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Discovery, characterization and engineering of ligases for amide synthesis

DOI: 10.1038/s41586-021-03447-w DOI Help

Authors: Michael Winn (The University of Manchester) , Michael Rowlinson (The University of Manchester) , Fanghua Wang (South China University of Technology; The University of Manchester) , Luis Bering (The University of Manchester) , Daniel Francis (The University of Manchester) , Colin Levy (The University of Manchester) , Jason Micklefield (The University of Manchester)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Nature , VOL 593 , PAGES 391 - 398

State: Published (Approved)
Published: May 2021
Diamond Proposal Number(s): 17773

Abstract: Coronatine and related bacterial phytotoxins are mimics of the hormone jasmonyl-L-isoleucine (JA-Ile), which mediates physiologically important plant signalling pathways1,2,3,4. Coronatine-like phytotoxins disrupt these essential pathways and have potential in the development of safer, more selective herbicides. Although the biosynthesis of coronatine has been investigated previously, the nature of the enzyme that catalyses the crucial coupling of coronafacic acid to amino acids remains unknown1,2. Here we characterize a family of enzymes, coronafacic acid ligases (CfaLs), and resolve their structures. We found that CfaL can also produce JA-Ile, despite low similarity with the Jar1 enzyme that is responsible for ligation of JA and L-Ile in plants5. This suggests that Jar1 and CfaL evolved independently to catalyse similar reactions—Jar1 producing a compound essential for plant development4,5, and the bacterial ligases producing analogues toxic to plants. We further demonstrate how CfaL enzymes can be used to synthesize a diverse array of amides, obviating the need for protecting groups. Highly selective kinetic resolutions of racemic donor or acceptor substrates were achieved, affording homochiral products. We also used structure-guided mutagenesis to engineer improved CfaL variants. Together, these results show that CfaLs can deliver a wide range of amides for agrochemical, pharmaceutical and other applications.

Journal Keywords: Biocatalysis; Biosynthesis; Jasmonic acid; Ligases; Sustainability

Diamond Keywords: Enzymes; Bacteria

Subject Areas: Biology and Bio-materials, Chemistry

Instruments: I03-Macromolecular Crystallography , I04-1-Macromolecular Crystallography (fixed wavelength)

Added On: 25/05/2021 13:10

Discipline Tags:

Catalysis Organic Chemistry Life Sciences & Biotech Health & Wellbeing Drug Discovery Pathogens Structural biology Chemistry Biochemistry

Technical Tags:

Diffraction Macromolecular Crystallography (MX)