Details zur Publikation

Kategorie Textpublikation
Referenztyp Zeitschriften
DOI 10.1021/acs.analchem.9b00978
Volltext Akzeptiertes Manuskript
Titel (primär) Quantifying a biocatalytic product from a few living microbial cells using microfluidic cultivation coupled to FT-ICR-MS
Autor Dusny, C.; Lohse, M.; Reemtsma, T.; Schmid, A.; Lechtenfeld, O.J.
Journal / Serie Analytical Chemistry
Erscheinungsjahr 2019
Department ANA; SOMA
Band/Volume 91
Heft 11
Seite von 7012
Seite bis 7018
Sprache englisch
UFZ Querschnittsthemen ProVIS;
Abstract he in vivo quantification of metabolic products from microbial single cells is one of the last grand challenges in (bio)analytical chemistry. To date, no label-free analytical concept exists that is powerful enough to detect or even quantify the minute amounts of secreted low molecular weight compounds produced by living and isolated single bacteria or yeast cells. Coupling microfluidic cultivation systems with ultrahigh resolution electrospray-ionization mass spectrometry with its exquisite sensitivity and specificity offers the prospect of single-cell product analysis and quantification, but has not been successfully implemented yet. We report an analytical framework that interfaces noninvasive microfluidic trapping and cultivation of a few bacterial single cells with the analysis of their catalytic products by Fourier-transform ion cyclotron resonance mass spectrometry (FT-ICR-MS). Cell trapping was performed with the microfluidic Envirostat platform for cultivating bacterial cells under continuous perfusion via negative dielectrophoresis (nDEP). A total of 1.5 μL of product-containing cell supernatant was sampled into microcapillaries using a dead volume-reduced world-to-chip interface. The samples were analyzed with a nanoESI ion source coupled to a FT-ICR-MS (limit of detection for lysine: 0.5 pg). As a biocatalytic model system, we analyzed few Corynebacterium glutamicum DM 1919 pSenLys cells that synthesized l-lysine from d-glucose. Secreted lysine was quantified from a few cells (down to 19). Single-cell specific lysine productivities were 2 and 10 fmol/cell/h. This demonstrates that coupling microfluidics and mass spectrometry (SIC-MS) now enables the quantification of catalytic products and extracellular metabolites from only a few living microbial cells.
dauerhafte UFZ-Verlinkung
Dusny, C., Lohse, M., Reemtsma, T., Schmid, A., Lechtenfeld, O.J. (2019):
Quantifying a biocatalytic product from a few living microbial cells using microfluidic cultivation coupled to FT-ICR-MS
Anal. Chem. 91 (11), 7012 - 7018