Heterologous Expression and Characteristics of Alkaline Polygalacturonate Lyase that Specifically Depolymerize Substrate with High Methyl Esterification Degree
目的:异源表达、纯化特异性降解高甲酯化果胶的碱性果胶酯裂解酶,研究其酶学性质。方法:采用PCR方法从枯草芽孢杆菌7-3-3中扩增到编码碱性果胶酯裂解酶的结构基因pel B,将去信号肽基因序列连接甲醇诱导型质粒p PIC9K,得到重组质粒p PIC9K-pel B,并转入毕赤酵母GS115进行表达,对该表达产物进行酶学性质分析。结果:纯化得到Pel B及其糖基化蛋白,糖基化不影响Pel B的酶活力。该酶可降解甲酯化程度为85%的果胶,比酶活达2 746 U/mg。在最适反应温度60℃,最适反应p H 9.0,45~65℃和p H 8.5~9.5范围具较高的酶活力。在40℃以下,p H 7.0~10.6的碱性范围,酶具有较高的稳定性。结论:Pel B蛋白对高度甲酯化的果胶底物具有高解聚能力,在碱性条件下稳定,提示其具有良好的应用价值。
【英文摘要】
Objective: To heterologously express and purify alkaline polygalacturonate lyase that specifically depolymerize substrate with high methyl esterification degree and to investigate its enzymatic properties, and provide the experimental basis for its development and application. Methods: Alkaline polygalacturonate lyase structural gene, pel B,was amplified from Bacillus subtilis 7-3-3 genome by PCR and discovered that it has close genetic relationship with Pel B from Bacillus subtilis BSn5 by constructing phy...
| 
推荐 
点击下列相关研究机构和相关文献作者，可以直接查到这些机构和作者被《中国知识资源总库》收录的其它文献，使您全面了解该机构和该作者的研究动态和历史。
