毕赤酵母异源表达β-葡聚糖酶酶学特性的研究  

Characterization of endo-β-1,4-glucanase heterologously expressed in Pichia pastoris

作　　者：王苑螈 王波[2] 王秀英[2,3] 申强 薛永[2] 李大伟[1,2] 彭日荷[2] 姚泉洪[2] 

机构地区：[1]上海海洋大学食品学院,上海201306 [2]上海市农业科学院生物技术研究所,上海201106 [3]上海师范大学生命与环境科学学院,上海200234 [4]无锡市农业技术推广总站,无锡214023

出　　处：《上海农业学报》2016年第1期10-14,共5页Acta Agriculturae Shanghai

基　　金：上海市农委重大项目(沪农科种字2013-8;沪农科种字2014-2)

摘　　要：根据酵母密码子的偏好性对来自某异源菌种的葡聚糖酶基因进行化学合成改造,用电转化方法整合到毕赤酵母基因组中进行表达,并对表达出的内切β-葡聚糖酶进行相关酶学特性研究。结果表明:酶的最适pH为4.5,最适温度为40℃C;酶的pH稳定性范围与缓冲溶液类型有关,为4.0—6.0;Al^(3+)对酶有激活作用,但Mn^(2+)和Cu^(2+)则会抑制酶的活性;此酶对羧甲基纤维素钠(CMC-Na)有较强的分解能力;胃蛋白酶对该酶有抑制作用。The β-1,4-glucanase gene from Gibberella zeae was re-synthesized chemically according to the yeast bias condon and tansformed into Pichia pastoris by electroporation.The expressed endo-β-glucanase was characterized,and the results showed that the optimal pH is 4.5 and the optimal temperature is 45℃.The pH stability of the enzyme was 4.0-6.0,which was related to the type of buffer solution.Al^（3＋） had activation on the enzyme,but Mn^（2＋） and Cu^（2＋） inhibited the enzyme activity.The substrate specificity on carboxylmethyl cellulose sodium salt（CMC-Na）was strong.Pepsin had an inhibiting effect on the enzyme.

关 键 词：Β-葡聚糖酶 毕赤酵母 酶学特性 DNS法 

分 类 号：S816.3[农业科学—饲料科学]