Hostname: page-component-586b7cd67f-2plfb Total loading time: 0 Render date: 2024-11-23T02:03:32.987Z Has data issue: false hasContentIssue false
  • English
  • Français

In vitro studies of the digestion of caprine whey proteins by human gastric and duodenal juice and the effects on selected microorganisms

Published online by Cambridge University Press:  08 March 2007

Hilde Almaas ,
Halvor Holm ,
Thor Langsrud ,
Ragnar Flengsrud  and
Gerd E. Vegarud
Hilde Almaas*
Affiliation:
Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, P. Box 1432-Ås, Norway
Halvor Holm
Affiliation:
University of Oslo, Department of Nutrition, P. Box 1046 Blindern, 0316 Oslo, Norway
Thor Langsrud
Affiliation:
Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, P. Box 1432-Ås, Norway
Ragnar Flengsrud
Affiliation:
Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, P. Box 1432-Ås, Norway
Gerd E. Vegarud
Affiliation:
Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, P. Box 1432-Ås, Norway
*
*Corresponding author: Hilde Almaas, fax +47 64965901, email email [email protected]
Rights & Permissions [Opens in a new window]

Abstract

Core share and HTML view are not available for this content. However, as you have access to this content, a full PDF is available via the ‘Save PDF’ action button.

The in vitro digestion of caprine whey proteins was investigated by a two-step degradation assay, using human gastric juice (HGJ) at pH 2·5 and human duodenal juice (HDJ) at pH 7·5. Different protein and peptide profiles were observed after the first (HGJ) and second (HDJ) enzymatic degradation. The minor whey proteins serum albumin, lactoferrin and Ig were rapidly degraded by HGJ, while α-lactalbumin (α-LA) and β-lactoglobulin (β-LG) were more resistant and survived both 30 and 45min of the enzymatic treatment. Further digestion with HDJ still showed intact β-LG, and the main part of α-LA also remained unchanged. The protein degradation by HGJ and HDJ was also compared with treatment by commercial enzymes, by using pepsin at pH 2·5, and a mixture of trypsin and chymotrypsin at pH 7·5. The two methods resulted in different caprine protein and peptide profiles. The digests after treatment with HGJ and HDJ were screened for antibacterial effects on some selected microorganisms, Escherichia coli, Bacillus cereus, Lactobacillus rhamnosus GG and Streptococcus mutans. Active growing cells of E. coli were inhibited by the digestion products from caprine whey obtained after treatment with HGJ and HDJ. Cells of B. cereus were inhibited only by whey proteins obtained after reaction with HGJ, while the products after further degradation with HDJ demonstrated no significant effect. Screenings performed on cells of Lb. rhamnosus GG and S. mutans all showed no signs of inhibition.

Keywords

Caprine whey proteinsDigestionGastric juiceDuodenal juiceAntibacterial
Type
Research Article
Information
British Journal of Nutrition , Volume 96 , Issue 3 , September 2006 , pp. 562 - 569
Copyright
Copyright © The Nutrition Society 2006

References

Arnold, RR, Brewer, M & Gautheir, J (1980) Bactericidal activity of human lactoferrin: sensitivity of a variety of microorganisms. Infect Immun 28, 893–898.Google Scholar
Arnold, RR, Cole, MF & McGhee, JR (1977) Bactericidal activity of human lactoferrin. Science 127, 263–265.Google Scholar
Bellamy, W, Takase, M, Wakabayashi, H, Kawase, K & Tomita, M (1992) Antibacterial spectrum of lactoferricin B, a potent bactericidal peptide derived from the N-terminal region of bovine lactoferrin. J Appl Bacteriol 73, 472–479.CrossRefGoogle ScholarPubMed
Bellamy, W, Wakabayashi, H, Takase, M, Kawase, K, Shimamura, S & Tomita, M (1993) Killing of candida-albicans by lactoferricin B, a potent antimicrobial peptide derived from the N-terminal region of bovine lactoferrin. Med Microbiol Immunol 182, 97–105.Google Scholar
Chatterton, DEW, Rasmussen, JT, Heegaard, CW, Sørensen, ES & Petersen, TE (2004) In vitro digestion of novel milk protein ingredients for use in infant formulas: research on biological functions. Trends Food Sci Technol 15, 373–383.Google Scholar
Dunn, BM (2002) Structure and mechanism of the pepsin-like family of aspartic peptidases. Chem Rev 102, 4431–4458.CrossRefGoogle ScholarPubMed
Farnaud, S & Ewans, RW (2003) Lactoferrin – a multifunctional protein with antimicrobial properties. Mol Immunol 40, 395–405.CrossRefGoogle ScholarPubMed
FitzGerald, RJ & Meisel, H (2003) Milk protein hydrolysates and bioactive peptides. In Advanced Dairy Chemistry, 3rd ed. Vol. 1, pp. 675–691 [Fox, PF and McSweeney, LH, editors]. New York: Kluwer Academi/lenum Publishers.Google Scholar
Guillou, H, Miranda, G & Pelisser, JP (1991) Hydrolysis of beta-casein by gastric proteases. I. Comparison of proteolytic action of bovine chymosin and pepsin A. Int J Pept Protein Res 37, 494–501.CrossRefGoogle ScholarPubMed
Holm, H, Hanssen, LE, Krogdahl, Å & Florholmen, J (1988) High and low inhibitor soybean meals affect human duodenal proteinase activity differently: in vivo comparison with bovine serum albumin. J Nutr 118, 515–520.Google Scholar
International Dairy Federation (1993) Milk – Determination of Nitrogen Content. Part 3, Block Digestion Method, Semi-micro Rapid Routine Method, IDF Standard 20B. Brussels: International Dairy Federation.Google Scholar
Kimura, M, Nam, M-S, Ohkouchi, Y, Kumura, H, Shimazaki, K-I & Yu, D-Y (2000) Antimicrobial peptide of Korean native goat lactoferrin and identification of the part essential for this activity. Biochem Biophys Res Commun 268, 333–336.Google Scholar
Kirkpatrick, CH, Green, I, Rich, RR & Schade, AL (1971) Inhibition of growth of candida-albicans by iron-unsaturated lactoferrin – relation of host-defence mechanism in chronic mucocutaneous candidiasis. J Infect Dis 124, 539.CrossRefGoogle Scholar
Korhonen, H, Marnilla, P & Gill, HS (2000a) Milk immunoglobulins and complement factors. Br J Nutr 84, 75–80.Google Scholar
Korhonen, H, Marnilla, P & Gill, HS (2000b) Bovine milk antibodies for health. Br J Nutr 84, 135–146.CrossRefGoogle ScholarPubMed
Krogdahl, Å & Holm, H (1979) Inhibition of human and rat pancreatic proteinases by crude and purified soybean proteinase inhibitors. J Nutr 109, 551–558.CrossRefGoogle ScholarPubMed
Kuwata, H, Yip, T-T, Tomita, M & Hutchens, TW (1998) Direct evidence of the generation in human stomach of an antimicrobial peptide domain (lactoferricin) from ingested lactoferrin. Biochim Biophys Acta 1429, 129–141.CrossRefGoogle ScholarPubMed
Laemmli, UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.Google Scholar
Meisel, H & Schlimme, E (1996) Bioactive peptides derived from milk proteins: ingredients for functional foods? Kieler Milchwirtschaftliche Forschungsberichte 48, 343–357.Google Scholar
Miranda, G, Mahé, M-F, Leroux, C & Martin, P (2004) Proteomic tools to characterize the protein fraction of Equidae milk. Proteomics 4, 2496–2509.CrossRefGoogle ScholarPubMed
Panyam, D & Kilara, A (1996) Enhancing the functionality of food proteins by enzymatic modification. Trends Food Sci Technol 7, 120–125.CrossRefGoogle Scholar
Pihlanto, A & Korhonen, H (2003) Advances in Food and Nutrition Research – Bioactive Peptides and Proteins, Vol. 47. Amsterdam: Elsevier Academic Press.Google ScholarPubMed
Pihlanto-Leppälä, A (2000) Bioactive peptides derived from bovine whey proteins: opoid and ace-inhibitory peptides. Trends Food Sci Technol 11, 347–356.CrossRefGoogle Scholar
Recio, I, Slangen, CJ & Visser, S (2000) Method for the production of antibacterial peptides from biological fluids at an ionic membrane – application to the isolation of nisin and caprine lactoferrin. Le Lait 80, 187–195.Google Scholar
Recio, I & Visser, S (2000) Antibacterial and binding characteristics of bovine, ovine and caprine lactoferrins: a comparative study. Int Dairy J 10, 597–605.CrossRefGoogle Scholar
Sanchez-Chiang, L, Cisternas, E & Ponce, O (1987) Partial purification of pepsins from adult and juvenile salmon fish – effect of NaCl on proteolytic activities. Comp Biochem Physiol 87, 793–797.Google Scholar
Schägger, H & von Jagow, G (1987) Tricine-sodium dodecyl sulfate-polyacrylamide gel electrotrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem 166, 368–379.Google Scholar
Scheele, G, Bartelt, D & Bieger, W (1981) Characterization of human exocrine pancreatic proteins by two-dimensional isoelectric focusin/odium dodecyl sulphate gel electrophoresis. Gastroenterology 80, 461–473.CrossRefGoogle Scholar
Soukka, T, Tenovuo, J & Lenanderlumikari, M (1992) Fungicidal effect of human lactoferrin against candida-albicans. FEMS Microbiol Lett 90, 223–228.CrossRefGoogle Scholar
Tomita, M, Bellamy, W & Takase, M (1991) Potent antibacterial peptides generated by pepsin digestion of bovine lactoferrin. J Dairy Sci 74, 4137–4142.Google Scholar
Tomita, M, Takase, M, Bellamy, W & Shimamura, S (1994) A review: the active peptide of lactoferrin. Acta Paediatr Jpn 36, 585–591.Google Scholar