Engineering Commons^™

Identification Of The [Fefe]-Hydrogenase Responsible For Hydrogen Generation In Thermoanaerobacterium Saccharolyticum And Demonstration Of Increased Ethanol Yield Via Hydrogenase Knockout, A. Joe Shaw, David A. Hogsett, Lee R. Lynd

Dartmouth Scholarship

Three putative hydrogenase enzyme systems in Thermoanaerobacterium saccharolyticum were investigated at the genetic, mRNA, enzymatic, and phenotypic levels. A four-gene operon containing two [FeFe]-hydrogenase genes, provisionally termed hfs (hydrogenase-Fe-S), was found to be the main enzymatic catalyst of hydrogen production. hfsB, perhaps the most interesting gene of the operon, contains an [FeFe]-hydrogenase and a PAS sensory domain and has several conserved homologues among clostridial saccharolytic, cellulolytic, and pathogenic bacteria. A second hydrogenase gene cluster, hyd, exhibited methyl viologen-linked hydrogenase enzymatic activity, but hyd gene knockouts did not influence the hydrogen yield of …