本文選題：環(huán)糊精酶　切入點：畢赤酵母　出處：《福建農(nóng)業(yè)學(xué)報》2017年01期 　論文類型：期刊論文

【摘要】：為實現(xiàn)環(huán)糊精酶在畢赤酵母中的高效表達,以優(yōu)化合成的環(huán)糊精酶基CGT2為基礎(chǔ),構(gòu)建組成型表達質(zhì)粒pGAPZαA-CGT2,運用電轉(zhuǎn)化方法將目的基因整合進酵母染色體,構(gòu)建環(huán)糊精酶酵母工程菌X33/pGAPZαA-CGT2,經(jīng)搖瓶發(fā)酵120h后,CGT2活力達0.21 U·mL~(-1);進一步對工程菌進行搖瓶發(fā)酵條件優(yōu)化,確定其最優(yōu)發(fā)酵條件為:pH6.5、28℃、200r·min~(-1)、每24h補加2.5%的甘油,120h后其胞外酶活力達到0.36U·mL~(-1),是優(yōu)化前的1.7倍,實現(xiàn)了環(huán)糊精酶在畢赤酵母中的組成型表達。
[Abstract]:In order to realize the efficient expression of cyclodextrin in Pichia pastoris, a constitutive expression plasmid pGAPZ 偽 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 偽 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，

本文編號：1632811