Engineering alternative butanol production platforms in heterologous bacteria

Metab Eng. 2009 Jul-Sep;11(4-5):262-73. doi: 10.1016/j.ymben.2009.05.003. Epub 2009 May 21.


Alternative microbial hosts have been engineered as biocatalysts for butanol biosynthesis. The butanol synthetic pathway of Clostridium acetobutylicum was first re-constructed in Escherichia coli to establish a baseline for comparison to other hosts. Whereas polycistronic expression of the pathway genes resulted in the production of 34 mg/L butanol, individual expression of pathway genes elevated titers to 200 mg/L. Improved titers were achieved by co-expression of Saccharomyces cerevisiae formate dehydrogenase while overexpression of E. coli glyceraldehyde 3-phosphate dehydrogenase to elevate glycolytic flux improved titers to 580 mg/L. Pseudomonas putida and Bacillus subtilis were also explored as alternative production hosts. Polycistronic expression of butanol biosynthetic genes yielded butanol titers of 120 and 24 mg/L from P. putida and B. subtilis, respectively. Production in the obligate aerobe P. putida was dependent upon expression of bcd-etfAB. These results demonstrate the potential of engineering butanol biosynthesis in a variety of heterologous microorganisms, including those cultivated aerobically.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Bacillus subtilis / genetics
  • Bacillus subtilis / metabolism
  • Butanols / metabolism*
  • Clostridium acetobutylicum / genetics
  • Clostridium acetobutylicum / metabolism
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Escherichia coli Proteins / metabolism
  • Gene Expression Regulation, Bacterial
  • Genes, Bacterial
  • Genetic Engineering / methods*
  • Genetic Enhancement / methods*
  • Glyceraldehyde 3-Phosphate / genetics
  • Protein Engineering / methods*


  • Butanols
  • Escherichia coli Proteins
  • Glyceraldehyde 3-Phosphate