环糊精酶基因在毕赤酵母中的组成型表达

发布时间：2018-03-19 04:21

  本文选题：环糊精酶　切入点：毕赤酵母　出处：《福建农业学报》2017年01期 　论文类型：期刊论文

【摘要】：为实现环糊精酶在毕赤酵母中的高效表达,以优化合成的环糊精酶基CGT2为基础,构建组成型表达质粒pGAPZαA-CGT2,运用电转化方法将目的基因整合进酵母染色体,构建环糊精酶酵母工程菌X33/pGAPZαA-CGT2,经摇瓶发酵120h后,CGT2活力达0.21 U·mL~(-1);进一步对工程菌进行摇瓶发酵条件优化,确定其最优发酵条件为:pH6.5、28℃、200r·min~(-1)、每24h补加2.5%的甘油,120h后其胞外酶活力达到0.36U·mL~(-1),是优化前的1.7倍,实现了环糊精酶在毕赤酵母中的组成型表达。
[Abstract]:In order to realize the efficient expression of cyclodextrin in Pichia pastoris, a constitutive expression plasmid pGAPZ 伪 A-CGT2 was constructed based on the optimized synthesis of cyclodextrin enzyme CGT2, and the target gene was integrated into yeast chromosome by electrotransformation. The strain X33 / pGAPZ 伪 A-CGT _ 2 was constructed, and the activity of CGT _ 2 reached 0.21 U 路mL ~ (-1) after 120 h of shaking flask fermentation. The optimum fermentation conditions were determined as follows: (1) pH 6.5 ~ (28 鈩，

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