Applied Microbiology and Biotechnology

, Volume 63, Issue 6, pp 698–704 | Cite as

Production of isoamyl acetate in ackA-pta and/or ldh mutants of Escherichia coli with overexpression of yeast ATF2

  • R. V. Vadali
  • C. E. Horton
  • F. B. Rudolph
  • G. N. BennettEmail author
  • K.-Y. San
Original Paper


The gene coding for alcohol acetyltransferase (ATF2), which catalyzes the esterification of isoamyl alcohol and acetyl coenzyme A (acetyl-CoA), was cloned from Saccharomyces cerevisiae and expressed in Escherichia coli. This genetically engineered strain of E. coli produced the ester isoamyl acetate when isoamyl alcohol was added externally to the cell culture medium. Various competing pathways at the acetyl-CoA node were inactivated to increase the intracellular acetyl-CoA pool and divert more carbon flux to the ester synthesis pathway. Several strains with deletions in the ackA-pta and/or ldh pathways and bearing the ATF2 on a high-copy-number plasmid were constructed and studied. Compared to the wild-type, ackA-pta and nuo mutants produced higher amounts of ester and an ackA-pta-ldh-nuo mutant lower amounts. Isoamyl acetate production correlated well with intracellular coenzyme A (CoA) and acetyl-CoA levels. The ackA-pta-nuo mutant had the highest intracellular CoA/acetyl-CoA level and hence produced the highest amount of ester (1.75 mM) during the growth phase under oxic conditions and during the production phase under anoxic conditions.


Anoxic Condition Isoamyl Isoamyl Alcohol Oxic Condition Isoamyl Acetate 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



This research was supported in part by grants from U.S. Department of Agriculture 2002–35505–11638), National Science Foundation (BES-0118815) and by the Robert A. Welch Foundation C-1268 (GNB) and C-1372 (FBR). Experiments comply with the current laws of the USA.


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Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • R. V. Vadali
    • 1
  • C. E. Horton
    • 2
  • F. B. Rudolph
    • 2
  • G. N. Bennett
    • 2
    Email author
  • K.-Y. San
    • 1
    • 3
  1. 1.Department of BioengineeringRice UniversityHoustonUSA
  2. 2.Department of Biochemistry and Cell BiologyRice UniversityHoustonUSA
  3. 3.Department of Chemical EngineeringRice UniversityHoustonUSA

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