Abstract
Escherichia coli MG1655 (DE3) with the ability to synthesize butanol from glycerol was constructed by metabolic engineering. The genes thil, adhe2, bcs operon (crt, bcd, etfB, etfA, and hbd) were cloned into the plasmid vectors, pETDuet-1 and pACYCDuet-1, then the two resulting plasmids, pACYC-thl-bcs and pET-adhe2, were transferred to E. coli, and the recombinant strain was able to synthesize up to 18.5 mg/L butanol on a glycerol-containing medium. After the glycerol transport protein gene GlpF was expressed, the butanol production was improved to 22.7 mg/L. The competing pathway of byproducts, such as ethanol, succinate, and lactate, was subsequently deleted to improve the 1-butanol production to 97.9 mg/L. Moreover, a NADH regeneration system was introduced into the E. coli, and finally a 154.0 mg/L butanol titer was achieved in a laboratory-scale shake-flask experiment.
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This work was supported by Open Funding Project of the State Key Laboratory of Bioreactor Engineering, P. R. China.
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Zhou, P., Zhang, Y., Wang, P. et al. Butanol production from glycerol by recombinant Escherichia coli . Ann Microbiol 64, 219–227 (2014). https://doi.org/10.1007/s13213-013-0654-5
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DOI: https://doi.org/10.1007/s13213-013-0654-5