2018 Springer Science+Business Media, LLC
Chemistry of Natural Compounds, Vol. 54, No. 3, May, 2018
ISOLATION AND CHARACTERIZATION OF ANTIMICROBIAL
PEPTIDES FROM Bos taurus BONE MARROW
U. Zh. Ishimov,
V. V. Maksimov,
and H. A. Aisa
Polypeptides from Bos taurus bone marrow (BBM) were purified for the first time using ultrafiltration and
two-stage ion-exchange chromatography. Extract of BBM contained 25 different polypeptides with molecular
masses in the range 1027.84–9916.32 Da. The water-soluble fraction of BBM contained large amounts of
charged polypeptides. The isolated peptides and proteins possessed dose-dependent antimicrobial activity
against pathogenic Candida albicans and Escherichia coli.
Keywords: bovine bone marrow, polypeptides, ion-exchange chromatography, electrophoresis in polyacrylamide
gel (PAAG), peptide charge, antimicrobial properties.
Drugs of animal origin are important components of Chinese traditional and folk medicine [1–12]. Currently, studies
of bone-marrow proteins and peptides are focused on treating diseases associated with it, e.g., its transplantation [7–9].
The compositions of proteins, lipids, carbohydrates, etc. in bovine bone marrow (BBM) were reported [10, 11]. It also
contained vitamins, various minerals, chondroitin, and mucopolysaccharides [2–4]. Chinese medicine reported hematopoietic
activity and anticoagulant properties for BBM and its use to quench thirst and treat leukorrhea . It is used as an additive to
herbal preparations, an analgesic and decongestant, and a treatment for damaged ligaments . In our opinion, the useful
properties of BBM polypeptides have not been studied enough . It is used mainly to feed animals . Therefore, it would
be interesting to expand the applications of inexpensive and ineffectively used BBM as a raw material for producing valuable
food products and drugs. According to the literature [5, 6], the protein–peptide composition of BBM is poorly studied.
Therefore, we determined the composition and biological properties of antimicrobial polypeptides from BBM of brown dairy
cows used in Xinjiang-Uyghur A. R., P. R. China.
The present work reports purification of the aqueous extract of BBM using precipitation by ammonium sulfate solution
of various degrees of saturation, ultrafiltration, and two-stage ion-exchange chromatography for studies of the protein–peptide
composition using PAAG electrophoresis and mass spectrometry. In addition, the antimicrobial activity of the isolated peptide
fractions was studied. Polypeptides were extracted from BBM by H
O and precipitated from the aqueous extract by 30, 50,
and 70% saturated (NH
solution. The polypeptide precipitates produced after each precipitation stage were separated
by centrifugation, dialyzed against distilled H
O, and lyophilized. Their yields and concentrations were determined. Fractions
BBM-30, BBM-50, and BBM-70 were produced in this manner and tested further for antimicrobial activity. The
isolated polypeptide fractions were separated according to acid–base properties by sequential ion-exchange
chromatography over columns of DEAE 23-Cellulose Servacel (DEAE-23) and TSKgel CM-650M Toyopearl (CM TSK).
1) Key Laboratory of Plants Resources and Chemistry of Arid Zone, Xinjiang Technical Institute of Physics
and Chemistry, Chinese Academy of Sciences, Beijing South Road 40-1, Urumqi 830011, Xinjiang, P. R. China,
e-mail: email@example.com; 2) Department of Nursing, Xinjiang Medical University, 830011, Urumqi, P. R. China,
Key Laboratory Basis of Xinjiang Indigenous Medicinal Plants Resource Utilization, Xinjiang Technical Institute of Physics
and Chemistry, Chinese Academy of Sciences, 830011, Urumqi, P. R. China; 3) University of Chinese Academy of Science,
100039, Beijing, P. R. China; 4) A. S. Sadykov Institute of Bioorganic Chemistry, Academy of Sciences of the Republic
of Uzbekistan, 100125, Tashkent, Uzbekistan. Translated from Khimiya Prirodnykh Soedinenii, No. 3, May–June, 2018,
pp. 446–449. Original article submitted November 10, 2017.