期刊论文

Expression and enzymological properties of XylB gene xylanase in Pichia pastoris

中文

食品科技

1005-9989

2018

43

5

1-9

10.13684/j.cnki.spkj.2018.05.002

2007AA05Z417:国家高技术研究发展计划（863计划） 2009AA03Z232:国家高技术研究发展计划（863计划）

本校为其他机构

目的：将来源于Thermotoga maritima的酸性耐高温木聚糖酶基因Xyn B通过密码子优化整合到毕赤酵母GS115基因组上,构建高效表达木聚糖酶Xyn B的毕赤酵母工程菌并研究其酶学性质。方法：通过分子克隆手段构建目的基因重组质粒以及二拷贝重组质粒,并将重组质粒导入毕赤酵母进行发酵表达,用DNS法测定发酵液酶活,并对酵母工程菌分泌的木聚糖酶进行一系列的酶学性质研究。结果：木聚糖酶的最适p H和温度分别为6.0和60℃;在p H46之间,稳定性较好,相对比酶活均在80%以上;在60℃和70℃保温2 h残留酶活分别为80%和70%左右;Co（2＋）对木聚糖酶活性有较明显的促进作用,Fe（2＋）、K＋和Zn（2＋）对酶的活性...

Objective：To integrate the acid hyperthermophilic xylanase gene Xyn B from Thermotoga maritima on the Pichia pastoris GS115 genome through codon optimization.The Pichia pastoris engineering bacteria of the highly expressed xylanase Xyn B was constructed and its enzymatic property was studied.Methods：The target gene recombination plasmid and dual-copy recombination plasmid was constructed via molecular clone.The recombination plasmid was imported into the pichia pastoris to make fermentation expression.Use DNS method to measure the enzyme acti...