Parker M W, Cornish A, Gossain V, Best D J
Eur J Biochem. 1987 Apr 1;164(1):223-7. doi: 10.1111/j.1432-1033.1987.tb11014.x.
Methanol dehydrogenase was purified from the obligate methanotroph, Methylosinus trichosporium OB3b, in two steps from disrupted biomass by aqueous two-phase partition and ion-exchange chromatography. Copartitioning of a cytochrome c was dependent upon the pH at which aqueous partition was carried out. The native enzyme has a Mr of 120,000, as determined by gel filtration chromatography, and consists of two identical subunits. The purified enzyme contained four electrophoretically distinct isoenzymes, with pI values of 6.3, 6.58, 6.63 and 6.88. The native enzyme has been crystallised in a form suitable for high-resolution X-ray crystallographic studies. The crystals diffract to better than 0.19 nm spacing and are relatively stable to irradiation with X-rays. The space group is P6(1)22 (or P6(5)22) with cell dimensions a = b = 10.21 nm, c = 29.32 nm and the crystal probably contains a single monomer in the asymmetric unit.
甲醇脱氢酶是从专性甲烷氧化菌甲基弯曲菌OB3b中纯化得到的，通过水相双相分配和离子交换色谱法，分两步从破碎的生物质中进行纯化。细胞色素c的共分配取决于进行水相分配时的pH值。通过凝胶过滤色谱法测定，天然酶的相对分子质量为120,000，由两个相同的亚基组成。纯化后的酶包含四种电泳性质不同的同工酶，其等电点分别为6.3、6.58、6.63和6.88。天然酶已结晶成适合高分辨率X射线晶体学研究的形式。这些晶体的衍射间距优于0.19 nm，并且对X射线照射相对稳定。空间群为P6(1)22（或P6(5)22），晶胞参数a = b = 10.21 nm，c = 29.32 nm，晶体的不对称单元中可能包含一个单体。
