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Ethanol production from biodiesel-derived crude glycerol by newly isolated Kluyvera cryocrescens

  • Bioenergy and Biofuels
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Abstract

The rapidly expanding market for biodiesel has increased the supply and reduced the cost of glycerol, making it an attractive sustainable feed stock for the fuel and chemical industry. Glycerol-based biorefinery is the microbial fermentation of crude glycerol to produce fuels and chemicals. A major challenge is to obtain microbes tolerant to inhibitors such as salts and organic solvents present in crude glycerol. Microbial screening was attempted to isolate novel strain capable of growing on crude glycerol as a sole carbon source. The newly isolated bacteria, identified as nonpathogenic Kluyvera cryocrescens S26 could convert biodiesel-derived crude glycerol to ethanol with high yield and productivity. The supplementation of nutrients such as yeast extract resulted in distinguished enhancement in cell growth as well as ethanol productivity under anaerobic condition. When glycerol fermentation is performed under microaerobic condition, there is also a remarkable improvement in cell growth, ethanol productivity and yield, compared with those under strict anaerobic condition. In batch fermentation under microaerobic condition, K. cryocrescens S26 produced 27 g/l of ethanol from crude glycerol with high molar yield of 80% and productivity of 0.61 g/l/h.

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References

  • Blankschien MD, Clomburg JM, Gonzalez R (2010) Metabolic engineering of Escherichia coli for the production of succinate from glycerol. Metab Eng 12:409–419

    Article  CAS  Google Scholar 

  • Brumm PJ, Hebeda RE (1998) Glycerol production in industrial alcohol fermentations. Biotechnol Lett 10:677–682

    Article  Google Scholar 

  • Çelik E, Ozbay N, Oktar N, Çalik P (2008) Use of biodiesel byproduct crude glycerol as the carbon source for fermentation processes by recombinant Pichia pastoris. Ind Eng Chem Res 47:2985–2990

    Article  Google Scholar 

  • Chen X, Xiu Z, Wang J, Zhang D, Xu P (2003) Stoichiometric analysis and experimental investigation of glycerol bioconversion to 1, 3-propanediol by Klebsiella pneumoniae under microaerobic conditions. Enzyme Microb Technol 33:386–394

    Article  CAS  Google Scholar 

  • Choi WJ (2008) Glycerol-based biorefinery for fuels and chemicals. Recent Pat Biotechnol 2:173–180

    Article  CAS  Google Scholar 

  • da Silva GP, Mack M, Contiero J (2009) Glycerol: a promising and abundant carbon source for industrial microbiology. Biotechnol Adv 27:30–39

    Article  Google Scholar 

  • de Graef MR, Alexeeva S, Snoep JL, Teixeira de Mattos MJ (1999) The steady-state internal redox state (NADH/NAD) reflects the external redox state and is correlated with catabolic adaptation in Escherichia coli. J Bacteriol 181:2351–2357

    Google Scholar 

  • Dharmadi Y, Murarka A, Gonzalez R (2006) Anaerobic fermentation of glycerol by Escherichia coli: a new platform for metabolic engineering. Biotechnol Bioeng 94:821–829

    Article  CAS  Google Scholar 

  • Durnin G, Clomburg J, Yeates Z, Alvarez PJJ, Zygourakis K, Campbell P, Gonzalez R (2009) Understanding and harnessing the microaerobic metabolism of glycerol in Escherichia coli. Biotechnol Bioeng 103:148–161

    Article  CAS  Google Scholar 

  • Gonzalez R, Murarka A, Dharmadi Y, Yazdani SS (2008) A new model for the anaerobic fermentation of glycerol in enteric bacteria: trunk and auxiliary pathways in Escherichia coli. Metab Eng 10:234–245

    Article  CAS  Google Scholar 

  • González-Pajuelo M, Andrade JC, Vasconcelos I (2004) Production of 1, 3-propanediol by Clostridium butyricum VPI 3266 using a synthetic medium and raw glycerol. J Ind Microbiol Biotechnol 31:442–446

    Article  Google Scholar 

  • Gupta A, Murarka A, Campbell P, Gonzalez R (2009) Anaerobic fermentation of glycerol in Paenibacillus macerans: metabolic pathways and environmental determinants. Appl Environ Microbiol 75:5871–5883

    Article  CAS  Google Scholar 

  • Herbert D, Phipps PJ, Strange RE (1971) Chemical analysis of microbial cells. In: Norris JM, Ribbons DW (eds) Methods in microbiology, vol 5b. Academic Press, London, pp 209–344

    Google Scholar 

  • Ibrahim MHA, Steinbüchel A (2010) Zobellella denitrificans strain MW1, a newly isolated bacterium suitable for poly(3-hydroxybutyrate) production from glycerol. J Appl Microbiol 108:214–225

    Article  CAS  Google Scholar 

  • Ito T, Nakashimada Y, Senba K, Matsui T, Nishio N (2005) Hydrogen and ethanol production from glycerol-containing wastes discharged after biodiesel manufacturing process. J Biosci Bioeng 100:260–265

    Article  CAS  Google Scholar 

  • Johnson DT, Taconi AT (2007) The glycerin glut: options for the value-added conversion of crude glycerol resulting from biodiesel production. Environ Prog 26:338–348

    Article  CAS  Google Scholar 

  • Karinen RS, Krause AOI (2006) New biocomponents from glycerol. Appl Catal Gen 306:128–133

    Article  CAS  Google Scholar 

  • Makri A, Fakas S, Aggelis G (2010) Metabolic activities of biotechnological interest in Yarrowia lipolytica grown on glycerol in repeated batch cultures. Bioresour Technol 101:2351–2358

    Article  CAS  Google Scholar 

  • Maleszka R, Wang PY, Schneider H (1982) Ethanol production from d-galactose and glycerol by Pachysolen tannophilus. Enzyme Microb Technol 4:349–352

    Article  CAS  Google Scholar 

  • Mantzouridou F, Naziri E, Tsimidou MZ (2008) Industrial glycerol as a supplementary carbon source in the production of β-carotene by Blakeslea trispora. J Agric Food Chem 56:2668–2675

    Article  CAS  Google Scholar 

  • Mazumdar S, Clomburg JM, Gonzalez R (2010) Escherichia coli strains engineered for homofermentative production of d-lactic acid from glycerol. Appl Environ Microbiol 76:4327–4336

    Article  CAS  Google Scholar 

  • Mu Y, Teng H, Zhang DJ, Wang W, Xiu ZL (2006) Microbial production of 1, 3-propanediol by Klebsiella pneumoniae using crude glycerol from biodiesel preparations. Biotechnol Lett 28:1755–1759

    Article  CAS  Google Scholar 

  • Munene CN, Kampen WH, Njapau H (2002) Effects of altering fermentation parameters on glycerol and bioethanol production from cane molasses. J Sci Food Agric 82:309–314

    Article  CAS  Google Scholar 

  • Murarka A, Dharmadi Y, Yazdani SS, Gonzalex R (2008) Fermentative utilization of glycerol by Escherichia coli and its implications for the production of fuels and chemicals. Appl Environ Microbiol 74:1124–1135

    Article  CAS  Google Scholar 

  • Nikel PI, Ramirez MC, Pettinari MJ, Méndez BS, Galvagno MA (2010) Ethanol synthesis from glycerol by Escherichia coli redox mutants expressing adhE from Leuconostoc mesenteroides. J Appl Microbiol 109:492–504

    CAS  Google Scholar 

  • Nissen TL, Hamann CW, Kielland-Brandt MC, Nielsen J, Villadsen J (2000) Anaerobic and aerobic batch cultivation of Saccharomyces cerevisiae mutants impaired in glycerol synthesis. Yeast 16:464–474

    Article  Google Scholar 

  • Pagliaro M, Ciriminna R, Kimura H, Rossi M, Pina CD (2007) From glycerol to value-added products. Angew Chem Int Ed 46:4434–4440

    Article  CAS  Google Scholar 

  • Papanikolaou S, Fakas S, Fick M, Chevalot I, Galiotou-Panayotou M, Komaitis M, Marc I, Aggelis G (2008) Biotechnological valorization of raw glycerol discharged after bio-diesel (fatty acid methyl esters) manufacturing process: production of 1,3-propanediol, citric acid and single cell oil. Biomass Bioenergy 32:60–71

    Article  CAS  Google Scholar 

  • Papanikolaou S, Fick M, Aggelis G (2004) The effect of raw glycerol concentration on the production of 1,3-propanediol by Clostridium butyricum. J Chem Technol Biotechnol 79:1189–1196

    Article  CAS  Google Scholar 

  • Petitdemange E, Dürr C, Andaloussi SA, Raval G (1995) Fermentation of raw glycerol to 1, 3-propanediol by new strains of Clostridium butyricum. J Ind Micribiol 15:498–502

    Article  CAS  Google Scholar 

  • Pyle DJ, Garcia RA, Wen Z (2008) Producing docosahexaenoic acid (DHA)-rich algae from biodiesel-derived crude glycerol: Effect of impurities on DHA production and algal biomass composition. J Agric Food Chem 56:3933–3939

    Article  CAS  Google Scholar 

  • Sakai S, Yagishita T (2007) Microbial production of hydrogen and ethanol from glycerol-containing wastes discharged from a biodiesel fuel production plant in a bioelectrochemical reactor with thionine. Biotechnol Bioeng 98:340–348

    Article  CAS  Google Scholar 

  • Scholten E, Renz T, Thomas J (2009) Continuous cultivation approach for fermentative succinic acid production from crude glycerol by Basfia succiniciproducens DD1. Biotechnol Lett 31:1947–1951

    Article  CAS  Google Scholar 

  • Shuler ML, Kargi F (2002) Bioprocess engineering: basic concepts. Prentice Hall, Upper Saddle River

    Google Scholar 

  • Temudo MF, Poldermans R, Kleerebezem R, van Loosdrecht MCM (2008) Glycerol fermentation by (open) mixed cultures: a chemostat study. Biotechnol Bioeng 100:1088–1098

    Article  CAS  Google Scholar 

  • Trinh CT, Srienc F (2009) Metabolic engineering of Escherichia coli for efficient conversion of glycerol to ethanol. Appl Environ Microbiol 75:6696–6705

    Article  CAS  Google Scholar 

  • Yazdani SS, Gonzalez R (2008) Engineering Escherichia coli for the efficient conversion of glycerol to ethanol and co-products. Metab Eng 10:340–351

    Article  CAS  Google Scholar 

  • Yu KO, Kim SW, Han SO (2010) Engineering of glycerol utilization pathway for ethanol production by Saccharomyces cerevisiae. Bioresour Technol 101:4157–4161

    Article  CAS  Google Scholar 

  • Zeng AP, Biebl H, Schlieker H, Deckwer WD (1993) Pathway analysis of glycerol fermentation by Klebsiella pneumoniae: regulation of reducing equivalent balance and product formation. Enzyme Microb Technol 15:770–779

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by Science and Engineering Research Council of Agency for Science, Technology and Research (A*STAR), Singapore, grant number ICES/07-173A02.

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Authors and Affiliations

  1. Institute of Chemical and Engineering Sciences, Agency for Science, Technology and Research (A*STAR), 1 Pesek Road, Jurong Island, Singapore

    Won Jae Choi, Maria Regina Hartono, Weng Heng Chan & Suan Siong Yeo

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  1. Won Jae Choi
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  2. Maria Regina Hartono
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  3. Weng Heng Chan
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  4. Suan Siong Yeo
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Correspondence to Won Jae Choi.

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Choi, W.J., Hartono, M.R., Chan, W.H. et al. Ethanol production from biodiesel-derived crude glycerol by newly isolated Kluyvera cryocrescens . Appl Microbiol Biotechnol 89, 1255–1264 (2011). https://doi.org/10.1007/s00253-010-3076-3

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