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Exploring the Factors which Result in Cytochrome P450 Catalyzed Desaturation Versus Hydroxylation

The cytochrome P450 family of monooxygenase enzymes have essential biological roles involving the selective oxidation of carbon‐hydrogen bonds. They can also catalyze other important metabolic reactions including desaturation to form alkenes. Currently the factors that control the partition between...

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Autores principales: Coleman, Tom, Doherty, Daniel Z., Zhang, Ting, Podgorski, Matthew N., Qiao, Ruihong, Lee, Joel H. Z., Bruning, John B., De Voss, James J., Zhou, Weihong, Bell, Stephen G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10100021/
https://www.ncbi.nlm.nih.gov/pubmed/36268769
http://dx.doi.org/10.1002/asia.202200986
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author Coleman, Tom
Doherty, Daniel Z.
Zhang, Ting
Podgorski, Matthew N.
Qiao, Ruihong
Lee, Joel H. Z.
Bruning, John B.
De Voss, James J.
Zhou, Weihong
Bell, Stephen G.
author_facet Coleman, Tom
Doherty, Daniel Z.
Zhang, Ting
Podgorski, Matthew N.
Qiao, Ruihong
Lee, Joel H. Z.
Bruning, John B.
De Voss, James J.
Zhou, Weihong
Bell, Stephen G.
author_sort Coleman, Tom
collection PubMed
description The cytochrome P450 family of monooxygenase enzymes have essential biological roles involving the selective oxidation of carbon‐hydrogen bonds. They can also catalyze other important metabolic reactions including desaturation to form alkenes. Currently the factors that control the partition between P450 hydroxylation and desaturation pathways are poorly defined. The CYP199A4 enzyme from the bacterium Rhodopseudomonas palustris HaA2 catalyzes the oxidation of 4‐ethyl‐ and 4‐isopropyl‐ benzoic acids with hydroxylation and desaturation occurring in significant quantities. Here we demonstrate that 4‐cyclopropylbenzoic acid is regioselectively hydroxylated by CYP199A4 at the benzylic carbon. In contrast, the oxidation of 4‐n‐propylbenzoic acid by CYP199A4 results in three major metabolites: an alkene from desaturation and two hydroxylation products at the benzylic (Cα) and Cβ carbons in similar quantities. Extending the length of the alkyl substituent resulted in 4‐n‐butylbenzoic acid being oxidized at the benzylic position (45%) and desaturated (55%). In contrast, 4‐isobutylbenzoic generated very little alkene (5%) but was hydroxylated at the benzylic position (54%) and at the tertiary Cβ position (41%). The oxidation of 4‐n‐propylbenzoic acid by the F298 V mutant of CYP199A4 occurred with no hydroxylation at Cβ and a significant increase in metabolites arising from desaturation (73%). The X‐ray crystal structures of CYP199A4 with each substrate revealed that they bind in the active site with the alkyl substituent positioned over the heme. However, the longer alkylbenzoic acids were bound in a different conformation as was 4‐n‐propylbenzoic acid in the F298 V mutant. Overall, the changes in metabolite distribution could be ascribed to bond strength differences and the position of the alkyl group relative to the heme.
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spelling pubmed-101000212023-04-14 Exploring the Factors which Result in Cytochrome P450 Catalyzed Desaturation Versus Hydroxylation Coleman, Tom Doherty, Daniel Z. Zhang, Ting Podgorski, Matthew N. Qiao, Ruihong Lee, Joel H. Z. Bruning, John B. De Voss, James J. Zhou, Weihong Bell, Stephen G. Chem Asian J Research Articles The cytochrome P450 family of monooxygenase enzymes have essential biological roles involving the selective oxidation of carbon‐hydrogen bonds. They can also catalyze other important metabolic reactions including desaturation to form alkenes. Currently the factors that control the partition between P450 hydroxylation and desaturation pathways are poorly defined. The CYP199A4 enzyme from the bacterium Rhodopseudomonas palustris HaA2 catalyzes the oxidation of 4‐ethyl‐ and 4‐isopropyl‐ benzoic acids with hydroxylation and desaturation occurring in significant quantities. Here we demonstrate that 4‐cyclopropylbenzoic acid is regioselectively hydroxylated by CYP199A4 at the benzylic carbon. In contrast, the oxidation of 4‐n‐propylbenzoic acid by CYP199A4 results in three major metabolites: an alkene from desaturation and two hydroxylation products at the benzylic (Cα) and Cβ carbons in similar quantities. Extending the length of the alkyl substituent resulted in 4‐n‐butylbenzoic acid being oxidized at the benzylic position (45%) and desaturated (55%). In contrast, 4‐isobutylbenzoic generated very little alkene (5%) but was hydroxylated at the benzylic position (54%) and at the tertiary Cβ position (41%). The oxidation of 4‐n‐propylbenzoic acid by the F298 V mutant of CYP199A4 occurred with no hydroxylation at Cβ and a significant increase in metabolites arising from desaturation (73%). The X‐ray crystal structures of CYP199A4 with each substrate revealed that they bind in the active site with the alkyl substituent positioned over the heme. However, the longer alkylbenzoic acids were bound in a different conformation as was 4‐n‐propylbenzoic acid in the F298 V mutant. Overall, the changes in metabolite distribution could be ascribed to bond strength differences and the position of the alkyl group relative to the heme. John Wiley and Sons Inc. 2022-11-11 2022-12-14 /pmc/articles/PMC10100021/ /pubmed/36268769 http://dx.doi.org/10.1002/asia.202200986 Text en © 2022 The Authors. Chemistry & An Asian Journal published by Wiley-VCH GmbH https://creativecommons.org/licenses/by-nc/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc/4.0/ (https://creativecommons.org/licenses/by-nc/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.
spellingShingle Research Articles
Coleman, Tom
Doherty, Daniel Z.
Zhang, Ting
Podgorski, Matthew N.
Qiao, Ruihong
Lee, Joel H. Z.
Bruning, John B.
De Voss, James J.
Zhou, Weihong
Bell, Stephen G.
Exploring the Factors which Result in Cytochrome P450 Catalyzed Desaturation Versus Hydroxylation
title Exploring the Factors which Result in Cytochrome P450 Catalyzed Desaturation Versus Hydroxylation
title_full Exploring the Factors which Result in Cytochrome P450 Catalyzed Desaturation Versus Hydroxylation
title_fullStr Exploring the Factors which Result in Cytochrome P450 Catalyzed Desaturation Versus Hydroxylation
title_full_unstemmed Exploring the Factors which Result in Cytochrome P450 Catalyzed Desaturation Versus Hydroxylation
title_short Exploring the Factors which Result in Cytochrome P450 Catalyzed Desaturation Versus Hydroxylation
title_sort exploring the factors which result in cytochrome p450 catalyzed desaturation versus hydroxylation
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10100021/
https://www.ncbi.nlm.nih.gov/pubmed/36268769
http://dx.doi.org/10.1002/asia.202200986
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