The Microbial Metabolism of Methanol

A methanol-oxidizing enzyme was extracted from a methanol-utilizing yeast, Kloeckera sp. No. 2201, and was found to be independent of the addition of a hydrogen acceptor. The enzyme, which was inducibly formed in methanol-grown cells, was purified by ammonium sulfate fractionation, DEAE-cellulose and Sephadex G–200 column chromatographies and crystallization with solid ammonium sulfate. The crystalline enzyme preparation gave a single band on polyacryamide disc gel electrophoresis and a single symmetrical peak in the ultracentrifuge. The molecular weight was calculated to be 570,000. The enzyme catalyzed the following reaction: and it was characterized to be an oxidase.With the crystalline preparation of alcohol oxidase of Kloeckera sp. No. 2201 grown on methanol as a sole carbon source, some properties of the enzyme were investigated. The visible absorption spectrum showed maxima at 373 mμ, 392 mμ and 461 mμ. which were bleached by an addition of substrate and were restored by oxygenation. The prosthetic...