Application of the Chitooligosaccharides and Fluorescence Polarization Technique for the Assay of Active Lysozyme in Hen Egg White

Application of the Chitooligosaccharides and Fluorescence Polarization Technique for the Assay of Active Lysozyme in Hen Egg White Научная публикация

Журнал

Biomolecules
ISSN: 2218-273X

Вых. Данные

Год: 2024, Том: 14, Номер: 12, Номер статьи : 1589, Страниц : DOI: 10.3390/biom14121589

Авторы

Mukhametova Liliya I. ¹ , Zherdev Dmitry O. ¹ , Eremin Sergei A. ¹ , Levashov Pavel A. ¹ , Siebert Hans-Christian ² , Tsvetkov Yury E. ³ , Yudina Olga N. ³ , Krylov Vadim B. ⁴ , Nifantiev Nikolay E. ³

Организации

1	Faculty of Chemistry, M.V. Lomonosov Moscow State University, Leninsky Gory 1/3, 119991 Moscow, Russia
2	RI-B-NT—Research Institute of Bioinformatics and Nanotechnology, Schauenburger Str. 116, 24118 Kiel, Germany
3	Laboratory of Glycoconjugate Chemistry, N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Leninsky Prospect 47, 119991 Moscow, Russia
4	Laboratory of Synthetic Glycovaccines, N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Leninsky Prospect 47, 119991 Moscow, Russia

This study describes the applicability of the fluorescence polarization assay (FPA) based on the use of FITC-labeled oligosaccharide tracers of defined structure for the measurement of active lysozyme in hen egg white. Depending on the oligosaccharide chain length of the tracer, this method detects both the formation of the enzyme-to-tracer complex (because of lectin-like, i.e., carbohydrate-binding action of lysozyme) and tracer splitting (because of chitinase activity of lysozyme). Evaluation of the fluorescence polarization dynamics enables simultaneous measurement of the chitinase and lectin activities of lysozyme, which is crucial for its detection in complex biological systems. Hen egg white lysozyme (HEWL), unlike human lysozyme (HL), formed a stable complex with the chitotriose tracer that underwent no further transformations. This fact allows for easy measurement of the carbohydrate-binding activity of the HEWL. The results of the lysozyme activity measurement for hen egg samples obtained through the FPA correlated with the results obtained using the traditional turbidimetry method. The FPA does not have the drawbacks of turbidimetry, which are associated with the need to use bacterial cells that cannot be precisely standardized. Additionally, FPA offers advantages such as rapid analysis, the use of compact equipment, and standardized reagents. Therefore, the new express technique for measuring the lysozyme activity is applicable for evaluating the complex biomaterial, including for the purposes of food product quality control.

Mukhametova L.I. , Zherdev D.O. , Eremin S.A. , Levashov P.A. , Siebert H-C. , Tsvetkov Y.E. , Yudina O.N. , Krylov V.B. , Nifantiev N.E.
Application of the Chitooligosaccharides and Fluorescence Polarization Technique for the Assay of Active Lysozyme in Hen Egg White
Biomolecules. 2024. V.14. N12. 1589 . DOI: 10.3390/biom14121589 WOS Scopus OpenAlex

≡ Web of science:	WOS:001387128300001
≡ Scopus:	2-s2.0-85213204911
≡ OpenAlex:	W4405324821