Open AccessPurification and Properties of a Methanol‐Oxidizing Enzyme in Pseudomonas C
Author(s) -
GOLDBERG Israel
Publication year - 1976
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1976.tb10225.x
Subject(s) - chemistry , methanol , formaldehyde , oxidizing agent , size exclusion chromatography , ammonium , enzyme , chromatography , enzyme assay , organic chemistry
A methanol‐oxidizing enzyme has been purified from Pseudomonas C, grown on methanol as a sole source for carbon and energy. The purification procedure involved ammonium sulphate precipitation, ion‐exchange chromatography and gel filtration and resulted in a yield of 35.4 %. Enzyme activity can be coupled to phenazine methosulfate and requires the presence of ammonium ions in the assay mixtures. The enzyme possesses a broad specificity for primary alcohols. Formaldehyde is also oxidized by the purified enzyme. The K m value for methanol is 15 μM. The optimum pH for the oxidation of both methanol and formaldehyde is about 10.4. The enzyme has a molecular weight of about 128000 and consists of two subunits each having a molecular weight of 60000.