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Steen EJ, Chan R, Prasad N, et al. Metabolic engineering of Saccharomyces cerevisiae for the production of n-butanol. Microb Cell Fact. 2008;7:36doi: 10.1186/1475-2859-7-36.
Steen, E. J., Chan, R., Prasad, N., Myers, S., Petzold, C. J., Redding, A., Ouellet, M., & Keasling, J. D. (2008). Metabolic engineering of Saccharomyces cerevisiae for the production of n-butanol. Microbial cell factories, 736. https://doi.org/10.1186/1475-2859-7-36
Steen, Eric J, et al. "Metabolic engineering of Saccharomyces cerevisiae for the production of n-butanol." Microbial cell factories vol. 7 (2008): 36. doi: https://doi.org/10.1186/1475-2859-7-36
Steen EJ, Chan R, Prasad N, Myers S, Petzold CJ, Redding A, Ouellet M, Keasling JD. Metabolic engineering of Saccharomyces cerevisiae for the production of n-butanol. Microb Cell Fact. 2008 Dec 03;7:36. doi: 10.1186/1475-2859-7-36. PMID: 19055772; PMCID: PMC2621116.
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Microb Cell Fact. 2008 Dec 03;7:36. doi: 10.1186/1475-2859-7-36.

Metabolic engineering of Saccharomyces cerevisiae for the production of n-butanol.

Microbial cell factories

Eric J Steen, Rossana Chan, Nilu Prasad, Samuel Myers, Christopher J Petzold, Alyssa Redding, Mario Ouellet, Jay D Keasling

Affiliations

  1. Joint BioEnergy Institute, 5885 Hollis Avenue, Emeryville, CA 94608, USA. [email protected].

PMID: 19055772 PMCID: PMC2621116 DOI: 10.1186/1475-2859-7-36

Abstract

BACKGROUND: Increasing energy costs and environmental concerns have motivated engineering microbes for the production of "second generation" biofuels that have better properties than ethanol.

RESULTS AND CONCLUSION: Saccharomyces cerevisiae was engineered with an n-butanol biosynthetic pathway, in which isozymes from a number of different organisms (S. cerevisiae, Escherichia coli, Clostridium beijerinckii, and Ralstonia eutropha) were substituted for the Clostridial enzymes and their effect on n-butanol production was compared. By choosing the appropriate isozymes, we were able to improve production of n-butanol ten-fold to 2.5 mg/L. The most productive strains harbored the C. beijerinckii 3-hydroxybutyryl-CoA dehydrogenase, which uses NADH as a co-factor, rather than the R. eutropha isozyme, which uses NADPH, and the acetoacetyl-CoA transferase from S. cerevisiae or E. coli rather than that from R. eutropha. Surprisingly, expression of the genes encoding the butyryl-CoA dehydrogenase from C. beijerinckii (bcd and etfAB) did not improve butanol production significantly as previously reported in E. coli. Using metabolite analysis, we were able to determine which steps in the n-butanol biosynthetic pathway were the most problematic and ripe for future improvement.

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