Characterization of Stress Tolerant High Potential Ethanol Producing Yeast from Agro-Industrial Waste
American Journal of BioScience
Volume 1, Issue 2, July 2013, Pages: 24-34
Received: May 13, 2013;
Published: Jul. 10, 2013
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Md. Fakruddin, Industrial Microbiology Laboratory, Institute of Food Science and Technology, Bangladesh Council of Scientific and Industrial Research, Dhaka, Bangladesh
Md. Ariful Islam, Department of Biotechnology, Brac University, Dhaka, Bangladesh
Md. Abdul Quayum, Department of Biotechnology, Brac University, Dhaka, Bangladesh
Monzur Morshed Ahmed, Industrial Microbiology Laboratory, Institute of Food Science and Technology, Bangladesh Council of Scientific and Industrial Research, Dhaka, Bangladesh
Nayuum Chowdhury, Department of Biotechnology, Brac University, Dhaka, Bangladesh
Bioethanol or biofuel as an alternative to fossil fuels has been expanded in the last few decades in the whole world. Use of bioethanol as a renewable transportation fuel will minimize the amounts of fossil-derived carbon dioxide (CO2) to the Earth’s atmosphere. Yeast is the most favorite organism for ethanol production because of its diverse substrate specificity and ease of production of ethanol under anaerobic condition. The main objective of this research work was to isolate & characterize stress tolerant, high potential ethanol producing yeast strains from agro industrial waste. In total 4 yeast isolates have been characterized on the basis of morphological and physico-chemical characters. Based on morphological appearance of vegetative cell under microscope, ascospore production, colony character and physico-chemical characters all the strains was identified to be Yeast. Phylogenetic identification by DNA sequencing confirmed that the strain P is Saccharomyces Unisporus, strain C is Saccharomyces cerevisiae, strain T is Saccharomyces cerevisiae & strain DB2 is Candida piceae. Most of the strains were thermotolerant, pH tolerant, ethanol tolerant as well as osmotolerant. They were resistant to cycloheximide at 0.0015g/100ml concentration, hydrogen peroxide (0.50%), Chloramphenicol (30µg/disc) but growth was inhibited in the presence of 1% acetic Acid. The strains P, C & T showed good Invertase activity & only the T strain was capable of producing killer toxin. They were capable of fermenting glucose, fructose, sucrose, amylose & trehalose. Ethanol producing capability of the strains was studied using sugarcane molasses as substrate. The bioethanol production capacity of the yeasts were found to be 15%, 14.5%, 12% & 8.15% for P, C, T & DB2 respectively at pH 6.0, 30oC temperature in media with 5.5% initial reducing sugar concentration in shaking condition. Pilot scale ethanol production by P strain was 13.10%, C strain 11.15%, T strain 9.80% & DB2 strains 7.85% at 60 hours. These strains could be potential for ethanol production from cane molasses.
Md. Ariful Islam,
Md. Abdul Quayum,
Monzur Morshed Ahmed,
Characterization of Stress Tolerant High Potential Ethanol Producing Yeast from Agro-Industrial Waste, American Journal of BioScience.
Vol. 1, No. 2,
2013, pp. 24-34.
Nadir, N., Mel, M.., Karim, M.I.A., Yunus, R.M. Comparison of sweet sorghum and cassava for ethanol production by using Saccharomyces cerevisiae.J. App. Sci.,2009; 9(17): 3068-3073
Ibeto CN, Ofoefule AU, Agbo KE. A global overview of biomass potentials for bioethanol production: A renewable alternative fuel. Trends App. Sci. Res.,2011; 6(5): 410-425
Jegannathan KR, Chan E, Ravindra P. Biotechnology in Biofuels- A cleaner technology. J. App. Sci.,2011; 11(13): 2421-2425
Chaudhury N, Qazi JI. Microbiological Saccharification and ethanol production from sugarcane bagasse.Biotechnol.,2006; 5(4): 517-521.
Noor AA, Hameed A, Bhatti KP, Tunio SA. Bio-ethanol fermentation by the bioconversion of sugar from Dates by Saccharomyces cerevisiae strains ASN-3 and HA-4. Biotechnol.,2003; 2(1): 8-17.
Somda MK, Savadogo A, Ouattara CAT, Ouattara AS, Traore AS. Improvement of Bioethanol production using amylasic properties from Bacillus licheniformis and yeasts strains fermentaiton for Biomass valorization. Asian J.Biotechnol.,2011a; 3(3): 254-261.
Somda MK, Savadogo A, Barro N, Thonart P, Traore AS. Effects of mineral salts in fermentation process using mango residues as carbon source for bioethanol production. Asian J.Biotechnol.,2011b; 3(1): 29-38.
Somda MK, Savadogo A, Ouattara CAT, Ouattara AS, Traore AS. Thermotolerant and Alcohol-tolerant yeasts targeted to optimize hydrolyzation from mango peel for high bioethanol production. Asian J.Biotechnol.,2011c; 3(1): 77-83.
Khongsay N, Laopaiboon L, Laopaiboon P. Growth and Batch fermentation of Saccharomyces cerevisiae on sweet sorghum stem juice under normal and very high gravity conditions. Biotechnol.,2010; 9(1): 9-16
Boboye B, Dayo-Owoyemi I. Evaluation of Dough Sensory Properties Impacted by Yeasts Isolated from Cassava. J. Appl. Sci.,2009; 9(4): 771-776.
Kurtzman CP, Robnett CJ, Ward JM, Bravton C, Gorelic P, Walsh TM(2005)Multigene phylogenetic analysis of pathogenic Candida species in the Kazachstania(Arxiozyma) telluriscomplex and description of their ascosporic states as Kazachstania bovina sp. nov., K. heterogenicasp. nov., K. pintolopesii sp. nov., and K. slooffiaesp. nov.J.Clin.Microbiol.,2005; 43(1): 101-111.
Kreger-Van Rij NJW. The Yeast a Taxonomic Study. New York: Elsevier Science Publishing Company,1984; pp. 1082.
Warren P, Shadomy L. Yeast fermentation broth base with carbohydrate and Durham tube. In: Manual of Clinical Microbiology.5th ed. Washington D.C., 1991.
Osman ME, Khattab OH, Hammad IA, El-Hussieny NI. Optimization of Bio-Fuel Production by Saccharomyces cerevisiae Isolated from Sugar Cane Bagasse. J. Am. Sci.,2011; 7(5):485-492
Willaert R, Viktor AN. Primary beer fermentation by immobilized yeast - a review on flavor formation and control strategies. J. Chem. Technol.Biotechnol.,2006; 81: 1353-1367.
Wayman M, Parekh SR. Microbiology of fermentation catalysts.In Biotechnology of Biomass Conversion. Milton Keynes: Open University press. 1990; pp. 75-100.
Ekunsanmi TJ, Odunfa SA. Ethanol tolerance, sugar tolerance and invertase activities of some yeasts strains isolated from steep water of fermenting cassava tubers. J. App.Bacteriol.,1990; 69: 672-675.
Kirby WWM, Bauer AW, Sherris JC. Antibiotic susceptibility testing by standardized single disk method. Am. J.Clin.Pathol.,1996; 45: 493-496.
Jimenez J, Benitez T. Characterization of wine yeasts for ethanol production. App.Microbiol.Biotechnol.,1986; 25: 150-154.
Bernfield P. Enzymes of starch degradation and synthesis. Adv.Enzymol.,1951; 12: 379-481.
Ribereau-Gayon P, Dubordieu D, Doneche B, Lonvaud A.Handbook of Enology: The Microbiology of Wine and Vinifications. JohnWiley and Sons, Chichester2000; 1:454.
Moslem MA,Bahkali AH, Abd-Elsalam KA, Wit PJ. An efficient method for DNA extraction from Cladosporioid fungi.Genet. Mol.Res., 2010;9: 2283-2291.
Fakruddin M, IslamS, AhmedMM, ChowdhuryA, HoqueMM. Development of Multiplex PCR (Polymerase Chain Reaction) Method for Detection of Salmonella spp. and Vibrio parahaemolyticus from Shrimp Samples of Bangladesh. Asian J. Biol. Sci.,2012a; 5(2): 76-85.
Borneman J, Hartin RJ. PCR primers that amplify fungal rRNA genes from environmental samples.Appl. Environ.Microbiol.,2000; 66:4356–4360.
Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. MEGA5: Molecular Evolutionary Genetics Analysis using Maximum Likelihood, Evolutionary Distance, and Maximum Parsimony Methods. Mol. Biol.Evol.,2011; 28: 2731-2739.
Fakruddin M, Quayum MA, Ahmed MM, Choudhury N. Analysis of key factors affecting ethanol production by Saccharomyces cerevisiae IFST-072011. Biotechnol.,2012b; 11(4): 248-252.
Miller GL. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal. Chem.,1959; 31:426-428.
Conway EJ.Micro-diffusion analysis and volumetric error. Crosby Lockwood and Son, London, 1939.
Lodder J. The yeasts: A Taxonomic study. NorthHoll and Publishing, Amsterdam, 1971.
Boekhout T, Kurtzman CP. Principles and methods used in yeast classification, and an overview of currently accepted yeast genera. In Wolf, K. Nonconventional Yeasts in Biotechnology: A Handbook. Springer-Verlag: Heidelberg. 1996; pp. 1-99.
Kurtzman CP, Fell JW.The Yeasts, a Taxonomic Study. Fourth Edition. Amsterdam: Elsevier Science Publishing Company.1997;pp-1055.
Gimeno CJ, Ljungdahl PO, Styles CA, Fink GR. Unipolar cell divisions in the yeast S. cerevisiae lead to filamentous growth: regulation by starvation and RAS. Cell,1992; 68: 1077–1090.
Ueno R, Urano N, Kimura S. Characterization of thermotolerant, fermentative yeasts from hot spring drainage. Fish Sci.,2001; 67: 138-145.
Hampsey M. A Review of Phenotypes in Saccharomyces cerevisiae. Yeast,1997; 13: 1099–1133.
Krems B, Charizanis C, Entian KD. Mutants of Saccharomyces cerevisiae sensitive to oxidative and osmotic stress. Curr.Genet.,1995; 27: 427–434
Saitou N, Nei M.The neighbor-joining method: A new method for reconstructing phylogenetic trees. Mol. Biol.Evol., 1987; 4:406-425.
Nei M, Kumar S. Molecular Evolution and Phylogenetics. Oxford University Press, New York, 2000.
Roehr M. The Biotechnology of Ethanol: Classical and Future Applications. Chichester: Wiley-VCH. 2001; pp.232.
Du Preez JC, Bosch MN, Prior BA.Temperature profiles of growth and ethanol tolerance of the xylose fermenting yeasts Candida shemataeand Pichiastipitis.App.Microbiol.Biotechnol.,1987; 25: 521-525.
Arshad M, Zia MA, Asghar M, Bhatti H. Improving bio-ethanol yield: Using virginiamycin and sodium flouride at a Pakistani distillery. Afr. J.Biotechnol.,2011; 10(53):11071-11074
Jones RK, Duncan HE, Payne GA, Leonard KJ.Factors influencing infection by Aspergillus flavus in silk-inoculated corn. Plant Disease,1980; 64: 859 - 863.
Lowes KF, Shearman CA, Payne J, McKenzie D, Archer DB, Merry RJ, Gasson MJ.Prevention of yeast spoilage in feed and food by the yeast mycocin HMK. Appl.Env.Microbiol. 2000; 66(3): 1066-1076.
Soares GAM, Sato HH. Characterization of the Saccharomyces cerevisiae Y500-4L killer toxin. Braz. J.Microbiol.,2000; 31: 291-297.