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Homologous overexpression of hydrogenase and glycerol dehydrogenase in Clostridium pasteurianum to enhance hydrogen production from crude glycerol

Sarma, Shyamali; Ortega, David; Minton, Nigel P.; Dubey, Vikash Kumar; Moholkar, Vijayanand S.

Authors

Shyamali Sarma

David Ortega

NIGEL MINTON nigel.minton@nottingham.ac.uk
Professor of Appliedmolecular Microbiology

Vikash Kumar Dubey

Vijayanand S. Moholkar vmoholkar@iitg.ernet.in@fax:913612582291



Abstract

This study reports engineering of a hypertransformable variant of C. pasteurianum for bioconversion of glycerol into hydrogen (H2). A functional glycerol-triggered hydrogen pathway was engineered based on two approaches: (1) increasing product yield by overexpression of immediate enzyme catalyzing H2 production, (2) increasing substrate uptake by overexpression of enzymes involved in glycerol utilization. The first strategy aimed at overexpression of hydA gene encoding hydrogenase, and the second one, through combination of overexpression of dhaD1 and dhaK genes encoding glycerol dehydrogenase and dihydroxyacetone kinase. These genetic manipulations resulted in two recombinant strains (hydA ++ /dhaD1K ++) capable of producing 97% H2 (v/v), with yields of 1.1 mol H2/mol glycerol in hydA overexpressed strain, and 0.93 mol H2/mol glycerol in dhaD1K overexpressed strain, which was 1.5 fold higher than wild type. Among two strains, dhaD1K ++ consumed more glycerol than hydA ++ which proves that overexpression of glycerol enzymes has enhanced glycerol intake rate.

Journal Article Type Article
Publication Date Jul 31, 2019
Journal Bioresource Technology
Print ISSN 0960-8524
Electronic ISSN 1873-2976
Publisher Elsevier
Peer Reviewed Peer Reviewed
Volume 284
Pages 168-177
APA6 Citation Sarma, S., Ortega, D., Minton, N. P., Dubey, V. K., & Moholkar, V. S. (2019). Homologous overexpression of hydrogenase and glycerol dehydrogenase in Clostridium pasteurianum to enhance hydrogen production from crude glycerol. Bioresource Technology, 284, 168-177. https://doi.org/10.1016/j.biortech.2019.03.074
DOI https://doi.org/10.1016/j.biortech.2019.03.074
Keywords Biohydrogen; Crude glycerol; Clostridium pasteurianum; Overexpression
Publisher URL https://www.sciencedirect.com/science/article/pii/S096085241930433X
Additional Information This article is maintained by: Elsevier; Article Title: Homologous overexpression of hydrogenase and glycerol dehydrogenase in Clostridium pasteurianum to enhance hydrogen production from crude glycerol; Journal Title: Bioresource Technology; CrossRef DOI link to publisher maintained version: https://doi.org/10.1016/j.biortech.2019.03.074; Content Type: article; Copyright: © 2019 Elsevier Ltd. All rights reserved.

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