重组枯草芽孢杆菌生产青霉素G酰化酶发酵条件的研究  

Studies on Fermentation Conditions for Penicillin G Acylase Production with a Recombinant Bacillus subtilis System

作　　者：徐志南[1] 陈秀奇[1] 陈新爱[1] 岑沛霖[1] 

机构地区：[1]浙江大学化学工程与生物工程学系,浙江杭州310027

出　　处：《高校化学工程学报》2003年第3期266-271,共6页Journal of Chemical Engineering of Chinese Universities

摘　　要：对产青霉素G酰化酶的温度诱导型重组枯草芽孢杆菌发酵条件进行了研究。结果表明,培养基的最佳氮源和碳源组成为:35gL-1牛肉膏、3gL-1葡萄糖和6gL-1淀粉;最佳诱导时机是细胞对数生长的中后期,诱导前应将pH调节至中性;最佳诱导条件为升高温度至50℃并维持4min;随后将温度降低到34℃进行重组蛋白的表达。在上述条件下,PGA的表达水平可以达到5.8UmL-1。SDS-PAGE电泳分析表明该重组菌能够将绝大部分表达的PGA分泌到细胞外,而且表达的PGA蛋白占有高比例(90%)。The fermentation conditions for penicillin G acylase production with a temperature-inducible recombinant Bacillus subtilis were studied. The medium should contain 35 g&middotL-1 of beef extract, 3 g&middotL-1 of glucose and 6 g&middotL-1 of starch. The temperature inducing should be carried out at the late stage of exponential growth phase and the pH should be regulated to 7.0. The optimal inducing temperature is 50°C and should last for 4 minutes. After inducing, the temperature should be lowered to 34°C and continuously cultured for another 7 hours. With the optimal conditions, the PGA activity as high as 5.8 U&middotmL-1 was reached. From SDS-PAGE analyses, the recombinant Bacillus subtilis MU 103 can secrete almost all of the expressed PGA into the medium, and more than 90% of total proteins in the supernatant is recombinant PGA.

关 键 词：青霉素G酰化酶 温度诱导 重组枯草杆菌 发酵条件 

分 类 号：TQ920.1[轻工技术与工程—发酵工程] Q55[生物学—生物化学]