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Liquid Chromatography and Mass Spectrometry Analysis of Isoprenoid Intermediates in Escherichia coli

  • Edward E. K. BaidooEmail author
  • George Wang
  • Chijioke J. Joshua
  • Veronica Teixeira Benites
  • Jay D. Keasling
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1859)

Abstract

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.

Key words

Isoprenoid intermediates Metabolomics LC-MS HILIC Mevalonate Deoxy-d-xylulose-5-phosphate 

Notes

Acknowledgments

The authors would like to acknowledge that this work was part of the DOE Joint BioEnergy Institute (http://www.jbei.org) supported by the US Department of Energy, Office of Science, Office of Biological and Environmental Research, through contract DE-AC02-05CH11231 between Lawrence Berkeley National Laboratory and the US Department of Energy.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Edward E. K. Baidoo
    • 1
    • 2
    Email author
  • George Wang
    • 1
    • 2
  • Chijioke J. Joshua
    • 1
    • 2
  • Veronica Teixeira Benites
    • 1
    • 2
  • Jay D. Keasling
    • 1
    • 2
    • 3
    • 4
    • 5
    • 6
  1. 1.Biological Systems and Engineering DivisionLawrence Berkeley National LaboratoryBerkeleyUSA
  2. 2.Joint BioEnergy InstituteEmeryvilleUSA
  3. 3.Department of Chemical EngineeringUniversity of CaliforniaBerkeleyUSA
  4. 4.Department of BioengineeringUniversity of CaliforniaBerkeleyUSA
  5. 5.Novo Nordisk Foundation Center for BiosustainabilityTechnical University of DenmarkKgs LyngbyDenmark
  6. 6.Center for Synthetic Biochemistry, Institute of Synthetic Biology ResearchShenzhen Institutes of Advanced TechnologiesShenzhenChina

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