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[嗜热栖热菌MSB8中极端耐热木聚糖酶B在大肠杆菌和毕赤酵母中的高效表达]

[High-level expression of an extreme-thermostable xylanase B from Thermotoga maritima MSB8 in Escherichia coli and Pichia pastoris].

作者信息

Yang Meng-hua, Li Ying, Guan Guo-hua, Jiang Zheng-qiang

机构信息

College of Biological Sciences, China Agricultural University, Beijing 100094, China.

出版信息

Wei Sheng Wu Xue Bao. 2005 Apr;45(2):236-40.

Abstract

The family 10 xylanase (XynB) from Thermotoga maritima MSB8 is extremely thermostable and great potential in the applications of various fields of industry. The gene mxynB(64) was amplified by the method of PCR with the template of the genomic DNA of Thermotoga maritima MSB8, and cloned into the expression vectors of Escherichia coli and Pichia pastoris respectively. Xylanase B(40kD) was successfully expressed by the two heterologous protein expression systems with high-level production. The recombinant protein of XynB expressed in Pichia pastoris showed extreme thermostability and pH stability, which was optimally active at 90 degrees C and quite stable over the pH range of pH 5.0-10.8 at 70 degrees C. After incubation of the enzyme at 100 degrees C for 30 min, XynB retained 70% higher residual activity. The recombinant XynB expressed in Pichia pastoris is of great use in a variety of industrial and agricultural applications.

摘要

来自嗜热栖热菌MSB8的家族10木聚糖酶(XynB)具有极高的热稳定性,在各个工业领域的应用中具有巨大潜力。采用PCR方法以嗜热栖热菌MSB8的基因组DNA为模板扩增基因mxynB(64),并分别克隆到大肠杆菌和巴斯德毕赤酵母的表达载体中。木聚糖酶B(40kD)通过这两种异源蛋白表达系统成功实现了高水平表达。在巴斯德毕赤酵母中表达的重组XynB蛋白表现出极高的热稳定性和pH稳定性,在90℃时活性最佳,在70℃下pH 5.0 - 10.8范围内相当稳定。该酶在100℃孵育30分钟后,XynB的残留活性仍高出70%。在巴斯德毕赤酵母中表达的重组XynB在各种工农业应用中具有重要用途。

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