From single-enzyme catalysis to multienzyme cascade： inspired from Professor Daniel I.C. Wang’s pioneer work in enzyme technology

doi:10.12211/2096-8280.2021-004

Abstract

Abstract:

As the founder and pioneer in the fields of bioengineering and biotechnology, Professor Daniel I. C. Wang at the Massachusetts Institute of Technology (MIT) had made tremendous contributions in many aspects in the up-, mid- and downstream of biochemical engineering for over 50 years. In the area of enzyme technology, he had impell several significant advances from the 1970s to 1990s, such as enzymatic digestion of fish protein, immobilization of methanol oxidase, cell-free multi-enzyme synthesis, and enzyme catalysis in organic medium, etc. From 2005 to 2015 through the Singapore-MIT Alliance Program, Prof. Wang had jointly supervised several PhD students with Prof. Li Zhi at the National University of Singapore, and achieved important contributions biocatalysis, including: 1) developed enzyme immobilization methods for fabrication of highly active and recyclable magnetic nano-biocatalysts; 2) explored P450 monooxygenase for asymmetric sulfur oxidation in aqueous phase-ionic liquid systems; 3) developed an NADPH regeneration system based on permeabilized whole cells; 4) successfully developed modular multi-enzyme cascade catalysis to synthesize high-value chiral compounds, which significantly expanded the realm of biocatalysis. Among them, multi-enzyme cascade catalysis has become a research hotspot in the field of enzyme technology, due to its several desirable features, including the possibility of retrosynthetic design of various synthetic routes, the facile one-pot synthesis of final products, the reduction of additional unit operations, the saving of inputs from manpower and materials, and the minimization of waste generation. In this account, we also review the latest progress of multi-enzyme cascades for the synthesis of chiral compounds (e.g., chiral amines, amino acids) and bulk chemicals (e.g., precursors for polymers), and discuss its future development directions. Last but not the least, we provide an outlook for integrating multi-enzyme cascades with synthetic biology, and thus assembling biochemical reactions together with quantitative analysis and engineering concepts, as advocated by Prof. Wang throughout his scientific career.

Key words: biochemical engineering, biocatalysis, enzyme technology, enzyme catalysis, enzyme cascades

摘要：

作为国际生物工程和生物技术领域的奠基人和业界泰斗，麻省理工学院王义翘教授几十年来的研究涵盖了生化工程的上、中、下游的众多方向。在酶技术领域，他在早期就完成了几项标志性的工作，例如鱼蛋白的酶消化、甲醇氧化酶的固定化、无细胞多酶合成、非水相酶催化等。从2005年开始，王教授在十年间通过新加坡-麻省理工联盟（Singapore-MIT Alliance）项目和新加坡国立大学李智教授联合指导了数名博士生，并在酶技术的几个子方向上取得了重要成果，包括：①开发了酶固定化的方法用于构建高活力可回收的磁性纳米生物催化剂；②探索了P450单加氧化酶在水相-离子液体体系中的不对称亚砜化反应；③开发了基于通透全细胞的辅酶NADPH再生系统；④成功开发了模块化的多酶级联催化合成高值手性化合物，显著拓展了多酶级联催化的复杂度。其中，多酶级联催化因其“一锅法”的合成特性，避免额外的单元操作，节省人力、物力的投入和废弃物的产生等特点，近年来已经成为酶技术领域的研究热点。在介绍王教授相关工作之余，本文还总结了多酶级联催化在合成手性化合物和大宗化学品方面的最新进展，讨论了其未来的发展方向，并就其进一步整合合成生物学以及王教授所践行的定量化和工程化的理念进行了展望。

关键词: 生化工程, 生物催化, 酶技术, 酶催化, 多酶级联反应

CLC Number:

Q814

Shuke WU, Yi ZHOU, Wen WANG, Wei ZHANG, Pengfei GAO, Zhi LI. From single-enzyme catalysis to multienzyme cascade： inspired from Professor Daniel I.C. Wang’s pioneer work in enzyme technology[J]. Synthetic Biology Journal, 2021, 2(4): 543-558.

吴淑可, 周颐, 王文, 张巍, 高鹏飞, 李智. 从单酶催化到多酶级联催化——从王义翘教授在酶技术领域的贡献说开去[J]. 合成生物学, 2021, 2(4): 543-558.

Figures/Tables 6

Fig. 1 Photos of Prof. Wang together with some of authors during the Singapore-MIT Alliance Program

Fig. 2 Four enzyme technology projects co-supervised by Prof. Wang through the Singapore-MIT Alliance Program

Fig. 3 Modular multi-enzyme cascade catalysis for the transformation of alkenes into three types of chiral molecules developed by Wu Shuke and co-supervised by Prof. Wang through the Singapore-MIT Alliance.EP—epoxidase; EH—epoxide hydrolase; ADH—alcohol dehydrogenase; ALDH—aldehyde dehydrogenase; ω-TA—ω-transaminase; AlaDH—alanine dehydrogenase; HO—hydroxy acid oxidase; α-TA—α-transaminase; CAT—catalase; GluDH—glutamate dehydrogenase

Fig. 4 Multi-enzyme cascades for converting secondary alcohols into chiral amines

Fig. 5 Multi-enzyme cascades for the production of chiral α-amino acids and β-amino acids

Fig. 6 Multi-enzyme cascades for the production of bulk chemicals

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[3]	Huibin WANG, Changli CHE, Song YOU. Recent advances of enzymatic synthesis of organohalogens catalyzed by Fe/αKG-dependent halogenases [J]. Synthetic Biology Journal, 2022, 3(3): 545-566.
[4]	Yujiao LOU, Jian XU, Qi WU. Progress of biocatalytic deuteration of inert carbon-hydrogen bonds [J]. Synthetic Biology Journal, 2022, 3(3): 530-544.
[5]	Lu YANG, Xudong QU. Application of imine reductase in the synthesis of chiral amines [J]. Synthetic Biology Journal, 2022, 3(3): 516-529.
[6]	Liangbin XIONG, Lu SONG, Yunqiu ZHAO, Kun LIU, Yongjun LIU, Fengqing WANG, Dongzhi WEI. Green biomanufacturing of steroids: from biotransformation to de novo synthesis by microorganisms [J]. Synthetic Biology Journal, 2021, 2(6): 942-963.
[7]	Faguang ZHANG, Ge QU, Zhoutong SUN, Jun′an MA. From chemical synthesis to biosynthesis: trends toward total synthesis of natural products [J]. Synthetic Biology Journal, 2021, 2(5): 674-696.
[8]	Heng TANG, Xin HAN, Shuping ZOU, Yuguo ZHENG. Application of multi-enzyme catalytic system in the synthesis of pharmaceutical chemicals [J]. Synthetic Biology Journal, 2021, 2(4): 559-576.
[9]	Yi-Heng ZHANG. Remembering Professor Daniel I.C. Wang’s contribution to biorefining and my perspective on the progress [J]. Synthetic Biology Journal, 2021, 2(4): 497-508.
[10]	Junting WANG, Xiaojia GUO, Qing LI, Li WAN, Zongbao ZHAO. Creation of non-natural cofactor-dependent methanol dehydrogenase [J]. Synthetic Biology Journal, 2021, 2(4): 651-661.
[11]	Zhiguo SU. Great impact of Professor Daniel I.C. Wang and BPEC on development of biochemical engineering [J]. Synthetic Biology Journal, 2021, 2(4): 470-481.