Development of a Rhodobacter capsulatus self-reporting model system for optimizing light-dependent, [FeFe]-hydrogenase-driven H2 production

Wecker, Matt S. A.; Beaton, Stephen E.; Chado, Robert A.; Ghirardi, Maria L.

doi:10.1002/bit.26076

Title: Development of a Rhodobacter capsulatus self-reporting model system for optimizing light-dependent, [FeFe]-hydrogenase-driven H₂ production

Journal Article · Wed Aug 17 00:00:00 EDT 2016 · Biotechnology and Bioengineering

DOI:https://doi.org/10.1002/bit.26076· OSTI ID:1339249

Wecker, Matt S. A. ^[1]; Beaton, Stephen E. ^[2]; Chado, Robert A. ^[2]; Ghirardi, Maria L. ^[3]

GeneBiologics, LLC, Boulder, CO (United States)
United States Air Force Academy, Colorado Springs, CO (United States)
National Renewable Energy Lab. (NREL), Golden, CO (United States)

ABSTRACT The photosynthetic bacterium Rhodobacter capsulatus normally photoproduces H ₂ as a by‐product of its nitrogenase‐catalyzed nitrogen‐fixing activity. Such H ₂ production, however, is expensive from a metabolic perspective, requiring nearly four times as many photons as the equivalent algal hydrogenase‐based system (Ghirardi et al., 2009 Photobiological hydrogen‐producing systems. Chem Soc Rev 38(1):52–61). Here, we report the insertion of a Clostridium acetobutylicum [FeFe]‐hydrogenase and its three attendant hydrogenase assembly proteins into an R. capsulatus strain lacking its native uptake hydrogenase. Further, this strain is modified to fluoresce upon sensing H ₂ . The resulting strain photoproduces H ₂ and self‐reports its own H ₂ production through fluorescence. This model system represents a unique method of developing hydrogenase‐based H ₂ production in R. capsulatus , may serve as a powerful system for in vivo directed evolution of hydrogenases and hydrogenase‐associated genes, and provides a means of screening for increased metabolic production of H ₂ . Biotechnol. Bioeng. 2017;114: 291–297. © 2016 Wiley Periodicals, Inc.

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

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: AC36-08GO28308

OSTI ID:: 1339249

Alternate ID(s):: OSTI ID: 1400579

Report Number(s):: NREL/JA-2700-66611

Journal Information:: Biotechnology and Bioengineering, Vol. 114, Issue 2; ISSN 0006-3592

Publisher:: WileyCopyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 3 works

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Title: Development of a Rhodobacter capsulatus self-reporting model system for optimizing light-dependent, [FeFe]-hydrogenase-driven H2 production

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Title: Development of a Rhodobacter capsulatus self-reporting model system for optimizing light-dependent, [FeFe]-hydrogenase-driven H₂ production