Complete integration of carbene-transfer chemistry into biosynthesis

Huang, Jing; Quest, Andrew; Cruz-Morales, Pablo; Deng, Kai; Pereira, Jose Henrique; Van Cura, Devon; Kakumanu, Ramu; Baidoo, Edward E. K.; Dan, Qingyun; Chen, Yan; Petzold, Christopher J.; Northen, Trent R.; Adams, Paul D.; Clark, Douglas S.; Balskus, Emily P.; Hartwig, John F.; Mukhopadhyay, Aindrila; Keasling, Jay D.

doi:10.1038/s41586-023-06027-2

Complete integration of carbene-transfer chemistry into biosynthesis

Journal Article · Wed May 03 00:00:00 EDT 2023 · Nature (London)

DOI:https://doi.org/10.1038/s41586-023-06027-2· OSTI ID:2469678

Huang, Jing ^[1]; Quest, Andrew ^[2]; Cruz-Morales, Pablo ^[3]; Deng, Kai ^[4]; Pereira, Jose Henrique ^[5]; ^[6]; Kakumanu, Ramu ^[7]; Baidoo, Edward E. K. ^[7]; Dan, Qingyun ^[7]; Chen, Yan ^[7]; ^[7]; ^[5]; ^[8]; Clark, Douglas S. ^[2]; ^[6]; ^[2]; ^[7]; ^[9]

Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States); Joint BioEnergy Institute (JBEI), Emeryville, CA (United States); University of California, Berkeley, CA (United States)
University of California, Berkeley, CA (United States); Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States); Joint BioEnergy Institute (JBEI), Emeryville, CA (United States); Technical Univ. of Denmark, Lyngby (Denmark)
Joint BioEnergy Institute (JBEI), Emeryville, CA (United States); Sandia National Lab. (SNL-CA), Livermore, CA (United States)
Joint BioEnergy Institute (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
Harvard Univ., Cambridge, MA (United States)
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States); Joint BioEnergy Institute (JBEI), Emeryville, CA (United States)
Joint BioEnergy Institute (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States); University of California, Berkeley, CA (United States)
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States); Joint BioEnergy Institute (JBEI), Emeryville, CA (United States); University of California, Berkeley, CA (United States); Technical Univ. of Denmark, Lyngby (Denmark); Shenzhen Institutes for Advanced Technologies (China)

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, in this paper, 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.

View Accepted Manuscript (DOE)

Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 2469678

Journal Information:: Nature (London), Journal Name: Nature (London) Journal Issue: 7960 Vol. 617; ISSN 0028-0836

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: English

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