Hydrogenase fusion protein for improved hydrogen production
AssigneeThe Board of Trustees of the Leland Stanford Junior University
InventorsPhillip Richard Smith, James R. Swartz
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Abstract
Compositions of a fusion protein comprising a spatially tethered ferredoxin-NADP-reductase (FNR) and an active [FeFe] hydrogenase, genetic sequences encoding such fusion proteins, and methods of use thereof are provided. The fusion proteins of the invention link an FNR polypeptide to an active [FeFe] hydrogenase through a polypeptide linker. The fusion protein facilitates improved electron transfer through a ferredoxin, and allows direct electron transfer from NADPH to the hydrogenase.
Claims13
A fusion protein comprising an active FeFe hydrogenase joined through a polypeptide linker to an active ferredoxin-NADP-reductase (FNR) in which the fusion protein, when provided in a cell lysate or a product of cell-free protein at a concentration of at least 0.64 μM synthesis is capable of catalyzing a volume production of H2 of at least about 5 mmol H2 L−1 hr−1 in a reaction mix comprising a sugar at a concentration of 5 mM and a ferredoxin at a concentration of 50 μM.
A fusion protein comprising an active FeFe hydrogenase joined through a polypeptide linker to an active ferredoxin-NADP-reductase (FNR) in which the fusion protein, when provided in a cell lysate or a product of cell-free protein at a concentration of at least 0.64 μM synthesis is capable of catalyzing a volume production of H2 of at least about 5 mmol H2 L−1 hr−1 in a reaction mix comprising a sugar at a concentration of 5 mM and a ferredoxin at a concentration of 50 μM, wherein the FeFe hydrogenase is Clostridium pasteurianum hydrogenase.
A fusion protein comprising an active FeFe hydrogenase joined through a polypeptide linker to an active ferredoxin-NADP-reductase (FNR) wherein the protein has an amino acid sequence set forth in any one of SEQ ID NO:
- 2, 4, 6, and 8.
Dependencies are inferred from the claim text.