文献类型：期刊文献

中文题名：Purification and Characterization of Three Alkaline Endopolygalacturonases from a Newly Isolated Bacillus gibsonii

英文题名：Purification and characterization of three alkaline endopolygalacturonases from a newly isolated Bacillus gibsonii

作者：Li, Zu-Ming; Jin, Bo; Zhang, Hong-Xun; Bai, Zhi-Hui; Xue, Wen-Tong; Li, Hong-Yu

通讯作者：Li, Z.-M.

机构：[1] Teacher's College, Beijing Union University, Beijing 100011, China; [2] School of Earth and Environmental Science, University of Adelaide, Adelaide, SA 5005, Australia; [3] Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China; [4] College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China

第一机构：北京联合大学师范学院

通讯机构：[1]Teacher's College, Beijing Union University, Beijing 100011, China|[1141711]北京联合大学师范学院;[11417]北京联合大学;

年份：2008

卷号：8

期号：4

起止页码：768-773

中文期刊名：过程工程学报

外文期刊名：Guocheng Gongcheng Xuebao/The Chinese Journal of Process Engineering

收录：CSTPCD;;EI(收录号：20083911603567);Scopus;北大核心:【北大核心2004】；CSCD:【CSCD2011_2012】；

基金：Beijing Municipal Commission of Education(No.20061D0502200295);National Natural Science Foundation of China(No.30600082);Australian Research Council International Linkage Fellowship(No.LX0560210)

语种：英文

中文关键词：净化方法;细菌;隔离方法;果胶酶

外文关键词：Alkaline endopolygalacturonases; Bacillus gibsonii; Enzyme activity; Purification

摘要：A newly isolated Bacillus gibsonii,designated as S-2(CGMCC1215),was cultivated for production of alkaline pectinases utilizing sugar beet pulp as growth substrate.Purification of three alkaline endopolygalacturonases(endoPGs)from the crude pectinases extract was carried out by ultra-filtration,ammonium sulphate fractionation and ion-exchange chromatography,and their enzyme activities characterized.The three purified alkaline endoPGs,designated as S-I,S-II,and S-III,had a molecular weight about38kDa as determined by SDS-PAGE.The Km value and optimal temperature for optimal enzyme activities of S-I,S-II and S-III were1.2mg/mL and60℃,0.9mg/mL and55℃,1.1mg/mL and60℃,respectively.Their best performances were given at an optimal pH10.5,and sodium polygalacturonate was found to be the best substrate.The isoelectric points of S-I,S-II and S-III were5.4,7.4,and8.2,respectively.Surfactants of Tween-80and Tween-20and metal ions such as Mg2+and Ca2+stimulated the activity of S-I,S-II and S-III,whereas S-III was inhibited by Ca2+,and Mn2+and Zn2+ions inhibited the activity of the three enzymes.
A newly isolated Bacillus gibsonii, designated as S-2 (CGMCC1215), was cultivated for production of alkaline pectinases utilizing sugar beet pulp as growth substrate. Purification of three alkaline endopolygalacturonases (endoPGs) from the crude pectinases extract was carried out by ultra-filtration, ammonium sulphate fractionation and ion-exchange chromatography, and their enzyme activities characterized. The three purified alkaline endoPGs, designated as S-I, S-II, and S-III, had a molecular weight about 38 kDa as determined by SDS-PAGE. The Km value and optimal temperature for optimal enzyme activities of S-I, S-II and S-III were 1.2 mg/mL and 60°C, 0.9 mg/mL and 55°C, 1.1 mg/mL and 60°C, respectively. Their best performances were given at an optimal pH 10.5, and sodium polygalacturonate was found to be the best substrate. The isoelectric points of S-I, S-II and S-III were 5.4, 7.4, and 8.2, respectively. Surfactants of Tween-80 and Tween-20 and metal ions such as Mg2+ and Ca2+ stimulated the activity of S-I, S-II and S-III, whereas S-III was inhibited by Ca2+, and Mn2+ and Zn2+ ions inhibited the activity of the three enzymes.