Abstract
For obtaining the enzyme source of acid phosphatase from Pholiota nameko, the mycelia were cultured at 25℃ for 30 d in darkness in a casamino acids glucose medium. It was suggested that mycelia possessed at least four acid phosphatase isozymes under the above growth condition. One of the isozymes was purified to be homogeneity by ammonium sulfate fractionation, DEAE Toyopearl anion exchange chromatography and DyeMatrex Red A affinity chromatography. Gel filtration analysis showed that the native molecule had a molecular weight of 44,000. The molecular weight on gel electrophoresis with SDS was 42,000, indicating that the native form of the enzyme was a monomer. The optimum pH and temperature of the enzyme were 5.9 and 40℃, respectively, and the isoelectric point of the enzyme was pH 5.1. The acid phosphatase showed high specificity towards glucose-1-phosphate among all substrates tested. Hg^<2+>, ammonium molybdate, and SDS inhibited the enzyme activity.
