Peptidases of the rumen bacterium Prevotella ruminicola

R. J. Wallace, N. McKain, G. A. Broderick, L. M. Rode, N. D. Walker, C. J. Newbold, J. Kopecny

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Abstract

Prevotella(formerlyBacteroides) ruminicolais a numerous rumen bacterium which plays a significant role in the metabolism of proteins and peptides in the rumen. Measurement of the hydrolysis of synthetic aminopeptidase substrates by sonicated extracts and whole cells of different species of rumen bacteria indicated thatP. ruminicolahad the greatest range and specific activity of dipeptidyl peptidases among the species tested.Streptococcus bovishydrolysed some dipeptidyl peptidase substrates to a lesser extent, and several species broke down Ala2-p-nitroanilide, includingRuminobacter amylophilus, Ruminococcusspp. andVeillonella parvula. Dipeptidyl peptidases, which cleave dipeptides from the amino-terminus of longer peptides, were much more active than aminopeptidases removing single amino acids inP. ruminicola. Ion-exchange chromatography of sonicated extracts ofP. ruminicolaM384 revealed at least four distinct activities: one hydrolysed Ala2-p-nitroanilide, ValAla-p-nitroanilide, Ala4and Ala5; another was an O2-sensitive activity hydrolysing GlyArg-4-methoxynapthylamide, ArgArg-4-methoxynaphthylamide, Gly5and ValGlySerGlu, similar to dipeptidyl peptidase type I DPP-1); a third hydrolysed GlyPro-p-nitroanilide and GlyPro-4-methoxynapthylamide and was similar to dipeptidyl peptidase type IV XDPP-4); a fourth broke down LysAla-4-methoxynaphthylamide. All of the enzymes, and particularly those active against Ala2-p-nitroanilide and GlyPro-p-nitroanilide, were inhibited by serine protease inhibitors, and all except DPP-4 were inhibited by EDTA. Both DPP-1 and the enzyme hydrolysing LysAla-4-methoxynaphthylamide were inhibited strongly by iodoacetate. DPP-4 was inhibited completely by diprotin A. Competitive inhibition experiments suggested that DPP-1 was less important than the other enzymes in the breakdown of peptide mixtures.
Original languageEnglish
Pages (from-to)35-42
Number of pages8
JournalAnaerobe
Volume3
Issue number1
DOIs
Publication statusPublished - Feb 1997

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Prevotella ruminicola
Dipeptidyl-Peptidases and Tripeptidyl-Peptidases
Rumen
Aminopeptidases
Peptide Hydrolases
diprotin A
Bacteria
Peptides
Enzymes
Cathepsin C
Prevotella
Iodoacetates
Dipeptidyl Peptidase 4
Serine Proteinase Inhibitors
Dipeptides
Ion Exchange Chromatography
Streptococcus
Cell Extracts
Edetic Acid
Hydrolysis

Cite this

Wallace, R. J., McKain, N., Broderick, G. A., Rode, L. M., Walker, N. D., Newbold, C. J., & Kopecny, J. (1997). Peptidases of the rumen bacterium Prevotella ruminicola. Anaerobe, 3(1), 35-42. https://doi.org/10.1006/anae.1996.0065

Peptidases of the rumen bacterium Prevotella ruminicola. / Wallace, R. J.; McKain, N.; Broderick, G. A.; Rode, L. M.; Walker, N. D.; Newbold, C. J.; Kopecny, J.

In: Anaerobe, Vol. 3, No. 1, 02.1997, p. 35-42.

Research output: Contribution to journalArticle

Wallace, RJ, McKain, N, Broderick, GA, Rode, LM, Walker, ND, Newbold, CJ & Kopecny, J 1997, 'Peptidases of the rumen bacterium Prevotella ruminicola', Anaerobe, vol. 3, no. 1, pp. 35-42. https://doi.org/10.1006/anae.1996.0065
Wallace RJ, McKain N, Broderick GA, Rode LM, Walker ND, Newbold CJ et al. Peptidases of the rumen bacterium Prevotella ruminicola. Anaerobe. 1997 Feb;3(1):35-42. https://doi.org/10.1006/anae.1996.0065
Wallace, R. J. ; McKain, N. ; Broderick, G. A. ; Rode, L. M. ; Walker, N. D. ; Newbold, C. J. ; Kopecny, J. / Peptidases of the rumen bacterium Prevotella ruminicola. In: Anaerobe. 1997 ; Vol. 3, No. 1. pp. 35-42.
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