title: Expression of the heparinase gene from Flavobacterium heparinum in Escherichia coli and its enzymatic properties.
abstract: Heparinase has an important application in the preparation of low-molecular-weight heparins by heparin enzymolysis. A heparinase gene from Flavobacterium heparinum was cloned and expressed in Escherichia coli BL21 in order to enhance its activity. The expressed heparinase was purified to homogeneity by a metal chelating affinity column and its enzymatic properties were evaluated. A maximal heparinase activity of 1061 IU/L toward the substrate heparin was achieved when the recombinant strain was induced with 0.5 mM isopropyl-β-D-thiogalactoside at 28 °C for 9 h. The optimal temperature and pH of heparinase were 30 °C and 7.0, respectively. The recombinant heparinase was heat-unstable and had a higher stability at pHs from 7.0 to 10.0. Observed activities of heparinase were the highest in the presence of Ca(2+) and Cu(2+) and the lowest in the presence of Mn(2+) and Pb(2+). These results lay a good foundation for the preparation of LMWHs by heparin enzymolysis.
authors.affiliation: College of Food Science and Biotechnology, Zhejiang Gongshang University, 149 Jiaogong Road, Hangzhou 310035, People's Republic of China.
authors.forename: Ping
authors.initials: P
authors.lastname: Yu
authors.forename: Yan
authors.initials: Y
authors.lastname: Wu
mesh_terms: D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D005417:Flavobacterium / Q000201:enzymology* / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D019763:Heparin Lyase / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D019763:Heparin Lyase