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Title: Expression of a clostridial [FeFe]-hydrogenase in Chlamydomonas reinhardtii prolongs photo-production of hydrogen from water splitting

The high oxygen (O 2) sensitivity of green algal [FeFe]-hydrogenases is a significant limitation for the sustained production of hydrogen gas (H 2) from photosynthetic water splitting. To address this limitation we replaced the native [FeFe]-hydrogenases with a more O 2-tolerant clostridial [FeFe]-hydrogenase CaI in Chlamydomonas reinhardtii strain D66ΔHYD ( hydA1 hydA2) that contains insertionally inactivated [FeFe]-hydrogenases genes. Expression and translocation of CaI in D66ΔHYD led to the recovery of H 2 photoproduction at ~ 20% of the rates of the wild-type parent strain D66. We show for the first time that a bacterial [FeFe]-hydrogenase can be expressed, localized and matured to a catalytically active form that couples to photosynthetic electron transport in the green alga C. reinhardtii. The lower rates of O 2 inactivation of CaI led to more sustained H 2 photoproduction when cultures were challenged with O 2 or kept under prolonged illumination at solar intensities. Lastly, these results provide new insights into the requisites for attaining photobiological H 2 production from water splitting using a more O 2-tolerant hydrogenase.
 [1] ;  [2] ;  [1] ;  [1] ;  [3] ;  [4] ;  [1] ;  [1]
  1. National Renewable Energy Lab. (NREL), Golden, CO (United States)
  2. National Renewable Energy Lab. (NREL), Golden, CO (United States); U. S. Dept. of Energy, Washington, D.C. (United States)
  3. Colorado School of Mines, Golden, CO (United States); Algenol, Fort Myers, FL (United States)
  4. Colorado School of Mines, Golden, CO (United States)
Publication Date:
Report Number(s):
Journal ID: ISSN 2211-9264
Grant/Contract Number:
Accepted Manuscript
Journal Name:
Algal Research
Additional Journal Information:
Journal Volume: 22; Journal ID: ISSN 2211-9264
Research Org:
National Renewable Energy Lab. (NREL), Golden, CO (United States)
Sponsoring Org:
USDOE Office of Energy Efficiency and Renewable Energy (EERE), Fuel Cell Technologies Office (EE-3F); USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
Country of Publication:
United States
08 HYDROGEN; hydrogen photoproduction; hydrogenase; heterologous expression; O2 tolerance
OSTI Identifier:
Alternate Identifier(s):
OSTI ID: 1396621