Engineering of an Escherichia coli Strain for the Production of 3-Methyl-1-Butanol

ABSTRACT 3-Methyl-1-butanol is a potential fuel additive or substitute. Previously this compound was identified in small quantities in yeast fermentation as one of the fusel alcohols. In this work, we engineered an Escherichia coli strain to produce 3-methyl-1-butanol from glucose via the host's amino acid biosynthetic pathways. Strain improvement with the removal of feedback inhibition and competing pathways increased the selectivity and productivity of 3-methyl-1-butanol. This work demonstrates the feasibility of production of 3-methyl-1-butanol as a biofuel and shows promise in using E. coli as a host for production.

[1]  L. Dijkhuizen,et al.  Rapid identification of target genes for 3-methyl-1-butanol production in Saccharomyces cerevisiae , 2006, Applied Microbiology and Biotechnology.

[2]  B. Pearson,et al.  A 13C Nuclear Magnetic Resonance Investigation of the Metabolism of Leucine to Isoamyl Alcohol in Saccharomyces cerevisiae * , 1997, The Journal of Biological Chemistry.

[3]  W. M. Haynes.  CRC Handbook of Chemistry and Physics , 1990 .

[4]  J. Keasling,et al.  Engineering a mevalonate pathway in Escherichia coli for production of terpenoids , 2003, Nature Biotechnology.

[5]  J. Liao,et al.  Non-fermentative pathways for synthesis of branched-chain higher alcohols as biofuels , 2008, Nature.

[6]  G. Stephanopoulos,et al.  Engineering Yeast Transcription Machinery for Improved Ethanol Tolerance and Production , 2006, Science.

[7]  R. C. Weast.  CRC Handbook of Chemistry and Physics , 1973 .

[8]  K. Shanmugam,et al.  Engineering Escherichia coli for efficient conversion of glucose to pyruvate. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[9]  B. Wanner,et al.  One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[10]  A. Salis,et al.  Biodiesel production from triolein and short chain alcohols through biocatalysis. , 2005, Journal of biotechnology.

[11]  G. Bennett,et al.  Production of isoamyl acetate in ackA-pta and/or ldh mutants of Escherichia coli with overexpression of yeast ATF2 , 2004, Applied Microbiology and Biotechnology.

[12]  Kevin M. Smith,et al.  Metabolic engineering of Escherichia coli for 1-butanol production. , 2008, Metabolic engineering.

[13]  E. Papoutsakis,et al.  Dynamics of Genomic-Library Enrichment and Identification of Solvent Tolerance Genes for Clostridium acetobutylicum , 2007, Applied and Environmental Microbiology.

[14]  W. R. Farmer,et al.  Improving lycopene production in Escherichia coli by engineering metabolic control , 2000, Nature Biotechnology.

[15]  G. Bennett,et al.  Heterologous expression of the Saccharomyces cerevisiae alcohol acetyltransferase genes in Clostridium acetobutylicum and Escherichia coli for the production of isoamyl acetate , 2003, Journal of Industrial Microbiology and Biotechnology.

[16]  H. Mori,et al.  Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection , 2006, Molecular systems biology.

[17]  H. Bujard,et al.  Independent and tight regulation of transcriptional units in Escherichia coli via the LacR/O, the TetR/O and AraC/I1-I2 regulatory elements. , 1997, Nucleic acids research.

[18]  K. Horikoshi,et al.  Enhanced production of isoamyl alcohol and isoamyl acetate by ubiquitination-deficient Saccharomyces cerevisiae mutants. , 2005, Cellular & molecular biology letters.

[19]  J. C. Liao,et al.  Engineered Synthetic Pathway for Isopropanol Production in Escherichia coli , 2007, Applied and Environmental Microbiology.

[20]  Stacy Cagle Davis,et al.  Transportation energy data book , 2008 .

[21]  J. Liao,et al.  Metabolic engineering of Escherichia coli for 1-butanol and 1-propanol production via the keto-acid pathways. , 2008, Metabolic engineering.

[22]  L. Ingram,et al.  Isolation and characterization of ethanol-tolerant mutants of Escherichia coli KO11 for fuel ethanol production , 1998, Journal of Industrial Microbiology and Biotechnology.