嗜酸乳杆菌细胞壁蛋白酶的提取优化.pdfVIP

260 2011, Vol. 32, No. 23 食品科学 ※生物工程 1 1,2, 李 敏 ，潘道东 * (1.南京师范大学 国家乳品加工技术研发分中心，江苏 南京 210097 ； 2.宁波大学生命科学与生物工程学院，浙江 宁波 315211) 为了获得较高酶活力的嗜酸乳杆菌细胞壁蛋白酶(CEP)，对嗜酸乳杆菌CEP 的提取方法进行优化。经过比 较超声波破碎法、Ca2+ -free 法、溶菌酶裂解 3 种方法，确认溶菌酶裂解法结合使用非离子清洁剂破壁是提取嗜酸 乳杆菌 CEP 的最佳方法。在单因素试验的基础上，采用正交试验对嗜酸乳杆菌CEP 的提取工艺进行优化，得出嗜 酸乳杆菌 CEP 的最佳提取方法：用裂解液(50mmol/L Tris-HCl 、100mmol/L NaCl、TritonX-100 添加量 1%、 1.5mg/mL 溶菌酶，pH 7.8)悬浮菌体(20mL/g) ，37 ℃保温3h ，4 ℃、4500r/min 离心20min ，取上清液。溶菌酶裂解 法提取CEP 过程中各因素对CEP 提取效果的影响依次为：溶菌酶质量浓度＞裂解pH 值＞TirtonX-100 添加量＞保温时间。 嗜酸乳杆菌；细胞壁蛋白酶(CEP) ；提取 Optimization of Extraction of Cell-envelope Proteinase from Lactobacillus acidophilus 1 1,2, LI Min ，PAN Dao-dong * (1.Branch of National Dairy Processing Technology Developing Center, Nanjing Normal University, Nanjing 210097, China； 2. Faculty of Life Science and Biotechnology, Ningbo University, Ningbo 315211, China) Abstract ：In this study, the extraction of cell-envelope proteinase (CEP) from Lactobacillus acidophilus was optimized. A comparison of ultrasonic treatment, Ca2+-free treatment and lysozyme lysis for their effectiveness in Lactobacillus acidophilus cell disruption was performed to determine lysozyme lysis as the best method among the three. Further, process conditions optimization for CEP extraction from Lactobacillus acidophilus by lysozyme lysis was carried out using orthogonal array design based on one-factor-at-a-time experiments. The best extraction results were obtained by 3 h incubation at 37 ℃ of Lacto