Combined Approach of Chromatographic Fractionation and Raman Spectroscopy for Metabolite Profiling of Enterobacter spp. Supernatant

Combined Approach of Chromatographic Fractionation and Raman Spectroscopy for Metabolite Profiling of Enterobacter spp. Supernatant Full article

Journal

International Journal of Molecular Sciences
ISSN: 1661-6596 , E-ISSN: 1422-0067

Output data

Year: 2026, Volume: 27, Number: 3, Article number : 1564, Pages count : DOI: 10.3390/ijms27031564

Tags

ESKAPE pathogens; bacterial secretome; Raman spectroscopy; chromatographic fractionation; virulence metabolites

Authors

Denisova Elizaveta ^1,2 , Avdyusheva Anastasia ² , Vasilieva Vera ² , Tyshchuk Elizaveta ^1,2 , Grebenkina Polina ^1,2 , Korenevsky Andrey ^1,2 , Chelibanov Ivan ^1,3 , Chelibanov Vladimir ³ , Totolian Areg ¹ , Kraeva Lyudmila ^1,4 , Sokolov Dmitry ^1,2

Affiliations

1	Saint-Petersburg Pasteur Institute, St. Petersburg 197101, Russia
2	Research Institute of Obstetrics and Gynecology Named After D.O. Ott, St. Petersburg 199034, Russia
3	«OPTEK» Joint Stock Company, St. Petersburg 199178, Russia
4	Military Medical Academy Named After S.M. Kirov, 6 Akademika Lebedeva Street, Building 6, St. Petersburg 194044, Russia

Funding (1)

Инициативная работа

The secretome of ESKAPE pathogens contains numerous bioactive molecules that play a key role in pathogenesis and the formation of an immunosuppressive microenvironment. However, analyzing this complex chemical composition presents significant methodological challenges. In this study, we propose a combined approach integrating chromatographic fractionation of cell-free supernatants with Raman spectroscopy to deconstruct the secretome of the clinically relevant Gram-negative pathogen—Enterobacter spp. Chromatographic separation of the Enterobacter spp. supernatant into seven fractions reduced spectral congestion and enabled identification of fraction 3 as having a unique metabolite profile, enriched in peptides (including tryptophan- and tyrosine-containing structures), nucleic acids, polysaccharides, and putative glutathione-like compounds. Notably, fraction 3 lacked markers of phenylalanine and sterol-like lipids, highlighting its distinct composition. Compared to conventional mass spectrometry and nuclear magnetic resonance, our hybrid strategy offers minimal sample preparation, preserves sample integrity for repeated analysis, avoids ionization bias, and is fully compatible with aqueous biological matrices—critical advantages for profiling labile or low-abundance metabolites in native secretomes. These findings demonstrate that the combination of preparative chromatography and Raman spectroscopy effectively resolves complex bacterial secretomes and identifies fractions potentially carrying key virulence or signaling functions.

Denisova E. , Avdyusheva A. , Vasilieva V. , Tyshchuk E. , Grebenkina P. , Korenevsky A. , Chelibanov I. , Chelibanov V. , Totolian A. , Kraeva L. , Sokolov D.
Combined Approach of Chromatographic Fractionation and Raman Spectroscopy for Metabolite Profiling of Enterobacter spp. Supernatant
International Journal of Molecular Sciences. 2026. V.27. N3. 1564 . DOI: 10.3390/ijms27031564 РИНЦ

Submitted:	Jan 22, 2026
Accepted:	Feb 3, 2026
Published print:	Feb 5, 2026

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