Optimation of Liquid Sugar Production on the Processing of Bioethanol From Arrowroot Tuber Starch (Maranta arundinaceae Linn) Using Aspergillus niger and Amyloglucosidase

Sawarni HasibuanEndrianur Rahman Zain

Department of Agroindustrial TechnologyFaculty of Agribusiness and Food TechnologyBogor Djuanda University

Introduction

Indonesia's energy needs continue to increase over time.

The use of alternative fuels as a substitute for fossil fuels should be implemented immediately.

Indonesia’s fuel needs in 2010 was 77.26 million KL, while total production amounted to 62.8 million KL, thereby incurred a deficit of 14.46 million KL.

Government Regulation.

Introduction

Government Regulation

2006

2006

2006

2005

Inpres Number 10 of 2005 on saving energy use, including the use of fuel.

Inpres Number 1 of 2006 about the use and supply of biofuel as an alternative fuel.

2005

2006

2006

2006

Presidential Regulation (Perpres) 2006 on National Energy Policy.

Inpres Number 5 of 2006 on renewable energy.

IntroductionTHE USE OF ALTERNATIVE FUELS SHOULD BE IMPLEMENTED IMMEDIATELY

BIOFUEL IS NOT SOMETHING NEW

DEVELOPMENT PROSPECTS

The use of alternative fuels must be implemented immediately especially in the form of liquid, because people are very familiar with the liquid fuel, one of which is bioethanol.

One example of the use of biofuel in ancient times was burning jatropha seed for lighting.

The arrowroot is a source of starch that can be decomposed into sugar and then fermented into bioethanol.

THE ARROWROOT TUBER PLANT IS ONE OF ALTERNATIVE SOURCE OF CARBOHYDRATE

Arrowroot can be processed into bioethanol as a substitute for premium gas.

Objectives

1. To determine the optimal timing of conversion of starch into sugar liquid sugar from arrowroot starch tubers.

2. To obtain the optimal concentration increase conditions of the Aspergillus niger and Amiloglukosidase starter on the production of liquid sugar arrowroot tubers.

3. To determine the efficiency of bioethanol production with the addition of arrowroot tubers Sacharomyces cerevisae.

The general objective of the research is to support the diversification of raw materials and alternative fuels especially bioethanol in Indonesia.

The Arrowroot Tuber Plant

Divisi : SpermatophytaSub Divisi : AngiospermaeKelas : MonocotyledoneaeOrdo : ZingiberlesFamili : MarantaceaeGenus : MarantaSpecies : Maranta arundinaceae L.

Area : Central Java and East Java

The shape is like torpedo about 20 - 40 cm, and diameter is about 2 - 5 cm.

Productivity in Malang about 12.5 – 30 ton/ha with average 20 ton/ha (150 Ha).

Government Program for arrowroot tuber plant about 18000 Ha

Table 1 Nutrient comparation of rice starch, white flour, and arrowroot tuber starch

No. Description Unit

Composition

Rice starch Wheat flour Arrowroot tuber starch

1 Calori Kal 360,0 365,0 355,0

2 Protein Gr. 6,8 8,9 0,7

3 Fat Gr. 0,7 1,3 0,2

4 Carbohydrate Gr. 78,9 77,3 85,2

5 Calsium Mg. 6,0 16,0 8,0

6 Phosphorus Mg. 140,0 106,0 22,0

7 Zinc Mg 0,8 1,2 1,5

8 Vitamin A Iu 0,0 0,0 0,0

9 Vitamin B1 Mg 0,12 0,12 0,09

10 Vitamin C Mg 0,0 0,0 0,0

11 Water Gr 13,0 12,0 13,6

12 Edible eating % 100,0 100,0 100,0

High purity arrowroot starch chemical industrial raw materials, cosmetics, liquid sugar, glue, a mixture of the capsule, and others.

Glucose Syrup

Liquid glucose is a solution with a viscosity of between 32-35 Be, produced by hydrolysis of starch with acid catalysts, enzymatic or a combination of both.

Hydrolysed starch can be derived from starch-containing materials such as corn, wheat, cassava, sweet potato (Maiden, 1970).

Starch hydrolysis process is basically the starch polymer chain termination (C6H12O6) into monosa-ccharide units (C6H12O6) (Meyer, 1978).

Enzymatic hydrolysis of starch is composed of two stages, namely the stage likuification and saccharification.

Hidrolysis of Starch

Enzym

Enzym

Microbe

Enzym for hidrolysis of starch

Enzym can isolated from flora, fauna, and microorganisme (Wirahadikusumah,1986).

The fungus can produce enzym i.e. Aspergilus, Penicillium, Mucor dan Rhizopus. Enzym that are produced such as amilase, invertase, protease dan pektinase (Lay B. & Hastowo, 1992).

Enzym alpha amilase (α- amilase)Enzym glukoamilase (AMG)

Aspergillus niger fungus

Aspergillus niger 20 kinds of enzym : α-amilase, β-amilase, maltase, laktase, trehalase, tanase, katalase, proteinase, polipepatidase, dipepatidase, renin, lipase, selulase, glucosa oxidase, glukosidase dehidrogenase, zymase, urease, inulase and melibiase (Tauber 1950).

Fermentation

The fermentation of sugar into ethanol, CO2, and by-products are divided into 3 parts:

1. The order of the reaction of glucose to gliseraldehid-3-phosphate which is the intermediate compound. This reaction using 2 mol ATP and not an oxidation reduction reaction.

2. Substrate level phosphorylation reaction (oxidation) that produces 4 moles of ATP and pyruvate.

3. Pyruvate reduction reaction becomes the main result of fermentation that is 2 moles of ethanol and carbon dioxide.

Microbes Produced Ethanol

Saccharomyces cerevisiae is a species of yeast that has a power conversion of sugar into ethanol is very high. Saccharomyces cerevisiae requires a temperature of 30oC and pH 4.0-4.5 in order to grow well. During fermentation there will be hot. If it is not waged cooling, the temperature will continue to rise so that the fermentation process is hampered (Oura, 1983). According to Harrison and Graham (1970), Saccharomyces cerevisiae, can be tolerant of a quite high alcohol (12-18% v / v), resistant to high sugar and remained active in the fermentation at 4-32oC.

Yeasts that are often used in ethanol fermentation industrial

are Saccharomyces cerevisiae, S. uvarium,

Schizosaccharomyces sp. and Kluyveromyces sp.

Alcohol

Ethanol (C2H5OH) is biochemistry fluid of fermentation of sugars from carbohydrate sources using the help of microorganisms. Bioethanol is made with raw materials such as sugar cane, palm juice, starchy ingredients such as corn and tubers, which form a fibrous branches of agricultural waste are still development in developed countries (Anonymous, 2000).

Materials:

1. Arrowroot flour1. Arrowroot flour

2. Aspergillus níger culture2. Aspergillus níger culture

3. Amyloglucosidase (AMG) 3. Amyloglucosidase (AMG)

4. Sacharomyces cereviseae yeast4. Sacharomyces cereviseae yeast

5. Potato Dextrose Agar (PDA) 5. Potato Dextrose Agar (PDA)

6. Distilled water 6. Distilled water

7. 70% alcohol, and 7. 70% alcohol, and

8. Other chemical materials for analysis. 8. Other chemical materials for analysis.

Methods

1

2

3

The preparation

The hydrolysis of arrowroot flour

The fermentation of arrowroot liquid sugar.

Flowchart of arrowroot flour processing

Arrowroot tuber

Water

Arrowroot flour

wastePeeling and washing

Slices

Drying60oC, 2 hours

Hammer mill

Heated(95 oC)

Starter

Aspergilus niger,3 ose

Tween 80, 0.1%

Aquades

Stirrer until gelatinization

Likuification(3 days)

Cooled (27 oC)

Sterilization(121oC, 15 min)

Flow chart of starter preparation

Arrowroot flour

Arrowroot flour

H2O

Heated 95oC

SterilizationT = 121oC, t = 15 min

Stirred until gelatinization

Cooled T = 27oC

HydrolysisT = 38oC, t = 9 hari

Starter A. niger 10%, 15%, 20%

Separation(sentrifiuge t = 15 min)

Sludge

Liquid sugar

Fermentation T= 28oC,pH=4.5,t= 6-11 days

Destilation Etanol

Analysis

Sacchromyces cerevisiae

Flowchart of process

Arrowroot flour

H2O

Heated 95oC

SterilizationT = 121oC, t = 15 min

Stirred until gelatinization

Cooled T = 27oC

HydrolysisT = 38oC, t = 9 hari

Starter AMG 5%, 7,5%, 10%

Separation(sentrifiuge t = 15 min)

Sludge

Liquid sugar

Fermentation T= 28oC,pH=4.5,t= 3 days

Destilation Etanol

Analysis

Sacchromyces cerevisiae

Flowchart of process

Experimental Design

Nested design with 2 treatments and 2 replications.

Yijk : Observation variable as a result of treatments of the i-th stage of factor A

with the j-th stage of factor B at the k-th repetitionμ : General mean Ai : Effects of treatment of the i-th stage of factor A (addition of starter i =

1, 2, 3)Bj : Effects of treatment of the j-th stage (fermentation time i = 1, 2, 3) of factor B

nested at the i-th stage of factor A (addition of starter i = 1, 2, 3)Єij : Experimental error

Yijk = μ + Ai + Bj(i) + Єijk

Preliminary Research

The chemical composition of arrowroot flour

Descriotion Average (%)

Water content 13.95

Ash content 1.00

Protein content 0.93

Fat content 1.79

Fiber content 4.56

Carbohydrate content 73.53

Carbohydrat content depends on the production process and cultivar used max. 85,2 % lower than cassava flour but higher than rice flour and wheat fluor .

1. After three days of fermentation, the substrate physically turned into a weak slurry with a blackish color

2. The blackish color on the substrate was due to the overgrowing of Aspergillus niger. Starter of Aspergillus

niger fungus

Preparation of Starter

Production of Starch Sugar Syrup

Figure 1 Average total sugar content of arrowroot glucose syrup with various concentrations of Aspergillus niger starters and lengths of fermentation.

6 9 110

2

4

6

8

10

12

14

10% 15% 20%

Length of fermentation (days)

Suga

r co

nten

t (%

)

Production of Arrowroot Sugar Syrup

Figure 2 Average total sugar content of arrowroot glucose syrup with AMG starter concentration of 1-10% and starch concentration of 20%-30%.

1% 5% 7.5% 10%0

5

10

15

20

25

30

20%

25%

30%

AMG enzyme concentration

Suga

r co

nten

t (%

)

Fermentation

Arrowroot sugar syrup

The average alcohol content of 6.62% with an average efficiency fermentation 65%.

Bioethanol

Conclusion

1. The optimum time to produce liquid sugar from arrowroot tuber starch using Aspergillus niger was 11 days, resulting in a total sugar content of 12%.

2. The use of commercial AMG was much more effective compared to the use of the Aspergillus niger starter. Sugar contents could reach 28.5% and shorter fermentation time, i.e. 24 hours.

3. Optimum conditions for Aspergillus niger in the production of liquid sugar from arrowroot tuber were concentration of 10%, 38oC, and 9 days; while AMG 10% and conversion was completed in 24 hours.

4. The fermentation time of 7 days, 270C and an addition of Saccharomyces cerevisiae produced an average alcohol level of 6%. The highest efficiency of fermentation of sugar into alcohol reached 65%.

Suggestion

If we will be using Aspergillus niger and distillation should be improved fermentation process, in order to produce bioethanol with 90-95% ethanol content.

Thank You