Engineering alternative butanol production platforms in heterologous bacteria |
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Authors: | David R. Nielsen Effendi Leonard Sang-Hwal Yoon Hsien-Chung Tseng Clara Yuan Kristala L. Jones Prather |
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Affiliation: | aDepartment of Chemical Engineering, 77 Massachusetts Institute of Technology, Room 66-458, Cambridge, MA 02139, USA;bSynthetic Biology Engineering Research Center (SynBERC), Massachusetts Institute of Technology, Cambridge, MA 02139, USA |
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Abstract: | 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. |
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Keywords: | Biofuel Butanol E. coli P. putida B. subtilis Tolerance Product inhibition |
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