Liquid chromatography and mass spectrometry analysis of isoprenoid intermediates in Escherichia coli

Edward E.K. Baidoo, George Wang, Chijioke J. Joshua, Veronica Teixeira Benites, Jay D. Keasling

Research output: Chapter in Book/Report/Conference proceedingBook chapterResearchpeer-review


Isoprenoids are a highly diverse group of natural products with broad application as high value chemicals and advanced biofuels. They are synthesized using two primary building blocks, namely, isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) that are generated via the mevalonate (MVA) or deoxy-d-xylulose-5-phosphate (DXP) pathways. Isoprenoid biosynthetic pathways are prevalent in eukaryotes, archaea, and bacteria. Measurement of isoprenoid intermediates via standard liquid chromatography–mass spectrometry (LC-MS) protocols is generally challenging because of the hydrophilicity and complex physicochemical properties of the molecules. In addition, there is currently no reliable analytical method that can simultaneously measure metabolic intermediates from MVA and DXP pathways, including the prenyl diphosphates. Therefore, we describe a robust hydrophilic interaction liquid chromatography time-of-flight mass spectrometry (HILIC-TOF-MS) method for analyzing isoprenoid intermediates from metabolically engineered Escherichia coli strains.
Original languageEnglish
Title of host publicationMicrobial Metabolomics
EditorsEdward E.K. Baidoo
Place of PublicationNew York
PublisherHumana Press
Publication date2019
ISBN (Print)978-1-4939-8756-6
ISBN (Electronic)978-1-4939-8757-3
Publication statusPublished - 2019
SeriesMethods in Molecular Biology


  • Isoprenoid intermediates
  • LC-MS
  • Metabolomics
  • Mevalonate
  • Deoxy-D-xylulose-5-phosphate

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