[Purification and properties of inulinase from Kluyveromyces sp. Y-85]

The crude endocellular inulinase from Kluyveromyces sp. Y-85 was purified to two components, designated as EI and EII, using PEG6000-phosphate buffer extraction, (NH4)2SO4 fractionation, DEAE chromatography and gel filtration (Protein-PAK); The crude exocellular inulinase from this strain was purified to Eexo by means of PEG6000-phosphate buffer extraction, double DEAE-Sephace chromatography, Sephadex G-150 gel filtration. EI, EII and Eexo were demonstrated to be homogeneous by Waters 650E protein purification system. Their molecular weights are 42 kD, 65 kD and 57 kD, respectively. All the inulinases were glycoproteins containing a saccharide (from 25% to 35%) and belonged to the endo-inulinase. In addition, EI, EII, Eexo were optimally reactive at pH4.6, 4.5, 4.6 and at 52 degrees C, 52 degrees C, 55 degrees C, respectively. Ag+, Hg2+ and PCMB inhibited these enzymes' activity strongly. The products of raw inulin extracted from Helianthus tuberosus hydrolyzed by these three enzymes were fructose (86.5%) and glycose (13.5%).