天津科技大学学报2016，Vol.31Issue(5)：8-13,6.DOI:10.13364/j.issn.1672-6510.20150217
碱性蛋白酶的异源表达及其在制备CPP中的应用
Heterologous Expression and Application of Gene Encoded Alkaline Protease in Preparation of CPP
摘要
Abstract
Alkaline protease is one of the most important enzymes in industrial enzymes.It plays a very important role in human daily life.Alkaline protease encoded gene(apr)was amplified from B.alcalophilus TCCC11006,genome.Then the recombined expression vector pHY-apr was successfully constructed and transformed into protease-deficient strain B.licheniformis H115.The alkaline protease gene was expressed in the recombined B.licheniformis H115.The highest alka-line protease activity was 3,199,U/mL in LB medium,which was 1.82,times of that of the starting strain.The fermentation cycle was decreased by 8,h.Then the hydrolyzing casein paramenters of recombined alkaline protease were optimized with single factor and orthogonal experiments.The optimum hydrolyzing time,temperature,pH and the amount of the enzyme were found to be 2,h,50,℃,9.5,and 4,000,U/g substrate.Under the optimal hydrolysis conditions,the yield and N/P mole ratio of the casein peptide phosphate(CPPs)prepared were 14.8%, and 8.47,respectively.Knowledge gained in the research can be a solid foundation for the application of alkaline protease in the manufacturing of functional food.关键词
碱性蛋白酶/异源表达/水解/酪蛋白磷酸肽Key words
alkaline protease/cloning expression/hydrolysis/CPP分类
化学化工引用本文复制引用
凌敏,董自星,李玉,叶松,王正祥,路福平..碱性蛋白酶的异源表达及其在制备CPP中的应用[J].天津科技大学学报,2016,31(5):8-13,6.基金项目
国家高技术研究发展计划(863计划)资助项目(2013AA102106-07) （863计划）
