Purification and Characterization of Two Endo-β-1,4-glucanases from Mollusca,Ampullaria crossean

作     者：Yan-Hong LI~(1#) Rui GUO~(3#) Qiu-Yu YIN~1 Ming DING~1 Si-Liang ZHANG~3 Gen-Jun XU~(1,2) Fu-Kun ZHAO~(1,2*) 1 Key Laboratory of Proteomics,Institute of Biochemistry and Cell Biology,Shanghai Institutes for Biological Sciences.Graduate School of the Chinese Academy of Sciences,Chinese Academy of Sciences,Shanghai 200031,China 2 College of Life Science,Zhejiang Sci-Tech University,Hangzhou 310018,China 3 Bioengineering Institute,East China University of Science and Technology,Shanghai 200233,China 

作者机构：Key Laboratory of Proteomics Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Graduate School of the Chinese Academy of Sciences Chinese Academy of SciencesShanghai 200031 China Bioengineering Institute East China University of Science and TechnologyShanghai 200233 China College of Life Science Zhejiang Sci-Tech UniversityHangzhou 310018 China 

出 版 物：《Acta Biochimica et Biophysica Sinica》 (生物化学与生物物理学报(英文版))

年 卷 期：2005年第37卷第10期

页      面：702-708页

核心收录：

学科分类：08[工学] 082203[工学-发酵工程] 0822[工学-轻工技术与工程] 

基　　金：This work was supported by the grants from the National Natural Science Foundation of China (No.30370336) the Major State Basic Research Development Program of China (No.2003CB716006 and No. 2004CB719702) the Creation Foundation from Shanghai Institute 

主　　题：cellulase endo-β-1,4-glucanase Ampullaria crossean purification substrate specificity 

摘      要：Two novel endo-β-1,4-glucanases,EG45 and EG27,were isolated from the gastric juice ofmollusca,Ampullaria crossean,by anion exchange,hydrophobic interaction,gel filtration and a secondround of anion exchange *** purified proteins EG45 and EG27 appeared as a single bandon sodium dodecylsulfate polyacrylamide gel electrophoresis with a molecular mass of 45 kDa and 27 kDa,*** optimum pH for CMC activity was 5.5 for EG45 and 4.4-4.8 for *** optimumtemperature range for EG27 was broad,between 50℃ and 60℃;for EG45 it was 50℃.The analysis on thestability of these two endo-β-1,4-glucanases showed that EG27 was acceptably stable at pH 3.0-11.0 evenwhen the incubation time was prolonged to 24 h at 30℃,whereas EG45 remained relatively stable at *** 85% of the activity of EG27 could be retained upon incubation at 60℃ for 24 ***,less than 10% residual activity of EG45 was detected at 50℃.Among different kinds of substrates,bothenzymes showed a high preference for carboxymethyl ***45,in particular,showed a carboxymethylcellulose hydrolytic activity of 146.5 IU/mg *** enzymes showed low activities to xylan (from oatspelt) and Sigmacell 101,and they were inactive to p-nitrophenyl-β-D-cellobioside,salicin and starch.