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响应面分析法优化α-环糊精的酶转化工艺
陈晓彤,黄立萍,孙 谧,郝建华
1.农业农村部极地渔业开发重点实验室 青岛海洋科学与技术试点国家实验室海洋药物与生物制品功能实验室 中国水产科学研究院黄海水产研究所 青岛 266071;2.上海海洋大学食品学院 上海 201306
摘要:
为获得较高的α-环糊精转化产率,采用单因素实验和响应面实验方法,筛选底物种类、底物浓度、加酶量、酶作用时间、作用温度和pH等多个单因素,对海洋芽孢杆菌Y112产的α-环糊精葡萄糖基转移酶产α-环糊精的条件进行了优化。然后利用Plackett-Burman实验筛选得到影响α-环糊精转化率的3个主要因素:底物浓度、温度和pH值。最终采用响应面分析法得到的最佳转化条件为马铃薯淀粉浓度为5%,加酶量200 U/g(淀粉),pH值为8.4,温度30℃,200 r/min反应6 h,α-环糊精转化率均值为28.67%,比优化前的产率提高了2.48倍。
关键词:  α-环糊精葡萄糖基转移酶  响应面分析法  α-环糊精  转化工艺
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Optimization of Conversion Process of α-Cyclodextrin with Enzyme Using Response Surface Methodology
CHEN Xiaotong1,2,3,4, HUANG Liping1,2,3,4, SUN Mi1,2,3, HAO Jianhua1,2,3
1.Key Laboratory of Polar Fisheries Development, Ministry of Agriculture and Rural Affairs;2.Laboratory for Marine Drugs and Bioproducts, Pilot National Laboratory for Marine Science and Technology (Qingdao);3.Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071;4.College of Food Sciences & Technology, Shanghai Ocean University, Shanghai 201306
Abstract:
In order to obtain a higher yield of α-cyclodextrin conversion, we use a single factor experiment and response surface experiment method, to optimize the production of α-cyclodextrin conditions by α-cyclodextrin glucosyltransferase. The enzyme is derived from strain Y112. First, the single factor method was used to screen six factors including substrate species, substrate concentration, enzyme content, reaction time, temperature, and pH. Second, these main variables affecting the conversion of α-cyclodextrin were evaluated with the Plackett-Burman test. The results suggested that substrate concentration, temperature, and pH were the most impactful factors. Finally, the optimal levels of the three factors were determined with central composite design and Design-Expert 8.05 software. The optimum conversion conditions were shown as follows: the concentration of potato starch was 5%, the amount of enzyme 200 U/g (starch), pH 8.4, 200 r/min, reacted at 30℃ for 6 h. Under the optimized conditions, the average value of α-CD conversion was 28.67%, which was an increase of 2.48 times compared with the pre-optimization yield.
Key words:  α-Cyclodextrin glucosyltransferase  Response surface analysis  α-Cyclodextrin  Conversion process