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Characterization of Cereulide Synthetase, a Toxin-Producing Macromolecular Machine
Cereulide synthetase is a two-protein nonribosomal peptide synthetase system that produces a potent emetic toxin in virulent strains of Bacillus cereus. The toxin cereulide is a depsipeptide, as it consists of alternating aminoacyl and hydroxyacyl residues. The hydroxyacyl residues are derived from...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4455996/ https://www.ncbi.nlm.nih.gov/pubmed/26042597 http://dx.doi.org/10.1371/journal.pone.0128569 |
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author | Alonzo, Diego A. Magarvey, Nathan A. Schmeing, T. Martin |
author_facet | Alonzo, Diego A. Magarvey, Nathan A. Schmeing, T. Martin |
author_sort | Alonzo, Diego A. |
collection | PubMed |
description | Cereulide synthetase is a two-protein nonribosomal peptide synthetase system that produces a potent emetic toxin in virulent strains of Bacillus cereus. The toxin cereulide is a depsipeptide, as it consists of alternating aminoacyl and hydroxyacyl residues. The hydroxyacyl residues are derived from keto acid substrates, which cereulide synthetase selects and stereospecifically reduces with imbedded ketoreductase domains before incorporating them into the growing depsipeptide chain. We present an in vitro biochemical characterization of cereulide synthetase. We investigate the kinetics and side chain specificity of α-keto acid selection, evaluate the requirement of an MbtH-like protein for adenylation domain activity, assay the effectiveness of vinylsulfonamide inhibitors on ester-adding modules, perform NADPH turnover experiments and evaluate in vitro depsipeptide biosynthesis. This work also provides biochemical insight into depsipeptide-synthesizing nonribosomal peptide synthetases responsible for other bioactive molecules such as valinomycin, antimycin and kutzneride. |
format | Online Article Text |
id | pubmed-4455996 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-44559962015-06-09 Characterization of Cereulide Synthetase, a Toxin-Producing Macromolecular Machine Alonzo, Diego A. Magarvey, Nathan A. Schmeing, T. Martin PLoS One Research Article Cereulide synthetase is a two-protein nonribosomal peptide synthetase system that produces a potent emetic toxin in virulent strains of Bacillus cereus. The toxin cereulide is a depsipeptide, as it consists of alternating aminoacyl and hydroxyacyl residues. The hydroxyacyl residues are derived from keto acid substrates, which cereulide synthetase selects and stereospecifically reduces with imbedded ketoreductase domains before incorporating them into the growing depsipeptide chain. We present an in vitro biochemical characterization of cereulide synthetase. We investigate the kinetics and side chain specificity of α-keto acid selection, evaluate the requirement of an MbtH-like protein for adenylation domain activity, assay the effectiveness of vinylsulfonamide inhibitors on ester-adding modules, perform NADPH turnover experiments and evaluate in vitro depsipeptide biosynthesis. This work also provides biochemical insight into depsipeptide-synthesizing nonribosomal peptide synthetases responsible for other bioactive molecules such as valinomycin, antimycin and kutzneride. Public Library of Science 2015-06-04 /pmc/articles/PMC4455996/ /pubmed/26042597 http://dx.doi.org/10.1371/journal.pone.0128569 Text en © 2015 Alonzo et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Alonzo, Diego A. Magarvey, Nathan A. Schmeing, T. Martin Characterization of Cereulide Synthetase, a Toxin-Producing Macromolecular Machine |
title | Characterization of Cereulide Synthetase, a Toxin-Producing Macromolecular Machine |
title_full | Characterization of Cereulide Synthetase, a Toxin-Producing Macromolecular Machine |
title_fullStr | Characterization of Cereulide Synthetase, a Toxin-Producing Macromolecular Machine |
title_full_unstemmed | Characterization of Cereulide Synthetase, a Toxin-Producing Macromolecular Machine |
title_short | Characterization of Cereulide Synthetase, a Toxin-Producing Macromolecular Machine |
title_sort | characterization of cereulide synthetase, a toxin-producing macromolecular machine |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4455996/ https://www.ncbi.nlm.nih.gov/pubmed/26042597 http://dx.doi.org/10.1371/journal.pone.0128569 |
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