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Engineered enzymes for the synthesis of pharmaceuticals and other high-value products

Nature Synthesis volume 3, pages 19–32 (2024)Cite this article

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Abstract

Catalysis has a central role in organic synthetic methodology, especially in stereoselective reactions. In many reactions, enantioselectivity is made possible through the use of chiral transition metal catalysts. For decades, enzymes had a minor role, but this changed with the advent of directed evolution in the 1990s. The experimental implementation of evolving stereoselective mutants convinced at least some chemists of the potential of enzymes in catalytic processes. Subsequently, efficient mutagenesis methods emerged, including the combinatorial active-site saturation test, iterative saturation mutagenesis and rational enzyme design, such as focused rational iterative site-specific mutagenesis, that led to the widely held belief that essentially any desired transformation is possible. In this Review, we introduce these mutagenesis strategies and then discuss individual cases of enzyme-catalysed syntheses of chiral therapeutic drugs and other value-added products as well as the associated reaction mechanisms. Also, the type of value-added product, the enzyme used and preferential mutagenesis method are discussed.

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Fig. 1: Summary of the protein engineering of enzymes that catalyse organic reactions.
Fig. 2: Regio- and stereoselective oxidation of C–H bonds catalysed by CYP monooxygenases or peroxygenases.
Fig. 3: Stereoselective ADHs reduce prochiral ketones.
Fig. 4: Mutants of BVMOs as catalysts in regioselective BV oxidations and stereoselective sulfoxidation reactions.
Fig. 5: Reprogramming the epoxide hydrolase LEH beyond the normal hydrolytic reaction mode.
Fig. 6: Lipase-catalysed hydrolysis of a racemic acid ethyl ester.
Fig. 7: Selective glycosylation leading to valorized products.
Fig. 8: Biocatalytic amine synthesis using transaminases.
Fig. 9: Synthesis of natural and unnatural terpenes by terpene synthases.

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Acknowledgements

This work was supported by the National Key Research and Development Program of China (grant no. 2019YFA0905100), the Tianjin Synthetic Biotechnology Innovation Capacity Improvement Project (grant no. TSBICIP-CXRC-009), the National Natural Science Foundation of China (grant nos. 32171268 and 32171474) and the Natural Science Foundation Applying System of Tianjin (grant no. 19JCQNJC09100). M.T.R. thanks the Max-Planck-Society for support. G.Q. thanks the Youth Innovation Promotion Association, CAS (2021175) for financial support.

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Authors and Affiliations

  1. Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China

    Manfred T. Reetz, Ge Qu & Zhoutong Sun

  2. Biocatalysis Section, Max-Planck-Institut für Kohlenforschung, Mülheim an der Ruhr, Germany

    Manfred T. Reetz

  3. Key Laboratory of Engineering Biology for Low-Carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China

    Ge Qu & Zhoutong Sun

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  1. Manfred T. Reetz
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M.T.R., Q.G. and Z.S. contributed to discussions and wrote the paper.

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Correspondence to Manfred T. Reetz, Ge Qu or Zhoutong Sun.

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Reetz, M.T., Qu, G. & Sun, Z. Engineered enzymes for the synthesis of pharmaceuticals and other high-value products. Nat. Synth 3, 19–32 (2024). https://doi.org/10.1038/s44160-023-00417-0

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