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- Title
Improving heterologous polyketide production in Escherichia coli by overexpression of an S-adenosylmethionine synthetase gene.
- Authors
Yong Wang; Boghigian, Brett A.; Pfeifer, Blaine A.
- 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.
- Subjects
STREPTOMYCES; ESCHERICHIA coli; ADENOSYLMETHIONINE; POLYKETIDES; GENE expression; GENETIC regulation
- Publication
Applied Microbiology & Biotechnology, 2007, Vol 77, Issue 2, p367
- ISSN
0175-7598
- Publication type
Article
- DOI
10.1007/s00253-007-1172-9