| 摘要: |
| 克隆并在大肠杆菌中表达的海栖热袍菌的 xyn B基因 ,其表达产物木聚糖酶 B的 C 末端带有 6×His标签 ,研究了这种基因重组酶的提纯方法。通过对粗酶提取液的热变性处理 ,Ni NTA亲和柱层析和离子柱层析 ,最终得到了电泳纯的木聚糖酶 B,提纯倍数 4 4.4 ,得率 11。SDS PAGE法测定木聚糖酶 B的相对分子质量为 4 2 ku,与理论推算值 4 2 333u相吻合 |
| 关键词: 海栖热袍菌 耐热木聚糖酶B 热处理 离子交换柱层析 提纯 |
| DOI： |
| 修订日期：2000-06-08 |
| 基金项目:联合国大学基金资助项目 |
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| Purification of an Extrem-thermostable Xylanase B From Thermotoga maritima |
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| Abstract: |
| The xyn B gene of Thermotoga maritima MSB8 was cloned and expressed in E.coli , the C terminal His tag was introduced, and purification methods of the recombinant enzyme were studied. The xylanase B was purified to homogeneity by heat treatment, affinity chromatography and ion exchange chromatography. The purified enzyme showed as a single protein band on SDS PAGE with a molecular weight of 42 ku, this is in good agreement with the molecular mass of enzyme deduced from the DNA sequence, 42 333 u. |
| Key words: Thermotoga maritima,thermostable xylanase B,heat treatment,Ni NTA affinity chromatography,ion exchange chromatography |
