合成生物学 ›› 2022, Vol. 3 ›› Issue (3): 545-566.DOI: 10.12211/2096-8280.2021-102

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Fe/α-酮戊二酸依赖型卤化酶在绿色卤化反应中的研究进展

王汇滨, 车昌丽, 游松   

  1. 沈阳药科大学生命科学与生物制药学院,辽宁 沈阳 110016
  • 收稿日期:2021-11-10 修回日期:2021-11-28 出版日期:2022-06-30 发布日期:2022-07-13
  • 通讯作者: 游松
  • 作者简介:王汇滨(1998—),男,硕士研究生。研究方向为天然产物生物合成机制。 E-mail:huibin0918@163.com|游松(1963—),男,博士,教授,博士生导师。研究方向为生物催化与生物转化。 E-mail:yousong206@aliyun.com
  • 基金资助:
    国家重点研发计划“绿色生物制造”项目(2021YFC2102004)

Recent advances of enzymatic synthesis of organohalogens catalyzed by Fe/αKG-dependent halogenases

Huibin WANG, Changli CHE, Song YOU   

  1. School of Life Sciences and Biopharmaceutical Sciences,Shenyang Pharmaceutical University,Shenyang 110016,Liaoning,China
  • Received:2021-11-10 Revised:2021-11-28 Online:2022-06-30 Published:2022-07-13
  • Contact: Song YOU

摘要:

将卤素原子引入药物、农药等小分子中可以有效提升其生物活性,并且碳卤键可用于有机合成后期功能化反应。Fe/α-酮戊二酸(α-ketoglutaric acid,αKG)依赖型卤化酶可以高立体选择性和区域选择性催化卤素原子引入到未经活化的sp3杂化碳中心。大自然是最伟大的化学家,本文遵循从学习自然到改造自然的逻辑顺序,首先介绍Fe/αKG依赖型卤化酶的发现历程,之后分别总结天然产物生物合成途径中的载体依赖型和独立型Fe/αKG依赖型卤化酶,进一步分析Fe/αKG依赖型卤化酶的结构特征以及基于蛋白质工程等方法改造扩展其底物谱并拓展新的反应类型,最后从新酶的挖掘与表征、酶催化活性的提升、酶区域选择性的控制、酶反应类型的拓展、人工生物合成途径的创建等5方面进行展望,丰富对Fe/αKG依赖型卤化酶的催化机制、底物范围和反应杂泛性的相关认识,为后续合成生物学的应用研究奠定酶学基础。

关键词: 卤化反应, Fe/α-酮戊二酸依赖型酶, 生物合成, 生物催化, 蛋白质工程

Abstract:

Synthetic biology is being developed as a bio-design and production platform through which more and more impressive products are synthesized. Enzymes, the cornerstone of synthetic biology, can catalyze diverse reactions, such as the concise synthesis of chiral alcohols and chiral amines. The biocatalytic halogenation reaction has gained research interest in recent years due to its mild reaction conditions and high selectivity. The insertion of halogen atoms into reaction agents such as drugs and agrochemicals can effectively enhance their biological activities, and the carbon-halogen bonds can be employed as a key building block for the late-stage functionalization reactions. Fe/α-ketoglutaric acid (αKG)-dependent halogenases can catalyze the insertion of halogen atoms into unactivated sp3-hybridized carbon centers with high stereoselectivity and regioselectivity based on the radical reaction mechanism. This review follows the logical sequence of learning from the nature, a powerful chemist. First, it introduces the discovery of Fe/αKG-dependent halogenases, and then summarizes the carrier protein-dependent and non-dependent Fe/αKG-dependent halogenases involved in the biosynthesis of natural products. Furthermore, it analyzes the structural characteristics of Fe/αKG-dependent halogenases, and address their substrate spectrum and novel reaction types expanded and created through protein engineering and other methods. Finally, the discovery and characterization of new enzymes, the improvement of their catalytic activities, the rational control of their regioselectivities, the expansion of tehir reaction types, and the creation of the artificial biosynthetic pathways are highlighted, expecting to enrich the knowledge on the catalytic mechanism, substrate scope and reaction promiscuity of Fe/αKG-dependent halogenases. Learning the cryptic chemistry mechanism hidden with natural products remains one of the hot topics in natural product chemistry. Beyond that, rational redesign and evolution of “supra-natural” product pathways will be emphasized for the purpose of discovering and developing novel lead compounds from an interdisciplinary point of view under the guidance of the synthetic biology. The review would lay a enzymology foundation for the related and subsequent research in synthetic biology.

Key words: halogenation reaction, Fe/αKG-dependent enzymes, biosynthesis, biocatalysis, protein engineering

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