Hostname: page-component-77c89778f8-7drxs Total loading time: 0 Render date: 2024-07-17T04:14:34.775Z Has data issue: false hasContentIssue false
  • English
  • Français

Effect of high hydrostatic pressure on the enzymic hydrolysis of β-lactoglobulin B by trypsin, thermolysin and pepsin

Published online by Cambridge University Press:  01 June 2009

Henrik Stapelfeldt ,
Per Hjort Petersen ,
Kristian Rotvig Kristiansen ,
Karsten Bruun Qvist  and
Leif H. Skibsted
Henrik Stapelfeldt
Affiliation:
Department of Dairy and Food Science, Royal Veterinary and Agricultural University, Rolighedsvej 30, DK-1958 Frederiksberg C, Denmark
Per Hjort Petersen
Affiliation:
Institute for Dairy Research, Rolighedsvej 30, DK-1958 Frederiksberg C, Denmark
Kristian Rotvig Kristiansen
Affiliation:
Institute for Dairy Research, Rolighedsvej 30, DK-1958 Frederiksberg C, Denmark
Karsten Bruun Qvist
Affiliation:
Institute for Dairy Research, Rolighedsvej 30, DK-1958 Frederiksberg C, Denmark
Leif H. Skibsted
Affiliation:
Department of Dairy and Food Science, Royal Veterinary and Agricultural University, Rolighedsvej 30, DK-1958 Frederiksberg C, Denmark
Get access Rights & Permissions [Opens in a new window]

Summary

Hydrolysis of β-lactoglobulin B (β-lg B) by pepsin, a process slow at ambient conditions, is facilitated at a moderately high hydrostatic pressure such as 300 MPa, corresponding to an apparent volume of activation ΔV# = −63 ml mol−1 at pH 2·5, 30 °C and Γ/2=0·16. Digestion of β-lg by trypsin and thermolysin is likewise enhanced by pressure, and the pressure effect has been traced to pressure denaturation of β-lg B, which by high-pressure fluorescence spectroscopy has been shown to have a large negative volume of reaction, ΔV° = −98 ml mol−1, at pH 6·7, 30 °C and Γ/2 = 0·16. Pressure denaturation is only slowly reversed following release of pressure and the enhanced digestibility is maintained at ambient pressure for several hours.

Type
Original Articles
Information
Journal of Dairy Research , Volume 63 , Issue 1 , February 1996 , pp. 111 - 118
Copyright
Copyright © Proprietors of Journal of Dairy Research 1996

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Balny, C. & Masson, P. 1993 Effects of high pressure on proteins. Food Reviews International 9 611–628CrossRefGoogle Scholar
Burstein, E. A., Vedenkina, N. S. & Ivkova, M. N. 1973 Fluorescence and the location of tryptophan residues in protein molecules. Photochemistry and Photobiology 18 263–279CrossRefGoogle ScholarPubMed
Cairoli, S., Iametti, S. & Bonomi, F. 1994 Reversible and irreversible modifications of β-lactoglobulin upon exposure to heat. Journal of Protein Chemistry 13 347–354CrossRefGoogle ScholarPubMed
Dufour, E., Hervé, G. & Haertlé, T. 1992 Limited proteolysis of bovine β-lactoglobulin with thermolysin under high pressure. In High Pressure and Biotechnology, pp. 147–150 (Eds Balny, C., Hayashi, R., Heremans, K. and Masson, P.). Paris: INSERMGoogle Scholar
Dufouk, E., Hervé, C. & Haertlé, T. 1995 Hydrolysis of β-lactoglobulin by thermolysin and pepsin under high hydrostatic pressure. Biopolymers 35 475–483Google Scholar
Dufour, E., Hui Bon Hoa, G. & Haertlé, T. 1994 High-pressure effects on β-lactoglobulin interactions with ligands studied by fluorescence. Biochimica et Biophysica Acta 1206 166–172CrossRefGoogle ScholarPubMed
Dumay, E. M., Kalichevsky, M. T. & Cheftel, J. C. 1994 High-pressure unfolding and aggregation of β- lactoglobulin and the baroprotective effects of sucrose. Journal of Agricultural and Food Chemistry 42 1861–1868CrossRefGoogle Scholar
Eftink, M. R. & Wasylevvski, Z. 1988 Pressure dependence of fluorescence quenching reactions in proteins. Biophysical Chemistry 32 121–130CrossRefGoogle Scholar
Hayashi, R., Kawamura, Y. & Kunugi, S. 1987 Introduction of high pressure to food processing: preferential proteolysis of β-lactoglobulin in milk whey. Journal of Food Science 52 1107–1108CrossRefGoogle Scholar
Heremans, K. 1982 High pressure effects on proteins and other biomolecules. Annual Review of Biophysics and Bioengineering 11 1–21CrossRefGoogle ScholarPubMed
Morild, E. 1981 The theory of pressure effects on enzymes. Advances in Protein Chemistry 34 93–166CrossRefGoogle ScholarPubMed
Neuman, R. C., Kauzmann, W. & Zrpp, A. 1973 Pressure dependence of weak acid ionization in aqueous buffers. Journal of Physical Chemistry 22 2687–2691CrossRefGoogle Scholar
Okamoto, M., Hayashi, R., Enomoto, A., Kaminogawa, S. & Yamauchi, K. 1991 High-pressure proteolytic digestion of food proteins: selective elimination of β-lactoglobulin in bovine milk whey concentrate. Agricultural and Biological Chemistry 55 1253–1257Google Scholar
Otte, J. A. H. J., Kristiansen, K. R., Zakora, M. & Qvist, K. B. 1994 Separation of individual whey proteins and measurements of α-lactalbumin and β-lactoglobulin by capillary zone electrophoresis. Netherlands Milk and Dairy Journal 48 81–97Google Scholar
Paladini, A. A. & Weber, G. 1981 Absolute measurements of fluorescence polarization at high pressures. Reviews of Scientific Instruments 52 419–427CrossRefGoogle Scholar
Privalov, P. L. 1979 Stability of proteins. Small globular proteins. Advances in Protein Chemistry 33 167–241CrossRefGoogle ScholarPubMed
Schmidt, D. G. & Poll, J. K. 1991 Enzymatic hydrolysis of whey proteins. Hydrolysis of α-lactalbumin and β-lactoglobulin in buffer solutions by proteolytic enzymes. Netherlands Milk and Dairy Journal 45 225–240Google Scholar
Schmidt, D. G. & Van Markwijk, B. W. 1993 Enzymic hydrolysis of whey proteins: influence of heat treatment of α-lactalbumin and β-lactoglobulin in the proteolysis by pepsin and papain. Netherlands Milk and Dairy Journal 47 15–22Google Scholar
Silva, J. L., Silvelra, C. F., Correia, A. & Pontes, L. 1992 Dissociation of a native dimer to a molten globule monomer. Effects of pressure and dilution on the association equilibrium of arc repressor. Journal of Molecular Biology 223 545–555CrossRefGoogle ScholarPubMed
Silva, J. L. & Weber, G. 1993 Pressure stability of proteins. Annual Review of Physical Chemistry 44 89–113CrossRefGoogle ScholarPubMed
Stapelfeldt, H. & Skibsted, L. H. 1994 Modification of β-lactoglobulin by aliphatic aldehydes in aqueous solution. Journal of Dairy Research 61 209–219CrossRefGoogle Scholar
Weber, G. 1986 Phenomenological description of the association of protein subunits subjected to conformational drift. Effects of dilution and of hydrostatic pressure. Biochemistry 25 3626–3631CrossRefGoogle ScholarPubMed
Weber, G. & Drickamer, H. G. 1983 The effect of high pressure upon proteins and other biomolecules. Quarterly Reviews of Biophysics 16 89–112CrossRefGoogle ScholarPubMed
Werbin, H. & McLaren, A. D. 1951 The effect of high pressure on the rates of proteolytic hydrolysis. II. Trypsin. Archives of Biochemistry and Biophysics 32 325–337CrossRefGoogle ScholarPubMed
Yu, H. -T., Colucci, W. J., McLaughlin, M. L. & Barkley, M. D. 1992 Fluorescence quenching in indoles by excited-state proton transfer. Journal of the American Chemical Society 114 8449–8454CrossRefGoogle Scholar