skip to main content
DOE PAGES title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Development of a Rhodobacter capsulatus self-reporting model system for optimizing light-dependent, [FeFe]-hydrogenase-driven H 2 production

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. 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. Furthermore, 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.

Authors:
 [1];  [2];  [2];  [3]
  1. GeneBiologics, LLC, Boulder, CO (United States)
  2. United States Air Force Academy, Colorado Springs, CO (United States)
  3. National Renewable Energy Lab. (NREL), Golden, CO (United States)
Publication Date:
Research Org.:
National Renewable Energy Lab. (NREL), Golden, CO (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1339249
Alternate Identifier(s):
OSTI ID: 1400579
Report Number(s):
NREL/JA-2700-66611
Journal ID: ISSN 0006-3592
Grant/Contract Number:  
AC36-08GO28308; AC36-08GC28308z
Resource Type:
Accepted Manuscript
Journal Name:
Biotechnology and Bioengineering
Additional Journal Information:
Journal Volume: 114; Journal Issue: 2; Journal ID: ISSN 0006-3592
Publisher:
Wiley
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS; R. capsulatus H2 sensor; high-throughput screening; photobiohydrogen; nitrogenase; H2 production

Citation Formats

Wecker, Matt S. A., Beaton, Stephen E., Chado, Robert A., and Ghirardi, Maria L. Development of a Rhodobacter capsulatus self-reporting model system for optimizing light-dependent, [FeFe]-hydrogenase-driven H2 production. United States: N. p., 2016. Web. doi:10.1002/bit.26076.
Wecker, Matt S. A., Beaton, Stephen E., Chado, Robert A., & Ghirardi, Maria L. Development of a Rhodobacter capsulatus self-reporting model system for optimizing light-dependent, [FeFe]-hydrogenase-driven H2 production. United States. doi:10.1002/bit.26076.
Wecker, Matt S. A., Beaton, Stephen E., Chado, Robert A., and Ghirardi, Maria L. Wed . "Development of a Rhodobacter capsulatus self-reporting model system for optimizing light-dependent, [FeFe]-hydrogenase-driven H2 production". United States. doi:10.1002/bit.26076. https://www.osti.gov/servlets/purl/1339249.
@article{osti_1339249,
title = {Development of a Rhodobacter capsulatus self-reporting model system for optimizing light-dependent, [FeFe]-hydrogenase-driven H2 production},
author = {Wecker, Matt S. A. and Beaton, Stephen E. and Chado, Robert A. and Ghirardi, Maria L.},
abstractNote = {The photosynthetic bacterium Rhodobacter capsulatus normally photoproduces H2 as a by-product of its nitrogenase-catalyzed nitrogen-fixing activity. Such H2 production, however, is expensive from a metabolic perspective, requiring nearly four times as many photons as the equivalent algal hydrogenase-based system. 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 H2. The resulting strain photoproduces H2 and self-reports its own H2 production through fluorescence. Furthermore, this model system represents a unique method of developing hydrogenase-based H2 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 H2.},
doi = {10.1002/bit.26076},
journal = {Biotechnology and Bioengineering},
number = 2,
volume = 114,
place = {United States},
year = {2016},
month = {8}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record

Save / Share:

Works referenced in this record:

Rewiring hydrogenase-dependent redox circuits in cyanobacteria
journal, February 2011

  • Ducat, D. C.; Sachdeva, G.; Silver, P. A.
  • Proceedings of the National Academy of Sciences, Vol. 108, Issue 10
  • DOI: 10.1073/pnas.1016026108

Redirection of Metabolism for Biological Hydrogen Production
journal, January 2007

  • Rey, F. E.; Heiniger, E. K.; Harwood, C. S.
  • Applied and Environmental Microbiology, Vol. 73, Issue 5
  • DOI: 10.1128/AEM.02565-06

Photobiological hydrogen-producing systems
journal, January 2009

  • Ghirardi, Maria Lucia; Dubini, Alexandra; Yu, Jianping
  • Chem. Soc. Rev., Vol. 38, Issue 1
  • DOI: 10.1039/B718939G

Metabolic engineering of Rhodobacter sphaeroides for improved hydrogen production
journal, April 2014


A simple hydrogenase-linked assay for ferredoxin and flavodoxin
journal, January 1979


Purification of a sixth ferredoxin from Rhodobacter capsulatus. Primary structure and biochemical properties
journal, June 1994


Four new derivatives of the broad-host-range cloning vector pBBR1MCS, carrying different antibiotic-resistance cassettes
journal, December 1995


Efficiency of ferredoxins and flavodoxins as mediators in systems for hydrogen evolution
journal, November 1980

  • Fitzgerald, M. P.; Rogers, L. J.; Rao, K. K.
  • Biochemical Journal, Vol. 192, Issue 2
  • DOI: 10.1042/bj1920665

High-throughput biosensor discriminates between different algal H 2 -photoproducing strains : Biosensor for Algal Hydrogen Photoproduction
journal, February 2014

  • Wecker, Matt S. A.; Ghirardi, Maria L.
  • Biotechnology and Bioengineering, Vol. 111, Issue 7
  • DOI: 10.1002/bit.25206

Truncated Photosystem Chlorophyll Antenna Size in the Green Microalga Chlamydomonas reinhardtii upon Deletion of the TLA3-CpSRP43 Gene
journal, October 2012

  • Kirst, Henning; Garcia-Cerdan, Jose Gines; Zurbriggen, Andreas
  • Plant Physiology, Vol. 160, Issue 4
  • DOI: 10.1104/pp.112.206672

Genetic Analysis of the Hox Hydrogenase in the Cyanobacterium Synechocystis sp. PCC 6803 Reveals Subunit Roles in Association, Assembly, Maturation, and Function
journal, November 2012

  • Eckert, Carrie; Boehm, Marko; Carrieri, Damian
  • Journal of Biological Chemistry, Vol. 287, Issue 52
  • DOI: 10.1074/jbc.M112.392407

Photobiological production of hydrogen gas as a biofuel
journal, June 2010


Design of a new biosensor for algal H2 production based on the H2-sensing system of Rhodobacter capsulatus
journal, August 2011

  • Wecker, Matt S. A.; Meuser, Jonathan E.; Posewitz, Matthew C.
  • International Journal of Hydrogen Energy, Vol. 36, Issue 17
  • DOI: 10.1016/j.ijhydene.2011.05.121

Maximizing the hydrogen photoproduction yields in Chlamydomonas reinhardtii cultures: The effect of the H2 partial pressure
journal, May 2012

  • Kosourov, Sergey N.; Batyrova, Khorcheska A.; Petushkova, Ekaterina P.
  • International Journal of Hydrogen Energy, Vol. 37, Issue 10
  • DOI: 10.1016/j.ijhydene.2012.01.082

Engineering a non-native hydrogen production pathway into Escherichia coli via a cyanobacterial [NiFe] hydrogenase
journal, July 2011


High yield single stage conversion of glucose to hydrogen by photofermentation with continuous cultures of Rhodobacter capsulatus JP91
journal, January 2013


Structure−Function Relationships of Alternative Nitrogenases
journal, January 1996


A new gene expression system based on a fructose-dependent promoter from Rhodobacter capsulatus
journal, July 1994


Sustained Photobiological Hydrogen Gas Production upon Reversible Inactivation of Oxygen Evolution in the Green Alga Chlamydomonas reinhardtii
journal, January 2000

  • Melis, Anastasios; Zhang, Liping; Forestier, Marc
  • Plant Physiology, Vol. 122, Issue 1
  • DOI: 10.1104/pp.122.1.127

Development of an In Vitro Compartmentalization Screen for High-Throughput Directed Evolution of [FeFe] Hydrogenases
journal, December 2010


Disrupting Calvin cycle phosphoribulokinase activity in Rhodopseudomonas palustris increases the H2 yield and specific production rate proportionately
journal, February 2016


Photosynthesis: a blueprint for solar energy capture and biohydrogen production technologies
journal, January 2005

  • Kruse, Olaf; Rupprecht, Jens; Mussgnug, Jan H.
  • Photochemical & Photobiological Sciences, Vol. 4, Issue 12
  • DOI: 10.1039/b506923h

Genetic and physical mapping of an hydrogenase gene cluster from Rhodobacter capsulatus
journal, February 1990

  • Colbeau, A.; Magnin, J. -P.; Cauvin, B.
  • MGG Molecular & General Genetics, Vol. 220, Issue 3
  • DOI: 10.1007/BF00391744

Characterization of an fdxN mutant of Rhodobacter capsulatus indicates that ferredoxin I serves as electron donor to nitrogenase
journal, November 1995

  • Jouanneau, Yves; Meyer, Christine; Naud, Isabelle
  • Biochimica et Biophysica Acta (BBA) - Bioenergetics, Vol. 1232, Issue 1-2
  • DOI: 10.1016/0005-2728(95)00106-X

Derepressive effect of NH 4+ on hydrogen production by deleting the glnA1 gene in Rhodobacter sphaeroides
journal, March 2010

  • Li, Xinfeng; Liu, Tong; Wu, Yongqiang
  • Biotechnology and Bioengineering, Vol. 106, Issue 4
  • DOI: 10.1002/bit.22722

Organization and regulation of genes encoding the molybdenum nitrogenase and the alternative nitrogenase in Rhodobacter capsulatus
journal, February 1996


Functional Studies of [FeFe] Hydrogenase Maturation in an Escherichia coli Biosynthetic System
journal, March 2006


New insights into the systems for heterologous synthesis and maturation of hydrogenases, the most efficient biohydrogen producers
journal, October 2013


The Difference a Se Makes? Oxygen-Tolerant Hydrogen Production by the [NiFeSe]-Hydrogenase from Desulfomicrobium baculatum
journal, October 2008

  • Parkin, Alison; Goldet, Gabrielle; Cavazza, Christine
  • Journal of the American Chemical Society, Vol. 130, Issue 40
  • DOI: 10.1021/ja803657d

Microalgae: a green source of renewable H2
journal, December 2000


Structure and evolution of a multidomain multiphosphoryl transfer protein
journal, June 1990