中南民族大学学报（自然科学版）Issue(3)：27-31,5.
组氨酸标签木聚糖酶的构建表达和酶特性鉴定
Construction and Characterization of a Thermostable Xylanase Mutant with 6 ×His-tag
摘要
Abstract
A thermostable xylanase mutant DSB from Thermomyces lanuginosus was expressed in Escherichia coli with a 6 × His-tag in C terminal.DSB with terminator changed to Glu was amplified by PCR and cloned into plasmid pET-22b(+), then transformed into E.coli BL21 ( DE3 ) .The cultivation was purified by ammonium sulfate precipitation and Ni-nitrilotriacetate affinity chromatography .The results from SDS-PAGE showed that xylanase DSB-His was electrophoretically homogeneous , with a molecular weight of approximately 23 kDa.The xylanase DSB showed optimal activity at 75℃and pH 6.5, was highly stable at pH 5.0~10.0, and retained more than 80%activity after incubation at 70℃for 30 min.关键词
耐热木聚糖酶/多聚组氨酸标签/定点突变/镍亲和层析/酶学特性Key words
thermostable xylanase/6 ×His-tag/site-directed mutagenesis/Ni-nitrilotriacetate affinity chromatography/enzyme characterization分类
医药卫生引用本文复制引用
熊海容,陈丰,王亚伟,张巍..组氨酸标签木聚糖酶的构建表达和酶特性鉴定[J].中南民族大学学报（自然科学版）,2014,(3):27-31,5.基金项目
武汉市科技攻关项目（SZY13005），中央高校基本科研业务费专项基金资助项目 （）
