SDS-Chitin-PAGE电泳法分析大葱叶几丁质酶特性  

Characterization of chitinase isoenzymes in Allium Fistulosum L leaves by Sodium dodecyl sulfate-Chitin-Polyacrylamide gel electrophoresis

作　　者：奚宇[1] 刘平[1] 王肖[1] 姜微波[1] 

机构地区：[1]中国农业大学食品科学与营养工程学院,北京100083

出　　处：《食品工业科技》2013年第7期153-157,共5页Science and Technology of Food Industry

基　　金：国家863项目资助(2008AA100803);国家级大学生创新训练项目资助(2011-2012causp13)

摘　　要：使用Sodium dodecyl sulfate-几丁质-聚丙烯胺酰胺凝胶电泳(SDS-Chitin-PAGE)方法分析了大葱葱叶几丁质酶的特性。在大葱叶组织中共检测到三种几丁质酶同工酶,分别命名为CH-A、CH-B、CH-C。分析结果表明,这三种几丁质酶具有SDS抗性,β-巯基乙醇对其活性也无显著影响。在SDS-Chitin-PAGE前,样品缓冲溶液与样品蛋白粗提液孵化处理的最适温度为24℃、时间为10～20min。凝胶电泳酶活分析表明,大葱葱叶中CH-A、CH-B的耐热温度可达60℃,CH-C的耐热温度达到70℃。葱叶中CH-A的最适pH为5.2～5.6,CH-B的最适pH为4.0～4.4,CH-C的最适pH为6.6～8.0。Allium Fistulosum L leaves was representatively selected to test their chitinase characteristics by the optimized SDS-Chitin- PAGE.The results showed that three chitinase isozymes, CH-A, CH- B and CH-C, were detected in Alliurn Fistulosum L leaves.Alliurn Fistulosum L had the very special characteristic of SDS resistance. β-mercaptoethanol had no significant effect on the chitinase activity.The optimal incubating temperature and time of the crude enzymes with the sample buffer were 24℃, 10-20rain before the electrophoresis.The activity of CH-A and CH-B could be observed up to 60℃,the activity of CH-C could be observed up to 70℃.The optimal pH ranges for CH-A,CH-B and CH-C were pH 5.2-5.6,pH 4.0-4.4,and pH 6.6-8.0,respectively.

关 键 词：几丁质酶大葱 酶学特性 聚丙烯酰胺凝胶原位显示电泳 

分 类 号：TS201.2[轻工技术与工程—食品科学]