Halogenases in biocatalysis: advances in mechanism elucidation, directed evolution, and green manufacturing

WANG Mingpeng; CHEN Lei; ZHAO Yiran; ZHANG Yimin; ZHENG Qifan; LIU Xinyang; WANG Yixue; WANG Qinhong

doi:10.12211/2096-8280.2024-091

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Halogenases in biocatalysis: advances in mechanism elucidation, directed evolution, and green manufacturing

Invited Review | 更新时间：2025-09-05

- Halogenases in biocatalysis: advances in mechanism elucidation, directed evolution, and green manufacturing
  封面论文
- Synthetic Biology Journal Vol. 6, Issue 4, Pages: 728-763(2025)
- 作者机构：
  
  1.曲阜师范大学生命科学学院，山东曲阜 273100
  2.中国科学院天津工业生物技术研究所，低碳合成工程生物学全国重点实验室，天津 300308
  3.国家合成生物技术创新中心，天津 300308
- 作者简介：
- 基金信息：
- DOI：10.12211/2096-8280.2024-091
  CLC： Q55
- Received：03 December 2024，
  
  Revised：2025-04-14，
  
  Published：31 August 2025
- 稿件说明：
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王明鹏，陈蕾，赵一冉，张祎慜，郑琪帆，刘馨阳，王毅学，王钦宏. 卤化酶在生物催化中的应用：机制解析、定向进化和绿色制造的进展［J］. 合成生物学， 2025， 6（4）： 728-763

WANG Mingpeng， CHEN Lei， ZHAO Yiran， ZHANG Yimin， ZHENG Qifan， LIU Xinyang， WANG Yixue， WANG Qinhong. Halogenases in biocatalysis： advances in mechanism elucidation， directed evolution， and green manufacturing［J］. Synthetic Biology Journal， 2025， 6（4）： 728-763
王明鹏，陈蕾，赵一冉，张祎慜，郑琪帆，刘馨阳，王毅学，王钦宏. 卤化酶在生物催化中的应用：机制解析、定向进化和绿色制造的进展［J］. 合成生物学， 2025， 6（4）： 728-763 DOI： 10.12211/2096-8280.2024-091.

WANG Mingpeng， CHEN Lei， ZHAO Yiran， ZHANG Yimin， ZHENG Qifan， LIU Xinyang， WANG Yixue， WANG Qinhong. Halogenases in biocatalysis： advances in mechanism elucidation， directed evolution， and green manufacturing［J］. Synthetic Biology Journal， 2025， 6（4）： 728-763 DOI： 10.12211/2096-8280.2024-091.

摘要

有机卤化物在医药和农业领域应用广泛，但其化学合成过程污染严重。卤化酶是实现化合物卤素修饰及功能改善的重要工具。与化学合成不同，卤化酶可以实现有机结构特定位置的精准卤化，并且反应条件温和，避免了苛刻反应条件以及有毒试剂的使用，其催化反应的过程符合绿色化学要求。本文综述了卤化酶在生物合成中的最新研究进展及其在工业生产中的潜在应用。首先，简要回顾了卤化酶的分类、结构特征及催化机制的研究现状，并介绍了相关领域的最新进展；其次，总结了近年来通过基因组挖掘、定向进化和合成生物学等技术发掘新酶资源、优化酶催化性能及扩展酶应用范围的策略；然后探讨了工程化卤化酶在药物、农药及其他生物活性物质合成中的具体应用案例；最后讨论了在机器学习迅速发展的背景下，卤化酶研究的未来发展趋势。

Abstract

Organic halides

which serve as critical structural motifs in pharmaceuticals

agrochemicals

and advanced materials

are typically synthesized using energy-intensive processes that involve toxic reagents and generate hazardous waste. In contrast

halogenases-nature’s biocatalytic tools-catalyze regio- and stereoselective halogenation under environmentally benign conditions

offering a paradigm shift towards sustainable chemistry. This review systematically consolidates recent breakthroughs in halogenase research

emphasizing mechanistic insights

engineering innovations

and scalable industrial applications. Halogenases are mechanistically classified into three major families: flavin-dependent enzymes that mediate electrophilic halogenation through

transient hypohalous acid intermediates; non-heme iron/α-ketoglutarate-dependent oxygenases that drive radical-based halogenation pathways; and

-adenosylmethionine (SAM)-dependent enzymes that facilitate rare nucleophilic halogenation. Cutting-edge structural biology techniques

enhanced by computational simulations

have elucidated dynamic substrate-enzyme interactions and transient catalytic states

facilitating the rational design of halogenases with tailored reactivity.

The integration of bioinformatics tools with high-throughput screening platforms has co

ncurrently accelerated the discovery of novel halogenases from underexplored microbial niches

revealing unprecedented catalytic diversity. To bridge natural enzymatic capabilities with industrial demands

interdisciplinary strategies are being deployed: Directed evolution optimizes activity and stability under non-native conditions; computational protein design rebuilds substrate-binding pockets for non-canonical substrates; and synthetic biology frameworks reconstruct halogenation pathways in engineered microbial hosts. These efforts collectively expand the biocatalytic toolbox

enabling precise halogenation of complex scaffolds

including aromatic systems

aliphatic chains

and heterocycles. In industrial contexts

enzymatic halogenation is gaining traction for synthesizing high-value compounds

ranging from antibiotic derivatives and antitumor agents to crop protection molecules

while circumventing the traditional reliance on heavy metal catalysts

extreme temperatures

and halogenated solvents. Emerging applications further extend to the functionalization of biomaterials and fine chemicals

underscoring the versatility of halogenases. Future advancements will likely harness machine learning algorithms to decode sequence-activity landscapes and predict multi-enzyme cascades for tandem halogenation-functional group interconversions. Such developments are in line with global sustainability agendas

positioning halogenases as key biocatalysts in the transition towards circular chemical economies. This review highlights the convergence of enzymology

systems biology

and green chemistry in unlocking the full potential of halogenases

paving the way for next-generation biomanufacturing.

关键词

Keywords

references

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