Improving heterologous polyketide production in <Emphasis Type="Italic">Escherichia coli</Emphasis> by overexpression of an <Emphasis Type="Italic">S</Emphasis>-adenosylmethionine synthetase gene |
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Authors: | Yong Wang Brett A Boghigian Blaine A Pfeifer |
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Institution: | (1) Department of Chemical and Biological Engineering, Tufts University, 4 Colby Street, Medford, MA 02155, USA;(2) State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, China |
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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
. 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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Keywords: | Polyketide Heterologous expression E coli 6-dEB S-adenosylmethionine synthetase |
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