胺转氨酶的蛋白质工程

Qinglong Meng, Carlos Ramírez-Palacios, Hein J. Wijma, D. Janssen
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摘要

蛋白质工程是一种强大且广泛应用的工具,用于调整酶的特性以满足特定应用的要求。一组有吸引力的生物催化剂是PLP依赖性胺转氨酶,其能够通过不对称催化将前手性酮转化为相应的手性胺。这些酶通常表现出高的对映选择性,并接受各种胺供体。这些胺转氨酶的实际应用可能受到酶不稳定性和其底物范围有限的阻碍。已经探索了各种提高胺转氨酶稳健性和重定向其底物特异性的策略,包括定向进化、合理设计和计算支持的工程。本文综述了所使用的方法和获得的结果,表明不同的策略可以以互补的方式使用,并可以扩大胺转氨酶在生物催化中的适用性。
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Protein engineering of amine transaminases
Protein engineering is a powerful and widely applied tool for tailoring enzyme properties to meet application-specific requirements. An attractive group of biocatalysts are PLP-dependent amine transaminases which are capable of converting prochiral ketones to the corresponding chiral amines by asymmetric catalysis. The enzymes often display high enantioselectivity and accept various amine donors. Practical applications of these amine transaminases can be hampered by enzyme instability and by their limited substrate scope. Various strategies to improve robustness of amine transaminases and to redirect their substrate specificity have been explored, including directed evolution, rational design and computation-supported engineering. The approaches used and results obtained are reviewed in this paper, showing that different strategies can be used in a complementary manner and can expand the applicability of amine transaminases in biocatalysis.
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