高产碱性蛋白酶纳豆芽胞杆菌的诱变筛选
首发时间:2021-04-08
摘要:为了获得一株高产碱性蛋白酶的纳豆芽孢杆菌菌株,将实验室原有的一株纳豆芽孢杆菌进行DES(硫酸二乙酯)和ARTP(常压室温等离子体)复合诱变,以添加脱脂奶粉的平板为筛选平板,根据透明圈绝对直径和透明圈直径相对菌株的相对直径为初筛判断指标进行筛选,并对初筛后的菌株进行摇瓶发酵复筛,和遗传稳定性实验。最终获得一株发酵产碱性蛋白酶菌株,发酵产碱性蛋白酶酶活力为116U mL-1的菌株,碱性蛋白酶酶活力比未诱变前提升45%。对该菌株进行摇瓶发酵条件单因素和正交优化实验,优化后,发酵产碱性蛋白酶酶活力为129U mL-1,比优化前提升10%以上。
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Mutation screening of Bacillus natto with high production of alkaline protease
Abstract:To obtain a strain capable of high alkaline protease production, the wild-type Bacillus natto strain stored in our laboratory was subjected to a combined mutagenesis, consisting of DES (diethyl sulfate) and ARTP (atmospheric pressure room temperature plasma). With skimmed milk powder used as the screening plate, the initial screening was performed based on the absolute diameter of the transparent circle and the relative diameter of the transparent circle relative to the strain. The repeat screening after the initial screening was made in shake-flasks, followed by the genetic stability test. With the combined mutation, a mutant with a high activity of alkaline protease was obtained. This mutated strain produced116 U mL-1 of alkaline protease, which was 45% higher than that of the parental strain. With optimization of single-fMutation screening of
Keywords: Bacillus natto alkaline protease compound mutagenesis fermentation optimization
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