title: High-Throughput Screening of Catalytic H2Production.
abstract: Hydrogenases, ferredoxins, and ferredoxin-NADP+reductases (FNR) are redox proteins that mediate electron metabolism in vivo, and are also potential components for biological H2production technologies. A high-throughput H2production assay device (H2PAD) is presented that enables simultaneous evaluation of 96 individual H2production reactions to identify components that improve performance. Using a CCD camera and image analysis software, H2PAD senses the chemo-optical response of Pd/WO3thin films to the H2produced. H2PAD-enabled discovery of hydrogenase and FNR mutants that enhance biological H2production is reported. From a library of 10 080 randomly mutated Clostridium pasteurianum [FeFe] hydrogenases, we found a mutant with nearly 3-fold higher H2production specific activity. From a library of 400 semi-randomly mutated Oryza sativa FNR, the top hit enabled a 60 % increase in NADPH-driven H2production rates. H2PAD can also facilitate elucidation of fundamental biochemical mechanisms within these systems.
authors.affiliation: Department of Chemical Engineering, Stanford University, Stanford, CA, 94305, USA.
authors.forename: Jamin
authors.initials: J
authors.lastname: Koo
authors.affiliation: Department of Chemical Engineering, Stanford University, Stanford, CA, 94305, USA.
authors.forename: Tim
authors.initials: T
authors.lastname: Schnabel
authors.affiliation: Department of Bioengineering, Stanford University, Stanford, CA, 94305, USA.
authors.forename: Sylvie
authors.initials: S
authors.lastname: Liong
authors.affiliation: Department of Chemical Engineering, Stanford University, Stanford, CA, 94305, USA.
authors.forename: Niklaus H
authors.initials: NH
authors.lastname: Evitt
authors.affiliation: Department of Chemical Engineering, Stanford University, Stanford, CA, 94305, USA.|Department of Bioengineering, Stanford University, Stanford, CA, 94305, USA.
authors.forename: James R
authors.initials: JR
authors.lastname: Swartz
mesh_terms: D055162:Biocatalysis; D005287:Ferredoxin-NADP Reductase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D057166:High-Throughput Screening Assays; D006859:Hydrogen / Q000737:chemistry / Q000378:metabolism*; D006864:Hydrogenase / Q000737:chemistry / Q000378:metabolism*; D009154:Mutation
chemical_list: D006859:Hydrogen; D006864:Hydrogenase; D005287:Ferredoxin-NADP Reductase
keywords: high-throughput screening; hydrogen; hydrogenase; metalloproteins; redox chemistry