Engineering alternative butanol production platforms in heterologous bacteria

David R. Nielsen, Effendi Leonard, Sang Hwal Yoon, Hsien Chung Tseng, Clara Yuan, Kristala L.Jones Prather

Research output: Contribution to journalArticlepeer-review

338 Scopus citations


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.

Original languageEnglish (US)
Pages (from-to)262-273
Number of pages12
JournalMetabolic Engineering
Issue number4-5
StatePublished - Jul 2009
Externally publishedYes


  • B. subtilis
  • Biofuel
  • Butanol
  • E. coli
  • P. putida
  • Product inhibition
  • Tolerance

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology


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