Effect of CaCl2 as activity stabilizer on purification of heparinase I from Flavobacterium heparinum |
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Authors: | Ma Xiaolai Wang Zunsheng Li Suxia Shen Qiong Yuan Qinsheng |
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Institution: | State Key Laboratory of Bioreactor Engineering, East China University of Science & Technology, 130 Meilong Road, P.O. Box 365, Shanghai 200237, PR China. |
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Abstract: | 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. |
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