufficient when histamine-rich foods were ingested. Therefore, it would be advisable to increase the supplemented DAO amount for a reliable reduction of exogenous histamine in the intestine.

55

P2 Enzymatic Production of Emulsifying Whey Protein Hydrolysates without the Need of Heat Inactivation

JACOB EWERT, ANJA LUZ, VERONIKA VOLK, TIMO STRESSLER, AND LUTZ FISCHERInstitute of Food Science and Biotechnology, Department of Biotechnology and Enzyme Science, University of HohenheimCorresponding author: [email protected]

Whey proteins represent 18-20 % of the total milk nitrogen content. Because of their excellent amino acid composition in view of the nutritional value, whey proteins evolved from a difficult to dispose by-product to a valued food ingredient. Nonetheless, one of the greatest drawbacks to the widespread application of whey proteins in food systems is their unreliable technofunctional behavior, especially in foams and emulsions. A tool to modify these properties is an enzymatic hydrolysis with peptidases. However, most studies covering the influence of the hydrolysis on whey proteins use a heating step (>65 °C) to inactivate the enzyme. This already leads to irreversible pro-duct changes, such as protein denaturation and an increased viscosity. Consequently, the heating and the effect of the hydrolysis influence the protein properties and their emulsifying behavior. As an alternative for a food-grade process, the enzymatic hydrolysis can be performed at an acidic pH(≤ 3) and at mild temperatures (≤45 °C). Therefore,peptidases, which are active at these conditions,

have to be applied. A suitable peptidase for such hydrolysis is Aspergillopepsin I (EC 3.4.23.18). Furthermore, this endopeptidase is favorable for a technofunctional product, due to its cleavage specificity for hydrophobic amino acids at the P1 and P1’ position. This can result in peptides, increased in their hydrophobicity and consequently a hydrolysate with improved interfacial behavior. In contrast, peptides with hydrophobic amino acids are generally perceived as bitter. A possibility to reduce the bitterness is the simultaneous ap-plication of exopeptidases during the hydrolysis. Therefore, by choosing the appropriate peptidase preparation including endo- and exopeptidases an improved technofunctional hydrolysate with re-duced bitterness can be produced.

In a recent study [manuscript in preparation], such peptidase preparations were biochemically inve-stigated and compared regarding a whey protein hydrolysis.

Poster Session Poster Session

56 5756

P3 Fungal Fermentation Induced Natural Flavor – an Insight from Bioeconomy

M. RIGLING, K. ZWINKMANN, A.K. NEDELE, F. FRIEDMAN, AND Y. ZHANGInstitute of Food Science and Biotechnology, Department of Flavor Chemistry, University of Hohenheim,Fruwirthstraße 12, 70599 Stuttgart, GermanyCorresponding author: [email protected]

Tea is after water the second most consumed beverage in the world. In legal terms ‘tea’ in-cludes leaves, buds, and stems of the tea plant Camellia sinensis L.O. Kuntze (Theaceae) and corresponding derived products. Nowadays, the annual consumption of tea reaches about 5 tril-lion cups (14 billion cups per day). Among them, ready-to-drink tea gains more popularity. During the process about 100,000 t of tea waste are pro-duced annually by the companies. The waste in general was burned or dumped instead of recy-cling utilization in the past. Since burning or dum-ping tea waste brings negative effects on the en-vironment, alternative applications of tea waste (e.g. an adsorption material for heavy metals, an available source for the extraction of caffeine, an ingredient of media for harvesting fruiting bodies of mushroom) are developed in the field of en-

vironmental protection, pharmacy, or food.Basidiomycetes as the highest developed fungi own enormous enzymatic systems for diverse bioflavor synthesis. Tea waste contains cellulose as well as trace amounts of crude protein, fat, and carotenoids. Bioflavor with high economic value could therefore be generated de novo or by transformation of precursors of tea waste. In our study, hedonic and intense aroma was observed in a series of tea waste after solid-state fermentation with basidiomycetes. Subsequently, interesting bioflavors were identified using the headspace solid phase microextraction (HS-SPME) in combination with a gas chromatography system equipped with a mass spectrometric de-tector and an olfactometry detection port (GC-MS-O).

57

P4 Bioflavor Generation by Basidiomycetes: Enhancing Flavor Attributes of Soy Drink

ANN-KATHRIN NEDELE, MERVAT ALMUHAMMAD, MARINA RIGLING, AND YANYAN ZHANGInstitute of Food Science and Biotechnology, Department of Flavor Chemistry, University of Hohenheim, Fruwirthstr. 12, 70599 Stuttgart, Germany Corresponding author: [email protected]

Soybean (Glycine max) is a legume which con-tains high amounts of proteins, vitamins, dietary fiber, and minerals. Soybean-based beverages are highly consumed in East Asia. In the last few decades, soy drink as a substitute for dairy milk have become popular especially for lactose intole-rant or milk allergic persons in western countries. However, its unpleasant green and beany off-fla-vor is a large challenge regarding the consumer acceptance in these countries. Although various physical, chemical, and microbial techniques have been applied for modification of aroma properties of soy drink, no research regarding the aroma en-hancement by use of the highest developed fungi is published so far. As ideal candidates in white biotechnology, edible basidiomycetes might be po-tent sources for enhancing aroma properties of soy drink by the generation of a series of pleasant na-

tural flavors, as their “volatile spectra” are closest to the ones of plants. Twenty-one different species of basidiomycetes were screened in a fermenta-tion system based on soy drink. Most of the se-lected basidiomycetes grew well and reduced or suppressed the undesired green and beany odor impression of soy drink. More importantly, highly hedonic flavors like bitter almond, nutty and fruity were perceived after the fermentation with speci-fic basidiomycetes. However, some fermented soy drink imparted unpleasant odor notes (painty, al-coholic, and yeasty) as well. The key odor-active compounds in the fermented soy drink with an in-teresting flavor were identified by means of stir bar sorptive extraction (SBSE) combined with a gas chromatography-mass spectrometry-olfactometry system(GC-MS-O).


58 5958

P5 Enzymatic Mitigation of 5-O-Chlorogenic Acid for an Improved Digestibilityof Coffee

M. SIEBERT, A. NIETER, AND R. G. BERGERInstitute of Food Chemistry, Gottfried Wilhelm Leibniz Universität Hannover,Callinstraße 5, 30167 Hannover, GermanyCorresponding author: [email protected]

References: A. Farah, M. C. Monteiro, V. Calado, A. S. Franca, L. C. Trugo (2006). Food Chem 98, 373-380.A. Nieter, S. Kelle, D. Linke, R. G. Berger (2017). N Biotechnol 37, 153-161.M. Rubach, R. Lang, G. Bytof, H. Stiebitz, I. Lantz, T. Hofmann, V. Somoza (2014). Mol Nutr Food Res 85, 1370-1373.M. Siebert, R. G. Berger, A. Nieter (2018). Food Chem 2018, 258, 124-128.

Coffee is one of the most popular beverages and consumed in large amounts all over the world. Nevertheless, sensitive people complain about heartburn and stomach irritation even after drin-king a small cup. The cause and mechanism of these symptoms are still unclear. Various sub-stances are under suspicion. Among them are chlorogenic acids and high molecular roasting products (Rubach et al. 2017). Chlorogenic acids are also an important factor of coffee quality. A low cup quality was related to a high 5-O -chlo-rogenic acid (5-CQA) concentration (Farah et al. 2006). A gentle roasting process reduces the chlorogenic acid content in coffee. However, roasting neither selectively removes chlorogenic acids, nor can roasting conditions be decoupled from the formation of desired volatile compounds and undesired risk compounds. Furthermore, a gentle roasting process leads to higher costs for the coffee industry. Enzymatic treatment of coffee presents a feasible approach to degrade chlorogenic acids without changing the roasting parameters. Nieter et al. (2017) characterized a p-coumaryl esterase from

Rhizoctonia solani (RspCAE) possessing the ability to hydrolyse 5-CQA, which is the most abundant chlorogenic acid in coffee. Treatment of roasted coffee powder with RspCAE led to a decline of up to 98 % of 5-CQA. HPLC-UV showed that simultaneously the caffeic acid concentration increased. Effects on aroma were determined by means of aroma extract dilution analysis. The aroma extracts were diluted step-wise 1:10 (v/v). Aroma active compounds with flavour dilution factors (FD-factors) between 1 and 10,000 were determined. Both extracts were similar regarding odour quality and intensity. FD-factors of treated and control extract differed in four volatile compounds (ethyl pyrazine, maltol, 1furfurylpyrrol, 2,5-dimethylphenol) only. Effect on flavour and taste was evaluated by sensory tests. No significant differences were perceived, and no off-flavour and offtaste was noted. Consequently, the enzyme treatment offers a technically feasible approach to improve the quality of coffee beverages by reducing 5-CQA concentration without significantly affecting the aroma and taste profile (Siebert et al. 2018).

59

P6 Regulating Sesquiterpene Production of the Basidiomycete Tyromyces floriformis in Submerged Cultures

M. MEYER, U. KRINGS, AND R. G. BERGERInstitute of Food Chemistry, Gottfried Wilhelm Leibniz Universität Hannover,Callinstraße 5, 30167 Hannover, GermanyCorresponding author: [email protected]

References:B. G. Abraham, R. G. Berger (1994). J Agric Food Chem 42, 2344 - 2348M. A. Fraatz, S. J. L. Riemer, R. Stöber, R. Kaspera, M. Nimtz, R. G. Berger, H. Zorn (2009). J Mol Cat B: Enzymatic 61, 202 – 207J. M. Jurado, O. Ballesteros, A. Alcázar, F. Pablos, M. J. Martín, J. L. Yílchez, A. Navalón (2007). Talanta 72, 506 – 511R. Kramer, W. R. Abraham (2012). Phytochem Rev 11, 15 - 37M. Parker, A. P. Pollnitz, D. Cozzolino, I. L. Francis, M. J. Herderich (2007). J Agric Food Chem 55, 5948-5955

Basidiomycetes are well known to produce and secrete a large variety of metabolites, among them potent volatile flavours (Abraham and Ber-ger 1994). Plant-like, but also many unique fungal sesquiterpenes constitute a highly diverse group of the fungal volatilome (Fraatz et al. 2009). While sesquiterpene biosynthesis is well characterized, pathway regulations are less clear (Kramer and Abraham 2012).The sesquiterpene producing basidiomycete Ty-romyces floriformis was grown in submerged cul-tures with different additives. Volatile production during cultivation was recorded using sequential stir bar or solvent extraction. Qualitative and quan-titative measurements of the concentrated extracts and stir bars were performed by TDS-GC- FID and TDS-GC-MS/O analyses.The aroma profile comprised more than 20 sesquiterpenoids. These were classified into products derived by 1,10 and 1,6 cyclation of the farnesyl pyrophosphate. Major product was the fruity α-ylangene inter alia known from Ylang-Ylang oil, cloves, grapes and wine (Jurado et al. 2007,

Parker et al. 2007). Unexpectedly, the addition of arabinoxylanes or other polysaccharides down-regulated the terpene yield. While the addition of the respective monosaccharides did not show the same effect, all examined dimers and oligomers of Arabinose and Xylose decreased the product yield. On contrary, lipids and lipid rich additives showed up-regulating effects, probably by filling up the acetate pool and facilitating product accumulation. Analyses of the biosynthetic background will follow.

Figure 1: Structure of the fruity flavour compound α-ylangene, the main product of T. floriformis.


60 6160

P7 Secretion of Active Lactose Hydrolyzing Enzymes in Y. lipolytica

PAUL SWIETALSKI, INES SEITL, AND LUTZ FISCHERDepartment of Biotechnology and Enzyme Science, Institute of Food Science and Biotechnology, University of Hohenheim, Stuttgart, GermanyCorresponding author: [email protected]

The use of various food relevant enzymes, like lactose hydrolyzing enzymes is a rapid increasing area. The growing demand for enzymes used in the food industry requires therefore the establis-hment of improved production organism. The di-morphic yeast Yarrowia lipolytica is a promising host for the production of heterologous proteins, due to its ability to produce high amounts of ac-tive extracellular enzymes.In this work three different lactose hydrolyzing enzymes with varying molecular weight (55 - 120 kDa) were heterologously expressed in Y. lipo-lytica to compare theirextracellular production in this yeast. The selected enzymes are originally derived from phylogenetic diverse microorganis-ms. To obtain a comparative approach, the three genes were codon optimized for Y. lipolytica and embedded in identical expression cassettes. For guiding the recombinant enzymes into the secre-tory pathway, a 22 amino acid long preXPR2 signal sequence was fused to the respective

N-terminus. The expression cassettes were inte-grated into the acid extracellular peptidase (AXP)locus of the Y. lipolytica PO1f strain using theCRISPR/Cas9 System. Recombinant Y. lipolyti-ca strains carrying one copy of either the small(55kDa) or large (120 kDa) lactose-hydrolyzingenzymes, were cultivated for 78h in 1 L multiforsbioreactors in YPD complex media. The completeamount of produced, recombinant enzymes wasanalyzed by intra- and extracellular activity mea-surement. Furthermore, the transcription rateof the individual target gene was determinedby quantitative real-time PCR. Finally, it was ob-served that the recombinant enzyme with smallest size was secreted by far the most. However, bothlarge enzymes with molecular weights around120 kDa were secreted by Y. lipolytica as well,although in quite low amounts. Based on this pro-of of principle, further investigations have to bedone to improve the secretory capacity of Y. lipo-lytica for large and complex enzymes.

61

P8 Enzyme Catalyzed Production of Short-Chain Fructooligosaccharides by a New Recombinant Fructosyltransferase in Batch and Packed Bed Enzyme Reactors

JAN PHILIPP BURGHARDT1,2, DOREEN GERLACH3, AND PETER CZERMAK1,2,3,4


2 Faculty of Biology and Chemistry, Justus Liebig University Giessen, Giessen, Germany3 Department of Bioresources, Fraunhofer Institute for Molecular Biology and Applied Ecology (IME), Giessen,

Germany 4 Department of Chemical Engineering, Kansas State University, Manhattan, USA Corresponding author: [email protected]

Short-chain fructooligosaccharides (scFOS) are a mixture of oligosaccharides consisting of sucrose molecules, linked with fructose units, until a po-lymerization degree of 3 to 5 additional fructose units. Due to their prebiotic effect in combination with a sweetness level of 30 to 60% as compared with sucrose, scFOS are used as additives in food and feed. At industrial-scale, FOS are produced from sucrose, catalyzed by fructosyltransferases (EC 2.4.1.9), which are present in commercial en-zyme mixes like Pectinex Ultra SP-L. Using the yeast Kluyveromyces lactis we established an ex-pression system for the recombinant production of a fructosyltransferase derived from Aspergillus terreus. The expression host K. lactis has already obtained the generally regarded as safe (GRAS) status for β-galactosidase production and was therefore chosen for the recombinant enzyme pro-duction. We cultivated K. lactis in an adapted che-mically defined medium at 5L scale. The secreted fructosyltransferase was concentrated after cell

removal with an 80 kDa ceramic membrane. The concentrated enzyme solution was applied for FOS production utilizing 600 g L-1 sucrose solution in a 50 mM phosphate-citrate buffer as substrate for the 20 min batch reaction in 1.5 mL micro reaction tubes. Optimization of enzymatic catalysis condi-tions applying a D-optimal design of experiments identified the conversion optima of 50°C at pH of 5 within the investigated region. Scale up of the en-zyme catalysis was performed in 100 mL enzyme reactors and the conversion efficiency of the free enzyme solution was compared with a packed-bed reactor containing immobilized fructosyltransfera-se. Three different epoxy resin carriers (Immobead 150P, Sepabeads SP70 and ECR8285) were used for the enzyme immobilization and the catalysis of FOS was compared with respect to the immobili-zation efficiency and the activity loss of the immo-bilized enzyme. Applying the epoxy resin carriers, we demonstrated a successful immobilization of catalytic active enzymes.


62 6363

P10 Enzymatic Modification of Steviol Glycosides by Fructosyltransferase andRhamnosidase for Improved Sweetness Levels

JAN PHILIPP BURGHARDT1,2, SEBASTIAN C. SPOHNER1*, DOREEN GERLACH3, AND PETER CZERMAK1,2,3,4


2 Faculty of Biology and Chemistry, Justus Liebig University Giessen, Giessen, Germany3 Department of Bioresources, Fraunhofer Institute for Molecular Biology and Applied Ecology (IME), Giessen, Germany

4 Department of Chemical Engineering, Kansas State University, Manhattan, USA* now BASF AG, Ludwigshafen, Germany


References: Spohner S C, D Zahn, V Schaum, H Quitmann, P Czermak: Recombinant α-L-Rhamnosidase from Aspergillus terreus in selec-tive trimming of α-L-Rhamnose from Steviol Glycosides, Journal of Molecular Catalysis B: Enzymatic 122:12, 248-254Spohner S C, P Czermak (2016): Enzymatic production of prebiotic fructo oligosteviol glycosides, Journal of Molecular Catalysis B: Enzymatic 131:79-84Spohner S C, P Czermak (2016): Heterologous expression of Aspergillus terreus fructosyltransferase in Kluyveromyces lactis, New Biotechnology, 33:4, 473-479

Steviol glycosides are natural sweet tasting con-stituents of Stevia rebaudiana. This plant grows naturally in the subtropical and tropical regions of western North America to South America. The main steviol glycoside in the leaves is stevioside with an amount of 5 % to 10 %. Further sweet compounds are rebaudioside A (2 - 4 %), dulco-side B (1 - 2 %) and dulcoside A (0.2 - 0.7 %). Purified steviol glycosides are a white to slightly yellow powder. They are heat-stable, pH-stable, do not ferment, and do not induce a glycemic re-sponse when ingested. In December 2011, steviol glycosides were approved for sale in Europe as sweeteners (E960). Even highly purified steviol glycosides still possess negative taste attributes such as bitterness or liquorice flavour aftertaste. The flavour notes become more prominent as the concentration increases. In this study a fungal

α-L-rhamnosidase was overexpressed in the me-thylotrophic yeast Pichia pastoris. The enzyme was applied to remove bitter tasting rhamnosyl residues from steviol glycosides. In a second approach, fructose molecules are transferred to rebaudioside A in two reaction steps to increase the sweetness level of rebaudioside A. For this purpose, an initial fructosylation of a terminal glucose unit in rebaudioside A is conducted with a native fructosyltransferase from the organism Microbacterium saccharophilum (DSM 28107). In the following step a fungal fructosyltransferase, which was overexpressed in the yeast Kluyver-omyces lactis, was applied to extend a fructo-se chain to the initial bound fructose unit. This approach with two enzymatic reaction steps de-monstrated the successful transfer of up to four fructose units to the rebaudioside A.

P9 Production of a New Recombinant Fructosyltransferase by Kluyveromyces Lactis for Prebiotic Fructooligosaccharide Synthesis

JAN PHILIPP BURGHARDT1,2, DOREEN GERLACH3, AND PETER CZERMAK1,2,3,4


2 Faculty of Biology and Chemistry, Justus Liebig University Giessen, Giessen, Germany3 Department of Bioresources, Fraunhofer Institute for Molecular Biology and Applied Ecology (IME), Giessen, Germany

4 Department of Chemical Engineering, Kansas State University, Manhattan, USA Corresponding author: [email protected]

In recent years, fructooligosaccharides (FOS) became attractive as sweeteners due to their prebiotic and low calorie properties. At industrial-scale, FOS are produced from sucrose, catalyzedby fructosyltransferases (EC 2.4.1.9), which are present in commercial enzyme mixes like Pectinex Ultra SP-L. Commercial enzyme cocktails contain a wide variety of different enzymes irrelevant for the FOS production. As alternative approach, the present study aims for the recombinant production of a fructosyltransferase, derived from Aspergillus terreus, by the yeast Kluyveromyces lactis. This yeast has already obtained the generally regarded as safe (GRAS) status for β-galactosidase production. Using K. lactis as expression system, we established a production platform secreting our target protein into the fermentation broth. The cultivation of K. lactis was performed at 30°C in both rich and chemically defined medium. Batch and fed-batch process mode was evaluated for the yeast cultivation to improve the final protein concentration.In the following purification step, the protein

solution was concentrated and afterwards dia-lyzed with potassium phosphate buffer using a ceramic ultrafiltration membrane. A major band of 80 kDa presented on SDS-PAGE corres-ponds to the target fructosyltransferase. The specificity of the enzyme was confirmed by successful production of 1-kestose, nystose and 1F-fructofranosyl-nystose applying a 2 M sucrose solution as substrate at 60°C. Produced FOS were analyzed by HPLC. Applying the purified enzyme yielded ~ 50% FOS in batch production. Increasing glucose concentration, released from sucrose during catalysis, caused fructosyltransferase inhibition. FOS solution from 100 mL batch scale was purified applying a thin ultrafiltration membrane to improve the ratio of FOS to remaining mono/-disaccharides. An abundant content of FOS in the sugar solution is crucial for the pursued prebiotic and low calorie quality. The operation mode of the enzymatic FOS production process will be further investigated to increase the FOS yield.


64 6565

P12 Selective Hydrolysis of Whey Proteins by Immobilized Trypsin in a Flow- Through Monolithic Reactor

Immobilized trypsin is a viable alternative to the free enzyme in solution for producing protein hydrolysates. To realize higher automation and throughput, an immobilized trypsin reactor (IMTR) at 8 mL scale, based on polymethacrylatemonolith with pore size 6 µm, was developed and used in a flow-through system. Optimization of operating parameters for this specific enzyme reactor could allow a better process control such that the desired hydrolysis can happen. Hence, the influences of flow rate, pH and ionic strength on the performance of IMTR were investigated to realize the selective hydrolysis of β-Lactoglobulin

(β-Lg) among whey proteins. The performance of IMTR was characterized using three descriptors: (i) efficiency, i.e. degree of hydrolysis (DH); (ii)the amount of depleted or remaining intact pro-teins, and (iii) the peptide profiles.Generally, higher flow rate contributed to a fastermass transfer so that the hydrolysis efficiencywas significantly improved. pH and ionic strengthsignificantly influenced the depletion rate ofβ-Lg, as well as the peptides profiles.The interesting result is that the immobilized tryp-sin in this case highly selectively hydrolyzed β-Lgthan α-lactalbumin at all the explored conditions.

P11 Utilization of Probiotic Bacillus coagulans in the Production of Fructo- oligosaccharides (FOS): Reduction of the Glycemic Index and Formation

of the Endospores

RONG FAN1 AND PETER CZERMAK1,2,3


2 Department of Bioresources, Fraunhofer Institute for Molecular Biology and Applied Ecology (IME), Giessen, Germany

3 Department of Chemical Engineering, Kansas State University, Manhattan, USA4 Faculty of Biology and Chemistry, Justus Liebig University Giessen, Giessen, Germany Corresponding author: [email protected]

Probiotics termed as functional food are gaining more and more interest of the food market. Lactobacilli, bifidobacteria, and Bacillus coagulans are frequently used probiotic strains, which have been demonstrated to modify and reinstate the intestinal flora, beneficially affec-ting the host health. This is mainly caused by altering the pH value, since they can convert diverse carbohydrates into lactic acid (LA). The probiotics are, therefore, often used to produce LA with anaerobic fermentation in food and chemical industry. In these microorganisms, Ba-cillus coagulans is a competitive candidate due to its thermophilic and spore-forming property. It can grow from 30 to 60°C and convert the carbohydrates via homolactic fermentation to L-lactic acid with high yield and optical purity.In our study, different operating modes andcarbon sources were applied to investigate

the fermentation of Bacillus coagulans. In the membrane bioreactor system, a stable continu-ous production of LA was achieved with the highest productivity of 8.6 g·L-1·h-1 and the high-est yield of 0.87 g·g-1 with pure glucose as a carbon source. This productivity is 2.5 times as high as the best overall productivity obtained in batch fermentations [1]. When the raw product of prebiotic fructooligosaccharides (FOS), which is a mixture of FOSs, residual sucrose, fructose and glucose [2], was applied as the carbon source, Bacillus coagulans showed a preference for the glucose than the other components, leading to a reduction of glucose in the FOS product and therefore to a reduction of the glycemic index of the mixture. Additionally, at the end of the fer-mentation, the population of the endospores achieved 1.2 × 108 mL-1 with the sporulation rate up to 81.8%.

References: [1] Fan R, M Ebrahimi, P Czermak (2017): Anaerobic membrane bioreactor for continuous lactic acid fermentation, Membranes 7:26,

http://dx.doi.org/10.3390/membranes7020026[2] Ur Rehman A, Z Kovacs, M Ebrahimi, H Quitmann, P Czermak (2016): Enzymatic production of fructooligosaccharides from

inexpensive and abundant substrates using a membrane reactor system, Separation Science and Technology, 51. pp. 1537-1545

References: Mao, Y., & Kulozik, U. (2018). Selective hydrolysis of whey proteins using a flow-through monolithic reactor with large pore size and immobilised trypsin. International Dairy Journal, 85, 96-104.

YUHONG MAO1 AND ULRICH KULOZIK1,2

1 Chair of Food and Bioprocess Engineering2 Institute of Food and Health (ZIEL)–Technology Unit, Technical University of Munich, Weihenstephaner Berg 1, D-85354, Freising-Weihenstephan, Germany Corresponding author: [email protected]


66 6767

P14 Open Source 3D Printed Add-On for Automated Elution Head-BasedHPTLC-MS

TIM T. HÄBE, DIMITRI FICHOU, AND GERTRUD E. MORLOCKChair of Food Science, Institute of Nutritional Science, and Interdisciplinary Research Center (IFZ), Justus Liebig University Giessen, Heinrich-Buff-Ring 26-32, 35392 Giessen, GermanyCorresponding author: [email protected]

The hyphenation of elution head-based high-performance thin-layer chromatography mass spectrometry (HPTLC-MS) still has to be operated manually in terms of plate positioning. Thus, an open-source 3D printed add-on is introduced [1], allowing an automatization of the TLC-MS Interface 2 from CAMAG. A newly developed software termed OC_manager controls the im-age-based assignment of elution zones and the automated positioning of the elution head on those zones. By the use of an automatic 6-port valve and four electronically controlled pneumatic valves, the elution and cleaning process can be adjusted individually and synchronized with the plate positioning and MS data acquisition. Mechanic movements were realized by three belt-driven carriages directly on the interface

base. The 6-port valve and all pneumatics were installed inside the device to preserve the compact footprint. The mean deviation (target shift) of the positioning was determined to be 160 µm for the ordered elution of 294 target zones of azophloxine on one HPTLC plate and to be 190 µm for the randomized elution order. The head cleaning control was improved, and remaining elution solvents and particles are completely transferred to the drawer. In combination with a controllable gas flow over the elution zone, cross-over contaminations or chromatogram dis-tortion by released layer particles or solvents are avoided. The presented automatization turns HPTLC-MS to be a highly efficient and repro-ducible hyphenation technique.

P13 Bioprofiling and Characterization to Bioquantification of Natural Antibio-tics by Direct Bioautography Linked to High-Resolution Mass Spectrometry

MARYAM JAMSHIDI-AIDJI AND GERTRUD E. MORLOCKChair of Food Science, Institute of Nutritional Science, and Interdisciplinary Research Center (IFZ), Justus Liebig University Giessen, Heinrich-Buff-Ring 26-32, 35392 Giessen, GermanyCorresponding author: [email protected]

Antibiotic resistance is a current challenge of public health and pharmaceutical industry. Hy-phenated planar chromatography (HPTLC-UV/Vis/FLD-EDA-HRMS) proved to be well-suited as a high-throughput bioanalytical tool that can contribute in this context. The Bacillus subtilis bioassay was directly applied in the chromato-gram to demonstrate the streamlined strategy from screening, characterization and identifica-tion to bioquantifi¬cation of natural antibiotics in root extracts of Salvia miltiorrhiza [1]. The sam-ple preparation was kept simple to let the sample extract as native as possible. The bioassay ap-plied in the chromatogram (bioautogram) eased the direct correlation of separated zones/peaks and effective zones.

Thus, the antimicrobial activity of the Salvia miltiorrhiza root extract was evident via the anti-microbial zones detected. These were characterized via chromatographic, spectroscopic and HRMS data. Inverse densitometric measurement was employed for bioquantification. The impor-

tance of two unknown antibiotics was specified via bioequivalency calculation. As a reference, cryptotanshinone was used. The overall antimi-crobial result obtained was referred to the acti-vity of two synthetic antibiotics, ciprofloxacin and marbofloxacin. These calculations were performed in a single run, meaning on the same plate.

Regular instrumentation was used. Hence, this strategy can be installed in every analytical labo-ratory without much microbiological effort. Any type of bacteria can be selected, depending on the effect of interest, and applied on the plate. Especially the linkage to microbiological assays with pathogenic bacteria will be of high relevance, in combination with HRMS/NMR/IR and bioquantification. A planar chromatographic approach for streamlined structure elucidation was recently reported [2]. The demonstrated potential of this bioprofiling can contribute to dis-covery of new antibiotics from natural sources.

References: [1] M. Jamshidi-Aidji, G. Morlock, Anal. Chem. 88 (2016) 10979−10986.[2] I. Yüce, G. Morlock, J. Chromatogr. A 1469 (2016) 120-127.

References: Häbe, T.T., Fichou, D. and Morlock, G.E., in submission.


68 69

List of Speakers

SCHEIBNER, KATRIN / KIEBIST, JANBrandenburg University of Technology (BTU) [email protected]

SCHMIDT, HERBERTUniversity of Hohenheim, [email protected]

SEBALD, KARINTechnical University of Munich (TUM) [email protected]

SPRENGER, GEORGUniversity of [email protected]

STRESSLER, TIMOUniversity of Hohenheim, [email protected]

WEISS, JOCHENUniversity of Hohenheim, [email protected]

XIE, MINGYONGNanchang [email protected]

XIONG, TAONanchang [email protected]

YAN, XINNanjing Agricultural [email protected]

YANG, RUIJIN Jiangnan [email protected]

ZHANG, CHENFuzhou [email protected]

ZHANG, HUIZhejiang [email protected]

ZHANG, YANYANUniversity of Hohenheim, [email protected]

ZHOU, PENGJiangnan [email protected]

ZORN, HOLGERJustus Liebig University Giessen [email protected]

List of Speakers

BUECHNER, KAITechnical University of Munich (TUM)[email protected]

BERGER, RALF G.University of [email protected]

BISPING, BERNWARDUniversity of [email protected]

CZERMAK, PETERUniversity of Appl. Sciences Mittelhessen, [email protected]

FENG, FENGQIN Zhejiang [email protected]

FISCHER, LUTZUniversity of Hohenheim, [email protected]

FISCHER, MARKUSUniversity of Hamburg [email protected]

FISCHER, ULRICHNeustadt Wine [email protected]

HINRICHS, JÖRGUniversity of Hohenheim, [email protected]

HU, JIELUNNanchang [email protected]

JAKOB, FRANKTechnical University of Munich (TUM)[email protected]

JIANG, BOJiangnan [email protected]

KULOZIK, ULRICHTechnical University of Munich (TUM)[email protected]

LIU, ZHIBINFuzhou [email protected]

MORLOCK, GERTRUD E.Justus Liebig University [email protected]

MU, WANMENGJiangnan [email protected]

NI, LIFuzhou University [email protected]

NIE, SHAOPINGNanchang [email protected]

OCHSENREITHER, KATRINKarlsruhe Institute of Technology (KIT)[email protected]

RUDAT, JENSKarlsruhe Institute of Technology (KIT)[email protected]

70 71

Authors & Co-Authors Index

AAlbermann, Christoph O8Almuhammad, Mervat P4

BBaumgärtner, Florian O8Becker, Thomas O24Berger, Ralf G. O33, P5-6Bisping, Bernward O2Bruins, Marieke Elisabeth O28Buechner, Kai O24Burghardt, Jan Philipp O16, P8-10

CChen, Liling O9Czermak, Peter O16, P8-11

DDelavault, André O10Ding, Qiao O26Dunkel, Andreas O25

EEbrahimi, Mehrdad O16Ewert, Jacob O18, P2

FFan, Rong P11Fang, Qingying O15Feng, Fengquin O7, ChairFeuerbaum, Stefanie O27Fichou, Dimitri P14Fischer, Lutz O14, O18,

P1-2, P7 Fischer, Markus O32Fischer, Ulrich O31Fraatz, Marco Alexander O34Franz, Charles M.A.P. O29Friedmann, Felisa P3

GGerlach, Doreen O16, P8-10Grüninger, Jens O10Guan, Qianqian O1, O5

HHäbe, Tim T. P14Han, Xin O3Hinrichs, Jörg O29Hofmann, Thomas O25Hofrichter, Martin O20Hollenbach, Rebecca O10Hu, Jielun O15, O26Huang, Danfei O26

JJakob, Frank O4Jamshidi-Aidji, Maryam P13Jiang, Bo O13, O17,

ChairJiang, M. O6

KKettner, Lucas P1Kiebist, Jan O20Krings, U. P6Kulozik, Ulrich O12, P12

LLi, Haishan O15Li, Pengfei O3Li, Qiqiong O15Liu, Changgen O5Liu, Zhibin O9, O23, O28Luz, Anja P2

Authors & Co-Authors Index

MMao, Yuhong P12Meyer, M. P6Morlock, Gertrud E. O30, P13, P14Mu, Wanmeng O17, ChairMüthing, Johannes O27

NNedele, Ann-Kathrin P3, P4Neve, Horst O29Ni, Li O9, O2 3, O28

Chair Nie, Qixing O15 Nie, Shaoping O1, O15, O26 Nieter, A. P5

OOchsenreither, Katrin O6

PPohlentz, Gottfried O27

QQian, X. O6

RRehmann, Amad Ur O16Rigling, Marina P3, P4Rudat, Jens O10

SSaile, Nadja O27Samtlebe, Meike O29Scheibner, Katrin O20Schmidt, Herbert O27Seitl, Ines P1, P7Siebenhaller, Sascha O10Siebert, Mareike P5Spohner, Sebastian C. P10Sprenger, Georg A. O8Stressler, Timo O18, P1, P2Swietalski, Paul P7Syldatk, Christoph O10

VVincken, Jean-Paul O28Vogel, Rudi F. O4Volk, Veronika P2

WWagner, Natalia O29Weiss, Jochen O22

XXiao, Yangsheng O5Xie, Mingyong O1, O15, O26,

ChairXiong, Tao O1, O5

YYan, Xin O19Yang, Ruijin O3Ye, Bin O19Yin, Junyi O26

ZZhang, Chen O9, O23Zhang, Hui O21Zhang, Wen O9Zhang, Wenbin O3Zhang, Wenli O17Zhang, Yanyan O34, P3, P4Zhao, Leizhen O19Zhao, Qiyan O3Zhou, Chaoyang O19Zhou, Peng O11, ChairZhu, Xiaoping O9Zorn, Holger O34, O35Zwinkmann, K. P3

72 73

NotesNotes

74 75

NotesNotes

Imprint Editor: University of Hohenheim, Research Center for Heath SciencesWollgrasweg 43, 70599 Stuttgart, +49 (0)711 459 24615, [email protected]://health.uni-hohenheim.dePrinted at the University of Hohenheim, KIM, September 2018Cover: University of Hohenheim

Documents

Program and Abstract Book