Highly efficient Aerococcus viridans L-alpha-glycerophosphate oxidase production in the presence of H2O2-decomposing agent: purification and kinetic characterization
Sa. Streitenberger et al., Highly efficient Aerococcus viridans L-alpha-glycerophosphate oxidase production in the presence of H2O2-decomposing agent: purification and kinetic characterization, APPL MICR B, 57(3), 2001, pp. 329-333
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.