Abstract

Because of their function of catalyzing the rearrangement of the carbon chains, thiolases have attracted increasing attentions over the past decades. The 3-ketoacyl-CoA thiolase (KAT) is a member of the thiolase, which is capable of catalyzing the Claisen condensation reaction between the two acyl-CoAs, thereby achieving carbon chain elongation. In this way, diverse value-added compounds might be synthesized starting from simple small CoA thioesters. However, most KATs are hampered by low stability and poor substrate specificity, which has hindered the development of large-scale biosynthesis. In this review, the common characteristics in the three-dimensional structure of KATs from different sources are summarized. Moreover, structure-guided rational engineering is discussed as a strategy for enhancing the performance of KATs. Finally, we reviewed the metabolic engineering applications of KATs for producing various energy-storage molecules, such as n-butanol, fatty acids, dicarboxylic acids, and polyhydroxyalkanoates.

Key points

• Summarize the structural characteristics and catalyzation mechanisms of KATs.

• Review on the rational engineering to enhance the performance of KATs.

• Discuss the applications of KATs for producing energy-storage molecules.

中文翻译：

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3-酮酰基-CoA硫解酶：一种工程化酶，用于化合物的碳链延伸。

摘要

由于它们具有催化碳链重排的功能，因此在过去的几十年中，硫磺酶引起了越来越多的关注。3-酮酰基-CoA硫解酶（KAT）是硫解酶的成员，它能够催化两个酰基-CoA之间的Claisen缩合反应，从而实现碳链延长。以这种方式，可以从简单的小CoA硫酯开始合成各种增值化合物。但是，大多数KAT受稳定性低和底物特异性差的困扰，这阻碍了大规模生物合成的发展。在这篇综述中，总结了来自不同来源的KAT的三维结构的共同特征。此外，讨论了结构引导的合理工程作为提高KAT性能的策略。最后，正丁醇，脂肪酸，二羧酸和聚羟基链烷酸酯。

关键点

•总结KAT的结构特征和催化机理。

•审查合理的工程设计以提高KAT的性能。

•讨论KAT在生产储能分子中的应用。