title: Redirecting reductant flux into hydrogen production via metabolic engineering of fermentative carbon metabolism in a cyanobacterium.
abstract: Some aquatic microbial oxygenic photoautotrophs (AMOPs) make hydrogen (H(2)), a carbon-neutral, renewable product derived from water, in low yields during autofermentation (anaerobic metabolism) of intracellular carbohydrates previously stored during aerobic photosynthesis. We have constructed a mutant (the ldhA mutant) of the cyanobacterium Synechococcus sp. strain PCC 7002 lacking the enzyme for the NADH-dependent reduction of pyruvate to D-lactate, the major fermentative reductant sink in this AMOP. Both nuclear magnetic resonance (NMR) spectroscopy and liquid chromatography-mass spectrometry (LC-MS) metabolomic methods have shown that autofermentation by the ldhA mutant resulted in no D-lactate production and higher concentrations of excreted acetate, alanine, succinate, and hydrogen (up to 5-fold) compared to that by the wild type. The measured intracellular NAD(P)(H) concentrations demonstrated that the NAD(P)H/NAD(P)(+) ratio increased appreciably during autofermentation in the ldhA strain; we propose this to be the principal source of the observed increase in H(2) production via an NADH-dependent, bidirectional [NiFe] hydrogenase. Despite the elevated NAD(P)H/NAD(P)(+) ratio, no decrease was found in the rate of anaerobic conversion of stored carbohydrates. The measured energy conversion efficiency (ECE) from biomass (as glucose equivalents) converted to hydrogen in the ldhA mutant is 12%. Together with the unimpaired photoautotrophic growth of the ldhA mutant, these attributes reveal that metabolic engineering is an effective strategy to enhance H(2) production in AMOPs without compromising viability.
authors.affiliation: Waksman Institute and Department of Chemistry & Chemical Biology, Rutgers University, Piscataway, New Jersey 08854, USA.
authors.forename: Kelsey
authors.initials: K
authors.lastname: McNeely
authors.forename: Yu
authors.initials: Y
authors.lastname: Xu
authors.forename: Nick
authors.initials: N
authors.lastname: Bennette
authors.forename: Donald A
authors.initials: DA
authors.lastname: Bryant
authors.forename: G Charles
authors.initials: GC
authors.lastname: Dismukes
mesh_terms: D000085:Acetates / Q000378:metabolism; D000409:Alanine / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002237:Carbohydrate Dehydrogenases / Q000172:deficiency; D002244:Carbon / Q000378:metabolism*; D002853:Chromatography, Liquid; D005818:Genetic Engineering; D006859:Hydrogen / Q000378:metabolism*; D019344:Lactic Acid / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D053858:Metabolic Networks and Pathways / Q000235:genetics*; D055442:Metabolome; D010084:Oxidation-Reduction; D019289:Pyruvic Acid / Q000378:metabolism; D019802:Succinic Acid / Q000378:metabolism; D046940:Synechococcus / Q000235:genetics* / Q000378:metabolism*
chemical_list: D000085:Acetates; D001426:Bacterial Proteins; D019344:Lactic Acid; D002244:Carbon; D006859:Hydrogen; D019289:Pyruvic Acid; D019802:Succinic Acid; D002237:Carbohydrate Dehydrogenases; C020669:Glycerate dehydrogenase; D000409:Alanine