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Characterization of microbial proteolytic enzymes in the rumen

Antonie van Leeuwenhoek Aims and scope Submit manuscript

Abstract

Up to 43% of the viable bacteria from the rumen of cows fed grass and concentrates grew on a medium containing casein as the main substrate. Proteolytic counts for a cow fed on straw and concentrates or for a hay-fed cow were lower than counts for cows fed grass and concentrates, both in absolute terms and in relation to the total anaerobic count.

In crude enzyme preparations derived from the rumen protozoa, amino acid arylamidase (leucine aminopeptidase)-like activity was the main proteolytic activity observed. In enzyme preparations extracted from the rumen bacteria in the presence of Triton X-100, trypsin-like activity was predominant. Amino acid arylamidase-and metal-chelating proteinase-like activity together with lower activities of carboxypeptidase A and B and a very low chymotrypsin-like activity were found as well.

Studies with enzyme inhibitors showed that the bacterial trypsin-like activity was largely of the cysteine-protease type in a hay-fed cow, but in addition comprised serine-protease activity in a cow fed grass and concentrates. Total proteolytic activity of the enzymes in the bacterial fraction and the spectrum of proteolytic enzymes were found to vary with the ration.

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References

  • Bergmeyer H. U. (ed.). 1974. Methoden der Enzymatischen Analyse, Band I, p. 1038–1044. — Verlag Chemie, Weinheim.

    Google Scholar 

  • Blackburn T. H. 1968. The protease liberated from Bacteriodes amylophilus strain h 18 by mechanical disintegration. — J. Gen. Microbiol. 53: 37–51.

    Google Scholar 

  • Bradford M. M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. — Anal. Biochem. 72: 248–254.

    Google Scholar 

  • Brock F. M., Forsberg C. W. and Buchanan-Smith J. G. 1982. Proteolytic activity of rumen microorganisms and effects of proteinase inhibitors. — Appl. Environ. Microbiol. 44: 561–569.

    Google Scholar 

  • Dehority B. A., and Grubb J. A. 1976. Basal medium for the selective enumeration of rumen bacteria utilizing specific energy sources. — Appl. Environ. Microbiol. 32: 703–710.

    Google Scholar 

  • Erlanger B. F., Edel F. and Cooper A. G. 1966. The action of chymotrypsin on two new chromogenic substrates. — Arch. Biochem. Biophys. 115: 206–210.

    Google Scholar 

  • Erlanger B. F., Kokowsky N. and Cohen W. 1961. The preparation and properties of two new chromogenic substrates of trypsin. — Arch. Biochem. Biophys. 95: 271–278.

    Google Scholar 

  • Hazlewoord G. P. and Edwards R. 1981. Proteolytic activities of a rumen bacterium, Bacteroides ruminicola R 8/4. — J. Gen. Microbiol. 125: 11–15.

    Google Scholar 

  • Henning P. A. and Van derWalt A. E. 1978. Inclusion of xylan in a medium for the enumeration of total culturable rumen bacteria. — Appl. Environ. Microbiol. 35: 1008–1011.

    Google Scholar 

  • Hobson P. N. and Wallace R. J. 1982. Microbial ecology and activities in the rumen: part II. — Crit. Rev. Microbiol. 9: 253–320.

    Google Scholar 

  • Hungate R. E. 1966. The Rumen and its Microbes. — Academic Press, New York.

    Google Scholar 

  • Kopecny J. and Wallace R. J. 1982. Cellular location and some properties of proteolytic enzymes of rumen bacteria. — Appl. Environ. Microbiol. 43: 1026–1033.

    Google Scholar 

  • Lee Y. P. and Takahashi T. 1966. An improved colorimetric determination of amino acids with the use of ninhydrin. — Anal. Biochem. 14: 71–77.

    Google Scholar 

  • Mahadevan S., Erfle J. D. and Sauer F. D. 1980. Degradation of soluble and insoluble proteins by Bacteroides amylophilus protease and by rumen microorganisms. — J. Anim. Sci. 50: 723–728.

    Google Scholar 

  • Mangan J. L. 1972. Quantitative studies on nitrogen metabolism in the bovine rumen. The rate of proteolysis of casein and ovalbumin and the release and metabolism of free amino acids. —Br. J. Nutr. 27: 261–283.

    Google Scholar 

  • Nagel W., Willig F., Peschke W. and Schmidt F. H. 1965. Über die Bestimmung von Trypsin und Chymotrypsin mit Aminosäure-p-nitroanilides. — Hoppe Seyler's Z. Physiol. Chem. 340: 1–10.

    Google Scholar 

  • Nugent, J. H. A. and Mangan, J. L. 1978. Rumen proteolysis of fraction 1 leaf protein, casein and bovine serum albumin. — Proc. Nutr. Soc. 37: 48a.

  • Nugent J. H. A. and Mangan J. L. 1981. Characteristics of the rumen proteolysis of fraction I (18S) leaf protein from lucerne (Medicago sativa L). — Br. J. Nutr. 46: 39–58.

    Google Scholar 

  • Tuppy H., Wiesbauer U. and Wintersberger E. 1962. Aminosäure-p-nitroanilide als Substrate für Aminopeptidasen und andere proteolytische Fermente. — Hoppe Seyler's Z. Physiol. Chem. 329: 278–288.

    Google Scholar 

  • Wachsmuth E. D., Fritze I. and Pfleiderer G. 1966. An aminopeptidase occurring in pig kidney. I. An improved method of preparation. Physical and enzymic properties. — Biochemistry 5: 169–174.

    Google Scholar 

  • Wallace R. J. and Kopecny J. 1983. Breakdown of diazotized proteins and synthetic substrates by rumen bacterial proteases. — Appl. Environ. Microbiol. 45: 212–217.

    Google Scholar 

  • Wohlt J. E., Sniffen C. J. and Hoover W. H. 1973. Measurement of protein solubility in common feedstuffs. — J. Dairy Sci. 56: 1052–1057.

    Google Scholar 

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Prins, R.A., van Rheenen, D.L. & van't Klooster, A.T. Characterization of microbial proteolytic enzymes in the rumen. Antonie van Leeuwenhoek 49, 585–595 (1983). https://doi.org/10.1007/BF00399852

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