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Integration of an [FeFe]-hydrogenase into the anaerobic metabolism of Escherichia coli

Lookup NU author(s): Dr Ciaran Kelly, Professor Tracy Palmer, Professor Frank Sargent

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This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).


Abstract

© 2015 The Author. Biohydrogen is a potentially useful product of microbial energy metabolism. One approach to engineering biohydrogen production in bacteria is the production of non-native hydrogenase activity in a host cell, for example Escherichia coli. In some microbes, hydrogenase enzymes are linked directly to central metabolism via diaphorase enzymes that utilise NAD+/NADH cofactors. In this work, it was hypothesised that heterologous production of an NAD+/NADH-linked hydrogenase could connect hydrogen production in an E. coli host directly to its central metabolism. To test this, a synthetic operon was designed and characterised encoding an apparently NADH-dependent, hydrogen-evolving [FeFe]-hydrogenase from Caldanaerobacter subterranus. The synthetic operon was stably integrated into the E. coli chromosome and shown to produce an active hydrogenase, however no H2 production was observed. Subsequently, it was found that heterologous co-production of a pyruvate::ferredoxin oxidoreductase and ferredoxin from Thermotoga maritima was found to be essential to drive H2 production by this system. This work provides genetic evidence that the Ca.subterranus [FeFe]-hydrogenase could be operating in vivo as an electron-confurcating enzyme.


Publication metadata

Author(s): Kelly CL, Pinske C, Murphy BJ, Parkin A, Armstrong F, Palmer T, Sargent F

Publication type: Article

Publication status: Published

Journal: Biotechnology Reports

Year: 2015

Volume: 8

Pages: 94-104

Print publication date: 01/12/2015

Online publication date: 19/10/2015

Acceptance date: 06/10/2015

Date deposited: 14/02/2019

ISSN (electronic): 2215-017X

Publisher: Elsevier B.V.

URL: https://doi.org/10.1016/j.btre.2015.10.002

DOI: 10.1016/j.btre.2015.10.002


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