METHOD FOR PRODUCING LOW MOLECULAR WEIGHT OF HEPARIN BY USING RECOMBINANT HEPARINASE CONTAINING A HISTIDINE OLIGOPEPTIDE

摘要

PURPOSE: A method for producing low molecular weight of heparin by using a recombinant heparinase containing a histidine oligopeptide is provided, thereby decreasing the immobilization time, storing the heparinase stably stored for a long time and reusing it. CONSTITUTION: The low molecular weight of heparin is produced by preparing an expression vector containing the structural gene of heparinase having the nucleotide sequence of SEQ ID NO: 1, which contains a gene coding histidine oligopeptide at 5'-terminal; transforming E. coli with the expression vector to produce a E. coli transformant; cultivating the E. coli transformant to produce heparinase containing a histidine oligopeptide; and reacting the heparinase with heparin at 20 to 28 deg.C and pH 5.0 to 8.0 to produce low molecular weight of heparin, in which the expression vector is pE16b-hepa2, the E. coli transformant is E. coli BL21(DE3)(KFCC-10903), and the heparinase is immobilized on a chelating supporter.