S. Tikhonov, N. Tikhonova, Angelika Popovskikh, M. Tikhonova
{"title":"Individual Peptides Extraction and Characteristics of Cow Colostrum Trypsin Hydrolysate with Biological Activity Assessment","authors":"S. Tikhonov, N. Tikhonova, Angelika Popovskikh, M. Tikhonova","doi":"10.29141/2500-1922-2022-7-4-10","DOIUrl":null,"url":null,"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.","PeriodicalId":7684,"journal":{"name":"Agro Food Industry Hi-tech","volume":"36 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2022-12-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Agro Food Industry Hi-tech","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.29141/2500-1922-2022-7-4-10","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q","JCRName":"Engineering","Score":null,"Total":0}
引用次数: 0
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.