两个活性位点残留物的突变将酸氨基醇特异性酸酶C转化为葡萄糖酸酶
Jianwen Feng1, Kimberly Stieglitz, Mary F Roberts
1Merkert Chemistry Center, Boston College, Chestnut Hill, Massachusetts 02467, USA.
Journal of the American Chemical Society
|January 30, 2004
概括
两个细菌酶突变创造了一个新的酸酶. 这种酶有效地分解葡萄糖-6-酸盐,与其原始的酸氨基醇特异性酸酶C形式不同.
科学领域:
- 生物化学 生物化学
- 酶学 是一种酶学.
- 分子生物学分子生物学
背景情况:
- 细菌酸丁氨基醇特异性酸酶C (PI-PLC) 酶通常会化酸.
- 葡萄糖-6-酸酶酶是不同的,并且具有特定的活性位点要求.
研究的目的:
- 为了研究细菌PI-PLC突变的酶活性和基质特异性.
- 探索通过向突变改变酶功能的潜力.
主要方法:
- 用于引入R69D和K115E突变到细菌PI-PLC中.
- 酶活性测定使用伊诺西-1-酸盐 (I-1-P) 和葡萄糖-6-酸盐 (G-6-P) 作为基质进行.
- 生物信息分析和能源最小化被用来合理化观察到的变化.
主要成果:
- 双重突变R69D/K115E与I-1-P相比,对G-6-P具有显著更高的酸酶活性.
- 单个突变R69D和K115E没有赋予显著的酸酶活性.
- 细菌PI-PLC没有显示已知葡萄糖-6-酸酶的序列同质性.
结论:
- 特定的突变可以将细菌PI-PLC转化为具有葡萄糖-6-酸酶活性的酶.
- 改变的基质特异性表明酶的催化机制和活性部位发生了显著的变化.
- 理性酶工程可以创建具有所需基质特性的新生物催化剂.
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