重组毕赤酵母生产耐高温α-淀粉酶发酵条件的优化 - 食品科学.pdf

266 2011, Vol. 32, No. 23 食品科学 ※生物工程 α- 1 , 2 1 , 2 , 1 1 1 秦亚楠 ，孟宪梅 * ，潘风光 * ，孙肖明 ，李凤祥 ，石彦忠 (1.吉林工商学院 粮油食品深加工重点实验室，吉林 长春 130062 ；2.吉林大学军需科技学院，吉林 长春 130062) 利用重组毕赤酵母诱导表达可产生耐高温 α- 淀粉酶，在摇瓶发酵基础上，从诱导时间、甲醇添加量、 pH 值、接种量 4 个因素考察发酵条件对耐高温 α- 淀粉酶酶活力的影响，结合四元线性回归正交试验设计研究方法 构建出甲醇诱导重组毕赤酵母生产耐高温 α- 淀粉酶的回归模型；优化最佳工艺参数为：诱导时间 96h 、甲醇添加 量 0.5% 、接种量 150mL/100mL、摇床转速200r/min 。确定最优工艺条件下，耐高温α- 淀粉酶酶活为 217.2U/mL 。 毕赤酵母；耐高温 α- 淀粉酶；发酵；优化 Optimization of Fermentation Conditions for Thermostable α-Amylase Production by Recombinant Pichia pastoris 1,2 1, 2, 1 1 1 QIN Ya-nan ，MENG Xian-mei *，PAN Feng-guang *，SUN Xiao-ming ，LI Feng-xiang ，SHI Yan-zhong (1. Key Laboratory of Oil and Foodstuffs Processing, Jilin Business and Technology College, Changchun 130062, China ； 2. College of Quartermaster Technology, Jilin University, Changchun 130062, China) Abstract ：The methanol-induced expression of thermostable α-amylase in a recombinant Pichia pastoris strain with the thermostable α-amylase gene of Bacillus licheniformis that had been constructed in our previous work was investigated during shake flask cultivation. A 4-variable linear regression design was used to explore the effects of 4 main cultivation conditions such as induction time, methanol amount, pH and inoculation amount on the expressed activity of thermostable α-amylase. A regression model describing the relationship between them was established. It was found that the optimal fermentation conditions were 96 h cultivation at an inoculation amount of 150 mL/100mL and a shake speed of 200 r/min in the presence o