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RESEARCH ARTICLE

 

Bioethanol Production by yeast fermentation Using Pomace Waste

 

T.C.Venkateswarulu1*, B. Bodaiah2, D. John Babu1, A. Venkata Naraya1, Y. Evangelin2

1Department of Biotechnology, Vignan’s University, Vadlamudi-522213, India

2Department of Biotechnology, Achrya Nagarjuna University, Guntur-522510, India

*Corresponding Author E-mail: venki_biotech327@yahoo.com

 

ABSTRACT:

The chemical analysis of pomaces revealed the presence of significant amounts of fermentable sugars that are retained in pomace after pressing of fruits or vegetables. Present study deals with bioethanol production by yeast fermentation using pomace waste. Seven different varieties of fruits (apple, grape, pineapple, pomegranate and orange) and vegetables (carrot and beetroot) pomaces were used. Individual pomace and combination of pomace extracts were fermented for ethanol production in which individual grape pomace extract gives highest yield of 2.6g ethanol, where as grape-apple, grape-pineapple and orange-grape combined pomace media produced a yield of 3.1, 3.0 and 3.0 g ethanol per 10g reducing sugars respectively which is 10-15% more bioethanol yield compared to regular medium for yeast strain.

 

KEY WORDS: Bioethanol, Yeast, Fruits, Vegetables, Pomace.

 

 

INTRODUCTION:

Bioethanol can be produced from a variety of feedstocks, including sugar substances, such as sugarcane juice and molasses, as well as starch-based materials such as wheat and corn1. During ethanol fermentation,  glucose and other sugars in medium are converted into ethanol and carbon dioxide2. Biofuels are generated from a wide range of feedstock sources, nearly all bioethanols worldwide now days are first generation biofuels, which are made from sugar or starch based feedstocks, mainly food crops.

 

Yeast is a unicellular fungus. It can be classified based on their ascospore, colony and its physiological characteristics. The most well-known and commercially significant yeasts that been primarily used for bioethanol production are the related species and strains of Saccharomyces cerevisiae3.

 

 

 

 

 

 

 

 

 

 

 

Received on 30.05.2015          Modified on 18.06.2015

Accepted on 24.06.2015        © RJPT All right reserved

Research J. Pharm. and Tech. 8(7): July, 2015; Page 841-844

DOI: 10.5958/0974-360X.2015.00137.7

 

Saccharomyces cerevisiae is also famously known as baker’s yeast. These organisms have long been utilized to ferment the sugars of rice, wheat, barley, and corn to produce alcoholic beverages and in the bakery industry4.Yeasts such as Kluveromyces maxianus, Clavispora sp., Schizosaccharomyces, Brettanomyces other species of Candida viz., Candida tenius, Candida tropicalis, Candida utilis, C. shehatae, Candida blankii, Candida friedrichii, Candida solani and Candida parapsilosis and species of Debaromyces viz., Debaromyces nepalensis,  Debaromyces polymorpha, Pichia stipitis, Candida shehatae and Pashysolan tannophilus  produce bioethanol from a variety of substrates  have tested 20 species of Candida, 21 strains of Saccharomyces and ethanol was the major product with most of the Candida stains. One yeast cell can ferment approximately its own weight of glucose in an hour. Under optimal conditions S. cerevisiae can produce up to 18 percent ethanol in fermentation broth5,6. Besides excellent in ethanol productivity, it also highly tolerance towards ethanol and the inhibitors found in the fermentation compared to other strains7.

 

In this study, the extracts of pomace waste were analyzed in order to look for the production of bioethanol. This analysis could be useful for selection of raw material for optimum production of bioethanol.

MATERIALS AND METHODS:

Ethanol production from individual pomace extracts

Ethanol production for individual pomace extracts was performed by taking 200 mL carrot extract, 181.8 mL of beet root extract, 125 mL of pomegranate extract, 105 mL of pineapple extract, 111 mL of apple extract, 117 mL of orange extract and 95 mL of grape extract obtained from their respective pomaces in different conical flasks. 2g of peptone and 2g of yeast extract was added to each of these flasks. 1% of actively growing yeast culture was added to all the flasks and incubated for fermentation.

 

Ethanol Production from mixed fruit pomace

The combinational raw materials effect on production of bioethanol was studied. In the first grouping 57mL of grape extract is kept as common in all flasks and 80 mL carrot extract, 72.5 mL beets extract, 50 mL pomegranate extract, 46.8 mL orange extract, 44.4 mL apple extract and 42.0 mL pineapple extracts were added to 6 different conical flasks. In second blend the apple extract volume 58mL kept as constant in all flasks with other sources like 80 mL carrot extract, 72.5 mL beets extract, 50 mL pomegranate extract, 46.8 mL orange extract, 44.4 mL apple extract and 42.0 mL pineapple extracts were added to 6 different conical flasks.

 

In third mixture 70 mL of orange extract is common in all flasks with combination of 80 mL carrot extract, 72.5 mL beets extract, 50 mL pomegranate extract, 38 mL grape extract, 44.4 mL apple extract and 42.0 mL pineapple extracts were added to 6 different conical flasks. In forth combination G-A used as constant volume in all flasks. The fermentation broth was prepared with 4g of reducing sugars obtained from each of grape – apple (G.A) extracts and 2g from other sources – carrot, Beets, Pomegranate and Orange by taking 38.0 mL of grape extract and 44.4 mL of apple extracts, 40.0 mL carrot extract, 36.2 mL beets extract, 25.0 mL pomegranate extract, 24.0 mL orange extract, and 21.0 mL pineapple extracts were added to 5 different conical flasks. In all combinations to the formulated fermentation medium of pomace extracts was additionally supplied with 2g peptone and 2g yeast extract and then all flasks were fermented with 1% overnight grown yeast culture.

 

RESULTS AND DISCUSSION:

Ethanol Production from individual sources

Bioethanol can be produced from a variety of feedstocks, including sugar substances, such as sugarcane juice and molasses. 2.1g of ethanol was obtained from carrot extract. 2.0g, 2.3g, 2.4g, 2.5g, 2.45g, 2.6g ethanol was produced from beet, pomegranate, pineapple, apple, orange and grape respectively. The optimum ethanol was produced from grape waste. Bar graph of ethanol Production from individual pomace extracts is shown in figure 1.

 

Fig.1 Ethanol production from individual pomace extracts

 

Ethanol Production from combinational sources

Ethanol produced from mixed pomace extract was estimated after distillation. 3.1g of ethanol was obtained from grape-apple combination extract, 2.4g, 2.5g, 2.65g, 3.0g and 3.0g ethanol was produced from other combinations like grape-carrot, grape-beet, grape-pomegranate, grape-pineapple and grape-orange respectively. (fig.2a) The conversion 31 % (w/w) was achieved when 57 mL of grape extract and 44.4 mL apple extracts was fermentation with selected yeast strain. In previous studies it was reported by Ajay Kumar Singh et al, produced BioEthanol from Banana peel by Simultaneous Saccharification  and Fermentation Process using cocultures Aspergillus niger and Saccharomyces cerevisiae8.

 

The quantity 2.8g of ethanol was obtained from grape combination extract. 2.4g, 2.5g, 2.6g, and 2.5g ethanol was produced from Carrot, Beet, Pomegranate, Pineapple and Orange respectively.(fig. 2b) Manoj et al., 2013 obtained 10.65 to 13% reducing sugars after treatment of apple pomace with α-amylase and cellulase. 4.15g (34 w/w) ethanol was produced from reducing sugars of apple pomace via simultaneous saccharification and fermentation with Saccharomyces cerevisiae9.

 

Hang et al., 1982 obtained a yield varied from 2.9 to 4.0g per 100g apple pomace by simultaneous saccharification and Saccharomyces cerevisiae fermentation10. Debajit et al., 2011 obtained 75g ethanol from 50g sucrose and 400g of apple pomace, rotten banana through anaerobic fermentation with Saccharomyces cerevisiae11.

2.8g of ethanol was obtained from orange combination extract. 2.0g, 2.5g, 2.6g, 2.7g and 2.2g ethanol was produced from Carrot, Beet, Pomegranate, Pineapple and Apple respectively.(fig.2c) Grape+ apple+ carrot extracts have been yielded 2.4g ethanol, grape+ apple+ beet root produced 2.5g ethanol, grape+ apple+ pomegranate yielded 2.6 g of ethanol, grape+ apple+ pineapple yielded 2.6g of ethanol and grape+ apple+ orange gave a yield of 2.5g ethanol. G.A. represents 38.0 mL of grape extract and 44.4 mL of apple extracts. (fig.2d)

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

CONCLUSION:

The maximum production of ethanol was produced from 57 mL grape pomace extract and 44.4 mL apple pomace extract through fermentation by    yeast strain is around 3.1g, where as the minimum production ethanol is 1.65g from extracts of 20mL carrot pomace and 36.2mL beet root.

 

REFERENCES:

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3.       Chandel, A.K., Chan, E.S., Rudravaram, R., Narasu M. L., Rao, L.V., and Ravindra, P.  Economics and Enviromental Impact of Bioethanol Production Technologies: an Appraisal. Biotechnology and Molecular Biology Review, 2(1),2007, 014-032.

4.       Tsuyoshi, N., Fudou, R., Yamanaka, S., Kozuki, M., Tamang, N., Thapa, S., and Tamang, J.P.  Identification of Yeast Strains Isolated from Marcha in Sikkim, a Microbial Starter of Amylolytic Fermentation. Food Microbiology, 99, 2005, 135-146.

5.       Abbi M, Kuhad RC, Singh , Bioconversion of pentose sugars to ethanol by free and immobilized cells of Candida shehatae (NCL- 3501): fermentation behaviour. Proc. Biochem., 31, 1996, 555-560.

6.       Gong CS, Claypool TA, McCracken KD, Mann CM, Ueng PP, Tsao GT , Conversion of pentoses by Yeast. Biotechnol. Bioeng., 25,1983, 85-104.

7.       Ahring, B.A. and Westermann , Coproduction of Bioethanol with Other Biofuels. Advanced Biochemical Engineering/ Biotechnology, 108, 2007, 289-302.

8.       Ajay Kumar Singh et al, BioEthanol Production from Banana peel by Simultaneous Saccharification  and Fermentation Process using cocultures Aspergillus niger and Saccharomyces cerevisiae, Int.J.Curr.Microbiol.App.Sci,3(5),2014,84-96.

9.       Manoj Kumar Mahawar, Anupama Singh, B. K. Kumbhar and Manvika Sehgal, Optimization of ethanol production from apple pomace through solid-state fermentation using enzymes and  yeasts combination through response surface methodology, Africal journal of agricultural research,8(24),2013, 3136-3145.

10.     Y. D. Hang, C. Y. Lee and E. E. Woodams, A Solid State Fermentation System for Production of Ethanol from Apple Pomace, J. Food Sci.,47(6),2006,1851–1852.

11.     Debajit Borah and Vimalendra Mishra, Production of Bio Fuel from Fruit Waste, International Journal of Advanced Biotechnology Research,1(1), 2011,71-74.