Project description:Several soil organisms are known to be capable of growth on caprolactam as soil carbon and nitrogen source, but the enzymes of the catabolic pathway have not been described. We isolated a caprolactam-degrading strain of Pseudomonas jessenni, and identified genes putatively involved in the 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 likely to be involved in later steps of the pathway. The identified caprolactamase consisted of 2 subunits and demonstrated high sequence identity to the 5-oxoprolinases. E. coli expressing this caprolactamase did not convert 5-oxoproline but was able to hydrolyze caprolactam to form 6-aminohexanoic acid in an ATP dependent manner. Characterization of the aminotransferase revealed that the enzyme deaminated 6-aminohexanoic acid to produce 6-oxohexanoate with pyruvate as amino acceptor. The amino acid sequence of the aminotransferase demonstrated high similarity to subgroup II ω-aminotransferases of the PLP fold type I proteins. Finally, analyses of the genome sequence demonstrated the presence of a caprolactam catabolism gene cluster consisting of all genes involved in the conversion of caprolactam to adipate.
Project description:ErfA is a transcription factor of Pseudomonas aeruginosa. We here define the genome-wide binding sites of ErfA by DAP-seq in Pseudomonas aeruginosa PAO1 and IHMA87, Pseudomonas chlororaphis PA23, Pseudomonas protegens CHA0 and Pseudomonas putida KT2440.
