Novel heparinases derived from Bacteroides stercoris HJ-15 which hydrolyze glycosaminoglycan a preparing method and a use thereof

摘要

PURPOSE: A novel heparinase capable of decomposing glycosaminoglycans(GAG), derived from human intestinal microorganism, a producing method thereof and the use thereof are provided, therefore the heparinase having improved substrate specificity and stability can be produced. CONSTITUTION: The heparinase capable of decomposing glycosaminoglycans(GAG) is isolated from Bacteroides stercoris HJ-15(KCCM-10096), wherein glycosaminoglycans(GAG) are heparin, heparan sulfate and acharan sulfate; the heparinase has 45 or 70 kDa of molecular weight and optimal NaCl concentration of 200 to 900 mM, optimal pH of 6.0 to 7.5 and optimal temperature of 35 to 55 deg. C or 40 to 45 deg. C; and the activity of heparinase is catalyzed by EDTA, Co2+, Cu2+, Ni2+, Ba2+ and Zn2+. The method for producing heparinase comprises the steps of: culturing Bacteroides stercoris in an appropriate medium; recovering the cultured cells and preparing cell extract; and subjecting the cell extract to chromatography, wherein the chromatography is selected from QAE-cellulose, DEAE-cellulose, CM-sephadex C-50, hydroxyapatite, CM-sephadex C-25 and Hi-Trap SP.