响应面法优化解淀粉芽孢杆菌发酵产凝乳酶的工艺条件.docVIP

响应面法优化解淀粉芽孢杆菌发酵产凝乳酶的工艺条件 李建涛收稿日期：2011-11-13基金资助：国家科技部“863”计划(2011AA100903); 收稿日期：2011-11-13 基金资助：国家科技部“863”计划(2011AA100903); “十一五”科技支撑计划(2009BADB9B06); 北京市科技计划(D10110504600000) 作者简介：李建涛(1986-)，女，硕士，研究方向为食品发酵工程 通讯作者：陈历俊 （1.大连工业大学，大连116034；2.北京三元食品股份有限公司，北京100085） 摘要：首先通过单因素实验分析了不同碳源、氮源、金属盐、磷源对解淀粉芽孢杆菌产凝乳酶的影响，然后在此基础上采用Box-Behnken设计对葡萄糖、酵母粉和CaCO3 三因素的最优组合进行了定量研究，建立并分析了各因素与凝乳酶活力关系的数学模型。结果表明，解淀粉芽孢杆菌产凝乳酶的最佳工艺条件：马铃薯浸粉0.5%，葡萄糖1.13%，酵母浸粉1.76%，CaCO3为0.32%（均为质量分数），在此最佳培养基条件下，凝乳酶活力可达436.8SU/mL，与理论预测值438.4SU/mL基本一致，优化后解淀粉芽孢杆菌产凝乳酶的活力比基础培养基提高了5.21倍。 关键词：解淀粉芽孢杆菌；凝乳酶；响应面法；优化 Response surface methodology in optimization of milk-clotting enzyme fermentation conditions by Bacillus amyloliquefaciens LI Jian-tao1，2，CHEN Li-jun2，JIANG Tie-min2 （1. Dalian Polytechnic University, Dalian 116034，China; 2. Beijing Sanyuan Foods Co. Ltd., Beijing 100085，China） Abstract: Firstly, four main factors, including different carbon sourcesnitrogen source, metal salt and P sources were analyzed by using single factor analysis method. Then, the influences of glucose, yeast extract powder and CaCO3 were studied according to the Box-Behnken design. One mathematical model was established by response surface methodology (RSM) to describe the relationships between the studied factors and milk-clotting activity(MCA). Based on the canonical, The result showed that the suitable fermentation conditions for milk-clotting enzyme production by Bacillus amyloliquefaciens was glucose 1.13%, yeast extract powder 1.76% and CaCO3 0.32%. Under above conditions, the MCA can reached 436.8Su/ml, which was 5.21 folds higher than that before optimization, and it was confirmed that the predicted and actual value of the MCA were identical. Key words: bacillus amyloliquefaciens; milk-clotting enzyme; responsesurface methodology；optimization 0 引言 凝乳酶是奶酪生产过程中使牛奶凝固的关键性酶，被广泛用于乳酪业。微生物凝乳酶是目前最有前途的发展方向[1]，近几年芽孢杆菌凝乳酶的文献报道较多，主要有枯草芽孢杆菌[2]、解淀粉芽孢杆菌[3]、蜡样芽孢杆菌[4]和地衣芽孢杆菌[5]，而高蛋白水解能力是细菌来源的凝乳酶成为商品凝乳酶的主要障碍，细菌体积小、生长周期短、成本低、发酵产物易于提取分离[6]，因此筛选凝乳