蜜蜂蜂蜜及其肽段的抑菌活性

S. Klychenkov, A. Kruchinina, L. Bichurina
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摘要

近年来,细菌耐药性问题越来越紧迫。由于病原微生物获得耐药性,医疗实践中使用的抗生素不断过时,因此越来越需要寻找各种性质的新型抗菌药物。这一领域的一个有前途的研究方向是研究蛋白质分子的抗菌活性。在过去的二十年中,积累的大量数据表明,抗菌肽在几乎所有生物体内广泛分布。本文研究了荞麦和混合两种蜂蜜中质量小于5kda的肽段的抑菌活性,以及相同蜂蜜样品的不同溶液的抑菌活性。以下方法依次用于多肽组分的分离和纯化:超滤-获得含有质量小于5 kDa的分子的溶液;在deae -纤维素上进行离子交换层析,从低分子量杂质(碳水化合物、色素、酚类等)中纯化肽段,在Sephadex G-25上进行凝胶过滤,从缓冲溶液中脱盐得到肽段。采用纸片扩散法对慢性和急性各种细菌感染患者的金黄色葡萄球菌、大肠杆菌、铜绿假单胞菌和肠炎沙门氏菌耐药菌株进行抑菌活性测定。在所研究的样品中,测定了酮态中多肽、碳水化合物和过氧化氢的浓度。结果表明,在蜂蜜溶液中,荞麦具有最高的抗菌活性,这与过氧化氢含量的增加有关。肽组分对试验培养物的拮抗活性研究表明,从所研究的蜂蜜样品中获得的低分子量肽对所用的微生物菌株没有抗菌作用。显然,这是由于所获得的馏分中缺乏低分子量抗菌肽。蜜蜂蜂蜜中可能完全不存在重量小于5kda的抗菌肽。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
ANTIBACTERIAL ACTIVITY OF BEE HONEY AND ITS PEPTIDE FRACTIONS
Recently, the problem of bacteriological resistance has become more and more urgent. The antibiotics used in medical practice are constantly becoming obsolete due to the acquisition of resistance by pathogenic microorganisms, therefore there is an ever-increasing need to search for new antimicrobial drugs of various nature. A promising line of research in this area is the study of the antibacterial activity of protein molecules. Over the past two decades, a lot of data has accumulated showing the wide distribution of antimicrobial peptides among almost all living organisms. This article studied the antibacterial activity of peptide fractions weighing less than 5 kDa obtained from honey of two varieties — buckwheat and mixed origin, as well as the similar activity of various solutions of the same honey samples. The following methods were sequentially applied for isolation and purification of peptide fractions: ultrafiltration — to obtain a solution containing molecules with a mass of less than 5 kDa; ion exchange chromatography on DEAE-cellulose for purification of the peptide fraction from low molecular weight impurities (carbohydrates, pigments, phenols, etc.) and gel filtration on Sephadex G-25 to desalt the obtained peptide fraction from the buffer solution. Determination of antibacterial activity was carried out using the disk diffusion method with antibiotic-resistant strains of Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Salmonella enteritis obtained from patients with various bacterial infections in chronic and acute forms. In the studied samples, the concentration of peptides, carbohydrates in ketosis and hydrogen peroxide was measured. The results obtained indicate that among the studied solutions of honey, buckwheat has the highest antibacterial activity, which is associated with an increased content of hydrogen peroxide. The study of the antagonistic activity of peptide fractions against test cultures showed that low molecular weight peptides obtained from the studied honey samples do not have an antibacterial effect against the used strains of microorganisms. Apparently, this is due to the absence of low molecular weight antimicrobial peptides in the obtained fractions. Probably, antimicrobial peptides weighing less than 5 kDa are completely absent in bee honey.
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