Fluidic production of molecular hydrogen using the biocatalyst hydrogenase

CHED 211

Dontarie M. Stallings, dstallings@georgiasouthern.edu, Kristin D’Antignac, and Kathleen Knappenberger. Department of Chemistry, Georgia Southern University, P. O. Box 8064, Statesboro, GA 30460
The enzyme hydrogenase, from the anaerobic bacterium Desulfovibrio desulfuricans, catalyzes the production of molecular hydrogen [2e- + 2H+ → H2] with a turnover rate of 10,400 H2 evolved / min per mg using dithionite-reduced methyl viologen as electron donor. In this research, Desulfovibrio desulfuricans hydrogenase was expressed with and without an altered Shine-Dalgarno sequence in the cyanobacterium Synechococcus PCC7942. The structural gene encoding hydrogenase was converted to that of the cat, chloramphenicol acetyltransferase, gene. Expression and activity of Desulfovibrio desulfuricans, altered structural gene versus wild type, was compared. Expression was confirmed by Western and Northern blot. H2 evolution was measured via Methyl Viologen assay.