Purification and characterization of a highly thermostable, oxygen-resistant, respiratory [NiFe]-hydrogenase from a marine, aerobic hydrogen-oxidizing bacterium Hydrogenovibrio marinus

Ki Suk Yoon, Keiichi Fukuda, Kiyoshi Fujisawa, Hirofumi Nishihara

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

The membrane-bound [NiFe]-hydrogenase from Hydrogenovibrio marinus (HmMBH) was purified homogeneously under anaerobic conditions. Its molecular weight was estimated as 110 kDa, consisting of a heterodimeric structure of 66 kDa and 37 kDa subunits. The purified enzyme exhibited high activity in a wide temperature range: 185 U/mg at 30 °C and 615 U/mg at 85 °C (the optimum temperature). The Km and kcat/Km values for H2 were, respectively, 12 μM and 8.58 × 107 M-1 s-1. The optimum reaction pH was 7.8, but its stability was particularly high at pH 4.0-7.0. Results show that HmMBH was remarkably thermostable and oxygen-resistant: its half-life was 75 h at 80 °C under H2, and more than 72 h at 4 °C under air. The air-oxidized HmMBH for 72 h showed only weak EPR signals of Ni-B, suggesting a structural feature in which the active center is not easily oxidized.

Original languageEnglish
Pages (from-to)7081-7088
Number of pages8
JournalInternational Journal of Hydrogen Energy
Volume36
Issue number12
DOIs
Publication statusPublished - Jun 1 2011
Externally publishedYes

Fingerprint

purification
bacteria
Purification
Bacteria
Hydrogen
Oxygen
air
oxygen
hydrogen
Air
half life
Paramagnetic resonance
enzymes
molecular weight
Enzymes
Molecular weight
membranes
Membranes
Temperature
temperature

All Science Journal Classification (ASJC) codes

  • Renewable Energy, Sustainability and the Environment
  • Fuel Technology
  • Condensed Matter Physics
  • Energy Engineering and Power Technology

Cite this

Purification and characterization of a highly thermostable, oxygen-resistant, respiratory [NiFe]-hydrogenase from a marine, aerobic hydrogen-oxidizing bacterium Hydrogenovibrio marinus. / Yoon, Ki Suk; Fukuda, Keiichi; Fujisawa, Kiyoshi; Nishihara, Hirofumi.

In: International Journal of Hydrogen Energy, Vol. 36, No. 12, 01.06.2011, p. 7081-7088.

Research output: Contribution to journalArticle

@article{fc970a882d3749b98c135868b388eeb5,
title = "Purification and characterization of a highly thermostable, oxygen-resistant, respiratory [NiFe]-hydrogenase from a marine, aerobic hydrogen-oxidizing bacterium Hydrogenovibrio marinus",
abstract = "The membrane-bound [NiFe]-hydrogenase from Hydrogenovibrio marinus (HmMBH) was purified homogeneously under anaerobic conditions. Its molecular weight was estimated as 110 kDa, consisting of a heterodimeric structure of 66 kDa and 37 kDa subunits. The purified enzyme exhibited high activity in a wide temperature range: 185 U/mg at 30 °C and 615 U/mg at 85 °C (the optimum temperature). The Km and kcat/Km values for H2 were, respectively, 12 μM and 8.58 × 107 M-1 s-1. The optimum reaction pH was 7.8, but its stability was particularly high at pH 4.0-7.0. Results show that HmMBH was remarkably thermostable and oxygen-resistant: its half-life was 75 h at 80 °C under H2, and more than 72 h at 4 °C under air. The air-oxidized HmMBH for 72 h showed only weak EPR signals of Ni-B, suggesting a structural feature in which the active center is not easily oxidized.",
author = "Yoon, {Ki Suk} and Keiichi Fukuda and Kiyoshi Fujisawa and Hirofumi Nishihara",
year = "2011",
month = "6",
day = "1",
doi = "10.1016/j.ijhydene.2011.03.049",
language = "English",
volume = "36",
pages = "7081--7088",
journal = "International Journal of Hydrogen Energy",
issn = "0360-3199",
publisher = "Elsevier Limited",
number = "12",

}

TY - JOUR

T1 - Purification and characterization of a highly thermostable, oxygen-resistant, respiratory [NiFe]-hydrogenase from a marine, aerobic hydrogen-oxidizing bacterium Hydrogenovibrio marinus

AU - Yoon, Ki Suk

AU - Fukuda, Keiichi

AU - Fujisawa, Kiyoshi

AU - Nishihara, Hirofumi

PY - 2011/6/1

Y1 - 2011/6/1

N2 - The membrane-bound [NiFe]-hydrogenase from Hydrogenovibrio marinus (HmMBH) was purified homogeneously under anaerobic conditions. Its molecular weight was estimated as 110 kDa, consisting of a heterodimeric structure of 66 kDa and 37 kDa subunits. The purified enzyme exhibited high activity in a wide temperature range: 185 U/mg at 30 °C and 615 U/mg at 85 °C (the optimum temperature). The Km and kcat/Km values for H2 were, respectively, 12 μM and 8.58 × 107 M-1 s-1. The optimum reaction pH was 7.8, but its stability was particularly high at pH 4.0-7.0. Results show that HmMBH was remarkably thermostable and oxygen-resistant: its half-life was 75 h at 80 °C under H2, and more than 72 h at 4 °C under air. The air-oxidized HmMBH for 72 h showed only weak EPR signals of Ni-B, suggesting a structural feature in which the active center is not easily oxidized.

AB - The membrane-bound [NiFe]-hydrogenase from Hydrogenovibrio marinus (HmMBH) was purified homogeneously under anaerobic conditions. Its molecular weight was estimated as 110 kDa, consisting of a heterodimeric structure of 66 kDa and 37 kDa subunits. The purified enzyme exhibited high activity in a wide temperature range: 185 U/mg at 30 °C and 615 U/mg at 85 °C (the optimum temperature). The Km and kcat/Km values for H2 were, respectively, 12 μM and 8.58 × 107 M-1 s-1. The optimum reaction pH was 7.8, but its stability was particularly high at pH 4.0-7.0. Results show that HmMBH was remarkably thermostable and oxygen-resistant: its half-life was 75 h at 80 °C under H2, and more than 72 h at 4 °C under air. The air-oxidized HmMBH for 72 h showed only weak EPR signals of Ni-B, suggesting a structural feature in which the active center is not easily oxidized.

UR - http://www.scopus.com/inward/record.url?scp=79955893150&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79955893150&partnerID=8YFLogxK

U2 - 10.1016/j.ijhydene.2011.03.049

DO - 10.1016/j.ijhydene.2011.03.049

M3 - Article

VL - 36

SP - 7081

EP - 7088

JO - International Journal of Hydrogen Energy

JF - International Journal of Hydrogen Energy

SN - 0360-3199

IS - 12

ER -