Isolation and Optimization of Ethanol Producing Bacteria from Natural Environments of Mazandaran Province in Iran

Document Type: Research Article

Authors

1 Department of Molecular and Cell Biology, Faculty of Science, University of Mazandaran, Babolsar, Iran

2 Department of Analytical Chemistry, Faculty of Chemistry, University of Mazandaran, Babolsar, Iran

Abstract

Ethanol producing bacteria are useful in industrial production of biofuel. There are interesting for screening of active bacteria from natural resources and introduce to biofuel industries. The present study aims to isolation of ethanol producing bacteria with characterization, optimization, and evaluation of their ethanol productivity. Samples from various fruits, plant saps and soils were screened for isolation of ethanol producing bacteria then evaluate to find the highest ethanol producer. Of all the 37 ethanol producing isolates, 6 highest producers were selected for characterization. Bacterial growth and ethanol production conditions were optimized based on pH, temperature, agitation, time and initial glucose concentration. Most isolates were occurred single or in pairs. All of isolates were motile and catalase positive but failed to hydrolyze gelatin and produce H2S. Among them, Zym6 was exhibited highest ethanol yield 6.28 gL-1 with optimum pH 6 and growth temperature 35 ˚C. In addition, Zym5 and Zym6 were exhibited highest ethanol yield 19.52 gL-1 and 18.75 gL-1 with xylose and tryptophan, respectively. Thus the optimum condition for ethanol production was a medium composed of pH 6, growth temperature 35 ˚C for 24-48 hours and xylose and tryptophan as carbon and nitrogen sources.

Keywords


Aggarwal NK, Niga P, Singh D, Yadav BS. 2001. Process optimization for the production of sugar for the bioethanol industry from sorghum, a non-conventional source of starch. Afr J Biotechnol 17: 411-415.

Brenner DJ, Krieg NR, Staley JT. 2004. Bergey's Manual of Systematic Bacteriology. 2nd ed., volume 2, Springer.

Chum HL, Overend RP 2001. Biomass and renewable fuels. Fuel Proc Technol 71: 187-195.

Desiniotis A, Kouvelis VN, Davenport K, Bruce D, Detter C, Tapia R, Han C, Goodwin LA, Woyke T, Kyrpides NC, Typas MA, Pappas KM. 2012. Complete genome sequence of the ethanol-producing Zymomonas mobilis subsp. mobilis centrotype ATCC 29191. J Bacteriol 194: 5966-5969.

Grootjen DRJ, Meijlink LHHM, Van der Lans RGJM, and Luyben KChAM. 1990. Cofermentation of glucose and xylose with immobilized Pichia stipites and Saccharomyces cerevisiae. Enzyme Microb Technol 12: 860-864.

Lee KJ, Scotinicki ML, Tribe DE, Rogers PL. 1981. The effects of temperature on the kinetics of ethanol production by strains of Zymomonas mobilis. Biotechnol Lett 3: 291-296.

Mohseni M, Ebrahimi H. 2013. Isolation, identification and optimization of ethanol producing bacteria from Saccharomyces-based fermentation process of alcohol industries in Iran. Bio J Microorg 2: 16-28.

Najafpour G, Younesi H, Syahidah Ku, Ismail K. 2004. Ethanol fermentation in an immobilized cell reactor using Saccharomyces cerevisiae. Bioresour Technol 92: 251-260.

Panesar PS, Marwaha SS, Rai R. 2000. Evaluation of ethanol production potential of Zymomonas mobilis strains. Asian Microb Biotechnol Environ 2: 15-19.

Panesar PS, Marwaha SS, Gill SS, Rai R. 2001. Screening of Zymomonas mobilis strains for ethanol production from molasses medium, J Chem Technol Biotechnol 41: 187-189.

Remize F, Roustan JL, Sablayrolles JM, Barre P, Dequin S. 1999. Glycerol overproduction by engineered Saccharomyces cerevisiae wine yeast strains leads to substantial changes in by-product formation and to a stimulation of fermentation rate in stationary phase. Appl Environ Microb 65: 143-149.

Rogers PL, Jeon YJ, Lee KJ, Lawford HG. 2007. Zymomonas mobilis for fuel ethanol and higher value products. Adv Biochem Eng Biotechnol 108: 263-288.

Rogers PL, Joachimsthal EL, Haggett KD. 1997. Ethanol from lignocellulosics: potential for a Zymomonas-based process. Australas Biotechnol 7: 304-309.

Rogers PL, Lee KJ, Skotnicki ML, Tribe DE. 1982. Ethanol production by Zymomonas mobilis. Microb React 23: 37-84.

Silva JP, Mussatto SI, Roberto IC. 2010. The Influence of Initial Xylose Concentration, Agitation, and Aeration on Ethanol Production by Pichia stipitis from Rice Straw Hemicellulosic Hydrolysate. Appl Biochem Biotechnol 162: 1306-1315.

Sobana P, Thirumalai V, Padma V, Vidhyadevi U. 2012. Cellulosic ethanol production by recombinant cellulolytic bacteria harbouring pdc and adh II genes of Zymomonas mobilis. Biotechnol Res Internat 10: 1-9.

Swings J, De Ley J. 1977. The biology of Zymomonas. Bacteriol Rev 41: 1-46.

Takeshi H, Tsuyoshi K, Kensuke F. 2012. Respiratory chain analysis of high ethanol producing Zymomonas mobilis mutants. Appl Environ Microbiol 10: 1-42.

Tao F, Miao JY, Shi GY, Zhang KC. 2005. Ethanol fermentation by an acid-tolerant Zymomonas mobilis under non-sterilized condition. Process Biochem 40: 183-187.

Wheals AE, Basso LC, Alves DM, Amorim H. V. 1999. Fuel ethanol after 25 years. Trends Biotechno 17: 482-487.

Wigmosta MS, Coleman AM, Skaggs RJ, Huesemann MH, Lane LJ. 2011. National microalgae biofuel production potential and resource demand. Water Resour Res 47: 1-13.

Yamashita Y, Kurosumi A, Sasaki C, Nakamura Y. 2008. Ethanol production from paper sludge by immobilized Zymomonas mobilis. Biochem Eng J 42: 314-319.

Yanase H, Miyawaki H, Sakurai M, Kawakami A, Matsumoto M, Haga K, Kojima M, Okamoto K. 2012. Ethanol production from wood hydrolysate using genetically engineered Zymomonas mobilis. Appl Microb Biotechnol 94: 1667-1678.

Yang Sh, Pan Ch, Tschaplinski TJ, Hurst GB, Engle NL, Zhou W, Dam Ph, Xu Y, Jr MR, Dice L, Johnson CM, Davison BH, Brown SD. 2013. System biology analysis of Zymomonas mobilis ZM4 ethanol stress responses. PLOS ONE 7: 1-14.