Highly efficient Aerococcus viridans L-alpha-glycerophosphate oxidase production in the presence of H2O2-decomposing agent: purification and kinetic characterization

Glycerophosphate oxidase was purified from Aerococcus viridans cells by pha se partitioning in Triton X-114, ammonium sulfate fractionation, FPLC ion-e xchange 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 lite rature for the same microorganism. The culture medium used in the present w ork is that commonly used for cultivation of this microorganism, except tha t an H2O2-decomposing enzyme was added. The addition of catalase to the gro wth medium had a clear effect on the growth rate. Furthermore, methylglyoxa l, a metabolite that is formed enzymatically from triose phosphates, was fo und to be an inactivator of glycerophosphate oxidase activity.