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Construction of a synthetic YdbK-dependent pyruvate:H2 pathway in Escherichia coli BL21(DE3)

Paper ID Volume ID Publish Year Pages File Format Full-Text
31911 44852 2009 9 PDF Available
Title
Construction of a synthetic YdbK-dependent pyruvate:H2 pathway in Escherichia coli BL21(DE3)
Abstract

A synthetic pyruvate:H2 pathway was constructed in Escherichia coli BL21(DE3) by co-expression of six proteins: E. coli YdbK, Clostridium pasteurianum [4Fe–4S]-ferredoxin, and Clostridium acetobutylicum HydF, HydE, HydG, and HydA. The effect of cofactor addition and host strain on H2 yield and fermentation product accumulation was studied, together with in vitro reconstitution of the entire pathway. The deletion of iscR and/or the addition of thiamine pyrophosphate to the medium enhanced the total and specific activity of recombinant YdbK and increased the yield of H2 per glucose. It was concluded that the introduced pathway outcompeted other pyruvate-consuming reactions, and that the ability to compete for pyruvate at least in part was determined by total YdbK activity. The results demonstrate the successful construction of a high-yielding H2 pathway in a microorganism that effectively does not synthesize any H2. The additional co-expression of Bacillus subtilis AmyE enabled starch-dependent H2 synthesis in minimal media.

Keywords
Escherichia coli BL21(DE3); Metabolic pathway construction; Pyruvate:ferredoxin-oxidoreductase; YdbK; Bacillus subtilis AmyE; Starch; H2
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Construction of a synthetic YdbK-dependent pyruvate:H2 pathway in Escherichia coli BL21(DE3)
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Publisher
Database: Elsevier - ScienceDirect
Journal: Metabolic Engineering - Volume 11, Issue 3, May 2009, Pages 139–147
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
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Full-text PDF Download
Online Support
Any Questions? feel free to contact us