PMID:21317262

Citation	McNeely, K, Xu, Y, Ananyev, G, Bennette, N, Bryant, DA and Dismukes, GC (2011) Synechococcus sp. strain PCC 7002 nifJ mutant lacking pyruvate:ferredoxin oxidoreductase. Appl. Environ. Microbiol. 77:2435-44
Abstract	The nifJ gene codes for pyruvate:ferredoxin oxidoreductase (PFOR), which reduces ferredoxin during fermentative catabolism of pyruvate to acetyl-coenzyme A (acetyl-CoA). A nifJ knockout mutant was constructed that lacks one of two pathways for the oxidation of pyruvate in the cyanobacterium Synechococcus sp. strain PCC 7002. Remarkably, the photoautotrophic growth rate of this mutant increased by 20% relative to the wild-type (WT) rate under conditions of light-dark cycling. This result is attributed to an increase in the quantum yield of photosystem II (PSII) charge separation as measured by photosynthetic electron turnover efficiency determined using fast-repetition-rate fluorometry (F(v)/F(m)). During autofermentation, the excretion of acetate and lactate products by nifJ mutant cells decreased 2-fold and 1.2-fold, respectively. Although nifJ cells displayed higher in vitro hydrogenase activity than WT cells, H(2) production in vivo was 1.3-fold lower than the WT level. Inhibition of acetate-CoA ligase and pyruvate dehydrogenase complex by glycerol eliminated acetate production, with a resulting loss of reductant and a 3-fold decrease in H(2) production by nifJ cells compared to WT cells. Continuous electrochemical detection of dissolved H(2) revealed two temporally resolved phases of H(2) production during autofermentation, a minor first phase and a major second phase. The first phase was attributed to reduction of ferredoxin, because its level decreased 2-fold in nifJ cells. The second phase was attributed to glycolytic NADH production and decreased 20% in nifJ cells. Measurement of the intracellular NADH/NAD(+) ratio revealed that the reductant generated by PFOR contributing to the first phase of H(2) production was not in equilibrium with bulk NADH/NAD(+) and that the second phase corresponded to the equilibrium NADH-mediated process.
Links	PubMed PMC3067432 Online version:10.1128/AEM.02792-10
Keywords	Acetates/metabolism; Darkness; Fermentation; Gene Knockout Techniques; Hydrogen/metabolism; Lactic Acid/metabolism; Light; NAD/metabolism; Oxidation-Reduction; Pyruvate Synthase/deficiency; Pyruvate Synthase/genetics; Pyruvic Acid/metabolism; Synechococcus/enzymology; Synechococcus/genetics; Synechococcus/growth & development
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
KLEPN:NIFJ		GO:0008901: ferredoxin hydrogenase activity	ECO:0000314:		F		Table 2. Hydrogenase activities.	complete CACAO 9136

References

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