發(fā)布時間：2018-03-24 15:57

  本文選題：葡萄糖氧化酶　切入點(diǎn)：黑曲霉　出處：《食品與生物技術(shù)學(xué)報》2017年09期

【摘要】：在畢赤酵母SMD1168中利用乙醇氧化酶AOX1強(qiáng)啟動子表達(dá)黑曲霉葡萄糖氧化酶(Glucose oxidase,GOD)。提取黑曲霉Aspergillus niger PCTC的基因組DNA,以此為模板進(jìn)行PCR擴(kuò)增獲得葡萄糖氧化酶基因,將目的基因插入到具有AOX1強(qiáng)啟動子的表達(dá)載體p PICZαA上,經(jīng)電轉(zhuǎn)化導(dǎo)入畢赤酵母SMD1168中。經(jīng)zeocin抗性平板初篩、搖床復(fù)篩以及SDS-PAGE蛋白質(zhì)電泳的檢測,獲得了一株產(chǎn)葡萄糖氧化酶活力的菌株,該株菌在30℃、200 r/min的培養(yǎng)條件下,經(jīng)體積分?jǐn)?shù)1.0%的甲醇誘導(dǎo)發(fā)酵1 d可獲得1.12 U/mL的酶活。對該菌株進(jìn)行了搖瓶產(chǎn)酶條件優(yōu)化,其最佳發(fā)酵條件為:在pH 5、30℃下經(jīng)體積分?jǐn)?shù)1.5%甲醇誘導(dǎo)7 d,酶活為32 U/mL。
[Abstract]:In Pichia pastoris SMD1168, the strong promoter of ethanol oxidase AOX1 was used to express glucose oxidase gene of Aspergillus Niger. The genomic DNA of Aspergillus niger PCTC was extracted, and the glucose oxidase gene was obtained by PCR amplification. The target gene was inserted into the expression vector p PICZ 偽 A with strong promoter of AOX1, and was transformed into Pichia pastoris SMD1168 by electroporation. The screening of zeocin resistance plate, shaking bed screening and SDS-PAGE protein electrophoresis were performed. A strain producing glucose oxidase activity was obtained. The strain was cultured at 30 鈩，

本文編號：1658999