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.

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.

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

• Journal Article Type: Article
• Acceptance Date: Mar 15, 2019
• Online Publication Date: Mar 16, 2019
• Publication Date: Jul 31, 2019
• Deposit Date: May 3, 2019
• Publicly Available Date: Mar 17, 2020
• Journal: Bioresource Technology
• Print ISSN: 0960-8524
• Electronic ISSN: 1873-2976
• Publisher: Elsevier
• Peer Reviewed: Peer Reviewed
• Volume: 284
• Pages: 168-177
• DOI: https://doi.org/10.1016/j.biortech.2019.03.074
• Keywords: Biohydrogen; Crude glycerol; Clostridium pasteurianum; Overexpression
• Public URL: https://nottingham-repository.worktribe.com/output/2018341
• 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.
• Contract Date: May 3, 2019