Computational redesign of enzymes for regio- and enantioselective hydroamination
Computational redesign of enzymes for regio- and enantioselective hydroamination
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用于区域和对映选择性氢胺化的酶的计算重新设计
DOI：
10.1038/s41589-018-0053-0
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发表时间：
2018-07-01
影响因子：
14.8
通讯作者：
Wu, Bian
中科院分区：
文献类型：
--
作者：
Li, Ruifeng;Wijma, Hein J.;Wu, Bian
创新生物催化工艺的引入为绿色化学的应用提供了广阔的前景。然而，由于催化性能有限，从自然界生物多样性中获得的酶往往需要通过耗时的实验室迭代进化来改进其所需的功能。在这里，我们描述了使用基于结构的计算酶设计将芽孢杆菌sp. YM55-1天冬氨酸酶（一种具有非常狭窄底物范围的酶）转化为一组互补的氢胺化生物催化剂。重新设计的酶催化氨对取代丙烯酸酯的不对称加成，提供对映纯脂肪、极性和芳香β-氨基酸，这些氨基酸是合成药物和生物活性化合物的宝贵基石。无需进一步优化实验室进化，重新设计的酶具有高达300 g/L浓度的底物耐受性，转化率高达99%，β-区域选择性>99%，产物对映体过量>99%。结果突出了计算设计的使用，以迅速适应酶的工业上可行的反应。
Introduction of innovative biocatalytic processes offers great promise for applications in green chemistry. However, owing to limited catalytic performance, the enzymes harvested from nature's biodiversity often need to be improved for their desired functions by time-consuming iterative rounds of laboratory evolution. Here we describe the use of structure-based computational enzyme design to convert Bacillus sp. YM55-1 aspartase, an enzyme with a very narrow substrate scope, to a set of complementary hydroamination biocatalysts. The redesigned enzymes catalyze asymmetric addition of ammonia to substituted acrylates, affording enantiopure aliphatic, polar and aromatic β-amino acids that are valuable building blocks for the synthesis of pharmaceuticals and bioactive compounds. Without a requirement for further optimization by laboratory evolution, the redesigned enzymes exhibit substrate tolerance up to a concentration of 300 g/L, conversion up to 99%, β-regioselectivity >99% and product enantiomeric excess >99%. The results highlight the use of computational design to rapidly adapt an enzyme to industrially viable reactions.