Characterization of the caprolactam degradation pathway in Pseudomonas jessenii using mass spectrometry-based proteomics
| Autor: | Marleen Otzen, Dick B. Janssen, Cyntia M. Palacio |
|---|---|
| Přispěvatelé: | Biotechnology |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Proteomics Omega aminostransferase Pseudomonas jessenii medicine.disease_cause Applied Microbiology and Biotechnology 03 medical and health sciences chemistry.chemical_compound Environmental Biotechnology Pseudomonas Gene cluster medicine Escherichia coli Omega aminotransferase Caprolactam Peptide sequence chemistry.chemical_classification biology Mass spectrometry General Medicine Metabolism biology.organism_classification 030104 developmental biology Enzyme chemistry Biochemistry Multigene Family Caprolactamase Biotechnology |
| Zdroj: | Applied Microbiology and Biotechnology Applied Microbiology and Biotechnology, 15, 6699-6711. SPRINGER |
| ISSN: | 1432-0614 0175-7598 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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