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The Role of Cytochrome P450 AbyV in the Final Stages of Abyssomicin C Biosynthesis |
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| Authors: | Andrew J Devine Dr Alice E Parnell Dr Catherine R Back Dr Nicholas R Lees Dr Samuel T Johns Ainul Z Zulkepli Rob Barringer Dr Katja Zorn Dr James E M Stach Prof Matthew P Crump Prof Martin A Hayes Dr Marc W van der Kamp Prof Paul R Race Prof Christine L Willis |
| Institution: | 1. School of Chemistry, University of Bristol, BS81TS Bristol, UK;2. School of Biochemistry, University of Bristol, BS81TD Bristol, UK;3. BioPharmaceuticals R&D, AstraZeneca, Pepparedsleden 1, 43183 Mölndal, Sweden;4. School of Natural and Environmental Sciences, Newcastle University, NE17RU Newcastle-upon-Tyne, UK |
| Abstract: | Abyssomicin C and its atropisomer are potent inhibitors of bacterial folate metabolism. They possess complex polycyclic structures, and their biosynthesis has been shown to involve several unusual enzymatic transformations. Using a combination of synthesis and in vitro assays we reveal that AbyV, a cytochrome P450 enzyme from the aby gene cluster, catalyses a key late-stage epoxidation required for the installation of the characteristic ether-bridged core of abyssomicin C. The X-ray crystal structure of AbyV has been determined, which in combination with molecular dynamics simulations provides a structural framework for our functional data. This work demonstrates the power of combining selective carbon-13 labelling with NMR spectroscopy as a sensitive tool to interrogate enzyme-catalysed reactions in vitro with no need for purification. |
| Keywords: | Antibiotics Biosynthesis P450 Enzymes Polyketides Structure Elucidation |