Publication

Characterization of the caprolactam degradation pathway in Pseudomonas jessenii using mass spectrometry-based proteomics

Otzen, M., Palacio, C. & Janssen, D. B., Aug-2018, In : Applied Microbiology and Biotechnology. 15, p. 6699-6711 13 p.

Research output: Contribution to journalArticleAcademicpeer-review

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.

Original languageEnglish
Pages (from-to)6699-6711
Number of pages13
JournalApplied Microbiology and Biotechnology
Volume15
Early online date31-May-2018
Publication statusPublished - Aug-2018

    Keywords

  • Caprolactam, Caprolactamase, Omega aminostransferase, Mass spectrometry, proteomics, Pseudomonas jessenii

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