1. Complete integration of carbene-transfer chemistry into biosynthesis.
- Author
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Huang, Jing, Quest, Andrew, Cruz-Morales, Pablo, Deng, Kai, Pereira, Jose, Van Cura, Devon, Kakumanu, Ramu, Baidoo, Edward, Dan, Qingyun, Chen, Yan, Petzold, Christopher, Northen, Trent, Adams, Paul, Clark, Douglas, Balskus, Emily, Hartwig, John, Mukhopadhyay, Aindrila, and Keasling, Jay
- Subjects
Azaserine ,Biological Products ,Multigene Family ,Styrene ,Cyclopropanes ,Coenzymes ,Biocatalysis ,Cytochrome P-450 Enzyme System - Abstract
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 reactions1. Although it was recently shown that carbene-transfer reactions can be performed in a cell and used for biosynthesis2,3, 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 in Streptomyces 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.
- Published
- 2023