8DSG

P411-PFA carbene transferase

Summary for 8DSG

• Entry DOI: 10.2210/pdb8dsg/pdb
• Descriptor: Cytochrome P450-BM3 variant P411-PFA, PROTOPORPHYRIN IX CONTAINING FE, 1,2-ETHANEDIOL, ... (6 entities in total)
• Functional Keywords: cytochrome p450, carbene transferase, cyanomethylase, fluoroalkylation, oxidoreductase
• Biological source: Priestia megaterium
• Total number of polymer chains: 4
• Total formula weight: 219841.37

Authors

Maggiolo, A.O.,Porter, N.J.,Zhang, J.,Arnold, F.H. (deposition date: 2022-07-22, release date: 2023-03-08, Last modification date: 2024-05-01)

Primary citation

Zhang, J.,Maggiolo, A.O.,Alfonzo, E.,Mao, R.,Porter, N.J.,Abney, N.,Arnold, F.H.
Chemodivergent C(sp 3 )-H and C(sp 2 )-H Cyanomethylation Using Engineered Carbene Transferases.
Nat Catal, 6:152-160, 2023

PubMed Abstract: The ubiquity of C-H bonds presents an attractive opportunity to elaborate and build complexity in organic molecules. Methods for selective functionalization, however, often must differentiate among multiple chemically similar and, in some cases indistinguishable, C-H bonds. An advantage of enzymes is that they can be finely tuned using directed evolution to achieve control over divergent C-H functionalization pathways. Here, we demonstrate engineered enzymes that effect a new-to-nature C-H alkylation with unparalleled selectivity: two complementary carbene C-H transferases derived from a cytochrome P450 from deliver an -cyanocarbene into the -amino C(sp)-H bonds or the -arene C(sp)-H bonds of -substituted arenes. These two transformations proceed via different mechanisms, yet only minimal changes to the protein scaffold (nine mutations, less than 2% of the sequence) were needed to adjust the enzyme's control over the site-selectivity of cyanomethylation. The X-ray crystal structure of the selective C(sp)-H alkylase, P411-PFA, reveals an unprecedented helical disruption which alters the shape and electrostatics in the enzyme active site. Overall, this work demonstrates the advantages of enzymes as C-H functionalization catalysts for divergent molecular derivatization.

PubMed: 36875868
DOI: 10.1038/s41929-022-00908-x

Experimental method

X-RAY DIFFRACTION (1.87 Å)