CAJ | 학술논문

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 pET －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 pH 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。
채용연건 Primer premier 5설계인물，종마류탈효고효균주 DCE01중극륭출일개과효매기인 Pel325，중조도표체재체 pET －28a 중，재 E．coli BL21（DE3）중성공표체。시험결과현시，매적활성수착저물（귤자과효）지화정도적증가이증가。포내매용지화정도≥85％적귤자과효작저물검측시매활최고（매활위37．5U ／ml），기최괄반응온도위55℃，최괄반응 pH 위8．0。해결과가위진일보발굴 DCE01균주적기인자원제공중요과학의거。
Primers were designed by Primer premier 5.Gene Pel325 was cloned from the whole ge-nome 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 BL21 (DE3).The results showed that the enzyme activity increases with the rise of the esterification degree of the substrates.The maximum intracellular enzyme activity appeared when use the pectin (from citrus fruit,≥85% esterified)as the substrates.The opti-mal condition for this enzyme was at 55℃,pH8.0.These results may provide scientific basis for explo-ring gene resources from the strain DCE01.