Enzymatic synthesis and chemical inversion provide both enantiomers of bioactive epoxydocosapentaenoic acids
Epoxy PUFAs are endogenous cytochrome P450 (P450) metabolites of dietary PUFAs. Although these metabolites exert numerous biological effects, attempts to study their complex biology have been hampered by difficulty in obtaining the epoxides as pure regioisomers and enantiomers. To remedy this, we sy...
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doaj-ff499f2b54d64dc29825dcd55a4ad17d2021-04-29T04:34:19ZengElsevierJournal of Lipid Research0022-22752018-11-01591122372252Enzymatic synthesis and chemical inversion provide both enantiomers of bioactive epoxydocosapentaenoic acidsMaris A. Cinelli0Jun Yang1Amy Scharmen2Joey Woodman3Lalitha M. Karchalla4Kin Sing Stephen Lee5Department of Pharmacology and Toxicology, Michigan State University, East Lansing, MI 48824Department of Entomology and Nematology University of California at Davis, Davis, CA 95616; UCD Comprehensive Cancer Center, University of California at Davis, Davis, CA 95616Department of Pharmacology and Toxicology, Michigan State University, East Lansing, MI 48824Department of Pharmacology and Toxicology, Michigan State University, East Lansing, MI 48824Department of Pharmacology and Toxicology, Michigan State University, East Lansing, MI 48824To whom correspondence should be addressed.; Department of Pharmacology and Toxicology, Michigan State University, East Lansing, MI 48824Epoxy PUFAs are endogenous cytochrome P450 (P450) metabolites of dietary PUFAs. Although these metabolites exert numerous biological effects, attempts to study their complex biology have been hampered by difficulty in obtaining the epoxides as pure regioisomers and enantiomers. To remedy this, we synthesized 19,20- and 16,17-epoxydocosapentaenoic acids (EDPs) (the two most abundant EDPs in vivo) by epoxidation of DHA with WT and the mutant (F87V) P450 enzyme BM3 from Bacillus megaterium. WT epoxidation yielded a 4:1 mixture of 19,20:16,17-EDP exclusively as (S,R) enantiomers. Epoxidation with the mutant (F87V) yielded a 1.6:1 mixture of 19,20:16,17-EDP; the 19,20-EDP fraction was ∼9:1 (S,R):(R,S), but the 16,17-EDP was exclusively the (S,R) enantiomer. To access the (R,S) enantiomers of these EDPs, we used a short (four-step) chemical inversion sequence, which utilizes 2-(phenylthio)ethanol as the epoxide-opening nucleophile, followed by mesylation of the resulting alcohol, oxidation of the thioether moiety, and base-catalyzed elimination. This short synthesis cleanly converts the (S,R)-epoxide to the (R,S)-epoxide without loss of enantiopurity. This method, also applicable to eicosapentaenoic acid and arachidonic acid, provides a simple, cost-effective procedure for accessing larger amounts of these metabolites.http://www.sciencedirect.com/science/article/pii/S0022227520309238chemoenzymatic synthesiscytochrome P450docosahexaenoic acidpolyunsaturated fatty acidslipids/chemistryepoxy fatty acids |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Maris A. Cinelli Jun Yang Amy Scharmen Joey Woodman Lalitha M. Karchalla Kin Sing Stephen Lee |
spellingShingle |
Maris A. Cinelli Jun Yang Amy Scharmen Joey Woodman Lalitha M. Karchalla Kin Sing Stephen Lee Enzymatic synthesis and chemical inversion provide both enantiomers of bioactive epoxydocosapentaenoic acids Journal of Lipid Research chemoenzymatic synthesis cytochrome P450 docosahexaenoic acid polyunsaturated fatty acids lipids/chemistry epoxy fatty acids |
author_facet |
Maris A. Cinelli Jun Yang Amy Scharmen Joey Woodman Lalitha M. Karchalla Kin Sing Stephen Lee |
author_sort |
Maris A. Cinelli |
title |
Enzymatic synthesis and chemical inversion provide both enantiomers of bioactive epoxydocosapentaenoic acids |
title_short |
Enzymatic synthesis and chemical inversion provide both enantiomers of bioactive epoxydocosapentaenoic acids |
title_full |
Enzymatic synthesis and chemical inversion provide both enantiomers of bioactive epoxydocosapentaenoic acids |
title_fullStr |
Enzymatic synthesis and chemical inversion provide both enantiomers of bioactive epoxydocosapentaenoic acids |
title_full_unstemmed |
Enzymatic synthesis and chemical inversion provide both enantiomers of bioactive epoxydocosapentaenoic acids |
title_sort |
enzymatic synthesis and chemical inversion provide both enantiomers of bioactive epoxydocosapentaenoic acids |
publisher |
Elsevier |
series |
Journal of Lipid Research |
issn |
0022-2275 |
publishDate |
2018-11-01 |
description |
Epoxy PUFAs are endogenous cytochrome P450 (P450) metabolites of dietary PUFAs. Although these metabolites exert numerous biological effects, attempts to study their complex biology have been hampered by difficulty in obtaining the epoxides as pure regioisomers and enantiomers. To remedy this, we synthesized 19,20- and 16,17-epoxydocosapentaenoic acids (EDPs) (the two most abundant EDPs in vivo) by epoxidation of DHA with WT and the mutant (F87V) P450 enzyme BM3 from Bacillus megaterium. WT epoxidation yielded a 4:1 mixture of 19,20:16,17-EDP exclusively as (S,R) enantiomers. Epoxidation with the mutant (F87V) yielded a 1.6:1 mixture of 19,20:16,17-EDP; the 19,20-EDP fraction was ∼9:1 (S,R):(R,S), but the 16,17-EDP was exclusively the (S,R) enantiomer. To access the (R,S) enantiomers of these EDPs, we used a short (four-step) chemical inversion sequence, which utilizes 2-(phenylthio)ethanol as the epoxide-opening nucleophile, followed by mesylation of the resulting alcohol, oxidation of the thioether moiety, and base-catalyzed elimination. This short synthesis cleanly converts the (S,R)-epoxide to the (R,S)-epoxide without loss of enantiopurity. This method, also applicable to eicosapentaenoic acid and arachidonic acid, provides a simple, cost-effective procedure for accessing larger amounts of these metabolites. |
topic |
chemoenzymatic synthesis cytochrome P450 docosahexaenoic acid polyunsaturated fatty acids lipids/chemistry epoxy fatty acids |
url |
http://www.sciencedirect.com/science/article/pii/S0022227520309238 |
work_keys_str_mv |
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