Purification and immobilization of dextranase

摘要: An enzymic characteristic of Novo dextranase was presented. In addition to a high dextranolytic activity (7,200 U/ml), the crude enzyme also contained small amounts protease, glucoamylase, polygalacturonase, carboxymethylcellulase, laminarinase and chitinase. A highly purified then simply separated from commercial preparation by column chromatographies on DEAE-Sepharose, CM-Sepharose, chromatofocussing Polybuffer Exchanger PBE-94. The recovered with an over 200-fold increase in specific yield 84%. final homogeneous, as observed during performance liquid chromatography (HPLC). Size-exclusion HPLC indicated that had molecular mass 35 kDa its isoelectric point, established chromatofocussing, 4.85. Analysis dextran break-down products represents endolytic mode action, isomaltose isomaltotriose were identified main reducing sugars hydrolysis. covalently coupled silanized porous glass beads modified glutaraldehyde (Carrier I) or carbodiimide II). It shown immobilization gave optimum pH temperature ranges 5.4 5.7 50 °C 60 °C, respectively. affinity substrate decreased factor more than 13 for immobilized Carrier I increased slightly (about 1.4-times) bound II.