Acyl-chain elongation drives ketosynthase substrate selectivity in trans-acyltransferase polyketide synthases
Type I modular polyketide synthases (PKSs), responsible for the biosynthesis of many biologically active agents, possess a ketosynthase (KS) domain within each module to catalyze chain elongation. Acylation of the KS active site Cys residue is followed by transfer to malonyl-acyl carrier protein,...
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| Format: | Article |
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Wiley-VCH Verlag
2015
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| Online Access: | https://eprints.nottingham.ac.uk/39789/ |
| _version_ | 1848795913868279808 |
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| author | Jenner, Matthew Afonso, José Pedro Bailey, Hannah R. Frank, Sarah Kampa, Annette Piel, Jörn Oldfield, Neil J. |
| author_facet | Jenner, Matthew Afonso, José Pedro Bailey, Hannah R. Frank, Sarah Kampa, Annette Piel, Jörn Oldfield, Neil J. |
| author_sort | Jenner, Matthew |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | Type I modular polyketide synthases (PKSs), responsible for the biosynthesis of many biologically active agents, possess a ketosynthase (KS) domain within each module to catalyze chain elongation.
Acylation of the KS active site Cys residue is followed by transfer to malonyl-acyl carrier protein, yielding an extended β-ketoacyl chain. To date, the precise contribution of KS selectivity in controlling product fidelity has been unclear. We submitted six KS domains from the trans-acyl transferase PKSs to a mass spectrometry-basedelongation assay, and identified higher substrat selectivity in the elongating step than in preceding acylation. A close correspondence between observed KS selectivity and that predicted by phylogenetic analysis was seen. Our findings provide insights into the mechanism of KS selectivity in this important group of PKSs, can serve as guidance for engineering, and show that targeted mutagenesis can be used to expand the repertoire of acceptable substrates. |
| first_indexed | 2025-11-14T19:39:39Z |
| format | Article |
| id | nottingham-39789 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T19:39:39Z |
| publishDate | 2015 |
| publisher | Wiley-VCH Verlag |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-397892020-05-04T17:03:10Z https://eprints.nottingham.ac.uk/39789/ Acyl-chain elongation drives ketosynthase substrate selectivity in trans-acyltransferase polyketide synthases Jenner, Matthew Afonso, José Pedro Bailey, Hannah R. Frank, Sarah Kampa, Annette Piel, Jörn Oldfield, Neil J. Type I modular polyketide synthases (PKSs), responsible for the biosynthesis of many biologically active agents, possess a ketosynthase (KS) domain within each module to catalyze chain elongation. Acylation of the KS active site Cys residue is followed by transfer to malonyl-acyl carrier protein, yielding an extended β-ketoacyl chain. To date, the precise contribution of KS selectivity in controlling product fidelity has been unclear. We submitted six KS domains from the trans-acyl transferase PKSs to a mass spectrometry-basedelongation assay, and identified higher substrat selectivity in the elongating step than in preceding acylation. A close correspondence between observed KS selectivity and that predicted by phylogenetic analysis was seen. Our findings provide insights into the mechanism of KS selectivity in this important group of PKSs, can serve as guidance for engineering, and show that targeted mutagenesis can be used to expand the repertoire of acceptable substrates. Wiley-VCH Verlag 2015-02-02 Article PeerReviewed Jenner, Matthew, Afonso, José Pedro, Bailey, Hannah R., Frank, Sarah, Kampa, Annette, Piel, Jörn and Oldfield, Neil J. (2015) Acyl-chain elongation drives ketosynthase substrate selectivity in trans-acyltransferase polyketide synthases. Angewandte Chemie International Edition, 54 (6). pp. 1817-1821. ISSN 1433-7851 http://onlinelibrary.wiley.com/doi/10.1002/anie.201410219/abstract;jsessionid=5CB1491BBFD74477E18B7BE69BBAD0D9.f02t03 doi:10.1002/anie.201410219 doi:10.1002/anie.201410219 |
| spellingShingle | Jenner, Matthew Afonso, José Pedro Bailey, Hannah R. Frank, Sarah Kampa, Annette Piel, Jörn Oldfield, Neil J. Acyl-chain elongation drives ketosynthase substrate selectivity in trans-acyltransferase polyketide synthases |
| title | Acyl-chain elongation drives ketosynthase substrate selectivity in trans-acyltransferase polyketide synthases |
| title_full | Acyl-chain elongation drives ketosynthase substrate selectivity in trans-acyltransferase polyketide synthases |
| title_fullStr | Acyl-chain elongation drives ketosynthase substrate selectivity in trans-acyltransferase polyketide synthases |
| title_full_unstemmed | Acyl-chain elongation drives ketosynthase substrate selectivity in trans-acyltransferase polyketide synthases |
| title_short | Acyl-chain elongation drives ketosynthase substrate selectivity in trans-acyltransferase polyketide synthases |
| title_sort | acyl-chain elongation drives ketosynthase substrate selectivity in trans-acyltransferase polyketide synthases |
| url | https://eprints.nottingham.ac.uk/39789/ https://eprints.nottingham.ac.uk/39789/ https://eprints.nottingham.ac.uk/39789/ |