Novel comprehensive multidimensional liquid chromatography approach for elucidation of the microbosphere of shikimate-producing Escherichia coli SP1.1/pKD15.071 strain
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Shikimic acid is a intermediate of aromatic amino acid biosynthesis and the preferred starting material for production of the most commonly prescribed anti-influenza drug, Tamiflu. Its six-membered carbocyclic ring is adorned with several chiral centers and various functionalities, making shikimic acid a valuable chiral synthon. When microbially-produced, in addition to shikimic acid, numerous other metabolites are exported out of the cytoplasm and accumulate in the culture medium. This extracellular matrix of metabolites is referred to as the microbosphere. Due to the high sample complexity, in this study, the microbosphere of shikimate-producing Escherichia coli SP1.1/pKD15.071 was analyzed by liquid chromatography and comprehensive two-dimensional liquid chromatography coupled to photodiode array and mass spectrometry detection. GC analysis of the trimethylsilyl derivatives was also carried out in order to support the elucidation of the selected metabolites in the microbosphere. The elucidation of the metabolic fraction of this bacterial strain might be of valid aid for improving, through genetic changes, the concentration and yield of shikimic acid synthesized from glucose.
KeywordsShikimic acid Metabolites Comprehensive two-dimensional liquid chromatography Microbosphere
The authors would like to thank Prof. John Frost and Dr. Karen Draths (Michigan State University, Lansing, MI, USA) for supplying the samples and helpful discussions.
The authors wish to thank the “University of Messina” for support through the “Research and Mobility” Project.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
- 1.Haslam E. Shikimic acid: metabolism and metabolites. New York: Wiley; 1993.Google Scholar
- 3.Kim CU, Lew W, Williams MA, Liu H, Zhang L, Swaminathan S, et al. Influenza neuraminidase inhibitors possessing a novel hydrophobic interaction in the enzyme active site: design, synthesis, and structural analysis of carbocyclic sialic acid analogues with potent anti-influenza activity. J Am Chem Soc. 1997;119:681–90.CrossRefPubMedGoogle Scholar
- 11.Cortes HJ. Multidimensional chromatography. Techniques and applications. New York: Marcel Dekker; 1990.Google Scholar
- 12.Mondello L, Lewis AC, Bartle KD. Multidimensional chromatography. New York: Wiley; 2002.Google Scholar
- 17.Julka S, Cortes HJ, Harfmann R, Bell B, Schweizer-Theobaldt A, Pursch M, et al. Quantitative characterization of solid epoxy resins using comprehensive two dimensional liquid chromatography coupled with electrospray ionization-time of flight mass spectrometry. Anal Chem. 2009;81:4271–9.CrossRefPubMedGoogle Scholar
- 25.Beccaria M, Costa R, Sullini G, Grasso E, Cacciola F, Dugo P, et al. Determination of the triacylglycerol fraction in fish oil by comprehensive liquid chromatography techniques with the support of gas chromatography and mass spectrometry data. Anal Bioanal Chem. 2015;407:5211–25.CrossRefPubMedGoogle Scholar
- 27.Montero L, Sánchez-Camargo AP, García-Canas V, Tanniou A, Stiger-Pouvreau V, Russo M, et al. Anti-proliferative activity and chemical characterization by comprehensive two-dimensional liquid chromatography coupled to mass spectrometry of phlorotannins from the brown macroalga Sargassum muticum collected on North-Atlantic coasts. J Chromatogr A. 2016;1428:115–25.CrossRefPubMedGoogle Scholar
- 28.Cacciola F, Giuffrida D, Utczas M, Mangraviti D, Beccaria M, Donato P, et al. Analysis of the carotenoid composition and stability in various overripe fruits by comprehensive two-dimensional liquid chromatography. LC-GC Eur. 2016;29:252–6.Google Scholar
- 33.Ouyang X, Leonards PEG, Tousova Z, Slobodnik J, de Boer J, Lamoree MH. Rapid screening of acetylcholinesterase inhibitors by effect-directed analysis using LC×LC fractionation, a high throughput in vitro assay, and parallel identification by time of flight mass spectrometry. Anal Chem. 2016;88:2353–60.CrossRefPubMedGoogle Scholar
- 35.Horsley LH, editor. Azeotropic data—III. Advances in chemistry series no. 166. Washington: American Chemical Society; 1973.Google Scholar
- 36.Moldoveanu SC, David V. Sample preparation in chromatography. J Chromatogr Libr. 2002;65:493–8.Google Scholar
- 38.Fekete S, Fekete J, Molnár I, Ganzler K. Rapid high performance liquid chromatography method development with high prediction accuracy, using 5 cm long narrow bore columns packed with sub 2 μm particles and design space computer modeling. J Chromatogr A. 2009;1216:7816–23.CrossRefPubMedGoogle Scholar