麻类脱胶高效菌株DCE01的Pel325基因克隆与表达  

Cloning and Expression of Gene Pel325 from Bast Fiber Degumming Strain DCE01

作　　者：李琦[1] 成莉凤[2] 曾洁[2] 李国高 刘正初[1,2] 

机构地区：[1]湖南农业大学植物保护学院,长沙410128 [2]中国农业科学院麻类研究所,长沙410205 [3]湖南利尔康生物股份有限公司,湖南岳阳414100

出　　处：《中国麻业科学》2015年第3期157-161,共5页Plant Fiber Sciences in China

基　　金：国家高技术发展计划(2012AA022209D);国家麻类产业技术体系建设专项(CARS-19-E24);国家农业科技创新工程(ASTIP-IBFC08)

摘　　要：采用软件Primer premier 5设计引物,从麻类脱胶高效菌株DCE01中克隆出一个果胶酶基因Pel325,重组到表达载体p ET-28a中,在E.coli BL21(DE3)中成功表达。试验结果显示,酶的活性随着底物(橘子果胶)酯化程度的增加而增加。胞内酶用酯化程度≥85%的橘子果胶作底物检测时酶活最高(酶活为37.5U/ml),其最适反应温度为55℃,最适反应p H为8.0。该结果可为进一步发掘DCE01菌株的基因资源提供重要科学依据。Primers were designed by Primer premier 5. Gene Pel325 was cloned from the whole gehome sequence of strain DCE01, which is an efficient strain for bast fiber degumming. Then the gene was linked to pET -28a and expressed in E. coli BI21 （DE3） . The results showed that the enzyme activity increases with the rise of the esteriflcation degree of the substrates. The maximum intracellular enzyme activity appeared when use the pectin （from citrus fruit, ≥ 85% esterified） as the substrates. The optimal condition for this enzyme was at 55℃, pH8.0. These results may provide scientific basis for exploring gene resources from the strain DCE01.

关 键 词：果胶酶 克隆 表达 麻类脱胶 Pel325基因 

分 类 号：S188[农业科学—农业基础科学]