Activating Compound | Comment | Organism | Structure |
---|---|---|---|
EDTA | 23.9% activation at 10 mM | Campylobacter jejuni |
Cloned (Comment) | Organism |
---|---|
gene hipO, expression of His6-tagged enzyme in Escherichia coli strain M15 | Campylobacter jejuni |
Protein Variants | Comment | Organism |
---|---|---|
D104N | site-directed mutagenesis, inactive mutant | Campylobacter jejuni |
D76N | site-directed mutagenesis, the mutant shows over 12fold reduced activity compared to the wild-type enzyme | Campylobacter jejuni |
E134Q | site-directed mutagenesis, inactive mutant | Campylobacter jejuni |
E135Q | site-directed mutagenesis, inactive mutant | Campylobacter jejuni |
H161A | site-directed mutagenesis, inactive mutant | Campylobacter jejuni |
H356A | site-directed mutagenesis, inactive mutant | Campylobacter jejuni |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
1,10-phenanthroline | 99% inhibition at 0.5 mM | Campylobacter jejuni | |
2-mercaptoethanol | 20% inhibition at 2 mM | Campylobacter jejuni | |
Ag+ | 98% inhibition at 2 mM | Campylobacter jejuni | |
Ca2+ | 55% inhibition at 2 mM | Campylobacter jejuni | |
Cd2+ | 65% inhibition at 2 mM | Campylobacter jejuni | |
Co2+ | 70% inhibition at 2 mM | Campylobacter jejuni | |
Cu2+ | 78% inhibition at 2 mM | Campylobacter jejuni | |
Fe2+ | - |
Campylobacter jejuni | |
additional information | no significant inhibition by E64, PMSF, and pepstatin at 0.5 mM | Campylobacter jejuni | |
Ni2+ | 65% inhibition at 2 mM | Campylobacter jejuni | |
Zn2+ | 35% inhibition at 2 mM | Campylobacter jejuni |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.159 | - |
N-benzoylglutamate | pH 7.5, 37°C, purified recombinant wild-type enzyme | Campylobacter jejuni | |
0.169 | - |
N-benzoylalanine | pH 7.5, 37°C, purified recombinant wild-type enzyme | Campylobacter jejuni | |
0.195 | - |
N-benzoylphenylalanine | pH 7.5, 37°C, purified recombinant wild-type enzyme | Campylobacter jejuni | |
0.303 | - |
N-benzoylmethionine | pH 7.5, 37°C, purified recombinant wild-type enzyme | Campylobacter jejuni | |
0.594 | - |
N-benzoylvaline | pH 7.5, 37°C, purified recombinant wild-type enzyme | Campylobacter jejuni | |
0.65 | - |
hippurate | pH 7.5, 37°C, native enzyme in cell extract | Campylobacter jejuni | |
0.764 | - |
hippurate | pH 7.5, 37°C, purified recombinant wild-type enzyme | Campylobacter jejuni | |
1.769 | - |
4-aminobenzoyl-glutamate | pH 7.5, 37°C, purified recombinant wild-type enzyme | Campylobacter jejuni | |
2.778 | - |
hippurate | pH 7.5, 37°C, recombinant mutant D76N | Campylobacter jejuni | |
3.377 | - |
N-acetylglycine | pH 7.5, 37°C, purified recombinant wild-type enzyme | Campylobacter jejuni | |
5.45 | - |
N-acetyl-L-alanine | pH 7.5, 37°C, purified recombinant wild-type enzyme | Campylobacter jejuni |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ba2+ | 10% activation at 10 mM | Campylobacter jejuni | |
K+ | 20% activation at 10 mM | Campylobacter jejuni | |
Mg2+ | 20% activation at 10 mM | Campylobacter jejuni | |
Zn2+ | 1.98 molecules of zinc per enzyme subunit molecule, metalloenzyme | Campylobacter jejuni |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
42400 | - |
4 * 42400, recombinant His6-tagged enzyme, SDS-PAGE | Campylobacter jejuni |
193000 | - |
recombinant His6-tagged enzyme, gel filtration | Campylobacter jejuni |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
hippurate + H2O | Campylobacter jejuni | - |
benzoate + glycine | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Campylobacter jejuni | - |
strain ATCC 43431, gene hipO | - |
Purification (Comment) | Organism |
---|---|
recombinant His6-tagged enzyme from Escherichia coli strain M15 by nickel affinity chromatography | Campylobacter jejuni |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
hippurate + H2O = benzoate + glycine | residues Asp76, Aps104, Glu134, Glu135, His161 and His356 are involved in zinc-binding and enzyme catalysis | Campylobacter jejuni |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
120 | - |
purified recombinant wild-type enzyme | Campylobacter jejuni |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-aminobenzoyl-glutamate + H2O | - |
Campylobacter jejuni | 4-aminobenzoate + glutamate | - |
? | |
hippurate + H2O | - |
Campylobacter jejuni | benzoate + glycine | - |
? | |
hippurate + H2O | i.e. N-benzoylglycine | Campylobacter jejuni | benzoate + glycine | - |
? | |
additional information | substrate specificity, the enzyme also shows metallocarboxypeptidase activity with carboxypeptidase substrates in vitro, displaying its highest activity against N-benzoyl-linked small aliphatic acids, overview, the enzyme does not hydrolyze typical substrates for aminopeptidases, lysine-p-nitroanilide, leucine-p-nitroanilide, or endopeptidases, N-benzoylarginine-p-nitroanilide, succinylphenylalanine-4-nitroanilide | Campylobacter jejuni | ? | - |
? | |
N-acetylalanine + H2O | - |
Campylobacter jejuni | acetate + alanine | - |
? | |
N-acetylglycine + H2O | - |
Campylobacter jejuni | acetate + glycine | - |
? | |
N-benzoylalanine + H2O | - |
Campylobacter jejuni | benzoate + alanine | - |
? | |
N-benzoylglutamate + H2O | - |
Campylobacter jejuni | benzoate + glutamate | - |
? | |
N-benzoylmethionine + H2O | - |
Campylobacter jejuni | benzoate + methionine | - |
? | |
N-benzoylphenylalanine + H2O | - |
Campylobacter jejuni | benzoate + phenylalanine | - |
? | |
N-benzoylvaline + H2O | - |
Campylobacter jejuni | benzoate + valine | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | the enzyme structure shows highly ordered alpha-helix and beta-sheet sequences | Campylobacter jejuni |
tetramer | 4 * 42400, recombinant His6-tagged enzyme, SDS-PAGE | Campylobacter jejuni |
Synonyms | Comment | Organism |
---|---|---|
hippurate hydrolase | - |
Campylobacter jejuni |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
- |
Campylobacter jejuni |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.052 | 2.1 | N-acetylglycine | pH 7.5, 37°C, purified recombinant wild-type enzyme | Campylobacter jejuni | |
0.17 | - |
N-benzoylglutamate | pH 7.5, 37°C, purified recombinant wild-type enzyme | Campylobacter jejuni | |
0.5 | - |
N-benzoylvaline | pH 7.5, 37°C, purified recombinant wild-type enzyme | Campylobacter jejuni | |
1.4 | - |
hippurate | pH 7.5, 37°C, purified recombinant wild-type enzyme | Campylobacter jejuni | |
2 | 8 | N-acetyl-L-alanine | pH 7.5, 37°C, purified recombinant wild-type enzyme | Campylobacter jejuni | |
2.32 | - |
hippurate | pH 7.5, 37°C, recombinant mutant D76N | Campylobacter jejuni | |
2.6 | - |
N-benzoylmethionine | pH 7.5, 37°C, purified recombinant wild-type enzyme | Campylobacter jejuni | |
5.95 | - |
N-benzoylphenylalanine | pH 7.5, 37°C, purified recombinant wild-type enzyme | Campylobacter jejuni | |
10.18 | - |
N-acetylglycine | pH 7.5, 37°C, purified recombinant wild-type enzyme | Campylobacter jejuni | |
13.33 | - |
N-benzoylvaline | pH 7.5, 37°C, purified recombinant wild-type enzyme | Campylobacter jejuni | |
14.75 | - |
4-aminobenzoyl-glutamate | pH 7.5, 37°C, purified recombinant wild-type enzyme | Campylobacter jejuni | |
22.12 | - |
N-acetyl-L-alanine | pH 7.5, 37°C, purified recombinant wild-type enzyme | Campylobacter jejuni | |
29.38 | - |
N-benzoylalanine | pH 7.5, 37°C, purified recombinant wild-type enzyme | Campylobacter jejuni | |
48.72 | - |
N-benzoylglutamate | pH 7.5, 37°C, purified recombinant wild-type enzyme | Campylobacter jejuni | |
57.13 | - |
N-benzoylmethionine | pH 7.5, 37°C, purified recombinant wild-type enzyme | Campylobacter jejuni | |
81 | - |
N-benzoylalanine | pH 7.5, 37°C, purified recombinant wild-type enzyme | Campylobacter jejuni | |
98.73 | - |
hippurate | pH 7.5, 37°C, purified recombinant wild-type enzyme | Campylobacter jejuni |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
- |
Campylobacter jejuni |