Computational thermostabilization of an enzyme
Computational thermostabilization of an enzyme
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DOI：
10.1126/science.1107387
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发表时间：
2005-05-06
期刊：
影响因子：
56.9
通讯作者：
Stoddard, BL
中科院分区：
文献类型：
--
作者：
Korkegian, A;Black, ME;Stoddard, BL
热稳定酶，同时保持其活性的工业或生物医学应用可能是困难的与传统的选择方法。我们描述了一种快速的计算方法，该方法确定了模型酶中的三个突变，这些突变使表观熔解温度T（m）增加10 ℃，在50 ℃时半衰期增加30倍，而催化效率没有降低。突变的影响是协同的，增加超过其单独影响的总和。重新设计的酶在需要其活性的条件下诱导增加的温度依赖性细菌生长速率，从而将分子和代谢工程结合起来。
Thermostabilizing an enzyme while maintaining its activity for industrial or biomedical applications can be difficult with traditional selection methods. We describe a rapid computational approach that identified three mutations within a model enzyme that produced a 10 degrees C increase in apparent melting temperature T(m) and a 30-fold increase in half-life at 50 degrees C, with no reduction in catalytic efficiency. The effects of the mutations were synergistic, giving an increase in excess of the sum of their individual effects. The redesigned enzyme induced an increased, temperature-dependent bacterial growth rate under conditions that required its activity, thereby coupling molecular and metabolic engineering.