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 language | English |
|---|---|
| Pages (from-to) | 7081-7088 |
| Number of pages | 8 |
| Journal | International Journal of Hydrogen Energy |
| Volume | 36 |
| Issue number | 12 |
| DOIs | |
| Publication status | Published - Jun 1 2011 |
| Externally published | Yes |
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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 journal › Article
}
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.
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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 -