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Open Access Original article

Bioconversion of glycerol to ethanol by a mutant Enterobacter aerogenes

RES Nwachukwu1, A Shahbazi25*, L Wang2, S Ibrahim3, M Worku4 and K Schimmel1

Author Affiliations

1 Energy and Environmental Systems, North Carolina A & T State University, Greensboro, NC, USA

2 Biological Engineering Program, North Carolina A & T State University, Greensboro, NC, USA

3 Food and Nutritional Sciences, North Carolina A & T State University, Greensboro, NC, USA

4 Department of Animal Sciences, North Carolina A & T State University, Greensboro, NC, USA

5 Biological Engineering Program, Sockwell Hall, North Carolina A & T State University, 1601 East Market Street, Greensboro, NC 27411, USA

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AMB Express 2012, 2:20  doi:10.1186/2191-0855-2-20

Published: 29 March 2012

Abstract

The main objective of this research is to develop, by adaptive evolution, mutant strains of Enterobacter aerogenes ATCC 13048 that are capable of withstanding high glycerol concentration as well as resisting ethanol-inhibition. The mutant will be used for high ethanol fermentation from glycerol feedstock. Ethanol production from pure (P-) and recovered (R-) glycerol using the stock was evaluated. A six-tube-subculture-generations method was used for developing the mutant. This involved subculturing the organism six consecutive times in tubes containing the same glycerol and ethanol concentrations at the same culture conditions. Then, the glycerol and/or ethanol concentration was increased and the six subculture generations were repeated. A strain capable of growing in 200 g/L glycerol and 30 g/L ethanol was obtained. The ability of this mutant, vis-à-vis the original strain, in utilizing glycerol in a high glycerol containing medium, with the concomitant ethanol yield, was assessed. Tryptic soy broth without dextrose (TSB) was used as the fermentation medium. Fermentation products were analyzed using HPLC.

In a 20 g/L glycerol TSB, E. aerogenes ATCC 13048 converted 18.5 g/L P-glycerol and 17.8 g/L R-glycerol into 12 and 12.8 g/L ethanol, respectively. In a 50 g/L P-glycerol TSB, it utilized only 15.6 g/L glycerol; but the new strain used up 39 g/L, yielding 20 g/L ethanol after 120 h, an equivalence of 1.02 mol ethanol/mol-glycerol. This is the highest ethanol yield reported from glycerol bioconversion. The result of this P-glycerol fermentation can be duplicated using the R-glycerol from biodiesel production.

Keywords:
Enterobacter aerogenes; Ethanol; Glycerol; Biodiesel; Fermentation