来源于盐惰菌属Halopiger xanaduensis的木聚糖酶基因在毕赤酵母中诱导表达及其酶学性质的研究  

Induced expression of xylanase gene from Halopiger xanaduensis in Pichia pastoris and its enzymatic properties

作　　者：于成野 韩红娟[1] 付晓燕[1] 游双红 孙淼 朱恒文 彭日荷[1] 姚泉洪[1] 

机构地区：[1]上海市农业科学院生物技术研究所,上海201106 [2]上海海洋大学食品学院,上海201306

出　　处：《上海农业学报》2017年第6期1-5,共5页Acta Agriculturae Shanghai

基　　金：上海市农业委员会重点项目(No.2011-1-8;2013D-8)

摘　　要：根据毕赤酵母的密码子偏爱性和木聚糖酶的蛋白序列设计并合成了木聚糖酶基因片段,与表达载体pYPX88连接,构建成分泌表达载体;构建的分泌表达载体经BglⅡ酶切后电击转化到毕赤酵母GS115中,筛选得到阳性克隆。研究了重组木聚糖酶的最适pH、pH稳定性、最适温度、温度稳定性以及部分金属离子、十二烷基硫酸钠(SDS)和二硫苏糖醇(DTT)对该酶性质的影响。结果表明:重组木聚糖酶最适pH为6.0,在pH 4.5—8.0时酶学性质稳定;最适温度为60℃在20-65℃时酶学性质稳定;金属离子Mn^(2+)与Cu^(2+)对该酶有抑制作用,化合物DTT对该酶有明显促进作用。The xylanase gene was designed and synthesized according to the codon preference of Pichia pastoris and xylanase protein sequence,which was connected with expression vector pYPX88 and constructed into secretory expression vector. The secretory expression vector was transformed into Pichia pastoris GS115 by electroporation after Bgl Ⅱ digestion,and the positive clones were screened out. The optimum pH,pH stability,optimum temperature and temperature stability of the recombinant xylanase,and the effects of some metal ions,sodium dodecyl sulfate(SDS),dithiothreitol( DTT)on the properties of the recombinant xylanase were studied.The results showed that the optimum pH of the recombinant xylanase was 6, and the enzyme properties were stable at pH 4. 5—8. 0; the optimum temperature was 60℃, and the enzyme properties were stable at 20—65℃; the metal ion Mn^(2+) and Cu^(2+)could inhibite the enzyme activity, and DTT coud promote the enzyme activity obviously.

关 键 词：木聚糖酶 毕赤酵母 酶学性质 

分 类 号：Q78[生物学—分子生物学]