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Characterization of the caprolactam degradation pathway in Pseudomonas jessenii using mass spectrometry-based proteomics
Some bacterial cultures are capable of growth on caprolactam as sole carbon and nitrogen source, but the enzymes of the catabolic pathway have not been described. We isolated a caprolactam-degrading strain of Pseudomonas jessenii from soil and identified proteins and genes putatively involved in cap...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Berlin Heidelberg
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6061476/ https://www.ncbi.nlm.nih.gov/pubmed/29850960 http://dx.doi.org/10.1007/s00253-018-9073-7 |
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author | Otzen, Marleen Palacio, Cyntia Janssen, Dick B. |
author_facet | Otzen, Marleen Palacio, Cyntia Janssen, Dick B. |
author_sort | Otzen, Marleen |
collection | PubMed |
description | Some bacterial cultures are capable of growth on caprolactam as sole carbon and nitrogen source, but the enzymes of the catabolic pathway have not been described. We isolated a caprolactam-degrading strain of Pseudomonas jessenii from soil and identified proteins and genes putatively involved in caprolactam metabolism using quantitative mass spectrometry-based proteomics. This led to the discovery of a caprolactamase and an aminotransferase that are involved in the initial steps of caprolactam conversion. Additionally, various proteins were identified that likely are involved in later steps of the pathway. The caprolactamase consists of two subunits and demonstrated high sequence identity to the 5-oxoprolinases. Escherichia coli cells expressing this caprolactamase did not convert 5-oxoproline but were able to hydrolyze caprolactam to form 6-aminocaproic acid in an ATP-dependent manner. Characterization of the aminotransferase revealed that the enzyme deaminates 6-aminocaproic acid to produce 6-oxohexanoate with pyruvate as amino acceptor. The amino acid sequence of the aminotransferase showed high similarity to subgroup II ω-aminotransferases of the PLP-fold type I proteins. Finally, analyses of the genome sequence revealed the presence of a caprolactam catabolism gene cluster comprising a set of genes involved in the conversion of caprolactam to adipate. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00253-018-9073-7) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-6061476 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Springer Berlin Heidelberg |
record_format | MEDLINE/PubMed |
spelling | pubmed-60614762018-08-09 Characterization of the caprolactam degradation pathway in Pseudomonas jessenii using mass spectrometry-based proteomics Otzen, Marleen Palacio, Cyntia Janssen, Dick B. Appl Microbiol Biotechnol Environmental Biotechnology Some bacterial cultures are capable of growth on caprolactam as sole carbon and nitrogen source, but the enzymes of the catabolic pathway have not been described. We isolated a caprolactam-degrading strain of Pseudomonas jessenii from soil and identified proteins and genes putatively involved in caprolactam metabolism using quantitative mass spectrometry-based proteomics. This led to the discovery of a caprolactamase and an aminotransferase that are involved in the initial steps of caprolactam conversion. Additionally, various proteins were identified that likely are involved in later steps of the pathway. The caprolactamase consists of two subunits and demonstrated high sequence identity to the 5-oxoprolinases. Escherichia coli cells expressing this caprolactamase did not convert 5-oxoproline but were able to hydrolyze caprolactam to form 6-aminocaproic acid in an ATP-dependent manner. Characterization of the aminotransferase revealed that the enzyme deaminates 6-aminocaproic acid to produce 6-oxohexanoate with pyruvate as amino acceptor. The amino acid sequence of the aminotransferase showed high similarity to subgroup II ω-aminotransferases of the PLP-fold type I proteins. Finally, analyses of the genome sequence revealed the presence of a caprolactam catabolism gene cluster comprising a set of genes involved in the conversion of caprolactam to adipate. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00253-018-9073-7) contains supplementary material, which is available to authorized users. Springer Berlin Heidelberg 2018-05-31 2018 /pmc/articles/PMC6061476/ /pubmed/29850960 http://dx.doi.org/10.1007/s00253-018-9073-7 Text en © The Author(s) 2018 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. |
spellingShingle | Environmental Biotechnology Otzen, Marleen Palacio, Cyntia Janssen, Dick B. Characterization of the caprolactam degradation pathway in Pseudomonas jessenii using mass spectrometry-based proteomics |
title | Characterization of the caprolactam degradation pathway in Pseudomonas jessenii using mass spectrometry-based proteomics |
title_full | Characterization of the caprolactam degradation pathway in Pseudomonas jessenii using mass spectrometry-based proteomics |
title_fullStr | Characterization of the caprolactam degradation pathway in Pseudomonas jessenii using mass spectrometry-based proteomics |
title_full_unstemmed | Characterization of the caprolactam degradation pathway in Pseudomonas jessenii using mass spectrometry-based proteomics |
title_short | Characterization of the caprolactam degradation pathway in Pseudomonas jessenii using mass spectrometry-based proteomics |
title_sort | characterization of the caprolactam degradation pathway in pseudomonas jessenii using mass spectrometry-based proteomics |
topic | Environmental Biotechnology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6061476/ https://www.ncbi.nlm.nih.gov/pubmed/29850960 http://dx.doi.org/10.1007/s00253-018-9073-7 |
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