IMPROVEMENT OF THE METHOD OF OBTAINING WATER-SOLUBLE PEPTIDES ISOLATED FROM M. DOMESTICA AND STUDY OF THEIR PROPERTIES

O. Larionova, L. S. Krylova, Y. Drevko, K. Smirnova
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Abstract

The article presents data on improvement of the method of obtaining fractions of water-soluble peptides by cold extraction from the biomass of Musca domestica larvae, their separation using dialysis membranes with sizes less than 3.5 kDa, 3.5-7 kDa, 7-14 kDa, more than 14 kDa, followed by a study by means of highly effective liquid chromatography. Retention time and chromatographic yield, protein content, antimicrobial properties, acute toxicity of 2 (BP2) protein fraction, which is the fraction isolated when using membranes of 3.5 - 7 kDa, were established. Therefore, this fraction has a mass of proteins in the range from 3.5 kDa to 7 kDa. The results of the antimicrobial activity of the analyzed protein fraction 2 to the following strains were obtained: B. cereus 8035, E. aerogenes ATCC-13048, E. coli Mordovia 21. It was found that the protein fraction 2 at a concentration of 10 mg/ml, 5 mg/ml, 2.5 mg /ml had antimicrobial activity against E. aerogenes ATCC-13048, E. coli Mordovia 21. Acute toxicity of this fraction was assessed on laboratory mice after a single intragastric and parenteral administration. The average lethal dose for intragastric and parenteral administration for 20% solution of protein fraction 2 could not be established, since the maximum possible doses for these types of administration did not lead to death of any animal. It was established that 20% solution of protein fraction 2 belongs to the 4th hazard class (substances of low hazard) according to the generally accepted hygienic classification.
家蝇水溶性肽提取方法的改进及其性质研究
本文改进了从家蝇幼虫生物量中冷提取水溶性肽的方法,采用小于3.5 kDa、3.5-7 kDa、7-14 kDa、大于14 kDa的透析膜进行分离,并采用高效液相色谱法进行了研究。测定了3.5 ~ 7 kDa膜分离得到的2 (BP2)蛋白组分的保留时间、层析收率、蛋白含量、抗菌性能和急性毒性。因此,该部分的蛋白质质量在3.5 kDa到7 kDa之间。分析所得蛋白片段2对蜡样芽孢杆菌8035、产气荚膜芽孢杆菌ATCC-13048、Mordovia大肠杆菌21的抑菌活性。结果表明,在10 mg/ml、5 mg/ml、2.5 mg/ml浓度下,蛋白组分2对产气大肠杆菌ATCC-13048、Mordovia大肠杆菌21均有抑菌活性。在单次灌胃和肠外给药后,对实验室小鼠进行了急性毒性评估。由于这两种给药方式的最大可能剂量未导致任何动物死亡,因此无法确定20%蛋白质组分2溶液的胃内和肠外平均致死剂量。根据公认的卫生分类,确定蛋白质组分2的20%溶液属于第4类危害物质(低危害物质)。
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