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

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.