Complete integration of carbene-transfer chemistry into biosynthesis
Complete integration of carbene-transfer chemistry into biosynthesis
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DOI：
10.1038/s41586-023-06027-2
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发表时间：
2023-05
期刊：
影响因子：
64.8
通讯作者：
Jing Huang;A. Quest;Pablo Cruz-Morales;Kai Deng;J. H. Pereira;Devon Van Cura;Ramu Kakumanu;E. Baidoo;Q. Dan;Yan Chen;C. Petzold;T. Northen;Paul D. Adams;D. Clark;E. Balskus;J. Hartwig;A. Mukhopadhyay;J. Keasling
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文献类型：
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作者：
Jing Huang;A. Quest;Pablo Cruz-Morales;Kai Deng;J. H. Pereira;Devon Van Cura;Ramu Kakumanu;E. Baidoo;Q. Dan;Yan Chen;C. Petzold;T. Northen;Paul D. Adams;D. Clark;E. Balskus;J. Hartwig;A. Mukhopadhyay;J. Keasling
生物合成是一种对环境无害且可再生的方法，可用于生产广泛的天然产品，在某些情况下，还可生产新产品。然而，生物学缺乏许多合成化学家可以获得的反应，这导致使用生物合成而不是合成化学时可获得的产品范围更窄。这种化学反应的一个主要例子是碳转移反应。虽然最近有研究表明，碳转移反应可以在细胞中进行并用于生物合成，但碳供体和非天然辅因子需要外源添加并运输到细胞中才能实现所需的反应，这阻碍了用这些反应扩大生物合成过程的成本效益。在这里，我们报告了通过细胞代谢获得重氮酯碳前体的途径，以及将非自然碳转移反应引入生物合成的微生物平台。α-重氮酯azazerine是通过表达一个生物合成基因簇inreptomyces albus合成的。细胞内生成的氮杂塞氨酸被用作另一种细胞内生成的分子苯乙烯的环丙烷化的碳供体。该反应由含有天然辅助因子的工程P450突变体催化，具有优异的非对映选择性和中等产量。我们的研究建立了一个可扩展的微生物平台，用于进行细胞内的非生物碳转移反应，以功能化一系列天然和新自然产物，并扩大了可以通过细胞代谢产生的有机产物的范围。
Biosynthesis is an environmentally benign and renewable approach that can be used to produce a broad range of natural and, in some cases, new-to-nature products. However, biology lacks many of the reactions that are available to synthetic chemists, resulting in a narrower scope of accessible products when using biosynthesis rather than synthetic chemistry. A prime example of such chemistry is carbene-transfer reactions. Although it was recently shown that carbene-transfer reactions can be performed in a cell and used for biosynthesis,, carbene donors and unnatural cofactors needed to be added exogenously and transported into cells to effect the desired reactions, precluding cost-effective scale-up of the biosynthesis process with these reactions. Here we report the access to a diazo ester carbene precursor by cellular metabolism and a microbial platform for introducing unnatural carbene-transfer reactions into biosynthesis. The α-diazoester azaserine was produced by expressing a biosynthetic gene cluster inStreptomyces albus. The intracellularly produced azaserine was used as a carbene donor to cyclopropanate another intracellularly produced molecule—styrene. The reaction was catalysed by engineered P450 mutants containing a native cofactor with excellent diastereoselectivity and a moderate yield. Our study establishes a scalable, microbial platform for conducting intracellular abiological carbene-transfer reactions to functionalize a range of natural and new-to-nature products and expands the scope of organic products that can be produced by cellular metabolism.