1. Homologous overexpression of hydrogenase and glycerol dehydrogenase in Clostridium pasteurianum to enhance hydrogen production from crude glycerol
- Author
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Shyamali Sarma, Vijayanand S. Moholkar, Nigel P. Minton, David Ortega, and Vikash Kumar Dubey
- Subjects
Glycerol ,0106 biological sciences ,Environmental Engineering ,Hydrogenase ,Overexpression ,Bioconversion ,Clostridium pasteurianum ,Bioengineering ,010501 environmental sciences ,01 natural sciences ,law.invention ,chemistry.chemical_compound ,Hyda ,law ,010608 biotechnology ,Waste Management and Disposal ,0105 earth and related environmental sciences ,Crude glycerol ,Clostridium ,chemistry.chemical_classification ,biology ,Renewable Energy, Sustainability and the Environment ,Wild type ,General Medicine ,biology.organism_classification ,Enzyme ,chemistry ,Biochemistry ,Glycerol dehydrogenase ,Recombinant DNA ,Biohydrogen ,Hydrogen ,Sugar Alcohol Dehydrogenases - 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.
- Published
- 2019