Shaofeng Hua, Shuben Li, Jiayin Xin, Jianzhong Niu, Chungu Xia, Haidong Tan, Wei Tang
State Key Laboratory for Oxo Synthesis and Selective Oxidation, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, China.
Biosci Biotechnol Biochem. 2007 Jan;71(1):122-9. doi: 10.1271/bbb.60402. Epub 2007 Jan 7.
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.
通过色谱法纯化甲烷单加氧酶羟化酶，并通过电泳和光谱法对其进行表征。通过凝胶过滤测定，羟化酶的分子量为201.3 kDa，而通过SDS-PAGE判断，其总分子量为234 kDa。结构研究表明，该酶为同型二聚体结构，由三个亚基组成，分别命名为α、β和γ，分子量分别为58 kDa、36 kDa和23 kDa。IEF分析表明该酶的pI为5.2。羟化酶的紫外-可见光谱在281 nm附近有一个吸收峰，在395 nm - 420 nm附近有一个弱肩峰，荧光光谱在341.3 nm处有一个发射峰。圆二色性测量表明，羟化酶主要由α-螺旋区域组成。最后，系统发育分析表明，该菌株与嗜甲基孢囊菌OB3b非常接近。
