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Crystal structure and site-directed mutagenesis of circular bacteriocin plantacyclin B21AG reveals cationic and aromatic residues important for antimicrobial activity
Plantacyclin B21AG is a circular bacteriocin produced by Lactiplantibacillus plantarum B21 which displays antimicrobial activity against various Gram-positive bacteria including foodborne pathogens, Listeria monocytogenes and Clostridium perfringens. It is a 58-amino acid cyclised antimicrobial pept...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Nature Publishing Group UK
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7562740/ https://www.ncbi.nlm.nih.gov/pubmed/33060678 http://dx.doi.org/10.1038/s41598-020-74332-1 |
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author | Gor, Mian-Chee Vezina, Ben McMahon, Róisín M. King, Gordon J. Panjikar, Santosh Rehm, Bernd H. A. Martin, Jennifer L. Smith, Andrew T. |
author_facet | Gor, Mian-Chee Vezina, Ben McMahon, Róisín M. King, Gordon J. Panjikar, Santosh Rehm, Bernd H. A. Martin, Jennifer L. Smith, Andrew T. |
author_sort | Gor, Mian-Chee |
collection | PubMed |
description | Plantacyclin B21AG is a circular bacteriocin produced by Lactiplantibacillus plantarum B21 which displays antimicrobial activity against various Gram-positive bacteria including foodborne pathogens, Listeria monocytogenes and Clostridium perfringens. It is a 58-amino acid cyclised antimicrobial peptide, with the N and C termini covalently linked together. The circular peptide backbone contributes to remarkable stability, conferring partial proteolytic resistance and structural integrity under a wide temperature and pH range. Here, we report the first crystal structure of a circular bacteriocin from a food grade Lactobacillus. The protein was crystallised using the hanging drop vapour diffusion method and the structure solved to a resolution of 1.8 Å. Sequence alignment against 18 previously characterised circular bacteriocins revealed the presence of conserved charged and aromatic residues. Alanine substitution mutagenesis validated the importance of these residues. Minimum inhibitory concentration analysis of these Ala mutants showed that Phe(8)Ala and Trp(45)Ala mutants displayed a 48- and 32-fold reduction in activity, compared to wild type. The Lys(19)Ala mutant displayed the weakest activity, with a 128-fold reduction. These experiments demonstrate the relative importance of aromatic and cationic residues for the antimicrobial activity of plantacyclin B21AG and by extension, other circular bacteriocins sharing these evolutionarily conserved residues. |
format | Online Article Text |
id | pubmed-7562740 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-75627402020-10-19 Crystal structure and site-directed mutagenesis of circular bacteriocin plantacyclin B21AG reveals cationic and aromatic residues important for antimicrobial activity Gor, Mian-Chee Vezina, Ben McMahon, Róisín M. King, Gordon J. Panjikar, Santosh Rehm, Bernd H. A. Martin, Jennifer L. Smith, Andrew T. Sci Rep Article Plantacyclin B21AG is a circular bacteriocin produced by Lactiplantibacillus plantarum B21 which displays antimicrobial activity against various Gram-positive bacteria including foodborne pathogens, Listeria monocytogenes and Clostridium perfringens. It is a 58-amino acid cyclised antimicrobial peptide, with the N and C termini covalently linked together. The circular peptide backbone contributes to remarkable stability, conferring partial proteolytic resistance and structural integrity under a wide temperature and pH range. Here, we report the first crystal structure of a circular bacteriocin from a food grade Lactobacillus. The protein was crystallised using the hanging drop vapour diffusion method and the structure solved to a resolution of 1.8 Å. Sequence alignment against 18 previously characterised circular bacteriocins revealed the presence of conserved charged and aromatic residues. Alanine substitution mutagenesis validated the importance of these residues. Minimum inhibitory concentration analysis of these Ala mutants showed that Phe(8)Ala and Trp(45)Ala mutants displayed a 48- and 32-fold reduction in activity, compared to wild type. The Lys(19)Ala mutant displayed the weakest activity, with a 128-fold reduction. These experiments demonstrate the relative importance of aromatic and cationic residues for the antimicrobial activity of plantacyclin B21AG and by extension, other circular bacteriocins sharing these evolutionarily conserved residues. Nature Publishing Group UK 2020-10-15 /pmc/articles/PMC7562740/ /pubmed/33060678 http://dx.doi.org/10.1038/s41598-020-74332-1 Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Gor, Mian-Chee Vezina, Ben McMahon, Róisín M. King, Gordon J. Panjikar, Santosh Rehm, Bernd H. A. Martin, Jennifer L. Smith, Andrew T. Crystal structure and site-directed mutagenesis of circular bacteriocin plantacyclin B21AG reveals cationic and aromatic residues important for antimicrobial activity |
title | Crystal structure and site-directed mutagenesis of circular bacteriocin plantacyclin B21AG reveals cationic and aromatic residues important for antimicrobial activity |
title_full | Crystal structure and site-directed mutagenesis of circular bacteriocin plantacyclin B21AG reveals cationic and aromatic residues important for antimicrobial activity |
title_fullStr | Crystal structure and site-directed mutagenesis of circular bacteriocin plantacyclin B21AG reveals cationic and aromatic residues important for antimicrobial activity |
title_full_unstemmed | Crystal structure and site-directed mutagenesis of circular bacteriocin plantacyclin B21AG reveals cationic and aromatic residues important for antimicrobial activity |
title_short | Crystal structure and site-directed mutagenesis of circular bacteriocin plantacyclin B21AG reveals cationic and aromatic residues important for antimicrobial activity |
title_sort | crystal structure and site-directed mutagenesis of circular bacteriocin plantacyclin b21ag reveals cationic and aromatic residues important for antimicrobial activity |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7562740/ https://www.ncbi.nlm.nih.gov/pubmed/33060678 http://dx.doi.org/10.1038/s41598-020-74332-1 |
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