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一种耐热精氨酸酶的克隆、表达及结晶

The cloning, expression and crystallisation of a thermostable arginase.

作者信息

Bewley M C, Lott J S, Baker E N, Patchett M L

机构信息

Department of Biochemistry, Massey University, Palmerston North, New Zealand.

出版信息

FEBS Lett. 1996 May 20;386(2-3):215-8. doi: 10.1016/0014-5793(96)00459-0.

Abstract

The gene for the thermostable arginase from the thermophilic bacterium 'Bacillus caldovelox' has been cloned and sequenced. Expression of recombinant arginase at high levels has been achieved in E. coli using an inducible T7 RNA polymerase-based system. A facile purification procedure incorporating a heat-treatment step yielded 0.2 g of recombinant arginase per litre of induced culture. The kinetic properties of the purified recombinant protein are essentially identical to the native enzyme. The recombinant protein has been crystallised and one crystal form is isomorphous to crystals of the native protein.

摘要

嗜热细菌“嗜热栖热芽孢杆菌”中热稳定精氨酸酶的基因已被克隆并测序。利用基于诱导型T7 RNA聚合酶的系统,已在大肠杆菌中实现了重组精氨酸酶的高水平表达。一种包含热处理步骤的简便纯化方法,每升诱导培养物可产生0.2克重组精氨酸酶。纯化后的重组蛋白的动力学特性与天然酶基本相同。重组蛋白已被结晶,其中一种晶体形式与天然蛋白的晶体同晶型。

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