Position: Home > Articles > Expression Purification and Characterization of a Thermostable Bifunctional β-xylosidase from Thermoga maritima
Hubei Agricultural Sciences
2016,55
(7)
1843-1847
极耐热双功能酶的表达纯化及性质研究
作 者：
王锐丽;王玉;薛业敏
单 位：
信阳农林学院生物与制药工程学院;南京师范大学金陵女子学院;焦作师范高等专科学院理工学院
关键词：
极耐热;双功能酶;海栖热袍菌(Thermoga maritima)β-木糖苷酶(Xyl);纯化;性质
摘 要：
将含海栖热袍菌(Thermoga maritima)β-木糖苷酶(Xyl)基因的重组质粒p ET-28a-xyl导入大肠杆菌BL21-Codon Plus(DE3)-RIL中,经诱导实现高效表达,经热处理和Ni2+亲和层析纯化达到电泳均一,并对纯化的重组酶Xyl的酶学性质进行研究。结果表明,重组Xyl同时具有木糖苷酶和阿拉伯糖酶活性,即为双功能酶。以对硝基苯酚-ɑ-阿拉伯呋喃糖苷(p NPAF)作为底物,重组酶Xyl的最适作用温度和最适p H分别为80~90℃和5.8,90℃的半衰期为1.0 h,在p H 7.0~7.8区间重组酶保持较高的稳定性。金属离子Mn2+对重组Xyl的双活性有促进作用,而Cu2+和SDS对酶的抑制作用最明显。重组Xyl能将低聚木糖中的木二糖和木三糖彻底分解为木糖。
译 名：
Expression Purification and Characterization of a Thermostable Bifunctional β-xylosidase from Thermoga maritima
作 者：
WANG Rui-li;WANG Yu;XUE Ye-min;Department of Biotechnology and Pharmaceutical Engineering ,Xinyang College of Agriculture and Forestry;Institute of Science Technology, Jiaozuo Teachers College;Jinling College,Nanjing Normal University;
关键词：
thermostable;;bifunctional enzyme;;β-xylosidase(Xyl) from Thermoga maritima;;purification;;characterization
摘 要：
The recombinant bifunctional β-xylosidase from Thermoga maritima was over-expressed from culture borth of gene engneering strain Escherichia coli BL21-Codon Plus(DE3)-RIL and purified through following steps:heat precipetation and nickel affinity chromatography. The purified recombinant β-xylosidase showed a single band on SDS polyacrylamide gel electrophoresis. With p-Nitrophenyl ɑ-L-arabinofuranoside(p NPAF) as substrate, the optimum activity was found at 80~90 ℃ and p H 5.8. The purified enzyme had a half-life of 1.0 h at 90 ℃,and retained over 80% of its activity after holding a p H ranging from 7.0 to 7.8. Metal ion Mn2+activated the enzyme,Cu2+and SDS showed various degrees of inhibitory effect on the recombinant Xyl. Xylobiose and xylotriose of Xylo-Oligosaccharide were hydrolized completely to xylose by the recombinant Xyl.
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