Insight into the broadened substrate scope of nitrile hydratase by static and dynamic structure analysis.
Insight into the broadened substrate scope of nitrile hydratase by static and dynamic structure analysis.
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通过静态和动态结构分析深入了解腈水合酶的拓宽底物范围
DOI：
10.1039/d2sc02319a
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发表时间：
2022-07-20
期刊：
影响因子：
8.4
通讯作者：
中科院分区：
文献类型：
--
作者：
狭窄的底物范围限制了酶的广泛工业应用。在这里，我们成功地扩大了底物范围的腈水合酶（NHase）通过突变的两个隧道入口残基的基础上合理的隧道计算。获得了两种变体，具有增加的比活性，特别是对庞大的底物。晶体结构分析表明，突变导致隧道入口扩大，这可能有利于衬底进入。更重要的是，分子动力学模拟表明，突变引入的反相关运动的区域周围的底物隧道和活性位点，这将促进底物的访问过程中的动态催化。此外，在其他NHases上相应的隧道入口残基上的突变也增强了它们对大体积底物的活性。这些结果不仅揭示了位于酶表面的残基是影响酶催化性能的关键因素，而且为深入了解酶底物范围拓宽提供了动力学证据。
The narrow substrate scope limits the wide industrial application of enzymes. Here, we successfully broadened the substrate scope of a nitrile hydratase (NHase) through mutation of two tunnel entrance residues based on rational tunnel calculation. Two variants, with increased specific activity, especially toward bulky substrates, were obtained. Crystal structure analysis revealed that the mutations led to the expansion of the tunnel entrance, which might be conducive to substrate entry. More importantly, molecular dynamics simulations illustrated that the mutations introduced anti-correlated movements to the regions around the substrate tunnel and the active site, which would promote substrate access during the dynamic process of catalysis. Additionally, mutations on the corresponding tunnel entrance residues on other NHases also enhanced their activity toward bulky substrates. These results not only revealed that residues located at the enzyme surface were a key factor in enzyme catalytic performance, but also provided dynamic evidence for insight into enzyme substrate scope broadening.
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影响因子：
2.9
作者：
Nelp MT;Astashkin AV;Breci LA;McCarty RM;Bandarian V
通讯作者：
Bandarian V
DOI：
10.3390/bioengineering8110188
发表时间：
2021-11-19
期刊：
Bioengineering (Basel, Switzerland)
影响因子：
--
作者：
Ittisoponpisan S;Jeerapan I
通讯作者：
Jeerapan I
影响因子：
14.9
作者：
HEMSLEY, A;ARNHEIM, N;GALAS, DJ
通讯作者：
GALAS, DJ
影响因子：
5
作者：
Hongsthong, A;Subudhi, S;Cheevadhanarak, S
通讯作者：
Cheevadhanarak, S
影响因子：
4.6
作者：
Meng, Shuaiqi;An, Ruipeng;Liu, Luo
通讯作者：
Liu, Luo