title: Purification and some properties of a novel microbial lactate oxidase.
abstract: Geotrichum candidum was found to produce a lactate oxidase. The enzyme was purified by gel filtration and ion-exchange chromatography. The purified lactate oxidase showed a molecular mass of 50 kDa under denaturing and about 400 kDa under non-denaturing conditions. Transmission electron microscopy analysis confirmed an octameric structure. FMN was found to be a cofactor for this enzyme. Polarographic studies confirmed an oxygen uptake by the lactate oxidase. The enzyme showed specificity towards the L isomer of lactate and did not oxidise pyruvate, fumarate, succinate, maleate and ascorbate. It was stable at alkaline pH and also for 15 min at 45 degrees C. The addition of glycerol and dextran 500,000 to the enzyme sample enhanced storage stability.
authors.affiliation: Gesellschaft für Biotechnologische Forschung, Braunschweig, Germany. HSZ@gbf-braunschweig.de
authors.forename: H
authors.initials: H
authors.lastname: Sztajer
authors.forename: W
authors.initials: W
authors.lastname: Wang
authors.forename: H
authors.initials: H
authors.lastname: Lünsdorf
authors.forename: A
authors.initials: A
authors.lastname: Stocker
authors.forename: R D
authors.initials: RD
authors.lastname: Schmid
mesh_terms: D000595:Amino Acid Sequence; D005486:Flavin Mononucleotide / Q000494:pharmacology; D017022:Flow Injection Analysis; D005848:Geotrichum / Q000201:enzymology*; D006899:Mixed Function Oxygenases / Q000187:drug effects / Q000302:isolation & purification* / Q000378:metabolism / Q000648:ultrastructure; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011048:Polarography; D011487:Protein Conformation; D017421:Sequence Analysis; D013053:Spectrophotometry; D013237:Stereoisomerism; D013379:Substrate Specificity
chemical_list: D005486:Flavin Mononucleotide; D006899:Mixed Function Oxygenases; C021497:lactate 2-monooxygenase