Redirecting reductant flux into hydrogen production via metabolic engineering of fermentative carbon metabolism in a cyanobacterium

Kelsey McNeely, Yu Xu, Nick Bennette, Donald A. Bryant, G. Charles Dismukes

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140 Scopus citations


Some aquatic microbial oxygenic photoautotrophs (AMOPs) make hydrogen (H2), 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 IdhA 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 IdhA 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 IdhA strain; we propose this to be the principal source of the observed increase in H2 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 IdhA mutant is 12%. Together with the unimpaired photoautotrophic growth of the IdhA mutant, these attributes reveal that metabolic engineering is an effective strategy to enhance H2 production in AMOPs without compromising viability.

Original languageEnglish (US)
Pages (from-to)5032-5038
Number of pages7
JournalApplied and environmental microbiology
Issue number15
StatePublished - Aug 2010

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Food Science
  • Applied Microbiology and Biotechnology
  • Ecology


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