title: Glycerol oxidase, a novel copper hemoprotein from Aspergillus japonicus. Molecular and catalytic properties of the enzyme and its application to the analysis of serum triglycerides.
abstract: Glycerol oxidase purified from Aspergillus japonicus AT 008 had Mr = 400,000 and contained 1 mol of protoheme IX and 2 g atoms of copper/mol of enzyme protein. The absorption maxima of the oxidized form were found at 557, 530, 420, 280, and 238 nm, and those of the reduced form at 557 and 430 nm. Anaerobic addition of glycerol to the enzyme produced both a shift of the Soret band from 420 to 410 nm and bleaching of the alpha and beta bands at 557 and 530 nm. The ESR spectrum of glycerol oxidase showed three major signals at g = 1.99, g = 2.00, and g = 2.02. The signals at g = 1.99 and g = 2.02 were diminished by the anaerobic addition of glycerol, and the three signals completely disappeared after the addition of either dithionite or diethyldithiocarbamate. Exposure of glycerol oxidase to a borate buffer of pH 10.0 resulted in activation of the enzyme with concomitant enhancement of the ESR signals at g = 1.99 and g = 2.02. Since glycerol oxidase acts predominantly on glycerol, the enzyme can be employed in a specific colorimetric assay for serum triglycerides in combination with lipoprotein lipase.
mesh_terms: D000596:Amino Acids / Q000032:analysis; D001230:Aspergillus / Q000201:enzymology*; D019411:Clinical Laboratory Techniques; D004578:Electron Spin Resonance Spectroscopy; D006801:Humans; D008970:Molecular Weight; D013053:Spectrophotometry; D013401:Sugar Alcohol Dehydrogenases / Q000378:metabolism*; D014280:Triglycerides / Q000097:blood*
chemical_list: D000596:Amino Acids; D014280:Triglycerides; D013401:Sugar Alcohol Dehydrogenases; C036131:glycerol oxidase