牛初乳中胰蛋白酶水解的肽

Q3 Economics, Econometrics and Finance Food Processing: Techniques and Technology Pub Date : 2023-03-21 DOI:10.21603/2074-9414-2023-1-2422
S. Tikhonov, N. Tikhonova, Kh. T. Tursunov, I. Danilova, V. Lazarev
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引用次数: 1

摘要

牛初乳含有生物活性物质,如免疫球蛋白、多肽和细胞因子,这使其成为许多功能性产品的合理组成部分。初乳肽也具有抗菌活性。这种生物利用度在蛋白水解酶发酵初乳期间增加。研究目的是描述从牛初乳胰蛋白酶水解上清液中分离的肽,并评价其抗菌和抗真菌性能。分离牛初乳胰蛋白酶水解产物上清液,离心3900 rpm,离心7 min,制备层析分离上清液。利用MALDI-TOF质谱仪测定其肽组成,利用Mascot数据库破译蛋白质序列。用硫酸铵沉淀蛋白质,用圆盘扩散法测定其对革兰氏阳性菌、革兰氏阴性菌和双极性真菌的抑菌活性。菌株在37℃的厚LB营养培养基上培养。对腹腔感染肠炎沙门氏菌92的Wistar大鼠进行了抑菌实验。牛初乳胰蛋白酶水解上清中发现4个多肽,其中1个为短肽,其余3个为多肽。分离得到的肽分子量分别为8.4、6.5、13.0和18 kDa。酶解产物对大肠杆菌和枯草芽孢杆菌具有杀菌作用,对白色念珠菌具有抗真菌活性。对感染肠炎沙门氏菌92的大鼠给予胰蛋白酶水解物,可促进其存活,降低LD50,使平均死亡天数从2天延长至4天。研究证实了初乳多肽的抗菌作用,并提示其具有免疫亲和性。从牛初乳胰蛋白酶水解上清液中获得的肽可作为抗菌产品的一部分推荐用于功能食品工业。
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Peptides of Trypsin Hydrolyzate in Bovine Colostrum
Bovine colostrum contains biologically active substances, e.g., immunoglobulins, peptides, and cytokines, which makes it a logical component of numerous functional products. Colostrum peptides also possess antimicrobial activity. This bioavailability increases during colostrum fermentation with proteolytic enzymes. The research objective was to describe peptides isolated from the trypsic hydrolyzate supernatant of bovine colostrum and to evaluate their antimicrobial and antifungal properties. The supernatant of trypsin hydrolyzate of bovine colostrum was isolated by centrifugation at 3900 rpm for 7 min. The supernatant was separated by preparative chromatography. Its peptide composition was determined on a MALDI-TOF mass spectrometer, while the protein sequences were deciphered using the Mascot database. Proteins were precipitated with ammonium sulfate, and the antimicrobial activity was measured by the disk-diffusion method against gram-positive and gram-negative bacteria and dipoloid fungi. Strains were cultivated on a thick LB nutrient medium at 37°C. The antimicrobial activity was defined experimentally on Wistar rats infected intraperitoneally with Salmonella enteritidis 92. The trypsin hydrolyzate supernatant of bovine colostrum revealed four peptides, one of which belonged to short peptides, while the remaining three belonged to polypeptides. The isolated peptides had different molecular weights of 8.4, 6.5, 13.0, and 18 kDa. The enzymatic hydrolyzate proved bactericidal against Escherichia coli and Bacillus subtilis and demonstrated antifungal activity against Candida albicans. When rats infected with S. enteritidis 92 were administered with trypsin hydrolysate, it promoted their survival, decreased LD50, and increased the mean day of death period from 2 to 4 days. The research proved the antimicrobial effect of colostrum peptides and suggested their immunotropic properties. The peptides obtained from the trypsin hydrolyzate supernatant of bovine colostrum can be recommended for functional food industry as part of antimicrobial products.
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来源期刊
Food Processing: Techniques and Technology
Food Processing: Techniques and Technology Engineering-Industrial and Manufacturing Engineering
CiteScore
1.40
自引率
0.00%
发文量
82
审稿时长
12 weeks
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