重组枯草芽孢杆菌PaE菌株的构建及碳代谢阻遏效应研究-食品科学.PDF

134 2014, Vol.35, No.23 食品科学 ※生物工程 重组枯草芽孢杆菌PaE菌株的构建及 碳代谢阻遏效应研究 * 李元召，孙俊良 ，梁新红，张永帅 （河南科技学院食品学院，河南新乡 453003 ） 摘  要： 168 α- amyE Paxo1 通过基因重组的方法，将枯草芽孢杆菌 的 淀粉酶基因 （除掉启动子）整合在 质粒上，然后 将重组表达质粒Paxo1-amyE导入枯草芽孢杆菌168菌株基因组中的半乳糖苷酶基因lacZ位点。通过转化筛选和重组 α- PaE 4 2 g/100 mL 基因分析，获得含有重组 淀粉酶基因的工程菌株 。经过添加 种不同的碳源发酵实验检测，在外加 α- 木糖诱导的情况下，重组菌株的 淀粉酶产量均有增加。而且重组菌株在一定程度上能够克服葡萄糖等还原性糖引 α- 起的碳代谢阻遏现象，提高了 淀粉酶产量。 α- 关键词： 淀粉酶；枯草芽孢杆菌；基因重组；木糖 Construction and Carbon Catabolite Repression of Recombinant Bacillus subtilis PaE * LI Yuan-zhao, SUN Jun-liang , LIANG Xin-hong, ZHANG Yong-shuai (School of Food Science, Henan Institute of Science and Technology, Xinxiang 453003, China) Abstract: Using gene recombination method, the alpha amylase gene amyE (without promoter) of Bacillus subtilis was integrated into Paxo1 plasmid and the galactosidase gene lacZ of B. subtilis strain 168 was introduced into the recombinant expression plasmid Paxo1-amyE. After transformation and screening analysis, genetically engineered strain PaE containing the recombinant alpha amylase gene was selected. With the addition of four different carbon sources, the alpha amylase production by the recombinant strain was promoted under the induction of 2 g/100 mL xylose. In addition, the recombinant strain could overcome the car