Unexpected Oxidation of a Depsipeptide Substrate Analogue in Crystalline Isopenicillin N Synthase

ChemBioChem ◽  
2006 ◽  
Vol 7 (2) ◽  
pp. 351-358 ◽  
Author(s):  
Adam Daruzzaman ◽  
Ian J. Clifton ◽  
Robert M. Adlington ◽  
Jack E. Baldwin ◽  
Peter J. Rutledge
Keyword(s):  
Substrate Analogue ◽  
Isopenicillin N

scholarly journals Active-site-mediated elimination of hydrogen fluoride from a fluorinated substrate analogue by isopenicillin N synthase

2004 ◽  
Vol 382 (2) ◽  
pp. 659-666 ◽  
Author(s):  
Annaleise R. GRUMMITT ◽  
Peter J. RUTLEDGE ◽  
Ian J. CLIFTON ◽  
Jack E. BALDWIN
Keyword(s):  
Lewis Acid ◽  
Hydrogen Fluoride ◽  
Mechanism Of Action ◽  
Active Site ◽  
Nmr Studies ◽  
X Ray ◽  
Substrate Analogue ◽  
Haem Iron ◽  
Active Site Region ◽  
Isopenicillin N

Isopenicillin N synthase (IPNS) is a non-haem iron oxidase that catalyses the formation of bicyclic isopenicillin N from δ-(L-α-aminoadipoyl)-L-cysteinyl-D-valine (ACV). In this study we report a novel activity for the iron of the IPNS active site, which behaves as a Lewis acid to catalyse the elimination of HF from the fluorinated substrate analogue, δ-(L-α-aminoadipoyl)-L-cysteinyl-D-β-fluorovaline (ACβFV). X-Ray crystallographic studies of IPNS crystals grown anaerobically with ACβFV reveal that the valinyl β-fluorine is missing from the active site region, and suggest the presence of the unsaturated tripeptide δ-(L-α-aminoadipoyl)-L-cysteinyl-D-isodehydrovaline in place of substrate ACβFV. 19F NMR studies confirm the release of fluoride from ACβFV in the presence of the active IPNS enzyme. These results suggest a new mode of reactivity for the IPNS iron centre, a mechanism of action that has not previously been reported for any of the iron oxidase enzymes.

Isopenicillin N Synthase Mediates Thiolate Oxidation to Sulfenate in a Depsipeptide Substrate Analogue: Implications for Oxygen Binding and a Link to Nitrile Hydratase?

2008 ◽  
Vol 130 (31) ◽  
pp. 10096-10102 ◽  
Author(s):  
Wei Ge ◽  
Ian J. Clifton ◽  
Jeanette E. Stok ◽  
Robert M. Adlington ◽  
Jack E. Baldwin ◽  
...  
Keyword(s):  
Nitrile Hydratase ◽  
Oxygen Binding ◽  
Substrate Analogue ◽  
Isopenicillin N

The crystal structure of an isopenicillin N synthase complex with an ethereal substrate analogue reveals water in the oxygen binding site

FEBS Letters ◽  
2013 ◽  
Vol 587 (16) ◽  
pp. 2705-2709 ◽  
Author(s):  
Ian J. Clifton ◽  
Wei Ge ◽  
Robert M. Adlington ◽  
Jack E. Baldwin ◽  
Peter J. Rutledge
Keyword(s):  
Crystal Structure ◽  
Binding Site ◽  
Oxygen Binding ◽  
Substrate Analogue ◽  
Isopenicillin N

Structural Studies on the Reaction of Isopenicillin N Synthase with a Sterically Demanding Depsipeptide Substrate Analogue

ChemBioChem ◽  
2009 ◽  
Vol 10 (12) ◽  
pp. 2025-2031 ◽  
Author(s):  
Wei Ge ◽  
Ian J. Clifton ◽  
Annaleise R. Howard-Jones ◽  
Jeanette E. Stok ◽  
Robert M. Adlington ◽  
...  
Keyword(s):  
Structural Studies ◽  
Substrate Analogue ◽  
Sterically Demanding ◽  
Isopenicillin N

scholarly journals Structural studies on the reaction of isopenicillin N synthase with the substrate analogue delta-(l-alpha-aminoadipoyl)-l-cysteinyl-d-alpha-aminobutyrate

2003 ◽  
Vol 372 (3) ◽  
pp. 687-693 ◽  
Author(s):  
Alexandra J. LONG ◽  
Ian J. CLIFTON ◽  
Peter L. ROACH ◽  
Jack E. BALDWIN ◽  
Christopher J. SCHOFIELD ◽  
...  
Keyword(s):  
Crystal Structures ◽  
Active Site ◽  
Structural Studies ◽  
Product Selectivity ◽  
Substrate Analogue ◽  
Modelling Studies ◽  
Key Residues ◽  
Haem Iron ◽  
Shed Light ◽  
Isopenicillin N

Isopenicillin N synthase (IPNS) is a non-haem iron(II) oxidase which catalyses the biosynthesis of isopenicillin N from the tripeptide δ-(l-α-aminoadipoyl)-l-cysteinyl-d-valine (ACV). Herein we report crystallographic studies to investigate the reaction of IPNS with the truncated substrate analogue δ-(l-α-aminoadipoyl)-l-cysteinyl-d-α-aminobutyrate (ACAb). It has been reported previously that this analogue gives rise to three β-lactam products when incubated with IPNS: two methyl penams and a cepham. Crystal structures of the IPNS–Fe(II)–ACAb and IPNS–Fe(II)–ACAb–NO complexes have now been solved and are reported herein. These structures and modelling studies based on them shed light on the diminished product selectivity shown by IPNS in its reaction with ACAb and further rationalize the presence of certain key residues at the IPNS active site.

The crystal structure of isopenicillin N synthase with a dipeptide substrate analogue

2013 ◽  
Vol 530 (1) ◽  
pp. 48-53 ◽  
Author(s):  
Adam Daruzzaman ◽  
Ian J. Clifton ◽  
Robert M. Adlington ◽  
Jack E. Baldwin ◽  
Peter J. Rutledge
Keyword(s):  
Crystal Structure ◽  
Substrate Analogue ◽  
Isopenicillin N

Crystallographic studies on the reaction of isopenicillin N synthase with an unsaturated substrate analogue

2003 ◽  
Vol 1 (9) ◽  
pp. 1455-1460 ◽  
Author(s):  
Jonathan M. Elkins ◽  
Peter J. Rutledge ◽  
Nicolai I. Burzlaff ◽  
Ian J. Clifton ◽  
Robert M. Adlington ◽  
...  
Keyword(s):  
Substrate Analogue ◽  
Isopenicillin N

The crystal structure of anlll-configured depsipeptide substrate analogue bound to isopenicillin N synthase

2010 ◽  
Vol 8 (1) ◽  
pp. 122-127 ◽  
Author(s):  
Wei Ge ◽  
Ian J. Clifton ◽  
Jeanette E. Stok ◽  
Robert M. Adlington ◽  
Jack E. Baldwin ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Substrate Analogue ◽  
Isopenicillin N

scholarly journals Activation modes in biocatalytic radical cyclization reactions

2021 ◽  
Author(s):  
Yuxuan Ye ◽  
Haigen Fu ◽  
Todd K Hyster
Keyword(s):  
Heme Iron ◽  
Radical Cyclization ◽  
Cytochrome P450 Monooxygenases ◽  
Radical Cyclizations ◽  
Cyclization Reactions ◽  
Complex Molecules ◽  
P450 Monooxygenases ◽  
Non Heme Iron ◽  
Essential Reactions ◽  
Isopenicillin N

Abstract Radical cyclizations are essential reactions in the biosynthesis of secondary metabolites and the chemical synthesis of societally valuable molecules. In this review, we highlight the general mechanisms utilized in biocatalytic radical cyclizations. We specifically highlight cytochrome P450 monooxygenases (P450s) involved in the biosynthesis of mycocyclosin and vancomycin, non-heme iron- and α-ketoglutarate-dependent dioxygenases (Fe/αKGDs) used in the biosynthesis of kainic acid, scopolamine, and isopenicillin N, and radical S-adenosylmethionine (SAM) enzymes that facilitate the biosynthesis of oxetanocin A, menaquinone, and F420. Beyond natural mechanisms, we also examine repurposed flavin-dependent ‘ene’-reductases (ERED) for non-natural radical cyclization. Overall, these general mechanisms underscore the opportunity for enzymes to augment and enhance the synthesis of complex molecules using radical mechanisms.

Isopenicillin N synthase: mechanistic studies

1990 ◽  
Vol 90 (7) ◽  
pp. 1079-1088 ◽  
Author(s):  
Jack E. Baldwin ◽  
Mark Bradley
Keyword(s):  
Mechanistic Studies ◽  
Isopenicillin N
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