Mutational Analyses of the Enzymes Involved in the Metabolism of Hydrogen by the Hyperthermophilic Archaeon Pyrococcus furiosus

Schut, Gerrit J.; Nixon, William J.; Lipscomb, Gina L.; Scott, Robert A.; Adams, Michael W.  W.

doi:10.3389/fmicb.2012.00163

Mutational Analyses of the Enzymes Involved in the Metabolism of Hydrogen by the Hyperthermophilic Archaeon Pyrococcus furiosus

Journal Article · Sun Jan 01 00:00:00 EST 2012 · Frontiers in Microbiology

DOI:https://doi.org/10.3389/fmicb.2012.00163· OSTI ID:1628061

Schut, Gerrit J. ^[1]; Nixon, William J. ^[2]; Lipscomb, Gina L. ^[2]; Scott, Robert A. ^[3]; Adams, Michael W. W. ^[2]

Univ. of Georgia, Athens, GA (United States). Dept. of Biochemistry and Molecular Biology; DOE/OSTI
Univ. of Georgia, Athens, GA (United States). Dept. of Biochemistry and Molecular Biology
Univ. of Georgia, Athens, GA (United States). Dept. of Biochemistry and Molecular Biology; Univ. of Georgia, Athens, GA (United States). Dept. of Chemistry

Pyrococcus furiosus grows optimally near 100°C by fermenting carbohydrates to produce hydrogen (H2) or, if elemental sulfur (S0) is present, hydrogen sulfide instead. It contains two cytoplasmic hydrogenases, SHI and SHII, that use NADP(H) as an electron carrier and a membrane-bound hydrogenase (MBH) that utilizes the redox protein ferredoxin. We previously constructed deletion strains lacking SHI and/or SHII and showed that they exhibited no obvious phenotype. This study has now been extended to include biochemical analyses and growth studies using the ΔSHI and ΔSHII deletion strains together with strains lacking a functional MBH (ΔmbhL). Hydrogenase activity in cytoplasmic extracts of various strains demonstrate that SHI is responsible for most of the cytoplasmic hydrogenase activity. The ΔmbhL strain showed no growth in the absence of S0, confirming the hypothesis that, in the absence of S0, MBH is the only enzyme that can dispose of reductant (in the form of H2) generated during sugar oxidation. Under conditions of limiting sulfur, a small but significant amount of H2 was produced by the ΔmbhL strain, showing that SHI can produce H2 from NADPH in vivo, although this does not enable growth of ΔmbhL in the absence of S0. We propose that the physiological function of SHI is to recycle H2 and provide a link between external H2 and the intracellular pool of NADPH needed for biosynthesis. This likely has a distinct energetic advantage in the environment, but it is clearly not required for growth of the organism under the usual laboratory conditions. The function of SHII, however, remains unknown.

View Accepted Manuscript (DOE)

Research Organization:: Univ. of Georgia, Athens, GA (United States)

Sponsoring Organization:: USDOE Office of Science (SC)

Grant/Contract Number:: FG02-08ER64690

OSTI ID:: 1628061

Journal Information:: Frontiers in Microbiology, Journal Name: Frontiers in Microbiology Vol. 3; ISSN 1664-302X

Publisher:: Frontiers Research FoundationCopyright Statement

Country of Publication:: United States

Language:: English

References (32)

A third type of hydrogenase catalyzing H2 activation Shima, Seigo; Thauer, Rudolf K. The Chemical Record, Vol. 7, Issue 1 https://doi.org/10.1002/tcr.20111	journal	January 2007
Pyrococcus furiosus sp. nov. represents a novel genus of marine heterotrophic archaebacteria growing optimally at 100°C Fiala, Gerhard; Stetter, Karl O. Archives of Microbiology, Vol. 145, Issue 1, p. 56-61 https://doi.org/10.1007/bf00413027	journal	June 1986
Metabolism in hyperthermophilic microorganisms Kelly, Robert M.; Adams, Michael W. W. Antonie van Leeuwenhoek, Vol. 66, Issue 1-3 https://doi.org/10.1007/bf00871643	journal	January 1994
Sugar utilization and its control in hyperthermophiles de Vos, W. M.; Kengen, Servé W. M.; Voorhorst, Wilfried G. B. Extremophiles, Vol. 2, Issue 3 https://doi.org/10.1007/s007920050061	journal	August 1998
An improved method for purification of the cold-insoluble globulin of human plasma (Clg) Chen, Anthony B.; Mosesson, Michael W. Analytical Biochemistry, Vol. 79, Issue 1-2 https://doi.org/10.1016/0003-2697(77)90388-8	journal	May 1977
Engineering hybrid genes without the use of restriction enzymes: gene splicing by overlap extension Horton, Robert M.; Hunt, Henry D.; Ho, Steffan N. Gene, Vol. 77, Issue 1, p. 61-68 https://doi.org/10.1016/0378-1119(89)90359-4	journal	April 1989
Continuous hydrogen production by the hyperthermophilic archaeon, Thermococcus kodakaraensis KOD1 Kanai, Tamotsu; Imanaka, Hiroyuki; Nakajima, Akihito Journal of Biotechnology, Vol. 116, Issue 3 https://doi.org/10.1016/j.jbiotec.2004.11.002	journal	March 2005
Hyperthermophilic redox chemistry: a re-evaluation Hagedoorn, Peter L.; Driessen, Martijn C. P. F.; van den Bosch, Marieke FEBS Letters, Vol. 440, Issue 3 https://doi.org/10.1016/s0014-5793(98)01466-5	journal	December 1998
A Variable-Temperature Direct Electrochemical Study of Metalloproteins from Hyperthermophilic Microorganisms Involved in Hydrogen Production from Pyruvate Smith, Eugene T.; Blamey, Jenny M.; Zhou, Zhi H. Biochemistry, Vol. 34, Issue 21 https://doi.org/10.1021/bi00021a030	journal	May 1995
Occurrence, Classification, and Biological Function of Hydrogenases: An Overview Vignais, Paulette M.; Billoud, Bernard Chemical Reviews, Vol. 107, Issue 10, p. 4206-4272 https://doi.org/10.1021/cr050196r	journal	October 2007
Analysis of histological and microRNA profiles changes in rabbit skin development Ding, Haisheng; Cheng, Guanglong; Leng, Jianjian Scientific Reports, Vol. 10, Issue 1 https://doi.org/10.1038/s41598-019-57327-5	journal	January 2020
Copaifera spp. oleoresins impair Toxoplasma gondii infection in both human trophoblastic cells and human placental explants Teixeira, Samuel Cota; de Souza, Guilherme; Borges, Bruna Cristina Scientific Reports, Vol. 10, Issue 1 https://doi.org/10.1038/s41598-020-72230-0	journal	September 2020
The function of the periplasmic Sud protein in polysulfide respiration of Wolinella succinogenes Klimmek, Oliver; Kreis, Vera; Klein, Carola European Journal of Biochemistry, Vol. 253, Issue 1 https://doi.org/10.1046/j.1432-1327.1998.2530263.x	journal	April 1998
Enzymes of hydrogen metabolism in Pyrococcus furiosus: P. furiosus enzymes of hydrogen metabolism Silva, Pedro J.; van den Ban, Eyke C. D.; Wassink, Hans European Journal of Biochemistry, Vol. 267, Issue 22 https://doi.org/10.1046/j.1432-1327.2000.01745.x	journal	November 2000
A simple energy-conserving system: Proton reduction coupled to proton translocation Sapra, R.; Bagramyan, K.; Adams, M. W. W. Proceedings of the National Academy of Sciences, Vol. 100, Issue 13, p. 7545-7550 https://doi.org/10.1073/pnas.1331436100	journal	June 2003
Hydrogenase of the hyperthermophile Pyrococcus furiosus is an elemental sulfur reductase or sulfhydrogenase: evidence for a sulfur-reducing hydrogenase ancestor. Ma, K.; Schicho, R. N.; Kelly, R. M. Proceedings of the National Academy of Sciences, Vol. 90, Issue 11 https://doi.org/10.1073/pnas.90.11.5341	journal	June 1993
Engineering Hyperthermophilic Archaeon Pyrococcus furiosus to Overproduce Its Cytoplasmic [NiFe]-Hydrogenase Chandrayan, Sanjeev K.; McTernan, Patrick M.; Hopkins, R. Christopher Journal of Biological Chemistry, Vol. 287, Issue 5, p. 3257-3264 https://doi.org/10.1074/jbc.m111.290916	journal	December 2011
Hydrogen production by hyperthermophilic and extremely thermophilic bacteria and archaea: mechanisms for reductant disposal Verhaart, Marcel R. A.; Bielen, Abraham A. M.; Oost, John van der Environmental Technology, Vol. 31, Issue 8-9 https://doi.org/10.1080/09593331003710244	journal	July 2010
KEGG for linking genomes to life and the environment Kanehisa, M.; Araki, M.; Goto, S. Nucleic Acids Research, Vol. 36, Issue Database https://doi.org/10.1093/nar/gkm882	journal	December 2007
SurR: a transcriptional activator and repressor controlling hydrogen and elemental sulphur metabolism in Pyrococcus furiosus Lipscomb, Gina L.; Keese, Annette M.; Cowart, Darin M. Molecular Microbiology, Vol. 71, Issue 2, p. 332-349 https://doi.org/10.1111/j.1365-2958.2008.06525.x	journal	January 2009
SurR regulates hydrogen production in Pyrococcus furiosus by a sulfur-dependent redox switch: SurR regulates H2 production in P. furiosus by a redox switch Yang, Hua; Lipscomb, Gina L.; Keese, Annette M. Molecular Microbiology, Vol. 77, Issue 5 https://doi.org/10.1111/j.1365-2958.2010.07275.x	journal	August 2010
Growth and energy conservation in batch cultures of Pyrococcus furiosus Kengen, ServÃ©W. M.; Stams, Alfons J. M. FEMS Microbiology Letters, Vol. 117, Issue 3 https://doi.org/10.1111/j.1574-6968.1994.tb06784.x	journal	April 1994
Natural Competence in the Hyperthermophilic Archaeon Pyrococcus furiosus Facilitates Genetic Manipulation: Construction of Markerless Deletions of Genes Encoding the Two Cytoplasmic Hydrogenases Lipscomb, Gina L.; Stirrett, Karen; Schut, Gerrit J. Applied and Environmental Microbiology, Vol. 77, Issue 7, p. 2232-2238 https://doi.org/10.1128/aem.02624-10	journal	February 2011
Relationship between Glycosyl Hydrolase Inventory and Growth Physiology of the Hyperthermophile Pyrococcus furiosus on Carbohydrate-Based Media Driskill, Lance E.; Kusy, Kevin; Bauer, Michael W. Applied and Environmental Microbiology, Vol. 65, Issue 3 https://doi.org/10.1128/aem.65.3.893-897.1999	journal	March 1999
Insights into the Metabolism of Elemental Sulfur by the Hyperthermophilic Archaeon Pyrococcus furiosus: Characterization of a Coenzyme A- Dependent NAD(P)H Sulfur Oxidoreductase Schut, G. J.; Bridger, S. L.; Adams, M. W. W. Journal of Bacteriology, Vol. 189, Issue 12, p. 4431-4441 https://doi.org/10.1128/jb.00031-07	journal	April 2007
Distinct Physiological Roles of the Three [NiFe]-Hydrogenase Orthologs in the Hyperthermophilic Archaeon Thermococcus kodakarensis Kanai, T.; Matsuoka, R.; Beppu, H. Journal of Bacteriology, Vol. 193, Issue 12 https://doi.org/10.1128/jb.01072-10	journal	April 2011
Reinvestigation of the Steady-State Kinetics and Physiological Function of the Soluble NiFe-Hydrogenase I of Pyrococcus furiosus van Haaster, D. J.; Silva, P. J.; Hagedoorn, P. -L. Journal of Bacteriology, Vol. 190, Issue 5 https://doi.org/10.1128/jb.01562-07	journal	December 2007
Deletion Strains Reveal Metabolic Roles for Key Elemental Sulfur-Responsive Proteins in Pyrococcus furiosus Bridger, S. L.; Clarkson, S. M.; Stirrett, K. Journal of Bacteriology, Vol. 193, Issue 23 https://doi.org/10.1128/jb.05445-11	journal	September 2011
Purification and Characterization of a Membrane-Bound Hydrogenase from the Hyperthermophilic Archaeon Pyrococcus furiosus Sapra, R.; Verhagen, M. F. J. M.; Adams, M. W. W. Journal of Bacteriology, Vol. 182, Issue 12, p. 3423-3428 https://doi.org/10.1128/jb.182.12.3423-3428.2000	journal	June 2000
Key Role for Sulfur in Peptide Metabolism and in Regulation of Three Hydrogenases in the Hyperthermophilic Archaeon Pyrococcus furiosus Adams, M. W. W.; Holden, J. F.; Menon, A. L. Journal of Bacteriology, Vol. 183, Issue 2 https://doi.org/10.1128/jb.183.2.716-724.2001	journal	January 2001
Minimal sulfur requirement for growth and sulfur-dependent metabolism of the hyperthermophilic archaeon Staphylothermus marinus Hao, Xiaolei; Ma, Kesen Archaea, Vol. 1, Issue 3 https://doi.org/10.1155/2003/626017	journal	January 2003
The complete genome sequence of Staphylothermus marinus reveals differences in sulfur metabolism among heterotrophic Crenarchaeota Anderson, Iain J.; Dharmarajan, Lakshmi; Rodriguez, Jason BMC Genomics, Vol. 10, Issue 1 https://doi.org/10.1186/1471-2164-10-145	journal	January 2009

Cited By (12)

High hydrostatic pressure adaptive strategies in an obligate piezophile Pyrococcus yayanosii Michoud, Grégoire; Jebbar, Mohamed Scientific Reports, Vol. 6, Issue 1 https://doi.org/10.1038/srep27289	journal	June 2016
Role of Mn ²⁺ and Compatible Solutes in the Radiation Resistance of Thermophilic Bacteria and Archaea Webb, Kimberly M.; DiRuggiero, Jocelyne Archaea, Vol. 2012 https://doi.org/10.1155/2012/845756	journal	January 2012
The modern “3G” age of archaeal molecular biology Robb, Frank T.; Lowe, Todd M.; Kelman, Zvi Frontiers in Microbiology, Vol. 3 https://doi.org/10.3389/fmicb.2012.00430	journal	January 2012
NADPH-generating systems in bacteria and archaea Spaans, Sebastiaan K.; Weusthuis, Ruud A.; van der Oost, John Frontiers in Microbiology, Vol. 6 https://doi.org/10.3389/fmicb.2015.00742	journal	July 2015
Overproduction of the membrane-bound [NiFe]-hydrogenase in Thermococcus kodakarensis and its effect on hydrogen production Kanai, Tamotsu; Simons, Jan-Robert; Tsukamoto, Ryohei Frontiers in Microbiology, Vol. 6 https://doi.org/10.3389/fmicb.2015.00847	journal	August 2015
Proteomic Insights into Sulfur Metabolism in the Hydrogen-Producing Hyperthermophilic Archaeon Thermococcus onnurineus NA1 Moon, Yoon-Jung; Kwon, Joseph; Yun, Sung-Ho International Journal of Molecular Sciences, Vol. 16, Issue 12 https://doi.org/10.3390/ijms16059167	journal	April 2015
Additional file 1 of Sulfur-cycling chemolithoautotrophic microbial community dominates a cold, anoxic, hypersaline Arctic spring Magnuson, Elisse; Altshuler, Ianina; Freyria, Nastasia J. figshare https://doi.org/10.6084/m9.figshare.24123771.v1	dataset	January 2023
Engineering Hydrogen Gas Production from Formate in a Hyperthermophile by Heterologous Production of an 18-Subunit Membrane-bound Complex Lipscomb, Gina L.; Schut, Gerrit J.; Thorgersen, Michael P. Journal of Biological Chemistry, Vol. 289, Issue 5 https://doi.org/10.1074/jbc.m113.530725	journal	December 2013
Two functionally distinct NADP ⁺ -dependent ferredoxin oxidoreductases maintain the primary redox balance of Pyrococcus furiosus Nguyen, Diep M. N.; Schut, Gerrit J.; Zadvornyy, Oleg A. Journal of Biological Chemistry, Vol. 292, Issue 35 https://doi.org/10.1074/jbc.m117.794172	journal	July 2017
Characterization of membrane-bound sulfane reductase: A missing link in the evolution of modern day respiratory complexes Wu, Chang-Hao; Schut, Gerrit J.; Poole, Farris L. Journal of Biological Chemistry, Vol. 293, Issue 43 https://doi.org/10.1074/jbc.ra118.005092	journal	September 2018
Molecular Hydrogen, a Neglected Key Driver of Soil Biogeochemical Processes Piché-Choquette, Sarah; Constant, Philippe Applied and Environmental Microbiology, Vol. 85, Issue 6 https://doi.org/10.1128/aem.02418-18	journal	January 2019
Carbohydrate Metabolism in Archaea: Current Insights into Unusual Enzymes and Pathways and Their Regulation Brasen, C.; Esser, D.; Rauch, B. Microbiology and Molecular Biology Reviews, Vol. 78, Issue 1 https://doi.org/10.1128/mmbr.00041-13	journal	March 2014

Similar Records

Novel and remarkably thermostable ferredoxin from the hyperthermophilic archaebacterium Pyrococcus furiosus

Journal Article · Thu Jun 01 00:00:00 EDT 1989 · Journal of Bacteriology; (USA) · OSTI ID:6995392

pH Homeostasis and Sodium Ion Pumping by Multiple Resistance and pH Antiporters in Pyrococcus furiosus

Journal Article · Sun Aug 15 20:00:00 EDT 2021 · Frontiers in Microbiology · OSTI ID:1850080

A mutant (‘lab strain’) of the hyperthermophilic archaeon Pyrococcus furiosus, lacking flagella, has unusual growth physiology

Journal Article · Wed Dec 03 19:00:00 EST 2014 · Extremeophiles · OSTI ID:1343557

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
Microbiology
Pyrococcus furiosus
anaerobe
energy metabolism
ferredoxin
hydrogenase
sulfur
thermophile

Mutational Analyses of the Enzymes Involved in the Metabolism of Hydrogen by the Hyperthermophilic Archaeon Pyrococcus furiosus

Citation Formats

References (32)

Cited By (12)

Similar Records

Related Subjects