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Expression systems for soluble metal-dependent formate dehydrogenase

Paper ID Volume ID Publish Year Pages File Format Full-Text
26478 43954 2015 9 PDF Available
Title
Expression systems for soluble metal-dependent formate dehydrogenase
Abstract

•We successfully established novel expression systems, which enables to produce soluble metal-dependent formate dehydrogenases more easily.•We successfully converted a membrane bound metal-dependent formate dehydrogenase from Escherichia coli into a soluble enzyme, and demonstrated that the newly created formate dehydrogenase can be purified under aerobic conditions.•We succeeded in the improvement of metal-dependent formate dehydrogenase by molecular evolution procedure.•We successfully create Desulfovibrio mutants expressing a soluble metal-dependent formate dehydrogenases tagged with Strep-tagII.

Molybdenum or tungsten-dependent formate dehydrogenases (FDH) can reduce carbon dioxide (CO2) to formate under ordinary conditions, and therefore, are considered promising catalysts for CO2 fixation. However, to our knowledge, no study on the modification of metal-dependent FDHs has been published, likely because of a lack of convenient expression systems for the recombinant enzymes. We attempted to establish an methodology for the preparation and modification of soluble oxygen-tolerant metal-dependent FDHs, for the following three strategies: (1) Escherichia coli FDH is converted from a membrane bound protein into a soluble protein by deleting the C-terminal membrane-anchor of small subunit (FdoH) and the whole membrane subunit (FdoI) and expressed homologously in E. coli; (2) originally soluble FDHs from Desulfovibrio are expressed heterologously in E. coli; and (3) Desulfovibrio FDHs are genetically engineered by the homologous gene recombination method and expressed homologously in Desulfovibrio. We successfully established the expression systems for (1) and (3), and succeeded in purification of the soluble FDHs with a tag by using affinity columns.

Keywords
Formate dehydrogenase; Recombinant protein; Expression system
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Expression systems for soluble metal-dependent formate dehydrogenase
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Photochemistry and Photobiology A: Chemistry - Volume 313, 1 December 2015, Pages 154–162
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
100% Money Back Guarantee
Full-text PDF Download
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Any Questions? feel free to contact us