The Role of Cytochrome P450 AbyV in the Final Stages of Abyssomicin C Biosynthesis

Andrew J Devine; Alice E Parnell; Catherine R Back; Nicholas R Lees; Samuel T Johns; Ainul Z Zulkepli; Rob Barringer; Katja Zorn; James E M Stach; Matthew P Crump; Martin A Hayes; Marc W van der Kamp; Paul R Race; Christine Willis

doi:10.1002/anie.202213053

The Role of Cytochrome P450 AbyV in the Final Stages of Abyssomicin C Biosynthesis

Andrew J Devine, Alice E Parnell, Catherine R Back, Nicholas R Lees, Samuel T Johns, Ainul Z Zulkepli, Rob Barringer, Katja Zorn, James E M Stach, Matthew P Crump, Martin A Hayes, Marc W van der Kamp, Paul R Race, Christine Willis

Research output: Contribution to journal › Article (Academic Journal) › peer-review

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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.

Original language	English
Journal	Angewandte Chemie - International Edition
Early online date	31 Oct 2022
DOIs	https://doi.org/10.1002/anie.202213053
Publication status	E-pub ahead of print - 31 Oct 2022

Bibliographical note

Funding Information:
This study was supported by BBSRC and EPSRC through the BrisSynBio Synthetic Biology Research Centre (BB/L01386X/1), BBSRC grants BB/T001968/1 and BB/M012107/1, PhD SWBio DTP studentships awarded to AD and RB (BBSRC, BB/T008741/1), studentships to NRL (EPSRC, EP/G036764/1), STJ (EPSRC, EP/L016494/1), AZZ (MARA) and a BBSRC David Phillips Fellowship to MvdK (BB/M026280/1). KZ is a Fellow of the AstraZeneca Postdoctoral Research Programme. The authors thank Dr. Christopher Williams for assistance with NMR data collection, Professor Ross Anderson for assistance in the collection of absorbance spectra and the staff of Diamond Light Source beamline I04‐1 for support with X‐ray diffraction data collection.

Publisher Copyright:
© 2022 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH.

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Devine, A. J., Parnell, A. E., Back, C. R., Lees, N. R., Johns, S. T., Zulkepli, A. Z., Barringer, R., Zorn, K., Stach, J. E. M., Crump, M. P., Hayes, M. A., van der Kamp, M. W., Race, P. R., & Willis, C. (2022). The Role of Cytochrome P450 AbyV in the Final Stages of Abyssomicin C Biosynthesis. Angewandte Chemie - International Edition. Advance online publication. https://doi.org/10.1002/anie.202213053

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title = "The Role of Cytochrome P450 AbyV in the Final Stages of Abyssomicin C Biosynthesis",

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.",

author = "Devine, {Andrew J} and Parnell, {Alice E} and Back, {Catherine R} and Lees, {Nicholas R} and Johns, {Samuel T} and Zulkepli, {Ainul Z} and Rob Barringer and Katja Zorn and Stach, {James E M} and Crump, {Matthew P} and Hayes, {Martin A} and {van der Kamp}, {Marc W} and Race, {Paul R} and Christine Willis",

note = "Funding Information: This study was supported by BBSRC and EPSRC through the BrisSynBio Synthetic Biology Research Centre (BB/L01386X/1), BBSRC grants BB/T001968/1 and BB/M012107/1, PhD SWBio DTP studentships awarded to AD and RB (BBSRC, BB/T008741/1), studentships to NRL (EPSRC, EP/G036764/1), STJ (EPSRC, EP/L016494/1), AZZ (MARA) and a BBSRC David Phillips Fellowship to MvdK (BB/M026280/1). KZ is a Fellow of the AstraZeneca Postdoctoral Research Programme. The authors thank Dr. Christopher Williams for assistance with NMR data collection, Professor Ross Anderson for assistance in the collection of absorbance spectra and the staff of Diamond Light Source beamline I04‐1 for support with X‐ray diffraction data collection. Publisher Copyright: {\textcopyright} 2022 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH.",

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AU - Devine, Andrew J

AU - Parnell, Alice E

AU - Back, Catherine R

AU - Lees, Nicholas R

AU - Johns, Samuel T

AU - Zulkepli, Ainul Z

AU - Barringer, Rob

AU - Zorn, Katja

AU - Stach, James E M

AU - Crump, Matthew P

AU - Hayes, Martin A

AU - van der Kamp, Marc W

AU - Race, Paul R

AU - Willis, Christine

N1 - Funding Information: This study was supported by BBSRC and EPSRC through the BrisSynBio Synthetic Biology Research Centre (BB/L01386X/1), BBSRC grants BB/T001968/1 and BB/M012107/1, PhD SWBio DTP studentships awarded to AD and RB (BBSRC, BB/T008741/1), studentships to NRL (EPSRC, EP/G036764/1), STJ (EPSRC, EP/L016494/1), AZZ (MARA) and a BBSRC David Phillips Fellowship to MvdK (BB/M026280/1). KZ is a Fellow of the AstraZeneca Postdoctoral Research Programme. The authors thank Dr. Christopher Williams for assistance with NMR data collection, Professor Ross Anderson for assistance in the collection of absorbance spectra and the staff of Diamond Light Source beamline I04‐1 for support with X‐ray diffraction data collection. Publisher Copyright: © 2022 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH.

PY - 2022/10/31

Y1 - 2022/10/31

N2 - 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.

AB - 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.

U2 - 10.1002/anie.202213053

DO - 10.1002/anie.202213053

M3 - Article (Academic Journal)

C2 - 36314667

SN - 1433-7851

JO - Angewandte Chemie - International Edition

JF - Angewandte Chemie - International Edition

ER -

The Role of Cytochrome P450 AbyV in the Final Stages of Abyssomicin C Biosynthesis

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