Purification and biochemical characterization of soluble methane monooxygenase hydroxylase from Methylosinus trichosporium IMV 3011.
Methane monooxygenase hydroxylase was purified by chromatography and characterized by electrophoresis and spectroscopy. The molecular mass of hydroxylase was 201.3 KDa as determined by gel filtration, whereas the total molecular mass was 234 KDa as judged by SDS-PAGE. Structure study indicated that the enzyme is a homodimer structure, consisting of three subunits, designated alpha, beta, and gamma, with molecular masses of 58 KDa, 36 KDa, and 23 KDa respectively. IEF analysis indicated that the enzyme has a pI of 5.2. The UV-Vis spectrum of hydroxylase revealed an absorption peak near 281 nm and a weak shoulder peak around 395 nm-420 nm, and a fluorescence spectrum revealed an emission peak at 341.3 nm. Circular dichroism measurement indicated that hydroxylase mainly consists of alpha-helical regions. Finally, phylogenetic analysis indicated that this strain is very close to Methylosinus trichosporium OB3b.
Shaofeng
Hua
H
State Key Laboratory for Oxo Synthesis and Selective Oxidation, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, China.
Shuben
Li
L
Jiayin
Xin
X
Jianzhong
Niu
N
Chungu
Xia
X
Haidong
Tan
T
Wei
Tang
T
D001483:Base Sequence; D055592:Biophysical Phenomena; D001703:Biophysics; D002942:Circular Dichroism; D007526:Isoelectric Point; D020602:Methylosinus trichosporium / Q000201:enzymology* / Q000235:genetics / Q000648:ultrastructure; D008855:Microscopy, Electron, Scanning; D010105:Oxygenases / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D010802:Phylogeny; D012995:Solubility
D010105:Oxygenases; C021555:methane monooxygenase