Purification and characterization of membrane-bound hydrogenase from a thermophilic hydrogen-oxidizing bacterium, Pseudomonas hydrogenothermophila strain TH-1.
A membrane-bound hydrogenase was purified aerobically by one step using a hydroxyapatite column after solubilization by acetone treatment from a thermophilic hydrogen-oxidizing bacterium, Pseudomonas hydrogenothermophila strain TH-1. The enzyme consists of two polypeptides of 63 and 31 kDa, respectively. The amino-terminal amino acid sequences of both subunits were homologous to membrane-bound type [Ni-Fe] hydrogenases from other origins. The thermostability under a hydrogen gas atmosphere is highly stable at 50 degrees C, which is the optimum temperature for the cell growth.
Ono
T
T
Department of Biotechnology, University of Tokyo, Japan.
Ishii
M
M
Yoon
K S
KS
Igarashi
Y
Y
Kodama
T
T
D000595:Amino Acid Sequence; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006859:Hydrogen / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D006864:Hydrogenase / Q000737:chemistry* / Q000302:isolation & purification / Q000502:physiology; D008565:Membrane Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011549:Pseudomonas / Q000737:chemistry* / Q000201:enzymology*; D012995:Solubility; D013053:Spectrophotometry; D013696:Temperature; D013997:Time Factors
D008565:Membrane Proteins; D006859:Hydrogen; D006864:Hydrogenase