T. pseudoethanolicus response to furfural stress - LC-MS/MS
Identification of the furfural detoxifying machinery in T. pseudoethanolicus (Tps) by LC-MS/MS based proteomics for engineering cellulolytic microbes involved in bioethanol production.
Sample Processing Protocol
Tps cells were grown in biological triplicate, proteins extracted via SDS/TCA/Urea modified FASP, and analyzed by 24-hr MudPIT LC-MS. Orbitrap XL was operated in DDA using a high mass accuracy in MS1 scans and LTQ in MS2 scans.
Data Processing Protocol
Spectra assiged to peptide sequences via SEQUEST/DTASelect. Spectra counts were measured, assigned nSpC values, and imported into JMP genomics to assign significant changers across conditions via ANOVA. Semiquantitative analysis with replication.
Clarkson SM, Hamilton-Brehm SD, Giannone RJ, Engle NL, Tschaplinski TJ, Hettich RL, Elkins JG. A comparative multidimensional LC-MS proteomic analysis reveals mechanisms for furan aldehyde detoxification in Thermoanaerobacter pseudethanolicus 39E. Biotechnol Biofuels. 2014 7(1):165 PubMed: 25506391