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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...

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Autores principales: Otzen, Marleen, Palacio, Cyntia, Janssen, Dick B.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2018
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.
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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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AT janssendickb characterizationofthecaprolactamdegradationpathwayinpseudomonasjesseniiusingmassspectrometrybasedproteomics