Effect of CaCl2 as activity stabilizer on purification of heparinase I from Flavobacterium heparinum

摘要

Heparinase I has been purified from E heparinum by a novel scheme with 10 mM CaCl2 added in crude extracts of cells. The enzyme was purified to apparent homogeneity through ammonium sulfate precipitation, Octyl-Sepharose chromatography, CM-52 chromatography, SP-650 chromatography, and Sephadex G-100 gel filtration chromatography. The specific activity of the purified enzyme was 90.33 U/mg protein with a purification fold of 185.1. The yield was 17.8%, which is higher than any previous scheme achieved. The molecular weight of the purified enzyme was 43 kDa with a pI of 8.5. It has an activity maximum at pH range of 6.4-7.0 and 41 degrees C. CaCl2 is a good stabilizer of the purified enzyme in liquid form toward either storaging at 4 degrees C or freezing-thawing. (c) 2006 Elsevier B.V. All rights reserved.