基因重组毕赤酵母产蛋清溶菌酶发酵工艺及表达条件的优化  

Fermentation technology and expression conditions optimization of recombinant Pichia pastoris producing egg white lysozyme

作　　者：宋增健 薛松 王向东[2] 林剑[1] SONG Zengjian;XUE Song;WANG Xiangdong;LIN Jian(College of Life Sciences,Yantai University,Yantai 264000,China;Institute of Applied Life Sciences,Shandong University,Jinan 250000,China)

机构地区：[1]烟台大学生命科学学院,山东烟台264000 [2]山东大学应用生命科学研究院,山东济南250000

出　　处：《中国酿造》2018年第10期20-24,共5页China Brewing

基　　金：国家海洋局海洋经济创新示范项目(HY2016K20)

摘　　要：以甲醇诱导型蛋清溶菌酶基因重组毕赤酵母(Pichia pastoris)NCY-2为研究菌株,选用麦芽汁为培养基,首先通过单因素试验,对培养基锥形瓶种类、原麦芽汁稀释度及外加氮源进行了优化,在此基础上,采用均匀设计试验,得到菌株NCY-2的最佳发酵工艺条件为发酵温度31℃、pH 5.5,转速220 r/min,接种量4%;蛋清溶菌酶最佳表达条件为诱导温度18℃、诱导pH 4.4、转速190 r/min、甲醇初始添加量为0.3%(之后每隔24 h分别添加体积分数1.0%、1.5%、2.0%,诱导72 h结束)。在此优化条件下,摇床水平发酵液中蛋清溶菌酶酶活力可达到962.4 U/m L。Using methanol inducible egg white lysozyme gene recombinant Pichia pastoris NCY-2 as research strain and wort as medium,the types of conical flask,the dilution degree of the original wort,and additional nitrogen source wort were optimized by single factor tests in flask level.On this basis,using the uniform design experiments,the optimal fermentation technology condition of strain NCY-2 was fermentation temperature 31℃,pH 5.5,rotation speed 220 r/min and inoculum 4%;and the optimal expression conditions of egg white lysozyme were obtained as follows:induce temperature 18℃,pH 4.4,rotation speed 190 r/min,initial methanol addition 0.3%(adding 1.0%,1.5%,2.0%after every 24 h,until 72 h).Under the optimal condition,the egg white lysozyme activity was 962.4 U/ml in the fermentation broth.

关 键 词：鸡蛋清溶菌酶 毕赤氏酵母 麦芽汁培养基 发酵工艺 表达条件 优化 

分 类 号：Q815[生物学—生物工程]