| 禾谷镰孢菌果胶裂解酶PelA的生物学功能分析 |
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| 引用本文： | 姚昇华,贾雷杰,张晓伟,张栋,许玲.禾谷镰孢菌果胶裂解酶PelA的生物学功能分析[J].植物生理学通讯,2014(3):243-252. |
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| 作者姓名： | 姚昇华 贾雷杰 张晓伟 张栋 许玲 |
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| 作者单位： | [1]华东师范大学生命科学学院,上海200241 [2]中国科学院上海生命科学研究院植物生理生态研究所,上海200032 |
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| 基金项目： | 国家基础研究项目（2011CB100702）和中国科学院知识创新项目（KSCX2-EW-N-06）. |
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| 摘 要： | 果胶裂解酶是果胶酶的重要成员之一，能够通过B-消除作用，催化降解（1—4）-α-D-聚半乳糖醛酸。本研究首次鉴定出禾谷镰孢茵中的一个果胶裂解酶PelA，并在体外检测了其生化特性。PelA基因（FGSG_02386）的cDNA被克隆并进行了原核表达，同时构建了单基因敲除突变体。将重组质粒pET-28a（＋）-PelA转化蛋白表达菌株大肠杆菌BL21表达目的蛋白，纯化后的蛋白具有果胶裂解酶活性。酶活性测定结果显示，该酶的活性受到外界环境因素如温度、钙离子浓度和pH条件的影响，其最适反应条件为50℃，CaCl：浓度0-5mmol·L^-1，pH8-5。在侵染小麦胚芽鞘时PelA单基因突变体的毒力较野生型菌株并没有出现明显变化。
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| 关 键 词： | 果胶裂解酶 禾谷镰孢菌 小麦胚芽鞘 |
Biological Function Analysis of A Pectate Lyase PelA in Fusarium graminearum |
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| YAO Sheng-Hua,JIA Lei-Jie,ZHANG Xiao-Wei,ZHANG Dong,XU Ling.Biological Function Analysis of A Pectate Lyase PelA in Fusarium graminearum[J].Plant Physiology Communications,2014(3):243-252. |
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| Authors: | YAO Sheng-Hua JIA Lei-Jie ZHANG Xiao-Wei ZHANG Dong XU Ling |
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| Institution: | 1.School of Life Sciences, East China Normal University, Shanghai 200241, China; 21nstitute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China) |
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| Abstract: | Pectate lyase can eliminate cleavage of （1→4）-α-D-galacturonan by β-elimination as a member of pectinases. This study identified a pectate lyase, PelA, in Fusarium graminearum for the first time, and its biochemical characteristics was verified in vitro. In this study, PelA was expressed in Escherichia coli and knock-out mutants were constructed. Biochemical analysis showed that PelA has pectate lyase activity in vitro. External environment such as temperature, calcium ion concentration and pH affected the enzyme activity, and enzyme activity reached the highest at 50 ℃, 0.5 rnmol·L^-1 CaCl2 and pH 8.5. PelA knock-out strains didn＇t show significant difference in virulence from wild-type strains during infection of wheat coleoptiles. |
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| Keywords: | pectate lyase Fusarium graminearum wheat coleoptile |
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