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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.
Authors:
 [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):
NREL/JA-2700-67527
Journal ID: ISSN 2211-9264
Grant/Contract Number:
AC36-08GO28308
Type:
Accepted Manuscript
Journal Name:
Algal Research
Additional Journal Information:
Journal Volume: 22; Journal ID: ISSN 2211-9264
Publisher:
Elsevier
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
Language:
English
Subject:
08 HYDROGEN; hydrogen photoproduction; hydrogenase; heterologous expression; O2 tolerance
OSTI Identifier:
1339351
Alternate Identifier(s):
OSTI ID: 1396621

Noone, Seth, Ratcliff, Kathleen, Davis, ReAnna, Subramanian, Venkataramanan, Meuser, Jonathan, Posewitz, Matthew C., King, Paul W., and Ghirardi, Maria L.. Expression of a clostridial [FeFe]-hydrogenase in Chlamydomonas reinhardtii prolongs photo-production of hydrogen from water splitting. United States: N. p., Web. doi:10.1016/j.algal.2016.12.014.
Noone, Seth, Ratcliff, Kathleen, Davis, ReAnna, Subramanian, Venkataramanan, Meuser, Jonathan, Posewitz, Matthew C., King, Paul W., & Ghirardi, Maria L.. Expression of a clostridial [FeFe]-hydrogenase in Chlamydomonas reinhardtii prolongs photo-production of hydrogen from water splitting. United States. doi:10.1016/j.algal.2016.12.014.
Noone, Seth, Ratcliff, Kathleen, Davis, ReAnna, Subramanian, Venkataramanan, Meuser, Jonathan, Posewitz, Matthew C., King, Paul W., and Ghirardi, Maria L.. 2016. "Expression of a clostridial [FeFe]-hydrogenase in Chlamydomonas reinhardtii prolongs photo-production of hydrogen from water splitting". United States. doi:10.1016/j.algal.2016.12.014. https://www.osti.gov/servlets/purl/1339351.
@article{osti_1339351,
title = {Expression of a clostridial [FeFe]-hydrogenase in Chlamydomonas reinhardtii prolongs photo-production of hydrogen from water splitting},
author = {Noone, Seth and Ratcliff, Kathleen and Davis, ReAnna and Subramanian, Venkataramanan and Meuser, Jonathan and Posewitz, Matthew C. and King, Paul W. and Ghirardi, Maria L.},
abstractNote = {The high oxygen (O2) sensitivity of green algal [FeFe]-hydrogenases is a significant limitation for the sustained production of hydrogen gas (H2) from photosynthetic water splitting. To address this limitation we replaced the native [FeFe]-hydrogenases with a more O2-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 H2 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 O2 inactivation of CaI led to more sustained H2 photoproduction when cultures were challenged with O2 or kept under prolonged illumination at solar intensities. Lastly, these results provide new insights into the requisites for attaining photobiological H2 production from water splitting using a more O2-tolerant hydrogenase.},
doi = {10.1016/j.algal.2016.12.014},
journal = {Algal Research},
number = ,
volume = 22,
place = {United States},
year = {2016},
month = {12}
}