Enzymatic reactions and microorganisms producing the various isomers of hydroxyproline
Enzymatic reactions and microorganisms producing the various isomers of hydroxyproline
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DOI：
10.1007/s00253-020-10603-1
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发表时间：
2020-04
影响因子：
5
通讯作者：
Ryotaro Hara;K. Kino
中科院分区：
文献类型：
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作者：
Ryotaro Hara;K. Kino
羟脯氨酸是一种重要的工业化合物，在医药、营养和化妆品工业中有广泛的应用。在8种可能的异构体中，反-4-羟基-L-脯氨酸被认为是最丰富的异构体(C-3或C-4上的羟基，顺式或反式，以及l-序式)。然而，很少有人注意到稀有的异构体，这可能是因为它们的可获得性有限。这篇简短的综述概述了微生物和酶工艺的最新进展，以开发各种羟基脯氨酸的实用生产策略。在这里，我们介绍了三种筛选策略，即基于活性、基于序列和基于代谢物的方法，允许识别具有不同产品特异性的不同脯氨酸羟基化酶。所有自然产生的羟脯氨酸异构体都可以通过使用适当的羟基酶以高度区域和立体选择性的方式产生。此外，相关羟基酶的晶体结构提供了许多关于它们的功能作用的信息。由于作用于游离脯氨酸的羟基酶属于依赖于2-羟基戊二酸的双加氧酶超家族，所以与羟基酶偶联的大肠杆菌的细胞代谢是合成2-氧基戊二酸的重要来源，而2-氧戊二酸作为1-脯氨酸羟化的辅助底物是不可缺少的。此外，微生物羟脯氨酸2-异构酶可以作为一种高度适应性的工具来将1-羟基脯氨酸转化为-羟脯氨酸。关键点·脯氨酸羟基酶是选择性-脯氨酸羟基化的有力工具。·工程大肠杆菌是生产羟脯氨酸的一个强大的平台。·羟基脯氨酸异构酶将1-羟基脯氨酸转化为-羟脯氨酸。
Hydroxyproline is an industrially important compound with applications in the pharmaceutical, nutrition, and cosmetic industries.trans-4-Hydroxy-l-proline is recognized as the most abundant of the eight possible isomers (hydroxy group at C-3 or C-4,cis- ortrans-configuration, andl- ord-form). However, little attention has been paid to the rare isomers, probably due to their limited availability. This mini-review provides an overview of recent advances in microbial and enzymatic processes to develop practical production strategies for various hydroxyprolines. Here, we introduce three screening strategies, namely, activity-, sequence-, and metabolite-based approaches, allowing identification of diverse proline-hydroxylating enzymes with different product specificities. All naturally occurring hydroxyproline isomers can be produced by using suitable hydroxylases in a highly regio- and stereo-selective manner. Furthermore, crystal structures of relevant hydroxylases provide much insight into their functional roles. Since hydroxylases acting on freel-proline belong to the 2-oxoglutarate-dependent dioxygenase superfamily, cellular metabolism ofEscherichia colicoupled with a hydroxylase is a valuable source of 2-oxoglutarate, which is indispensable as a co-substrate inl-proline hydroxylation. Further, microbial hydroxyproline 2-epimerase may serve as a highly adaptable tool to convertl-hydroxyproline intod-hydroxyproline.Key points• Proline hydroxylases serve as powerful tools for selectivel-proline hydroxylation.• Engineered Escherichia coli are a robust platform for hydroxyproline production.• Hydroxyproline epimerase convertsl-hydroxyproline intod-hydroxyproline.