Improved 13C metabolic flux analysis in Escherichia coli metabolism: application of a high resolution MS (GC-EI-QTOF) for comprehensive assessment MS/MS fragments

J Ind Microbiol Biotechnol. 2023 Nov 13:kuad039. doi: 10.1093/jimb/kuad039. Online ahead of print.ABSTRACTGas chromatography-tandem mass spectrometry with electron ionization (GC-EI-MS/MS) provides rich information on stable-isotope labeling for 13C-metabolic flux analysis (13C-MFA). To pave the way for the routine application of tandem MS data for metabolic flux quantification, we aimed to compile a comprehensive library of GC-EI-MS/MS fragments of tert-butyldimethylsilyl (TBDMS) derivatized proteinogenic amino acids. First, we established an analytical workflow which combines high resolution gas chromatography-quadrupole time-of-flight mass spectrometry (GC-EI-QTOFMS) and fully 13C-labeled biomass to identify and structurally elucidate tandem MS amino acid fragments. Application of the high-mass accuracy MS procedure resulted into the identification of 129 validated precursor-product ion pairs of 13 amino acids with 30 fragments being accepted for 13C-MFA. The practical benefit of the novel tandem MS data was demonstrated by a proof-of-concept study which confirmed the importance of the compiled library for high resolution 13C-MFA.PMID:37960978 | DOI:10.1093/jimb/kuad039
Source: Journal of Industrial Microbiology and Biotechnology - Category: Microbiology Authors: Source Type: research