A quantum chemical study of the omega-transaminase reaction mechanism
2015 (English)In: Organic and biomolecular chemistry, ISSN 1477-0520, E-ISSN 1477-0539, Vol. 13, no 31, 8453-8464 p.Article in journal (Refereed) Published
omega-Transaminases are valuable tools in biocatalysis due to their stereospecificity and their broad substrate range. In the present study, the reaction mechanism of Chromobacterium violaceum omega-transaminase is investigated by means of density functional theory calculations. A large active site model is designed based on the recent X-ray crystal structure. The detailed energy profile for the half-transamination of (S)-1-phenylethylamine to acetophenone is calculated and the involved transition states and intermediates are characterized. The model suggests that the amino substrate forms an external aldimine with the coenzyme pyridoxal-5'-phosphate (PLP), through geminal diamine intermediates. The external aldimine is then deprotonated in the rate-determining step, forming a planar quinonoid intermediate. A ketimine is then formed, after which a hemiaminal is produced by the addition of water. Subsequently, the ketone product is obtained together with pyridoxamine-5'-phosphate (PMP). In the studied half-transamination reaction the ketone product is kinetically favored. The mechanism presented here will be valuable to enhance rational and semi-rational design of engineered enzyme variants in the development of omega-transaminase chemistry.
Place, publisher, year, edition, pages
The Royal Society of Chemistry , 2015. Vol. 13, no 31, 8453-8464 p.
IdentifiersURN: urn:nbn:se:su:diva-120490DOI: 10.1039/c5ob00690bISI: 000358733100011OAI: oai:DiVA.org:su-120490DiVA: diva2:852851
FunderSwedish Research CouncilKnut and Alice Wallenberg Foundation