Purification and Characterization of Internal Invertase in Rhodosporidum toruloides Mating Type a Cells

The internal invertase of Rhodosporidium toruloids mating type a cells was purified to a single band on SDS-PAGE from cell-free extract by acid precipitation, ion exchange chromatogaphy andgel filtration. The determined molecular weight of he purified enzyme was about 95,000 by gel filtration and 100,000 daltons on SDS-polyacryamide gel electrophoresis. This enzyme didn't show any activity change by several metal ions except 15.4% decrease by {TEX}$Mn^{2+}${/TEX} and was strongly inhibited by 2-mercaptoethanol and SDS. The invertase maintained its activity at high level until 70℃, but inactivated at 80℃ almost completely. The optimal temperature and pH of the enzyme were about 60℃ and pH 5.0, respectively. The stable pH range of invertase was narrow from pH 3.0 to 6.0. The Km value and isoelectric point of enzyme were 3.4×{TEX}$10^{3}${/TEX} M, pH 4.4, respectively.