文献类型：期刊文献

英文题名：Effect of CaCl2 as activity stabilizer on purification of heparinase I from Flavobacterium heparinum

作者：Ma, Xiaolai[1]; Wang, Zunsheng[1,2]; Li, Suxia[1]; Shen, Qiong[1]; Yuan, Qinsheng[1]

机构：[1] State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China; [2] Department of Biology, Shenyang Normal University, Shenyang, 110034, China

年份：2006

卷号：843

期号：2

起止页码：209

外文期刊名：Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences

收录：EI(收录号：20064410212795)

语种：英文

外文关键词：Bacteria - Cell culture - Chromatography - pH effects - Precipitation (chemical) - Purification

摘要：Heparinase I has been purified from F. 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 °C. CaCl2 is a good stabilizer of the purified enzyme in liquid form toward either storaging at 4 °C or freezing-thawing. ? 2006 Elsevier B.V. All rights reserved.