BIOTECHNOLOGIE ALIMENTAIRE AU MAROC · food biotechnology in morocco a. tantaoui el araki (sup’agro, casablanca) biotechnologies alimentaires au maroc: entre la recherche scientifique

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MICROBIOD 2009

International Conference Microbial Biotechnology for

DevelopmentNovember 02-05; 2009 Marrakesh – MOROCCO

FOOD BIOTECHNOLOGY IN MOROCCO

A. TANTAOUI EL ARAKI(SUP’AGRO, Casablanca)

BIOTECHNOLOGIES ALIMENTAIRES AU MAROC: ENTRE LA RECHERCHE SCIENTIFIQUE ET LA REALITE DE L’INDUSTRIE

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INTRODUCTION

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Food Biotechnology in Morocco

Introduction

- Fermented food products popular in Moroccan alimentary tradition;

- Industrialization: introduction of new products, imitation of traditional ones;

- Scientific research: characterization of traditional products, attempts to monitor their technology for industrial use

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Food Biotechnology in Morocco

Content

1- Tradition2- Industry

3- Scientific Research

4- Tradition and industry: a failed union5- Research and industry: an aborted

relationship

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1- TRADITION

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- Traditional Fermented Dairy Products : Lben, Smen, Jben;

- Traditional Bread: starter = dough from the day before;

- Fermented Plant Products: olives, vegetables, lemon, etc.

Food Biotechnology in Morocco 1- Tradition

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« Chekoua » (Goat skin) for lben fermentation

and churning

Raw Whole Milk

Lben

Spontaneous Fermentation

Churning

Water

Traditional

butter

http://www.agadez-niger.com/photos_niger/autour_agadez_04.jpg

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« Khabia » (Earthenwar jar) for

lben fermentation

Wooden tool used for churning

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Moroccan Jben

Raw Whole Milk

SpontaneousCoagulation

Draining

Jben

Whey

http://images.google.co.ma/imgres?imgurl=http://www.saveursetcuisinedumaroc.com/images/stories/imgs/le_jben.jpg&imgrefurl=http://www.saveursetcuisinedumaroc.com/content/view/111/1/lang,fr/&usg=__Qdpdb9PsUJlgvW2_TSrAn2okctE=&h=128&w=180&sz=8&hl=fr&start=2&tbnid=6x6GvX_bYkZxPM:&tbnh=72&tbnw=101&prev=/images%3Fq%3Djben%26gbv%3D2%26hl%3Dfr

http://www.bedfordcheeseshop.com/images/1692__10112003171147.jpg

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Old Smen ( Hayl )

Smen in an

earthenwar jar

smen

Traditional Butter

Butter working& salting

Maturation (1) (3-6 months)

Smen

Maturation (2) (up to 6 years)

Smen « Hayl »

http://images.google.co.ma/imgres?imgurl=http://www.sitefeminin.com/image_article_big/img1816.jpg&imgrefurl=http://www.sitefeminin.com/recettes-1816-Beurre_rinc%25C3%25A9_Smen_baldi_ou_hayl.xhtml&usg=__ULhx4DteULfZDwbRU6FXKleecBk=&h=337&w=450&sz=121&hl=fr&start=9&tbnid=BI0nackseLKTeM:&tbnh=95&tbnw=127&prev=/images%3Fq%3Dsmen%26gbv%3D2%26hl%3Dfr

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Traditional Dough

Oven Cooking

Traditional Bread

Cooking on Charcoal

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2- INDUSTRY

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Food Biotechnology in Morocco 2- Industry

- Baker’s yeast: 2 plants SOMADIR (Casablanca, El-Jadida ); & 1 LESAFFRE Maroc (Fès);

- Brewery: plants in Casablanca, Marrakech, Fès, Tanger, Salé, etc.

- Wine production: various locations;

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- Fermented milk: yogurt, « raïbi », others (probiotics with Bifidobacterium, Lactobacillus acidophilus, etc.)

- Cheese:

. fresh: great number of plants;

. ripened: FROMITAL (Agadir), SOFRAM (Casablanca); Fromagerie du Québec (Fès);

. processed: BEL Maroc (Tanger), FDD (El-Jadida), MARGAFRIQUE (Casablanca).

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- Fermented olives: Meknès, Marrakech, etc. (mainly for export)

- Capers- Vinegar

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3- SCIENTIFIC RESEARCH

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Food Biotechnology in Morocco 3- Scientific Research

3.1- Characterization of the traditional products

3.2- Attempts to monitor their technology for industrial use

3.3- « Biological » Control of Food Hygienic Quality

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3.1- Characterization of the traditional products

3.1.1- Lben

Preliminary study: traditional technology, composition, physicochemical and microbiological characterististics

(Tantaoui-Elaraki et al., Le Lait, 1983, 63, 230-45)

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Food Biotechnology in Morocco 3- Scientific Research 3.1- Characterization of the traditional products

3.1.1- Lben

Physicochemical Characterization: average values (pH, acidity, lactose, fat content, chlorides, flavor compounds); very wide variability

(Boubekri et al., Le Lait, 1984, 64, 436-47)

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3.1.1- LbenHygienic Quality:

Aboundance of:

- Total and Fecal Coliforms;

- Enterococci;

- Yeasts and Moulds.

(Benkerroum et al., MAN, 1984, 2, 199-206)

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3.1.2- SmenChemical study:

- wide variability of fat, water and non fat dry matter contents;

- profile of fatty acids similar to butter’s;

- important lipolysis (free fatty acids);

- week oxidation.

(El Marrakchi et al., Le Lait, 1986, 66, 117-33):

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3.1.2- SmenMicrobial Flora: I- natural flora and flora of hygienic significance:

- disappearence of Coliforms and Enterococci;

- absence of Salmonella and presumably enterotoxigenic Staphylococci;

- proeminence of Bacillus (tolerance to salt and free fatty acids).

(El Marrakchi et al., Le Lait, 1988, 68, 205-18)

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3.1.2- SmenMicrobial flora : II- Lipolytic and Caseolytic Flora

- Staphylococcus cohnii, Bacillus cereus, Aeromonas hydrophila

- Good lipolytic activity (on tweens, tributyrin, butyric fat)

(El Marrakchi et al., Le Lait, 1988, 68, 333-48)

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Food Biotechnology in Morocco 3- Scientific Research 3.2- Attempts to monitor their technology for industrial use

3.2.1- LbenPreparation of Pasteurized Moroccan Lben with selected bacterial strains

- definition of organoleptic criteria (acidity, taste and aroma);

- identification of LAB responsible for fermentation:

. Lactococcus lactis, Lc. diacetylactis, . Leuconostoc lactis, Ln. cremoris

(Tantaoui-Elaraki et al., Actes Inst. Agron. Vét., 1983, 3, 49-58)

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3.2.1- Lben

- pasteurized milk inoculted with starters made from selected bacteria;

- product obtained: satisfactory (hygienic and organoleptic properties).

(Tantaoui-Elaraki et al., Actes Inst. Agron. Vét., 1983, 3, 49-58)

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3.2.2- SmenStudy of the lipolysis

- Industrial pasteurized butter with Candida zeylanoides lipase or conidia of Penicillium roquefortii

- Product obtained: free fatty acids profile (GC/MS) similar to smen profile

- organoleptic analysis: satisfactoryFaid et al., Actes Inst. Agron. Vét., 1991, 11, 21-5


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3.2.2- SmenProduction from pasteurized industrialbutter with lipolytic microorganismsStaphylococcus xylosus, S. simulans, Lactobacillus plantarum, Penicillium roquefortii, Debaryomyces hansenii, Geotrichum candidum)

Faid et al., MAN, 1991, 9, 279-86

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Production from pasteurized industrialbutter with lipases:

- type VII of Candida zeylanoides,- type XIII of Rhizopus arrhyzus,

- type XI of Pseudomonas sp.

Satisfactory results with D. hansenii, P. roquefortii & type VII lipase)

Faid et al., MAN, 1991, 9, 279-86

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3.2.2- SmenPilot Scale Production

- pasteurized butter + lipase EC.3.1.1.3 (type VII) from C. zeylanoides

- GC/MS analysis : free fatty acids profile satisfactory

- Organoleptic tests: satisfactory

Faid et al., J. Soc. Dairy Technol., 1993, 46, 9-11

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3.3- « Biological » control of food hygienic quality

Escherichia coli

Staphylococcus aureus

Listeria monocytogenes

Some bacteria harmful to foods and consumers

http://images.google.co.ma/imgres?imgurl=http://jpkc.njau.edu.cn/spwswx/imgbank/tuku/Coccoid%2520Bacterium,%2520Staphylococcus%2520aureus%2520(division)%2520(SEM%2520x23,000).jpg&imgrefurl=http://jpkc.njau.edu.cn/spwswx/cankao/ShowArticle.asp%3FArticleID%3D314&usg=__xZGG_mtLV3FhxI94kYn8WLDorW4=&h=394&w=500&sz=35&hl=fr&start=8&tbnid=_INN92JEQkSQdM:&tbnh=102&tbnw=130&prev=/images%3Fq%3Dstaphylococcus%2Baureus%26gbv%3D2%26hl%3Dfr

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Food Biotechnology in Morocco 3- Scientific Research 3.3- « Biological » control of food hygienic quality

Microbial destruction in food requires:

- heat treatments (energy consuming, nutritional and organoleptic side effects);

- chemical preservatives (sanitary and organoleptic side effects).

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Other means:

- Use of antagonistic microorganisms- Use of bacteriocins

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Lactococcus lactis Streptococcus

thermophilus

Lactobacillus bulgaricus

Lactobacillus curvatus

http://www.enitac.fr/cerpat/saucissons/photos/18.jpg

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3.3.1- Screening for bacteriocin producing bacteriaA Moroccan lactic acid bacterium with high utilization potential in the fermentation of milk and meat products

- bacteriocin producing Lactobacillus lactis subsp. lactis isolated from traditional lben,

- active lyophilized form for industrial use

(Benkerroum, IAV Info, 2002, 9)

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3.3.1-Screening for bacteriocin producing strainsInhibitory activity of Lactobacillus curvatus CWBI-B28 on Listeria monocytogenes and ST2 verotoxin producing Escherichia coli O157

- L. curvatus isolated from crude meat- Inhibits L. monocytogenes and E. coli

pathogenic strain(Ghalfi et al., Afric. J. Biotech., 2006, 5, 2303-6)

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3.3.1-Screening for bacteriocin producing strainsScreening for bacteriocin producing lactic acid bacteria from various Moroccan food products and partial characterization of the presumed bacteriocins

- Bac+ Enterococcus, Lactococcus and Streptococcus strains isolated;

- Inhibition of Listeria monocytogenes (+ other Gram positive and Gram negative bacteria) by all Bac+ strains

(Benkerroum et al., Biotechnology, 2007, 6, 481-8)

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3.3.1-Screening for bacteriocin producing strainsIsolation of bacteriocin producing lactic acid bacteria

- Production of bacteriocins by lactic acid bacteria

- Active against Listeria innocua and Escherichia coli

(Yaakoubi et al., J. Rapid Meth. Autom. Microbiol., 2009, 17, 32-45)

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3.3.2- Applications on dairy products

Inhibition of Listeria monocytogenes in Moroccan Jben

- Jben processed from milk contaminated with L. monocytogenes and inoculated with a bacteriocin producing L. lactis

- 107 CFU/ml: 2,7 log decrease within 30 hours- 104 CFU/ml: total elimination within 24 hours

(Benkerroum et al., J. Appl. Microbiol., 2000, 89, 960-8)

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3.3.2- Applications on dairy productsBiocontrol of Listeria monocytogenes in lben by an in situ produced Lactococcus lactis bacteriocin :

- strater Bac+ L. lactis and L. diacetylactis: L. monocytogenes under detectable limit within 24h

- control: survival 6 days at 7°C(Benkerroum et al., Int. J. Dairy Technol., 2002, 55, 145-51)

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3.3.2- Applications on dairy productsBehaviour of Listeria monocytogenes in yogurt fermented with a bacteriocin producing thermophilic starter:

- Milk contaminated with L. monocytogenes (103 CFU/ml)

- Starter: Bac+ Streptococcus thermophilus and Bac- Lactobacillus bulgaricus

- L. monocytogenes under detectable limit within 24h

(Benkerroum et al., J. Food Prot., 2002, 65, 799-805)

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3.3.2- Applications on dairy productsBehaviour of Staphylococcus aureus in yogurt fermented with a bacteriocin producing thermophilic starter:

- Milk contaminated with S. aureus (106 CFU/ml)

- Starter: Bac+ Streptococcus thermophilus and Bac- Lactobacillus bulgaricus

- S. aureus: survival 10 days at 22°C even with Bac+ starter

(Benkerroum et al., J. Food Prot., 2002, 65, 799-805)

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3.3.2- Applications on meat productsProtection of dry sausage against Listeria monocytogenes by Lactobacillus curvatus and Lactococcus lactis subsp. lactis.

- Usual starter + L. monocytogenes (102-103 CFU/g)

- Lb. curvatus: no surviving Listeria after 8 days

- Lc. lactis: no surviving Listeria after 15 days(Benkerroum et al., J. Appl. Microbiol., 2005, 98, 56-63)

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3.3.2- Applications on meat products Behaviour of Listeria monocytogenes in Merguez (crude sausage) in presence of a bacteriocin producing Lactococcus

- L. monocytogenes added at 106 CFU/g- More drastic decrease with Bac+ L. lactis

than with Bac- L. lactis- Nitrites (0,4%) reduce Listeria decrease.

(Benkerroum et al.,Meat Sci., 2003, 63, 479-84)

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3.3.2- Applications on meat products

Bacteriocin producing Lactobacillus curvatus as protecting agent in bacon

- L. monocytogenes added (106 CFU/g)

- Lb curvatus: decrease of Listeria under detectable limit within 1 week (without nitrites); within 2 weeks (with nitrites)

(Ghalfi et al., Food Sci. Technol. Int., 2006, 12, 325-33)

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3.3.3- Application on fish

Inactivation of Listeria monocytogenes by bacteriocin producing Lactobacillus curvatus in smoked salmon

- partially purified bacteriocin added, or produced in situ: decrease of Listeria under detectable limit, then further growth

(Ghalfi et al., J. Food Prot., 2006, 69, 1066-71)

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3.3.3- Application on fishInactivation of Listeria monocytogenes by bacteriocin producing Lactobacillus curvatus in smoked salmon

- bioactive wrapping: slower decrease, no further growth (22 days)

- bacteriocin adsorbed on bacterial cells : total inactivation (3 days), no further growth (22 days)

(Ghalfi et al., J. Food Prot., 2006, 69, 1066-71)

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4 – TRADITION AND INDUSTRY: A FAILED

UNION

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Food Biotechnology in Morocco 4- Tradition and industry: a failed union

- Industrial products bearing traditional names

- Do not satisfy the specifications- main products: lben et smen

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4.1- Lben

- general processing scheme of products called «lben» or «leben» or «l’ben»:

. pasteurized skimmilk (hygienic goal);

. often inoculated with imported starters;

- wide diversity of physico-chemical and organoleptic characteristics;

- often far from traditional lben characteristics

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Food Biotechnology in Morocco 4- Tradition and industry: a failed union4.1- Lben

Que dit la législation?Lben: lait fermenté ayant subi une fermentation lactique à l’aide de bactéries sélectionnées; acidité minimale 70°D; extrait sec dégraissé 80g/l

What about regulation?Lben: milk fermented with selected lactic acid bacteria; minimal acidity 0.7% (lactic acid); non fat dry matter 80g/l

Décret n°2-00-425 du 10 ramadan 1421 (7 décembre 2000) relatif au contrôle de la production et de la commercialisation du lait et produits laitiers

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4.2- Smen

- general processing scheme of products called smen :

. Industrial pasteurized butter (often imported);

. Salted . Conditioned in hermetically sealed glass or

plastic jars.

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Food Biotechnology in Morocco 4- Tradition and industry: a failed union4.1- Smen

Que dit la législation?Smen: produit obtenu à partir du lait, de la crème ou du beurre pasteurisé ou non, par salage et par l’action de lipases et/ou de l’action de bactéries lipolytiques

sélectionnées…

Maximum 16% d’eau et 5% de sel; indice d’acide minimal de 10.

What about regulation?Smen: product obtained from milk, cream or butter (pasteurized or not) by salting and by the action of lipases and/or

selected lipolytical bacteria…

Maximum content: 16% water, 5% salt; minimum acidity index 10

Arrêté viziriel du 21 rebia I 1340 (22 novembre 1921) relatif à la vente des beurres, saindoux, huiles et matières grasses alimentaires, tel qu’il a été modifié et complété

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5- RESEARCH AND INDUSTRY: AN ABORTED

RELATIONSHIP

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Food Biotechnology in Morocco 5- Research and industry: an aborted relationship

- Research and industry: potential partners that ignore each other;

- Some cooperation attempts that failed;

- Real opportunities.

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Food Biotechnology in Morocco 5- Research and industry: an aborted relationship

Exemple of cooperation possibility

Baker’s yeast Saccharomyces cerevisiae : amylasic activity nil or very weak; Solution: baking enhancers (améliorants de panification)

. S. diastaticus (in Moroccan traditional bread dough): high amylasic activity

. Possibilities: starter with a mixture of the 2 species; genetic recombination between the 2 species.

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CONCLUSION

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Food Biotechnology in Morocco Conclusion

- Food industry in Morocco is to develop- Traditional products more and more

appreciated by the consumers- Industry has to respect these products

specifications- Industry could take benefit from scientific

potentialities to succed in binding tradition to modernity

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Aknowledgements

Prof. Mostapha EL HADDADProf. Mohamed CHAOUFI

For correcting English translation

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BIOTECHNOLOGIE ALIMENTAIRE AU MAROC · food biotechnology in morocco a. tantaoui el araki (sup’agro, casablanca) biotechnologies alimentaires au maroc: entre la recherche scientifique