Abstract

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.

Keywords

Colostrum, milk protein, enzyme, hydrolysis, antimicrobial activity, antifungal activity, biologically active substances

Contribution

All authors provided critical feedback and helped shape the research, analysis and manuscript.

CONFLICTS OF INTEREST

The authors declared no conflict of interests regarding the publication of this article.

REFERENCES

1. Główka N, Woźniewicz M. Potential use of Colostrum bovinum supplementation in athletes – A review. Acta Scientiarum Polonorum Technologia Alimentaria. 2019;18(2):115–123. https://doi.org/10.17306/J.AFS.2019.0654
2. Van Hese I, Goossens K, Vandaele L, Opsomer G. Invited review: MicroRNAs in bovine colostrum – Focus on their origin and potential health benefits for the calf. Journal of Dairy Science. 2020;103(1):1–15. https://doi.org/10.3168/jds.2019-16959
3. Sukhikh SA, Astakhova LA, Golubcova YuV, Lukin AA, Prosekova EA, Milent`eva IS, et al. Functional dairy products enriched with plant ingredients. Foods and Raw Materials. 2019;7(2):428–438. http://doi.org/10.21603/2308-4057-2019-2-428-438
4. Kharitonov VD, Asafov VA, Iskakova EL, Tankova NL, Halavach TM, Kurchenko VP. Quality control of colostrum and protein calf milk replacers. Food Processing: Techniques and Technology. 2021;51(1):188–195. https://doi.org/10.21603/2074-9414-2021-1-188-195.
5. Chandwe K, Kelly P. Colostrum therapy for human gastrointestinal health and disease. Nutrients. 2021;13(6). https://doi.org/10.3390/nu13061956
6. Menchetti L, Traina G, Tomasello G, Casagrande-Proietti P, Leonardi L, Barbato O, et al. Potential benefits of colostrum in gastrointestinal diseases. Frontiers in Bioscience-Scholar. 2016;8(2):331–351. https://doi.org/10.2741/S467
7. Playford RJ, Weiser MJ. Bovine colostrum: Its constituents and uses. Nutrients. 2021;13(1). https://doi.org/10.3390/nu13010265
8. Sanctuary MR, Kain JN, Chen SY, Kalanetra K, Lemay DG, Rose DR, et al. Pilot study of probiotic/colostrum supplementation on gut function in children with autism and gastrointestinal symptoms. PLoS ONE. 2019;14(1). https://doi.org/10.1371/journal.pone.0210064
9. Jørgensen ALW, Juul-Madsen HR, Stagsted J. Colostrum and bioactive, colostral peptides differentially modulate the innate immune response of intestinal epithelial cells. Journal of Peptide Science. 2010;16(1):21–30. https://doi.org/10.1002/psc.1190
10. Korhonen HJ. Production and properties of health-promoting proteins and peptides from bovine colostrum and milk. Cellular and Molecular Biology. 2013;59(1):12–24.
11. Novoselova MV, Prosekov AYu. Technological options for the production of lactoferrin. Foods and Raw Materials. 2016;4(1):90–101. http://doi.org/10.21179/2308-4057-2016-1-90-101
12. Koroleva OV, Agarkova EYu, Botina SG, Nikolaev IV, Ponomareva NV, Melnikova EI, et al. Functional properties of fermented milk products with whey protein hydrolysates. Dairy Industry. 2013;(11):52–55. (In Russ.).
13. Pradeep H, Najma U, Aparna HS. Milk peptides as novel multi-targeted therapeutic candidates for SARS-CoV2. The Protein Journal. 2021;40(3):310–327. https://doi.org/10.1007/s10930-021-09983-8
14. Rezaei Ahvanooei MR, Norouzian MA, Vahmani P. Beneficial Effects of vitamins, minerals, and bioactive peptides on strengthening the immune system against COVID-19 and the role of cow's milk in the supply of these nutrients. Biological Trace Element Research. 2021;200(11):4664–4677. https://doi.org/10.1007/s12011-021-03045-x
15. Kütt M-L, Stagsted J. Caseins from bovine colostrum and milk strongly bind piscidin-1, an antimicrobial peptide from fish. International Journal of Biological Macromolecules. 2014;70:364–372. https://doi.org/10.1016/j.ijbiomac.2014.06.063
16. Shteinfer-Kuzmine A, Amsalem Z, Arif T, Zooravlov A, Shoshan-Barmatz V. Selective induction of cancer cell death by VDAC1-based peptides and their potential use in cancer therapy. Molecular Oncology. 2018;12(7):1077–1103. https://doi.org/10.1002/1878-0261.12313
17. Ko S-C, Kim D, Jeon Y-J. Protective effect of a novel antioxidative peptide purified from a marine Chlorella ellipsoidea protein against free radical-induced oxidative stress. Food and Chemical Toxicology. 2012;50(7):2294–2302. https://doi.org/10.1016/j.fct.2012.04.022
18. Poirier K, Van Esch H, Friocourt G, Saillour Y, Bahi N, Backer S, et al. Neuroanatomical distribution of ARX in brain and its localisation in GABAergic neurons. Molecular Brain Research. 2004;122(1):35–46. https://doi.org/10.1016/j.molbrainres.2003.11.021
19. Morgan AJ, Riley LG, Sheehy PA, Wynn PC. The influence of protein fractions from bovine colostrum digested in vivo and in vitro on human intestinal epithelial cell proliferation. Journal of Dairy Research. 2014;81(1):73–81. https://doi.org/10.1017/S0022029913000654
20. Birkemo GA, O'Sullivan O, Ross RP, Hill C. Antimicrobial activity of two peptides casecidin 15 and 17, found naturally in bovine colostrum. Journal of Applied Microbiology. 2009;106(1):233–240. https://doi.org/10.1111/j.1365-2672.2008.03996.x
21. Sears KT, Tennant SM, Reymann MK, Simon R, Konstantopoulos N, Blackwelder WC, et al. Bioactive immune components of anti-diarrheagenic enterotoxigenic Escherichia coli hyperimmune bovine colostrum products. Clinical and Vaccine Immunology. 2017;24(8). https://doi.org/10.1128/CVI.00186-16
22. Anderson RC, Dalziel JE, Haggarty NW, Dunstan KE, Gopal PK, Roy NC. Short communication: Processed bovine colostrum milk protein concentrate increases epithelial barrier integrity of Caco-2 cell layers. Journal of Dairy Science. 2019;102(12):10772–10778. https://doi.org/10.3168/jds.2019-16951
23. Playford RJ, Weiser MJ, Marchbank T. Methods to improve efficacy of orally administered bioactive peptides using bovine colostrum as an exemplar. PLoS ONE. 2021;16(6). https://doi.org/10.1371/journal.pone.0253422