Project PXD009664

PRIDE Assigned Tags:
Biological Dataset
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Summary

Title

Proteome analysis of B. bassiana

Description

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.

Contact

Jan Klenk, JK
Bernhard Hauer, Institute of Biochemistry and Technical Biochemistry, University of Stuttgart, Germany ( lab head )

Submission Date

04/05/2018

Publication Date

12/06/2019

Instrument

LTQ Orbitrap Elite

Software

Not available

Modification

Carbamidomethyl
Oxidation

Quantification

Not available

Experiment Type

Gel-based experiment

Assay count

3

Publication

    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

Assay

Showing 1 - 3 of 3 results
# Accession Title Proteins Peptides Unique Peptides Spectra Identified Spectra View in Reactome
1 91721 BB_Dic (F018288).mzid 3318 14469 4495 6572 0
2 91722 BB_Glk (F018289).mzid 3465 21550 7219 9802 0
3 91720 BB_Phen (F018292).mzid 3219 18079 5765 8156 0