Biochemical and molecular characterization of the [NiFe] hydrogenase from the hyperthermophilic archaeon, Thermococcus litoralis

G. Rákhely, Zhi H. Zhou, Michael W W Adams, K. Kovács

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Thermococcus litoralis is a hyperthermophilic archaeon that grows at temperatures up to 98 °C by fermentative metabolism and reduces elemental sulfur (S°) to H2S. A [NiFe] hydrogenase, responsible for H2S or H2 production, has been purified and characterized. The enzyme is composed of four subunits with molecular mass 46, 42, 34 and 32 kDa. Elemental analyses gave approximate values of 22 Fe, 22 S and 1 Ni per hydrogenase. EPR spectra at 70 and 5 K indicated the presence of four or five [4Fe-4S] and one [2Fe- 2S] type clusters. The optimal temperature for both H2 evolution and oxidation, using artificial electron carriers, was around 80 °C. The operon encoding the T. litoralis enzyme is composed of four genes forming one transcriptional unit, and transcription is not regulated by S°. An unusual transcription-initiation site is located 139 bp upstream from the translational start point. Sequence analyses indicated the presence of new putative nucleotide-binding domains. Upstream from the hydrogenase operon, ORFs probably encoding a molybdopterin oxidoreductase enzyme have been identified. Based on sequence, biochemical and biophysical analyses, a model of the enzyme and the pathway of electron flow during catalysis is proposed.

Original languageEnglish
Pages (from-to)1158-1165
Number of pages8
JournalEuropean Journal of Biochemistry
Volume266
Issue number3
DOIs
Publication statusPublished - Dec 15 1999

Fingerprint

Thermococcus
Archaea
Hydrogenase
Enzymes
Operon
Electrons
Temperature
Transcription Initiation Site
Molecular mass
Transcription
Catalysis
Sulfur
Metabolism
Open Reading Frames
Paramagnetic resonance
Sequence Analysis
Oxidoreductases
Nucleotides
Genes
Oxidation

Keywords

  • [NiFe] hydrogenase
  • Hydrogen evolution
  • Hyperthermophilic
  • Nucleotide- binding domains
  • Thermo-coccus litoralis

ASJC Scopus subject areas

  • Biochemistry

Cite this

Biochemical and molecular characterization of the [NiFe] hydrogenase from the hyperthermophilic archaeon, Thermococcus litoralis. / Rákhely, G.; Zhou, Zhi H.; Adams, Michael W W; Kovács, K.

In: European Journal of Biochemistry, Vol. 266, No. 3, 15.12.1999, p. 1158-1165.

Research output: Contribution to journalArticle

@article{3f3b8595e7014c41bb16eb6f5ca6e4be,
title = "Biochemical and molecular characterization of the [NiFe] hydrogenase from the hyperthermophilic archaeon, Thermococcus litoralis",
abstract = "Thermococcus litoralis is a hyperthermophilic archaeon that grows at temperatures up to 98 °C by fermentative metabolism and reduces elemental sulfur (S°) to H2S. A [NiFe] hydrogenase, responsible for H2S or H2 production, has been purified and characterized. The enzyme is composed of four subunits with molecular mass 46, 42, 34 and 32 kDa. Elemental analyses gave approximate values of 22 Fe, 22 S and 1 Ni per hydrogenase. EPR spectra at 70 and 5 K indicated the presence of four or five [4Fe-4S] and one [2Fe- 2S] type clusters. The optimal temperature for both H2 evolution and oxidation, using artificial electron carriers, was around 80 °C. The operon encoding the T. litoralis enzyme is composed of four genes forming one transcriptional unit, and transcription is not regulated by S°. An unusual transcription-initiation site is located 139 bp upstream from the translational start point. Sequence analyses indicated the presence of new putative nucleotide-binding domains. Upstream from the hydrogenase operon, ORFs probably encoding a molybdopterin oxidoreductase enzyme have been identified. Based on sequence, biochemical and biophysical analyses, a model of the enzyme and the pathway of electron flow during catalysis is proposed.",
keywords = "[NiFe] hydrogenase, Hydrogen evolution, Hyperthermophilic, Nucleotide- binding domains, Thermo-coccus litoralis",
author = "G. R{\'a}khely and Zhou, {Zhi H.} and Adams, {Michael W W} and K. Kov{\'a}cs",
year = "1999",
month = "12",
day = "15",
doi = "10.1046/j.1432-1327.1999.00969.x",
language = "English",
volume = "266",
pages = "1158--1165",
journal = "FEBS Journal",
issn = "1742-464X",
publisher = "Wiley-Blackwell",
number = "3",

}

TY - JOUR

T1 - Biochemical and molecular characterization of the [NiFe] hydrogenase from the hyperthermophilic archaeon, Thermococcus litoralis

AU - Rákhely, G.

AU - Zhou, Zhi H.

AU - Adams, Michael W W

AU - Kovács, K.

PY - 1999/12/15

Y1 - 1999/12/15

N2 - Thermococcus litoralis is a hyperthermophilic archaeon that grows at temperatures up to 98 °C by fermentative metabolism and reduces elemental sulfur (S°) to H2S. A [NiFe] hydrogenase, responsible for H2S or H2 production, has been purified and characterized. The enzyme is composed of four subunits with molecular mass 46, 42, 34 and 32 kDa. Elemental analyses gave approximate values of 22 Fe, 22 S and 1 Ni per hydrogenase. EPR spectra at 70 and 5 K indicated the presence of four or five [4Fe-4S] and one [2Fe- 2S] type clusters. The optimal temperature for both H2 evolution and oxidation, using artificial electron carriers, was around 80 °C. The operon encoding the T. litoralis enzyme is composed of four genes forming one transcriptional unit, and transcription is not regulated by S°. An unusual transcription-initiation site is located 139 bp upstream from the translational start point. Sequence analyses indicated the presence of new putative nucleotide-binding domains. Upstream from the hydrogenase operon, ORFs probably encoding a molybdopterin oxidoreductase enzyme have been identified. Based on sequence, biochemical and biophysical analyses, a model of the enzyme and the pathway of electron flow during catalysis is proposed.

AB - Thermococcus litoralis is a hyperthermophilic archaeon that grows at temperatures up to 98 °C by fermentative metabolism and reduces elemental sulfur (S°) to H2S. A [NiFe] hydrogenase, responsible for H2S or H2 production, has been purified and characterized. The enzyme is composed of four subunits with molecular mass 46, 42, 34 and 32 kDa. Elemental analyses gave approximate values of 22 Fe, 22 S and 1 Ni per hydrogenase. EPR spectra at 70 and 5 K indicated the presence of four or five [4Fe-4S] and one [2Fe- 2S] type clusters. The optimal temperature for both H2 evolution and oxidation, using artificial electron carriers, was around 80 °C. The operon encoding the T. litoralis enzyme is composed of four genes forming one transcriptional unit, and transcription is not regulated by S°. An unusual transcription-initiation site is located 139 bp upstream from the translational start point. Sequence analyses indicated the presence of new putative nucleotide-binding domains. Upstream from the hydrogenase operon, ORFs probably encoding a molybdopterin oxidoreductase enzyme have been identified. Based on sequence, biochemical and biophysical analyses, a model of the enzyme and the pathway of electron flow during catalysis is proposed.

KW - [NiFe] hydrogenase

KW - Hydrogen evolution

KW - Hyperthermophilic

KW - Nucleotide- binding domains

KW - Thermo-coccus litoralis

UR - http://www.scopus.com/inward/record.url?scp=0033573148&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033573148&partnerID=8YFLogxK

U2 - 10.1046/j.1432-1327.1999.00969.x

DO - 10.1046/j.1432-1327.1999.00969.x

M3 - Article

C2 - 10583413

AN - SCOPUS:0033573148

VL - 266

SP - 1158

EP - 1165

JO - FEBS Journal

JF - FEBS Journal

SN - 1742-464X

IS - 3

ER -