Purification, crystallisation and preliminary X‐ray diffraction characterisation of methanol dehydrogenase from Methylosinus trichosporium OB3b

Abstract

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 cystallised 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 P6122 (or P6522) with cell dimensions a=b= 10.21 nm, c= 29.32 nm and the crystal probably contains a single monomer in the asymmetric unit. Copyright © 1987, Wiley Blackwell. All rights reserved