SDS-PAGE电泳法分析绿豆芽几丁质酶特性  

Characterization of Chitinase Isoenzymes of Mung Bean Sprout by Sodium Dodecyl Sulfate-chitin-polyacrylamide Gel Electrophoresis

作　　者：刘平[1] 李雯妮[1] 奚宇[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：《农产品加工（创新版）（中）》2013年第5期1-4,共4页Farm Products Processing

基　　金："863"计划项目"蔬菜绿色供应链技术创新与产品研制"(2008AA100803)

摘　　要：采用Sodium dodecyl sulfate(SDS)-几丁质-聚丙烯酰胺凝胶电泳(SDS-Chitin-PAGE)方法分析绿豆芽(子叶和下胚轴)几丁质酶的特性。在绿豆芽子叶和绿豆芽下胚轴中共检测到5个几丁质酶同工酶,分别将绿豆芽子叶中3个几丁质酶同工酶命名为CH-1,CH-2和CH-3,绿豆芽下胚轴的2个同工酶命名为CH-4和CH-5。对绿豆芽而言,在SDS-Chitin-PAGE前,样品缓冲溶液与样品蛋白粗提液混合的最适温度为24℃,时间为10 min。样品缓冲溶液中添加β-巯基乙醇会极大地降低绿豆芽(子叶和下胚轴)中几丁质酶同工酶活性。绿豆芽子叶中CH-1和CH-2,绿豆芽下胚轴中CH-4的耐受温度达80℃,绿豆芽子叶中CH-3,绿豆芽下胚轴中CH-5的耐受温度也可达到70℃。绿豆芽子叶中CH-1的最适pH值为4.0～4.4,CH-2在pH值4.0～8.0的活性变化不大,CH-3的最适pH值为4.4～5.2。绿豆芽下胚轴中CH-4的最适pH值为5.2～5.6,CH-5的最适pH值为4.4～5.2。Mung bean sprout is selected to test their chitinase characteristics by the optimized SDS-Chitin-PAGE. The results show that： Three chitinase isozymes are named as CH-1, CH-2 and CH-3 in mung bean sprout cotyledon and two chitinase isozymes, CH-4 and CH-5, are detected in the mung bean sprout hypocotyl. 13 -mercaptoethanol decreases chitinase isozyme activity in mung bean sprout. For mung bean sprout, optimal temperature of buffer solutions and crude enzymes is 24 ℃ and the time is 10~20 min before SDS-Chitin-PAGE. The heat resistant temperatures of CH-1, CH-2 and CH-3 in mung bean sprout cotyledon are up to 80, 80 and 70 ℃, respectively. The heat resistant temperatures of CH-4 and CH-5 in the mung bean sprout hypocotyl are 80 and 70 ℃, respectively. The optimal pH ranges of CH-1, CH-2 and CH-3 are pH 4.0~4.4, pH 4.0-8.0 and pH 4.4-5.2, respectively. The optimal pH ranges of CH--4 and CH-5 in the mung bean sprout hypocotyl are pH 5.2~5.6 and 4.4~5.2, respectively.

关 键 词：几丁质酶 绿豆芽 酶学特性 聚丙烯酰胺凝胶原位显示电泳 

分 类 号：Q814[生物学—生物工程]