title: Use of dye affinity chromatography for the purification of Aerococcus viridans lactate oxidase.
abstract: Lactate oxidase was purified from Aerococcus viridans (A. viridans) by dye affinity chromatography and FPLC ion exchange chromatography. The lactate oxidase could be purified by comparatively simple procedures, the purification achieved from a crude extract of A. viridans was 41-fold with a specific activity of 143 units/(mg of protein). The purified enzyme was a L-lactate oxidase, which catalyses the conversion of L-lactate in the presence of molecular oxygen to pyruvate and H(2)O(2). This purified lactate oxidase showed an apparent molecular mass of 48,200 in SDS-PAGE and the native molecular weight, as estimated by FPLC gel filtration, was 187,300. This molecular weight indicates that lactate oxidase exists in tetrameric form after gel filtration. To differing degrees, all the triazine dyes tested were inhibitors of lactate oxidase, solutions of free triazine dyes showing an inhibition mechanism which was both time- and pH-dependent.
authors.affiliation: Department of Biochemistry and Molecular Biology-A, Faculty of Biology, University of Murcia, Campus de Espinardo, E-30071 Murcia, Spain.
authors.forename: Sergio A
authors.initials: SA
authors.lastname: Streitenberger
authors.forename: José A
authors.initials: JA
authors.lastname: López-Más
authors.forename: Alvaro
authors.initials: A
authors.lastname: Sánchez-Ferrer
authors.forename: Francisco
authors.initials: F
authors.lastname: García-Carmona
mesh_terms: D002846:Chromatography, Affinity / Q000379:methods*; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004396:Coloring Agents / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D006899:Mixed Function Oxygenases / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D008970:Molecular Weight; D013289:Streptococcaceae / Q000201:enzymology*
chemical_list: D004396:Coloring Agents; D006899:Mixed Function Oxygenases; C021497:lactate 2-monooxygenase