Delineation of the complete reaction cycle of a natural Diels-Alderase.

Autor:	Maschio L; School of Biochemistry, University Walk, University of Bristol BS8 1TD UK., Back CR; School of Biochemistry, University Walk, University of Bristol BS8 1TD UK., Alnawah J; School of Chemistry, University of Bristol Cantock's Close BS8 1TS UK Chris.Willis@bristol.ac.uk.; Department of Chemistry, King Faisal University, College of Science Al-Ahsa 31982 Saudi Arabia., Bowen JI; School of Chemistry, University of Bristol Cantock's Close BS8 1TS UK Chris.Willis@bristol.ac.uk., Johns ST; School of Biochemistry, University Walk, University of Bristol BS8 1TD UK., Mbatha SZ; School of Chemistry, University of Bristol Cantock's Close BS8 1TS UK Chris.Willis@bristol.ac.uk., Han LC; School of Chemistry, University of Bristol Cantock's Close BS8 1TS UK Chris.Willis@bristol.ac.uk., Lees NR; School of Chemistry, University of Bristol Cantock's Close BS8 1TS UK Chris.Willis@bristol.ac.uk., Zorn K; Discovery Sciences, Biopharmaceuticals R&D, AstraZeneca Pepparedsleden 1 431 83 Mölndal Sweden., Stach JEM; School of Natural and Environmental Sciences, Newcastle University NE1 7RU UK paul.race1@newcastle.ac.uk., Hayes MA; Discovery Sciences, Biopharmaceuticals R&D, AstraZeneca Pepparedsleden 1 431 83 Mölndal Sweden., van der Kamp MW; School of Biochemistry, University Walk, University of Bristol BS8 1TD UK., Pudney CR; Department of Biology and Biochemistry, University of Bath Claverton Down BA2 7AY UK., Burston SG; School of Biochemistry, University Walk, University of Bristol BS8 1TD UK., Willis CL; School of Chemistry, University of Bristol Cantock's Close BS8 1TS UK Chris.Willis@bristol.ac.uk., Race PR; School of Natural and Environmental Sciences, Newcastle University NE1 7RU UK paul.race1@newcastle.ac.uk.
Jazyk:	angličtina
Zdroj:	Chemical science [Chem Sci] 2024 Jun 24; Vol. 15 (29), pp. 11572-11583. Date of Electronic Publication: 2024 Jun 24 (Print Publication: 2024).
DOI:	10.1039/d4sc02908a
Abstrakt:	The Diels-Alder reaction is one of the most effective methods for the synthesis of substituted cyclohexenes. The development of protein catalysts for this reaction remains a major priority, affording new sustainable routes to high value target molecules. Whilst a small number of natural enzymes have been shown capable of catalysing [4 + 2] cycloadditions, there is a need for significant mechanistic understanding of how these prospective Diels-Alderases promote catalysis to underpin their development as biocatalysts for use in synthesis. Here we present a molecular description of the complete reaction cycle of the bona fide natural Diels-Alderase AbyU, which catalyses formation of the spirotetronate skeleton of the antibiotic abyssomicin C. This description is derived from X-ray crystallographic studies of AbyU in complex with a non-transformable synthetic substrate analogue, together with transient kinetic analyses of the AbyU catalysed reaction and computational reaction simulations. These studies reveal the mechanistic intricacies of this enzyme system and establish a foundation for the informed reengineering of AbyU and related biocatalysts. Competing Interests: The authors declare that they have no conflicts of interest. (This journal is © The Royal Society of Chemistry.)
Databáze:	MEDLINE
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