Individual Peptides Extraction and Characteristics of Cow Colostrum Trypsin Hydrolysate with Biological Activity Assessment

S. Tikhonov, N. Tikhonova, Angelika Popovskikh, M. Tikhonova
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Abstract

The research concerned the trypsin peptides analysis of colostrum hydrolyzate. The authors isolated seven individual peptides: TT1, TT2, TT3, T(1), T1.1, T1.2, mpT. They determined the molecular weight of the peptides and the amino acid sequence in the peptide chain by mass spectrometry; peptides – using the international database Protein NCBI. The mpT peptide contains the largest number of amino acids – 49. The T1.2 peptide is similar to the membrane-active peptide BAMA Bos taurus, which opens prospects for its use as a part of antibacterial agents or as a BAS carrier. In colostrum hydrolyzate the T1 peptide has the T highest concentration (2.04 mg/ml). It consists of 17 amino acids and matches the peptide NCI_CGAP_Brn23 cDNA Homo sapiens clone, which is similar to TR: O35085 ARX HOMEOPROTEIN. The homeobox protein containing the development gene of a PRD-like class is related to the T(1) peptide. This gene in humans is located on the X chromosome and is involved in the central nervous system and pancreas development. According to the results, the researchers isolated peptides from colostrum hydrolyzate, and identified some of them. Still a man understudies the biological role of most of the determined peptides, and has not proved its safety. In this regard, the study aimed at assessing the hydrolyzate cytotoxicity using the Neutral Red Assay method on the HEK293 cell line during a 24-hour incubation. Cytotoxicity in the first, second and third measurements at a concentration of 0.1 mg/ml was: −3.3; −1.0 and 0.20; at a concentration of 0.3 mg/ml: −1.3; −3.5 and −8.7; at 0.5 mg/ml: −2.6; −0.6 and 0.4. The results indicate no toxicity. The authors studied the antimicrobial and antifungal hydrolyzate activity. Colostrum hydrolyzate demonstrates antimicrobial activity against E. coli and B. subtilis bacteria and antifungal activity against C. albicans. The results assert that the peptides are of clinical importance and can serve as a template for the anti-infective drug development.
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牛初乳胰蛋白酶水解产物的单肽提取、特性及生物活性评价
本研究涉及初乳水解产物胰蛋白酶肽的分析。作者分离出7个肽:TT1、TT2、TT3、T(1)、T1.1、T1.2、mpT。他们用质谱法测定了肽的分子量和肽链上的氨基酸序列;肽-使用国际数据库蛋白质NCBI。mpT肽含有最多的氨基酸——49个。T1.2肽与膜活性肽BAMA Bos taurus相似,这为其作为抗菌药物的一部分或作为BAS载体开辟了前景。初乳水解液中T1肽T浓度最高(2.04 mg/ml)。它由17个氨基酸组成,与肽NCI_CGAP_Brn23 cDNA智人克隆相匹配,与TR: O35085 ARX HOMEOPROTEIN相似。含有prd样类发育基因的同源盒蛋白与T(1)肽有关。人类的这种基因位于X染色体上,参与中枢神经系统和胰腺的发育。根据研究结果,研究人员从初乳水解物中分离出多肽,并对其中一些进行了鉴定。然而,大多数已确定的多肽的生物学作用尚未得到充分研究,其安全性尚未得到证实。因此,本研究旨在利用中性红法对HEK293细胞系进行24小时孵育,以评估水解产物的细胞毒性。在浓度为0.1 mg/ml时,第一、第二和第三次测量的细胞毒性为:−3.3;−1.0和0.20;浓度为0.3 mg/ml时:−1.3;−3.5和−8.7;0.5 mg/ml:−2.6;−0.6和0.4。结果表明无毒性。研究了水解产物的抗菌和抗真菌活性。初乳水解产物对大肠杆菌和枯草芽孢杆菌具有抗菌活性,对白色念珠菌具有抗真菌活性。结果表明,这些肽具有重要的临床意义,可作为抗感染药物开发的模板。
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来源期刊
Agro Food Industry Hi-tech
Agro Food Industry Hi-tech 工程技术-生物工程与应用微生物
CiteScore
0.29
自引率
0.00%
发文量
0
审稿时长
6-12 weeks
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