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Insights into E. coli Cyclopropane Fatty Acid Synthase (CFAS) Towards Enantioselective Carbene Free Biocatalytic Cyclopropanation.
Iman Omar ,
Michele Crotti ,
Chuhan Li ,
Krisztina Pisak,
Blazej Czemerys,
Salvatore Ferla ,
Aster van Noord,
Caroline E. Paul ,
Kersti Karu ,
Cagakan Ozbalci ,
Ulrike Eggert ,
Richard Lloyd,
Sarah M. Barry ,
Daniele Castagnolo
Angewandte Chemie International Edition
Swansea University Author: Salvatore Ferla
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© 2024 The Authors. This is an open access article under the terms of the Creative Commons Attribution License.
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DOI (Published version): 10.1002/anie.202403493
Abstract
Cyclopropane fatty acid synthases (CFAS) are a class of S-adenosylmethionine (SAM) dependent methyltransferase enzymes able to catalyse the cyclopropanation of unsaturated phospholipids. Since CFAS enzymes employ SAM as a methylene source to cyclopropanate alkene substrates, they have the potential...
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ISSN: | 1433-7851 1521-3773 |
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2024
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v2 66386 2024-05-09 Insights into E. coli Cyclopropane Fatty Acid Synthase (CFAS) Towards Enantioselective Carbene Free Biocatalytic Cyclopropanation. d4c62248f510e3b221916989a7bbe6a6 0000-0002-5918-9237 Salvatore Ferla Salvatore Ferla true false 2024-05-09 MEDS Cyclopropane fatty acid synthases (CFAS) are a class of S-adenosylmethionine (SAM) dependent methyltransferase enzymes able to catalyse the cyclopropanation of unsaturated phospholipids. Since CFAS enzymes employ SAM as a methylene source to cyclopropanate alkene substrates, they have the potential to be mild and more sustainable biocatalysts for cyclopropanation transformations than current carbene based approaches. This work describes the characterisation of E. coli CFAS enzyme (ecCFAS) and its exploitation in the stereoselective biocatalytic synthesis of cyclopropyl lipids. ecCFAS was found to convert phosphatidylglycerol (PG) to methyl dihydrosterculate 1 from in up to 58% conversion and 73% ee and the absolute configuration (9S,10R) was established. Substrate tolerance of ecCFAS was found to be correlated with the electronic properties of phospholipid headgroups and for the first time ecCFAS was found to catalyse cyclopropanation of both phospholipid chains to form dicyclopropanated products. In addition, mutagenesis and in-silico experiments were carried out to identify the enzyme residues with key roles in catalysis and to provide structural insights into the lipid substrate preference of ecCFAS. Finally, the biocatalytic synthesis of methyl dihydrosterculate 1 and its deuterated analogue was also accomplished combining pure ecCFAS with the SAM regenerating AtHMT enzyme in presence of CH3I and CD3I. Journal Article Angewandte Chemie International Edition 0 Wiley 1433-7851 1521-3773 14 6 2024 2024-06-14 10.1002/anie.202403493 COLLEGE NANME Medical School COLLEGE CODE MEDS Swansea University Another institution paid the OA fee Biotechnology and Biological Sciences Research Council. Grant Numbers: BB/M009513/1, BIDS3000037082, POC-045, R116824, BB/P019811/1 Chinese Government Scholarship. Grant Number: 202108310078 H2020 Marie Skłodowska-Curie Actions. Grant Number: 101027045 H2020 European Research Council. Grant Number: 949910 2024-06-27T15:38:27.0000222 2024-05-09T15:23:09.4818876 Faculty of Medicine, Health and Life Sciences Swansea University Medical School - Pharmacy Iman Omar 0000-0002-4216-5394 1 Michele Crotti 0000-0003-3274-9393 2 Chuhan Li 0000-0001-5989-0093 3 Krisztina Pisak 4 Blazej Czemerys 5 Salvatore Ferla 0000-0002-5918-9237 6 Aster van Noord 7 Caroline E. Paul 0000-0002-7889-9920 8 Kersti Karu 0000-0002-3974-0271 9 Cagakan Ozbalci 0000-0002-7572-5511 10 Ulrike Eggert 0000-0003-0932-5525 11 Richard Lloyd 12 Sarah M. Barry 0000-0001-8188-2153 13 Daniele Castagnolo 0000-0002-7517-5732 14 66386__30772__01c5effe77b145cb901e73dec438b44e.pdf 66386.VoR.pdf 2024-06-27T15:36:24.3514976 Output 8912595 application/pdf Version of Record true © 2024 The Authors. This is an open access article under the terms of the Creative Commons Attribution License. true eng http://creativecommons.org/licenses/by/4.0/ |
title |
Insights into E. coli Cyclopropane Fatty Acid Synthase (CFAS) Towards Enantioselective Carbene Free Biocatalytic Cyclopropanation. |
spellingShingle |
Insights into E. coli Cyclopropane Fatty Acid Synthase (CFAS) Towards Enantioselective Carbene Free Biocatalytic Cyclopropanation. Salvatore Ferla |
title_short |
Insights into E. coli Cyclopropane Fatty Acid Synthase (CFAS) Towards Enantioselective Carbene Free Biocatalytic Cyclopropanation. |
title_full |
Insights into E. coli Cyclopropane Fatty Acid Synthase (CFAS) Towards Enantioselective Carbene Free Biocatalytic Cyclopropanation. |
title_fullStr |
Insights into E. coli Cyclopropane Fatty Acid Synthase (CFAS) Towards Enantioselective Carbene Free Biocatalytic Cyclopropanation. |
title_full_unstemmed |
Insights into E. coli Cyclopropane Fatty Acid Synthase (CFAS) Towards Enantioselective Carbene Free Biocatalytic Cyclopropanation. |
title_sort |
Insights into E. coli Cyclopropane Fatty Acid Synthase (CFAS) Towards Enantioselective Carbene Free Biocatalytic Cyclopropanation. |
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d4c62248f510e3b221916989a7bbe6a6 |
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d4c62248f510e3b221916989a7bbe6a6_***_Salvatore Ferla |
author |
Salvatore Ferla |
author2 |
Iman Omar Michele Crotti Chuhan Li Krisztina Pisak Blazej Czemerys Salvatore Ferla Aster van Noord Caroline E. Paul Kersti Karu Cagakan Ozbalci Ulrike Eggert Richard Lloyd Sarah M. Barry Daniele Castagnolo |
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10.1002/anie.202403493 |
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Wiley |
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Faculty of Medicine, Health and Life Sciences |
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Swansea University Medical School - Pharmacy{{{_:::_}}}Faculty of Medicine, Health and Life Sciences{{{_:::_}}}Swansea University Medical School - Pharmacy |
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description |
Cyclopropane fatty acid synthases (CFAS) are a class of S-adenosylmethionine (SAM) dependent methyltransferase enzymes able to catalyse the cyclopropanation of unsaturated phospholipids. Since CFAS enzymes employ SAM as a methylene source to cyclopropanate alkene substrates, they have the potential to be mild and more sustainable biocatalysts for cyclopropanation transformations than current carbene based approaches. This work describes the characterisation of E. coli CFAS enzyme (ecCFAS) and its exploitation in the stereoselective biocatalytic synthesis of cyclopropyl lipids. ecCFAS was found to convert phosphatidylglycerol (PG) to methyl dihydrosterculate 1 from in up to 58% conversion and 73% ee and the absolute configuration (9S,10R) was established. Substrate tolerance of ecCFAS was found to be correlated with the electronic properties of phospholipid headgroups and for the first time ecCFAS was found to catalyse cyclopropanation of both phospholipid chains to form dicyclopropanated products. In addition, mutagenesis and in-silico experiments were carried out to identify the enzyme residues with key roles in catalysis and to provide structural insights into the lipid substrate preference of ecCFAS. Finally, the biocatalytic synthesis of methyl dihydrosterculate 1 and its deuterated analogue was also accomplished combining pure ecCFAS with the SAM regenerating AtHMT enzyme in presence of CH3I and CD3I. |
published_date |
2024-06-14T15:38:26Z |
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1803025495467491328 |
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11.017776 |