PRIDE Assigned Tags:Biological Dataset
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Proteome analysis of B. bassiana
The enzymes involved (P450s) remain mostly hidden when using microbial cell factories. In this respect, we aimed to identify P450s, which are responsible for the oxyfunctionalization of DCF (1) and (R)-2-phenoxypropionic acid (POPS) in B. bassiana using omics studies.
Sample Processing Protocol
A 3-day PEG broth preculture was used to inoculate 100 mL PEG broth with 1/75 volume of preculture. The biotransformations were started after 24 h incubation with the addition of 0.5 g L-1 of the substrates or glucose as a reference. The reactions were stopped after first products were detected (48 h for (R)-2-phenoxypropionic acid (POPS) and 68 h for diclofenac (1)). The cells were then lysed as described above and the proteins of the samples analyzed and diluted as a whole cell suspension on a 10% and a 15% SDS-polyacrylamide gel.The proteome analysis was carried out at the mass spectrometry service unit in Hohenheim (Germany, group of Dr. Pfannstiel). For this purpose, the proteins were isolated from the SDS gels, trypsin-digested and the resulting peptide fragments were analyzed by QTRAP LC-MS / MS.
Data Processing Protocol
Data processing was done using MASCOT software. Identification of the proteins was based on a global comparison with the protein database NCBI.
Klenk JM, Kontny LH, Escobedo-Hinojosa W, Nebel BA, Hauer B. Oxyfunctionalization of non-steroidal anti-inflammatory drugs by filamentous-fungi. J Appl Microbiol. 2019 PubMed: 31173436