8FBC

Crystal structure of P450T2

Summary for 8FBC

• Entry DOI: 10.2210/pdb8fbc/pdb
• Descriptor: Cytochrome P450, PROTOPORPHYRIN IX CONTAINING FE (3 entities in total)
• Functional Keywords: p450, biosynthetic protein
• Biological source: metagenome
• Total number of polymer chains: 2
• Total formula weight: 89863.86

Authors

Pereira, J.H.,Huang, J.,Keasling, J.,Adams, P.D. (deposition date: 2022-11-29, release date: 2023-05-10, Last modification date: 2024-05-22)

Primary citation

Huang, J.,Quest, A.,Cruz-Morales, P.,Deng, K.,Pereira, J.H.,Van Cura, D.,Kakumanu, R.,Baidoo, E.E.K.,Dan, Q.,Chen, Y.,Petzold, C.J.,Northen, T.R.,Adams, P.D.,Clark, D.S.,Balskus, E.P.,Hartwig, J.F.,Mukhopadhyay, A.,Keasling, J.D.
Complete integration of carbene-transfer chemistry into biosynthesis.
Nature, 617:403-408, 2023

PubMed 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 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 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.

PubMed: 37138074
DOI: 10.1038/s41586-023-06027-2

Experimental method

X-RAY DIFFRACTION (1.53 Å)