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Appl Microbiol Biotechnol. 2010 Apr;86(4):1155-64. doi: 10.1007/s00253-009-2401-1. Epub 2010 Jan 14.

3-Methyl-1-butanol production in Escherichia coli: random mutagenesis and two-phase fermentation.

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Department of Chemical and Biomolecular Engineering, University of California, Los Angeles, CA 90095, USA.


Interest in producing biofuels from renewable sources has escalated due to energy and environmental concerns. Recently, the production of higher chain alcohols from 2-keto acid pathways has shown significant progress. In this paper, we demonstrate a mutagenesis approach in developing a strain of Escherichia coli for the production of 3-methyl-1-butanol by leveraging selective pressure toward L-leucine biosynthesis and screening for increased alcohol production. Random mutagenesis and selection with 4-aza-D,L-leucine, a structural analogue to L-leucine, resulted in the development of a new strain of E. coli able to produce 4.4 g/L of 3-methyl-1-butanol. Investigation of the host's sensitivity to 3-methyl-1-butanol directed development of a two-phase fermentation process in which titers reached 9.5 g/L of 3-methyl-1-butanol with a yield of 0.11 g/g glucose after 60 h.

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