Purification and characterization of extracellular inulinase from a marine yeast Pichia guilliermondii and inulin hydrolysis by the purified inulinase
The extracellular inulinase in the supernatant of cell culture of the marine yeast Pichia guilliermondii 1 was purified to homogeneity with a 7.2-fold increase in inulinase activity as compared to that in the supernatant by ultrafiltrafion,concentration,gel filtration chromatography(SephadexTM G-75)and anion-exchange chromatography(DEAE-Sepharose Fast Flow Anion-Exchange).The molecular mass of the purified enzyme was estimated to be 50.0 kDa.The optimal pH and temperature of the purified enzyme were 6.0 and 60 ℃,respectively.The enzyme was activated by Mn2+,Ca2+,K+,Li+,Na+,Fe3+,Fe2+,Cu2+,and Co2+.However,Mg2+,Hg2+ and Ag+ acted as inhibitors in decreasing activity of the purified inulinase.The enzyme was strongly inhibited by Phenylmethanesulphonyl fluoride(PMSF),iodoacetic acid,EDTA,and 1,10-phenanthroline.The Km and Vmax values of the purified enzyme for inulin were 21.1 mg ml-1 and 0.082 mg min-1,respectively.A large amount of monosaccharides were detected after the hydrolysis of inulin.
Inulinase Marine yeasts Purification Pichia guilliermondii
F.Gong Z.Chi J.Sheng J.Li
Unesco Chinese Center of Marine Biotechnology,Ocean University of China
国内会议
青岛
英文
67-79
2007-08-01（万方平台首次上网日期，不代表论文的发表时间）