Purification and properties of a type-II-like dipeptidyl peptidase from the ruminal peptidolytic bacterium, Prevotella albensis M384

H.-D. Kim, N. D. Walker, N. McKain, R. J. Wallace

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

A dipeptidyl peptidase which hydrolyses the synthetic dipeptidyl peptidase (DPP) substrate, Ala2- p -nitroanilide, was purified 193-fold from the ruminal peptidolytic bacterium, Prevotella albensis M384. The enzyme was a homodimer of molecular mass 91 kDa. Its activity against Ala2- p -nitroanilide had optimal pH and temperature of 7.2 and 40°C respectively. Enzyme activity was inhibited by the serine protease inhibitors, PMSF and dichloroisocoumarin, but not by inhibitors of other categories of proteases. Synthetic substrates for DPP-1 (GlyArg- p -nitroanilide, GlyArg-4-methoxy-naphthylamide), DPP-3 (ArgArg-4-methoxynaphthylamide) and DPP-4 (GlyPro-4-methoxynaphthylamide) or for leucine or alanine aminopeptidase were not hydrolysed, nor were di- or tripeptides. N-Acetyl-Ala2- p -nitroanilide was not hydrolysed. Oligopeptides with Ala, Ile, Ser or Val adjacent to the N-terminal amino acid were all hydrolysed, while peptides with basic or acidic residues in the same position were not. The purified DPP from P. albensis is therefore most similar in its catalytic properties to mammalian DPP-2.
Original languageEnglish
Pages (from-to)307-313
Number of pages7
JournalAnaerobe
Volume7
Issue number6
DOIs
Publication statusPublished - Dec 2001

Fingerprint

dipeptidyl peptidase II
Dipeptidyl-Peptidases and Tripeptidyl-Peptidases
Prevotella
Bacteria
Dipeptidyl Peptidase 4
CD13 Antigens
Leucyl Aminopeptidase
Oligopeptides
Serine Proteinase Inhibitors
Dipeptides
Enzymes
Peptide Hydrolases
Amino Acids

Cite this

Purification and properties of a type-II-like dipeptidyl peptidase from the ruminal peptidolytic bacterium, Prevotella albensis M384. / Kim, H.-D.; Walker, N. D.; McKain, N.; Wallace, R. J.

In: Anaerobe, Vol. 7, No. 6, 12.2001, p. 307-313.

Research output: Contribution to journalArticle

@article{1363b44192024610bbff3e39471fa34d,
title = "Purification and properties of a type-II-like dipeptidyl peptidase from the ruminal peptidolytic bacterium, Prevotella albensis M384",
abstract = "A dipeptidyl peptidase which hydrolyses the synthetic dipeptidyl peptidase (DPP) substrate, Ala2- p -nitroanilide, was purified 193-fold from the ruminal peptidolytic bacterium, Prevotella albensis M384. The enzyme was a homodimer of molecular mass 91 kDa. Its activity against Ala2- p -nitroanilide had optimal pH and temperature of 7.2 and 40°C respectively. Enzyme activity was inhibited by the serine protease inhibitors, PMSF and dichloroisocoumarin, but not by inhibitors of other categories of proteases. Synthetic substrates for DPP-1 (GlyArg- p -nitroanilide, GlyArg-4-methoxy-naphthylamide), DPP-3 (ArgArg-4-methoxynaphthylamide) and DPP-4 (GlyPro-4-methoxynaphthylamide) or for leucine or alanine aminopeptidase were not hydrolysed, nor were di- or tripeptides. N-Acetyl-Ala2- p -nitroanilide was not hydrolysed. Oligopeptides with Ala, Ile, Ser or Val adjacent to the N-terminal amino acid were all hydrolysed, while peptides with basic or acidic residues in the same position were not. The purified DPP from P. albensis is therefore most similar in its catalytic properties to mammalian DPP-2.",
author = "H.-D. Kim and Walker, {N. D.} and N. McKain and Wallace, {R. J.}",
note = "Spares in Box V.",
year = "2001",
month = "12",
doi = "10.1006/anae.2001.0396",
language = "English",
volume = "7",
pages = "307--313",
journal = "Anaerobe",
issn = "1075-9964",
publisher = "Academic Press Inc.",
number = "6",

}

TY - JOUR

T1 - Purification and properties of a type-II-like dipeptidyl peptidase from the ruminal peptidolytic bacterium, Prevotella albensis M384

AU - Kim, H.-D.

AU - Walker, N. D.

AU - McKain, N.

AU - Wallace, R. J.

N1 - Spares in Box V.

PY - 2001/12

Y1 - 2001/12

N2 - A dipeptidyl peptidase which hydrolyses the synthetic dipeptidyl peptidase (DPP) substrate, Ala2- p -nitroanilide, was purified 193-fold from the ruminal peptidolytic bacterium, Prevotella albensis M384. The enzyme was a homodimer of molecular mass 91 kDa. Its activity against Ala2- p -nitroanilide had optimal pH and temperature of 7.2 and 40°C respectively. Enzyme activity was inhibited by the serine protease inhibitors, PMSF and dichloroisocoumarin, but not by inhibitors of other categories of proteases. Synthetic substrates for DPP-1 (GlyArg- p -nitroanilide, GlyArg-4-methoxy-naphthylamide), DPP-3 (ArgArg-4-methoxynaphthylamide) and DPP-4 (GlyPro-4-methoxynaphthylamide) or for leucine or alanine aminopeptidase were not hydrolysed, nor were di- or tripeptides. N-Acetyl-Ala2- p -nitroanilide was not hydrolysed. Oligopeptides with Ala, Ile, Ser or Val adjacent to the N-terminal amino acid were all hydrolysed, while peptides with basic or acidic residues in the same position were not. The purified DPP from P. albensis is therefore most similar in its catalytic properties to mammalian DPP-2.

AB - A dipeptidyl peptidase which hydrolyses the synthetic dipeptidyl peptidase (DPP) substrate, Ala2- p -nitroanilide, was purified 193-fold from the ruminal peptidolytic bacterium, Prevotella albensis M384. The enzyme was a homodimer of molecular mass 91 kDa. Its activity against Ala2- p -nitroanilide had optimal pH and temperature of 7.2 and 40°C respectively. Enzyme activity was inhibited by the serine protease inhibitors, PMSF and dichloroisocoumarin, but not by inhibitors of other categories of proteases. Synthetic substrates for DPP-1 (GlyArg- p -nitroanilide, GlyArg-4-methoxy-naphthylamide), DPP-3 (ArgArg-4-methoxynaphthylamide) and DPP-4 (GlyPro-4-methoxynaphthylamide) or for leucine or alanine aminopeptidase were not hydrolysed, nor were di- or tripeptides. N-Acetyl-Ala2- p -nitroanilide was not hydrolysed. Oligopeptides with Ala, Ile, Ser or Val adjacent to the N-terminal amino acid were all hydrolysed, while peptides with basic or acidic residues in the same position were not. The purified DPP from P. albensis is therefore most similar in its catalytic properties to mammalian DPP-2.

U2 - 10.1006/anae.2001.0396

DO - 10.1006/anae.2001.0396

M3 - Article

VL - 7

SP - 307

EP - 313

JO - Anaerobe

JF - Anaerobe

SN - 1075-9964

IS - 6

ER -