Application of multi-enzyme catalytic system in the synthesis of pharmaceutical chemicals

doi:10.12211/2096-8280.2021-028

Abstract

Abstract:

The multi-enzyme catalysis system has become a hotspot in the field of bio-catalysis in recent years. Due to the catalytic process's controllability and the ease of downstream separation, more and more intracellular metabolic pathways are being developed and applied to produce fine chemicals in vitro. With the gradual maturity of multi-enzyme catalytic system construction technology, it will bring a broad application prospect for the manufacture of chemical products and pharmaceutical products in the future. In this regard, we review recently published examples of multi-enzyme catalysis, compare relevant design principles. Thermodynamics and kinetics of the reaction process and synthesis path analysis are used to design a bio-catalysis path. The critical enzymes are then mined in the pathway. By combining various assembly strategies, enzymes with different functions are cascaded into a whole structure and function unit to form a "substrate channel." Then intermediate loss and side reaction was reduced during the efficient bioconversion process from simple substrates to complex products. This method has the advantages of improving reaction efficiency, high regional selectivity and stereoselectivity, and also reducing environmental impact. Thus, biocatalysts are now widely used to produce valuable commercial chemicals, pharmaceuticals, and fuels. The target products could be obtained by the bio-catalysis process in vivo or in vitro. High-value products that are difficult to synthesize, or in an extremely complex synthesis pathway, or a very costly synthesis process by non-biological methods can now be produced in microbial hosts. To match the corresponding products, various genes in the host cell need to be optimized and fine-tuned. However, in vitro bio-catalysis can avoid these limitations. By adding a variety of enzymes to the reaction system, the product can be obtained after completing the catalysis at one time. Thus, industrial bio-fabrication has emerged as an attractive and economically viable alternative to conventional large-scale chemical synthesis. The multi-enzyme catalysis system in the synthesis of pharmaceutical chemicals (such as antibiotics, drugs for anti-cancer, cardiovascular disease treatment, liver disease treatment and psychiatric treatment) and various active ingredients (such as D-gluconic acid, terpenoids, 5-aminolevulinic acid) were discussed. The problems existing in the multi-enzyme catalytic system and the possible solutions are also summarized.

Key words: biocatalysis, multi-enzyme system, retrosynthetic analysis, cell-free, substrate channel, immobilization

摘要：

多酶催化体系成为近年来生物催化领域的研究热点。由于过程可控性及下游易分离的特性，越来越多的体外多酶催化体系已成功构建，部分体系还可耦合化学催化步骤，应用于精细化学品的合成。随着多酶催化体系构建技术的逐步成熟，将会给未来化工与医药类产品的生物制造带来广阔的应用前景。本文介绍了多酶催化体系的相关设计原则，通过对反应过程和合成路径进行热力学和动力学分析，设计高价值产物的生物合成路径，挖掘路径中的关键酶，结合各类新型组装策略将功能各异的酶级联组装成一个结构和功能整体，形成“底物通道”，减少中间体损失和降低副反应，实现从简单底物向复杂产物的高效生物转化。系统分析了多酶催化体系在医药化学品（如抗生素、抗癌药物、心血管疾病治疗药物、肝病治疗药物和精神疾病治疗药物及各类活性成分如D-葡萄糖二酸、萜类化合物和5-氨基乙酰丙酸）合成中的应用实例，并总结多酶催化体系仍存在的问题及可能的解决方法。

关键词: 生物催化, 多酶催化体系, 逆合成分析, 无细胞, 底物通道, 固定化

CLC Number:

Q814.9

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.

汤恒, 韩鑫, 邹树平, 郑裕国. 多酶催化体系在医药化学品合成中的应用[J]. 合成生物学, 2021, 2(4): 559-576.

Figures/Tables 16

Fig. 1 Substrate channel in a two-step metabolic pathway［7］

Fig. 2 Assembly technology of multi-enzyme catalytic system［32］

Fig. 3 Comparison of two deacetyl-7-aminocephalosporanic acid synthetic routes［42］

Fig. 4 Synthesis of D-2-aminobutyric acid catalyzed by multi-enzyme［50］

Fig. 5 Multienzyme catalyzed synthesis pathway of enterococcin in vitro［54］

Fig. 6 Synthesis of L-tyrosine derivatives by one-pot multi-enzyme cascade catalysis［66］

Fig. 7 One-pot multi-enzyme cascade production route of deoxyguanosine-2'-monophosphate［70］

Fig. 8 Atorvastatin side chain precursor production route［75］

Fig. 9 Key intermediate synthesis pathway of ursodeoxycholic acid［82］

Fig. 10 (R)-1-Phenyl-1,2-ethanediol were produced by in vitro multi-enzyme catalytic de-racemization system［87］

Fig. 11 Production of D-gluconic acid by multi-enzyme catalysis from glucose［90］

Fig. 12 Production of monoterpenes from glucose by multi-enzyme catalysis［102］

Fig. 13 Synthesis of 5-aminolevulinic acid catalyzed by cell-free multi-enzyme［107］

Fig. 14 Schematic representation of multi-enzyme detection using Cy5-LEVLVP-QD nanoprobe［109］

Fig. 15 Illustration of preparation of SWCNT-(PDDA/ALP)n-PDDA-PSS-Ab2 bioconjugates［110］

Fig. 16 General scheme of CRISPR/Cas-directed programmable assembly of multienzyme complexes on a DNA scaffold［111］

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