Effect of CaCl2 as activity stabilizer on purification of heparinase I from Flavobacterium heparinum.
Heparinase I has been purified from F. heparinum by a novel scheme with 10mM CaCl(2) 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. CaCl(2) is a good stabilizer of the purified enzyme in liquid form toward either storaging at 4 degrees C or freezing-thawing.
Ma
Xiaolai
X
State Key Laboratory of Bioreactor Engineering, East China University of Science & Technology, 130 Meilong Road, P.O. Box 365, Shanghai 200237, PR China.
Wang
Zunsheng
Z
Li
Suxia
S
Shen
Qiong
Q
Yuan
Qinsheng
Q
D002122:Calcium Chloride / Q000494:pharmacology*; D002850:Chromatography, Gel / Q000379:methods*; D004795:Enzyme Stability / Q000187:drug effects; D005417:Flavobacterium / Q000201:enzymology*; D019763:Heparin Lyase / Q000187:drug effects / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D013696:Temperature
D019763:Heparin Lyase; D002122:Calcium Chloride