Isozymes of Methanol Oxidase in a Methanol-Utilizing Yeast, Pichia methanolica IAM 12901

概要

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Methanol oxidases (MODs) were investigated in some methanol-utilizing yeasts. Pichia methanolica IAM 12901 and IAM 12481 (=Pichia cellobiosa) produced 9 forms of methanol oxidase on electrophoresis when grown on methanol but 1 form when grown on glycerol or pectin. Other strains-Candida boidinii IAM 12875,Pichia angusta IAM 12897,and Pichia pastoris IAM 12903-produced one band on electrophoresis when grown on methanol. Methanol oxidases with the smallest and largest Rm (relative mobility)-MOD A and MOD B-were purified from methanol-grown cells; another methanol oxidase (MOD P) was purified from pectin-grown cells of P. methanolica IAM 12901. All the purified enzymes were octamers composed of subunits with a molecular mass of 70,000 Da. MOD A and MOD P were mostly identical with each other in terms of their enzymatic characteristics, but MOD B differed from the other two with respect to substrate specificity, the prosthetic group, and the apparent K_m for methanol. Southern analysis of the genomic DNA of P. methanolica IAM 12901 was carried out with the partial methanol oxidase gene as a probe. The genomic DNA of this yeast showed the presence of 2 methanol oxidase genes. It was concluded that the 9 forms of MOD in P. methanolica were isozymes of a chimera octamer consisting of 2 subunits encoded by different methanol oxidase genes.
社団法人日本生物工学会の論文
1996-06-25