3-Methyl-1-butanol production in Escherichia coli: random mutagenesis and two-phase fermentation.
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.
Connor
Michael R
MR
Department of Chemical and Biomolecular Engineering, University of California, Los Angeles, CA 90095, USA.
Cann
Anthony F
AF
Liao
James C
JC
D056804:Biofuels / Q000382:microbiology*; D004926:Escherichia coli / Q000378:metabolism*; D005285:Fermentation; D007930:Leucine / Q000378:metabolism; D016296:Mutagenesis; D000439:Pentanols / Q000378:metabolism*; D012641:Selection, Genetic
D056804:Biofuels; D000439:Pentanols; C029683:isopentyl alcohol; D007930:Leucine