Isolation and purification method of low molecular weight peptides from bee products using chromatographic methods

Sergey V.  Klychenkov; Anastasiya D.  Kruchinina

doi:10.17308/sorpchrom.2023.23/10998

Sergey V. Klychenkov Penza State University, Penza, Russia https://orcid.org/0000-0003-4748-1334
Anastasiya D. Kruchinina Penza State University, Penza, Russia https://orcid.org/0000-0002-2620-527X

DOI: https://doi.org/10.17308/sorpchrom.2023.23/10998

Keywords: royal jelly peptides, drone brood peptides, bee products, royal jelly, drone brood.

Abstract

The study of the biological activity of various peptide substances is one of the modern research areas of biochemistry. Over the past few decades, a large number of reports in the scientific literature about the presence of a certain effect of bee products on various biological systems were published, while the number of peptides revealed in royal jelly and drone brood is increasing every year. This resulted in the need to develop a convenient and productive method for the isolation and purification of the peptide fraction from bee products, which was the goal of this study. As a result, the method for isolation of the fraction of low molecular weight peptides (with the molecular weight less than 5 kDa) from royal jelly and drone brood was developed and tested. This method includes ultrafiltration for the separation of the solution molecules according to the molecular weight parameter, ion-exchange chromatography on DEAE-cellulose for purification of peptides from low molecular weight impurities and gel filtration on Sephadex G-25 for the removal salts of buffer solutions. The proposed method for the isolation and purification of peptides can be easily scaled and automated, which allows the use of the proposed method under industrial conditions. The control of the peptide composition in comparison of the original filtrate and the purified fraction was carried out using high performance liquid chromatography. As a result of using this method, an acceptable degree of purification is achieved, and the purified peptides were obtained in an aqueous solution suitable both for further assessment of biological activity and for the investigation of the composition by various physicochemical methods. Initial data on the physicochemical properties of purified peptides were also obtained: Five out of nine groups of royal jelly peptides and four out of eleven groups of peptides in the composition of drone brood contained amino acids with aromatic radicals; at pH=10, majority of royal jelly peptides were anionic, and majority of drone brood peptides were cationic.

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Author Biographies

Sergey V. Klychenkov, Penza State University, Penza, Russia

senior lecturer, dep. of General Biology and Biochemistry, Penza State University, Pedagogical Institute named afer V. G. Belinsky, Penza, Russia, e-mail 79048510599@ya.ru

Anastasiya D. Kruchinina, Penza State University, Penza, Russia

Associate Professor, Department of General Biology and Biochemistry, Ph.D., Penza State University, Pedagogical Institute named after V. G. Belinsky, Penza, Russia, e-mail a.d.kruchinina@mail.ru

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