Abstract
An S-adenosylmethionine synthetase gene (metK) from Streptomyces spectabilis was cloned into an expression plasmid under the control of an inducible T7 promoter and introduced into a strain of Escherichia coli (BAP1(pBP130/pBP144)) capable of producing the polyketide product 6-deoxyerythronolide B (6-dEB). The metK coexpression in BAP1(pBP130/pBP144) improved the specific production of 6-dEB from 10.86 to 20.08 mg l−1 \( {\text{OD}}^{{ - 1}}_{{600}} \). In an effort to probe the reason for this improvement, a series of gene deletion and expression experiments were conducted based on a metK metabolic pathway that branches between propionyl-CoA (a 6-dEB precursor) and autoinducer compounds. The deletion and expression studies suggested that the autoinducer pathway had a larger impact on improved 6-dEB biosynthesis. Supporting these results were experiments demonstrating the positive effect conditioned media (the suspected location of the autoinducer compounds) had on 6-dEB production. Taken together, the results of this study show an increase in heterologous 6-dEB production concomitant with heterologous metK gene expression and suggest that the mechanism for this improvement is linked to native autoinducer compounds.
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Acknowledgements
We thank Prof. Yi-guang Wang (Institute of Medical Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College) for the provision of metK gene, the Tufts University BREEM Program (started through National Institutes of Health #R25 GM073177-01) for supporting BAB, and the Tufts University Faculty Research Awards Committee. YW and BAB have contributed equally to this work.
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Wang, Y., Boghigian, B.A. & Pfeifer, B.A. Improving heterologous polyketide production in Escherichia coli by overexpression of an S-adenosylmethionine synthetase gene. Appl Microbiol Biotechnol 77, 367–373 (2007). https://doi.org/10.1007/s00253-007-1172-9
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DOI: https://doi.org/10.1007/s00253-007-1172-9