枯草芽孢杆菌工程菌产耐酸性高温α-淀粉酶发酵条件的优化  

Optimization of Fermentation Conditions for Acid-resistant and Heat-stable α-Amylase Production with Engineered Strain Bacillus subtilis

作　　者：胡博[1,2] 刘逸寒[1,2] 徐艳静[1,2] 薄嘉鑫 路福平[1,2] 

机构地区：[1]工业发酵微生物教育部重点实验室 [2]天津科技大学生物工程学院,天津300457

出　　处：《天津科技大学学报》2012年第6期1-6,共6页Journal of Tianjin University of Science & Technology

基　　金：国家自然科学基金资助项目(31101219)

摘　　要：利用基因工程手段获得的产耐酸性高温α-淀粉酶基因枯草芽孢杆菌工程菌株pWB-amyd/WB600,通过单因素筛选及正交实验进行发酵培养基优化,得到的最佳配方为(g/L):玉米粉20,蛋白胨30,CaCl20.5,Na2HPO48.同时对实验室摇瓶条件下液体发酵的主要影响因素初始pH、接种量、装液量、转速、温度等进行探讨,确定了最佳培养条件:37,℃、pH 6.5、200,r/min摇床培养36,h,接种量为2%,装液量为30,mL/250,mL.在此优化条件下,耐酸性高温α-淀粉酶活力达到3,980,U/mL,是未优化条件下的2.1倍.Single factor experiments and orthogonal experiments were adopted to optimize the liquid fermentation medium and conditions of the engineered strain Bacillus subtilis pWB-amyd/WB600, which expresses acid-resistant and heat-stable t^-amylase. The results indicated that the constituents of the medium, the filling volume of shaking flask, the rotation speed and the temperature were the major factors that affected the yield of a-amylase. The composition of the best medium was 20 g/L maizena, 30 g/L peptone, 0.5 g/L CaC12, and 8 g/L Na2HPO4. The optimal fermentation conditions were the combina- tion of 37 ℃ temperature, 6.5 initial pH, shaking at 200 r/min for 36 h, 2% of inoculum volume and 30 mL/250 mL medium volume. The enzyme activity of a-amylase is 3980 U/mL in the optimized conditions, which is 2.1 times of the enzyme ac- tivity level in unoptimized conditions.

关 键 词：枯草芽孢杆菌工程菌 耐酸性高温α-淀粉酶 培养基优化 正交实验 发酵条件优化 

分 类 号：Q814.4[生物学—生物工程]