The dual effect of a ferredoxin-hydrogenase fusion protein in vivo: successful divergence of the photosynthetic electron flux towards hydrogen production and elevated oxygen tolerance

Eilenberg, Haviva; Weiner, Iddo; Ben-Zvi, Oren; Pundak, Carmel; Marmari, Abigail; Liran, Oded; Wecker, Matt S.; Milrad, Yuval; Yacoby, Iftach

doi:10.1186/s13068-016-0601-3

Title: The dual effect of a ferredoxin-hydrogenase fusion protein in vivo: successful divergence of the photosynthetic electron flux towards hydrogen production and elevated oxygen tolerance

Journal Article · Tue Aug 30 00:00:00 EDT 2016 · Biotechnology for Biofuels

DOI:https://doi.org/10.1186/s13068-016-0601-3· OSTI ID:1626974

Eilenberg, Haviva ^[1]; Weiner, Iddo ^[1]; Ben-Zvi, Oren ^[1]; Pundak, Carmel ^[1]; Marmari, Abigail ^[1]; Liran, Oded ^[1]; Wecker, Matt S. ^[2]; Milrad, Yuval ^[1];

^[1]

Tel Aviv Univ., Ramat Aviv (Israel). The George S. Wise Faculty of Life Science. Dept. of Molecular Biology and Ecology of Plants
GeneBiologics, LLC, Boulder, CO (United States); National Renewable Energy Lab. (NREL), Golden, CO (United States)

Background: Hydrogen photo-production in green algae, catalyzed by the enzyme [FeFe]-hydrogenase (HydA), is considered a promising source of renewable clean energy. Yet, a significant increase in hydrogen production efficiency is necessary for industrial scale-up. We have previously shown that a major challenge to be resolved is the inferior competitiveness of HydA with NADPH production, catalyzed by ferredoxin-NADP+-reductase (FNR). In this work, we explored the in vivo hydrogen production efficiency of Fd-HydA, where the electron donor ferredoxin (Fd) is fused to HydA and expressed in the model organism Chlamydomonas reinhardtii. Results: We show that once the Fd-HydA fusion gene is expressed in micro-algal cells of C. reinhardtii, the fusion enzyme is able to intercept photosynthetic electrons and use them for efficient hydrogen production, thus supporting the previous observations made in vitro. We found that Fd-HydA has a ~4.5-fold greater photosynthetic hydrogen production rate standardized for hydrogenase amount (PHPRH) than that of the native HydA in vivo. Furthermore, we provide evidence suggesting that the fusion protein is more resistant to oxygen than the native HydA. Conclusions: The in vivo photosynthetic activity of the Fd-HydA enzyme surpasses that of the native HydA and shows higher oxygen tolerance. Therefore, our results provide a solid platform for further engineering efforts towards efficient hydrogen production in microalgae through the expression of synthetic enzymes

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Research Organization:: National Renewable Energy Laboratory (NREL), Golden, CO (United States)

Sponsoring Organization:: USDOE Office of Science (SC)

Grant/Contract Number:: AC36-08GO28308

OSTI ID:: 1626974

Journal Information:: Biotechnology for Biofuels, Vol. 9, Issue 1; ISSN 1754-6834

Publisher:: BioMed CentralCopyright Statement

Country of Publication:: United States

Language:: English

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Related Subjects

59 BASIC BIOLOGICAL SCIENCES
Biotechnology & Applied Microbiology
Energy & Fuels
H2 production
Ferredoxin
Hydrogenase
Oxygen sensitivity
Fusion enzyme
Chlamydomonas reinhardtii

of The dual effect of a ferredoxin-hydrogenase fusion protein in vivo: successful divergence of the photosynthetic electron flux towards hydrogen production and elevated oxygen tolerance Eilenberg, Haviva; Weiner, Iddo; Ben-Zvi, Oren https://doi.org/10.6084/m9.figshare.10037153 The current record is supplemented by this image	image	January 2019
of The dual effect of a ferredoxin-hydrogenase fusion protein in vivo: successful divergence of the photosynthetic electron flux towards hydrogen production and elevated oxygen tolerance Eilenberg, Haviva; Weiner, Iddo; Ben-Zvi, Oren https://doi.org/10.6084/m9.figshare.10037153.v1 The current record is supplemented by this image	image	January 2019
of The dual effect of a ferredoxin-hydrogenase fusion protein in vivo: successful divergence of the photosynthetic electron flux towards hydrogen production and elevated oxygen tolerance Eilenberg, Haviva; Weiner, Iddo; Ben-Zvi, Oren https://doi.org/10.6084/m9.figshare.10037156 The current record is supplemented by this image	image	January 2019
of The dual effect of a ferredoxin-hydrogenase fusion protein in vivo: successful divergence of the photosynthetic electron flux towards hydrogen production and elevated oxygen tolerance Eilenberg, Haviva; Weiner, Iddo; Ben-Zvi, Oren https://doi.org/10.6084/m9.figshare.10037156.v1 The current record is supplemented by this image	image	January 2019
of The dual effect of a ferredoxin-hydrogenase fusion protein in vivo: successful divergence of the photosynthetic electron flux towards hydrogen production and elevated oxygen tolerance Eilenberg, Haviva; Weiner, Iddo; Ben-Zvi, Oren https://doi.org/10.6084/m9.figshare.10037159 The current record is supplemented by this image	image	January 2019
of The dual effect of a ferredoxin-hydrogenase fusion protein in vivo: successful divergence of the photosynthetic electron flux towards hydrogen production and elevated oxygen tolerance Eilenberg, Haviva; Weiner, Iddo; Ben-Zvi, Oren https://doi.org/10.6084/m9.figshare.10037159.v1 The current record is supplemented by this image	image	January 2019
of The dual effect of a ferredoxin-hydrogenase fusion protein in vivo: successful divergence of the photosynthetic electron flux towards hydrogen production and elevated oxygen tolerance Eilenberg, Haviva; Weiner, Iddo; Ben-Zvi, Oren https://doi.org/10.6084/m9.figshare.10037162 The current record is supplemented by this image	image	January 2019
of The dual effect of a ferredoxin-hydrogenase fusion protein in vivo: successful divergence of the photosynthetic electron flux towards hydrogen production and elevated oxygen tolerance Eilenberg, Haviva; Weiner, Iddo; Ben-Zvi, Oren https://doi.org/10.6084/m9.figshare.10037162.v1 The current record is supplemented by this image	image	January 2019
of The dual effect of a ferredoxin-hydrogenase fusion protein in vivo: successful divergence of the photosynthetic electron flux towards hydrogen production and elevated oxygen tolerance Eilenberg, Haviva; Weiner, Iddo; Ben-Zvi, Oren https://doi.org/10.6084/m9.figshare.10037165 The current record is supplemented by this image	image	January 2019
of The dual effect of a ferredoxin-hydrogenase fusion protein in vivo: successful divergence of the photosynthetic electron flux towards hydrogen production and elevated oxygen tolerance Eilenberg, Haviva; Weiner, Iddo; Ben-Zvi, Oren https://doi.org/10.6084/m9.figshare.10037165.v1 The current record is supplemented by this image	image	January 2019
of The dual effect of a ferredoxin-hydrogenase fusion protein in vivo: successful divergence of the photosynthetic electron flux towards hydrogen production and elevated oxygen tolerance Eilenberg, Haviva; Weiner, Iddo; Ben-Zvi, Oren https://doi.org/10.6084/m9.figshare.10037168 The current record is supplemented by this image	image	January 2019
of The dual effect of a ferredoxin-hydrogenase fusion protein in vivo: successful divergence of the photosynthetic electron flux towards hydrogen production and elevated oxygen tolerance Eilenberg, Haviva; Weiner, Iddo; Ben-Zvi, Oren https://doi.org/10.6084/m9.figshare.10037168.v1 The current record is supplemented by this image	image	January 2019
of The dual effect of a ferredoxin-hydrogenase fusion protein in vivo: successful divergence of the photosynthetic electron flux towards hydrogen production and elevated oxygen tolerance Eilenberg, Haviva; Weiner, Iddo; Ben-Zvi, Oren https://doi.org/10.6084/m9.figshare.10037171 The current record is supplemented by this image	image	January 2019
of The dual effect of a ferredoxin-hydrogenase fusion protein in vivo: successful divergence of the photosynthetic electron flux towards hydrogen production and elevated oxygen tolerance Eilenberg, Haviva; Weiner, Iddo; Ben-Zvi, Oren https://doi.org/10.6084/m9.figshare.10037171.v1 The current record is supplemented by this image	image	January 2019
of The dual effect of a ferredoxin-hydrogenase fusion protein in vivo: successful divergence of the photosynthetic electron flux towards hydrogen production and elevated oxygen tolerance Eilenberg, Haviva; Weiner, Iddo; Ben-Zvi, Oren https://doi.org/10.6084/m9.figshare.10037174 The current record is supplemented by this image	image	January 2019
of The dual effect of a ferredoxin-hydrogenase fusion protein in vivo: successful divergence of the photosynthetic electron flux towards hydrogen production and elevated oxygen tolerance Eilenberg, Haviva; Weiner, Iddo; Ben-Zvi, Oren https://doi.org/10.6084/m9.figshare.10037174.v1 The current record is supplemented by this image	image	January 2019
The dual effect of a ferredoxin-hydrogenase fusion protein in vivo: successful divergence of the photosynthetic electron flux towards hydrogen production and elevated oxygen tolerance Eilenberg, Haviva; Weiner, Iddo; Ben-Zvi, Oren https://doi.org/10.6084/m9.figshare.c.3597584 The current record is supplemented by this collection	collection	January 2016
The dual effect of a ferredoxin-hydrogenase fusion protein in vivo: successful divergence of the photosynthetic electron flux towards hydrogen production and elevated oxygen tolerance Eilenberg, Haviva; Weiner, Iddo; Ben-Zvi, Oren https://doi.org/10.6084/m9.figshare.c.3597584.v1 The current record is supplemented by this collection	collection	January 2016