title: Highly efficient Aerococcus viridans L-alpha-glycerophosphate oxidase production in the presence of H2O2-decomposing agent: purification and kinetic characterization.
abstract: Glycerophosphate oxidase was purified from Aerococcus viridans cells by phase partitioning in Triton X-114, ammonium sulfate fractionation, FPLC ion-exchange chromatography and FPLC hydrophobic-interaction chromatography. The purification achieved from a crude extract of A. viridans was 38-fold with a 32% recovery of activity. Under the growth conditions used, A. viridans strain CECT 978 proved to be an excellent glycerophosphate-oxidase producer, with enzyme production 2,800-fold greater than that described in the literature for the same microorganism. The culture medium used in the present work is that commonly used for cultivation of this microorganism, except that an H2O2-decomposing enzyme was added. The addition of catalase to the growth medium had a clear effect on the growth rate. Furthermore, methylglyoxal, a metabolite that is formed enzymatically from triose phosphates, was found to be an inactivator of glycerophosphate oxidase activity.
authors.affiliation: Department of Biochemistry and Molecular Biology-A, Faculty of Biology, University of Murcia, Spain.
authors.forename: S A
authors.initials: SA
authors.lastname: Streitenberger
authors.forename: J A
authors.initials: JA
authors.lastname: López-Mas
authors.forename: A
authors.initials: A
authors.lastname: Sánchez-Ferrer
authors.forename: F
authors.initials: F
authors.lastname: García-Carmona
mesh_terms: D002374:Catalase / Q000378:metabolism / Q000494:pharmacology*; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D005993:Glycerolphosphate Dehydrogenase / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification*; D006861:Hydrogen Peroxide / Q000032:analysis / Q000378:metabolism*; D007700:Kinetics; D011765:Pyruvaldehyde / Q000378:metabolism; D013289:Streptococcaceae / Q000201:enzymology*
chemical_list: D011765:Pyruvaldehyde; D006861:Hydrogen Peroxide; D005993:Glycerolphosphate Dehydrogenase; D002374:Catalase