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Literature summary extracted from

  • Byun, T; Tang, M.; Sloma, A.; Brown, K.M.; Marumoto, C.; Fujii, M.; Blinkovsky, A.M.
    Aminopeptidase from Sphingomonas capsulata (2001), J. Biol. Chem., 276, 17902-17907.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.4.11.2 DNA and amino acid sequence determination and analysis, expression in Escherichia coli strain XL1-Blue Novosphingobium capsulatum
3.4.11.22 DNA and amino acid sequence determination and analysis, expression in Bacillus subtilis and Escherichia coli Novosphingobium capsulatum

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.4.11.2 1,10-phenanthroline
-
 Novosphingobium capsulatum
3.4.11.2 additional information no inhibition by EDTA and PMSF Novosphingobium capsulatum
3.4.11.2 Zn2+ slight inhibition, as ferrocyanide, iodate, and phosphate, Ki value depends on solubility of the salt Novosphingobium capsulatum
3.4.11.22 ferrocyanide
-
 Novosphingobium capsulatum
3.4.11.22 iodate
-
 Novosphingobium capsulatum
3.4.11.22 additional information no inhibition by EDTA and PMSF Novosphingobium capsulatum
3.4.11.22 o-phenanthroline 96% inhibition Novosphingobium capsulatum
3.4.11.22 phosphate
-
 Novosphingobium capsulatum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.4.11.2 6.7
-
 L-Ala-2-naphthylamide pH 7.5, 43°C Novosphingobium capsulatum
3.4.11.2 14
-
 L-Ala-4-nitroanilide pH 7.5, 43°C Novosphingobium capsulatum
3.4.11.22 6.7
-
 Ala beta-naphthylamide pH 7.5, 21°C, native enzyme Novosphingobium capsulatum
3.4.11.22 14
-
 Ala-4-nitroanilide pH 7.5, 21°C, native enzyme Novosphingobium capsulatum

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.4.11.2 extracellular the enzyme is secreted Novosphingobium capsulatum
-
-
3.4.11.22 extracellular secretion to culture medium Novosphingobium capsulatum
-
-
3.4.11.22 additional information not in the periplasm Novosphingobium capsulatum
-
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.4.11.2 Zn2+ the enzyme contains the zinc binding motif HEXXH Novosphingobium capsulatum
3.4.11.22 additional information no detectable content of calcium and cobalt Novosphingobium capsulatum
3.4.11.22 Zinc 1 atom per enzyme molecule, contains the HEXXH motif Novosphingobium capsulatum

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.4.11.2 66000
-
 x * 66000, SDS-PAGE Novosphingobium capsulatum
3.4.11.22 66000
-
 x * 66000, SDS-PAGE Novosphingobium capsulatum

Organism

EC Number Organism UniProt Comment Textmining
3.4.11.2 Novosphingobium capsulatum
-
-
-
3.4.11.2 Novosphingobium capsulatum IFO 12533
-
-
-
3.4.11.22 Novosphingobium capsulatum
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.4.11.2 native enzyme from culture broth supernatant by ultrafiltration, ion exchange chromatography, and gel filtration Novosphingobium capsulatum
3.4.11.22 native from culture supernatant Novosphingobium capsulatum

Reaction

EC Number Reaction Comment Organism Reaction ID
3.4.11.22 Release of an N-terminal amino acid, preferably a neutral or hydrophobic one, from a polypeptide. Aminoacyl-arylamides are poor substrates Gly-Gly bond is preferably hydrolyzed Novosphingobium capsulatum
a

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.4.11.2 105
-
 purified enzyme Novosphingobium capsulatum
3.4.11.22 105
-
 purified native enzyme, substrate Ala-4-nitroanilide Novosphingobium capsulatum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.4.11.2 Gly-4-nitroanilide + H2O 14% of the activity with L-Ala-4-nitroanilide Novosphingobium capsulatum Gly + 4-nitroaniline
-
 ?
3.4.11.2 L-Ala-2-naphthylamide + H2O
-
 Novosphingobium capsulatum L-Ala + 2-naphthylamine
-
 ?
3.4.11.2 L-Ala-4-nitroanilide + H2O best substrate Novosphingobium capsulatum L-Ala + 4-nitroaniline
-
 ?
3.4.11.2 L-Asp-4-nitroanilide + H2O 14% of the activity with L-Ala-4-nitroanilide Novosphingobium capsulatum L-Asp + 4-nitroaniline
-
 ?
3.4.11.2 L-Glu-4-nitroanilide + H2O 12% of the activity with L-Ala-4-nitroanilide Novosphingobium capsulatum L-Glu + 4-nitroaniline
-
 ?
3.4.11.2 L-Leu-4-nitroanilide + H2O 27% of the activity with L-Ala-4-nitroanilide Novosphingobium capsulatum L-Leu + 4-nitroaniline
-
 ?
3.4.11.2 L-Lys-4-nitroanilide + H2O 3% of the activity with L-Ala-4-nitroanilide Novosphingobium capsulatum L-Lys + 4-nitroaniline
-
 ?
3.4.11.2 L-Met-4-nitroanilide + H2O 24% of the activity with L-Ala-4-nitroanilide Novosphingobium capsulatum L-Met + 4-nitroaniline
-
 ?
3.4.11.2 additional information no activity with 4-nitroanilides of Pro, Ile, Val, and Phe Novosphingobium capsulatum ?
-
 ?
3.4.11.22 Ala-4-nitroanilide + H2O best Xaa-4-nitroanilide substrate Novosphingobium capsulatum Ala + 4-nitroaniline
-
 ?
3.4.11.22 Ala-4-nitroanilide + H2O best Xaa-4-nitroanilide substrate Novosphingobium capsulatum IFO 12533 Ala + 4-nitroaniline
-
 ?
3.4.11.22 Ala-Ala-Pro-Tyr-Lys-amide + H2O
-
 Novosphingobium capsulatum Ala + Ala-Pro-Tyr-Lys-amide
-
 ?
3.4.11.22 Ala-beta-naphthylamide + H2O
-
 Novosphingobium capsulatum Ala + beta-naphthylamine
-
 ?
3.4.11.22 Asp-4-nitroanilide + H2O
-
 Novosphingobium capsulatum Asp + 4-nitroaniline
-
 ?
3.4.11.22 Glu-4-nitroanilide + H2O
-
 Novosphingobium capsulatum Glu + 4-nitroaniline
-
 ?
3.4.11.22 Glu-Ala-Pro-Tyr-Lys-amide + H2O
-
 Novosphingobium capsulatum Glu + Ala-Pro-Tyr-Lys-amide
-
 ?
3.4.11.22 Gly-4-nitroanilide + H2O
-
 Novosphingobium capsulatum Gly + 4-nitroaniline
-
 ?
3.4.11.22 Gly-Ala-Pro-Tyr-Lys-amide + H2O best pentapeptide substrate Novosphingobium capsulatum Gly + Ala-Pro-Tyr-Lys-amide
-
 ?
3.4.11.22 Leu-4-nitroanilide + H2O
-
 Novosphingobium capsulatum Leu + 4-nitroaniline
-
 ?
3.4.11.22 Leu-4-nitroanilide + H2O
-
 Novosphingobium capsulatum IFO 12533 Leu + 4-nitroaniline
-
 ?
3.4.11.22 Leu-Ala-Pro-Tyr-Lys-amide + H2O
-
 Novosphingobium capsulatum Leu + Ala-Pro-Tyr-Lys-amide
-
 ?
3.4.11.22 leucine enkephalin + H2O recombinant enzyme, release of Tyr, Gly, Phe and Leu Novosphingobium capsulatum ?
-
 ?
3.4.11.22 Lys-4-nitroanilide + H2O low activity Novosphingobium capsulatum Lys + 4-nitroaniline
-
 ?
3.4.11.22 Lys-4-nitroanilide + H2O low activity Novosphingobium capsulatum IFO 12533 Lys + 4-nitroaniline
-
 ?
3.4.11.22 Met-4-nitroanilide + H2O
-
 Novosphingobium capsulatum Met + 4-nitroaniline
-
 ?
3.4.11.22 Met-4-nitroanilide + H2O
-
 Novosphingobium capsulatum IFO 12533 Met + 4-nitroaniline
-
 ?
3.4.11.22 additional information no activity with 4-nitroanilides of Pro, Ile, Val, and Phe Novosphingobium capsulatum ?
-
 ?
3.4.11.22 additional information no activity with 4-nitroanilides of Pro, Ile, Val, and Phe Novosphingobium capsulatum IFO 12533 ?
-
 ?
3.4.11.22 Pro-Ala-Pro-Tyr-Lys-amide + H2O
-
 Novosphingobium capsulatum Pro + Ala-Pro-Tyr-Lys-amide
-
 ?

Subunits

EC Number Subunits Comment Organism
3.4.11.2 ? x * 66000, SDS-PAGE Novosphingobium capsulatum
3.4.11.22 ? x * 66000, SDS-PAGE Novosphingobium capsulatum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.4.11.2 43
-
-
 Novosphingobium capsulatum
3.4.11.22 43
-
-
 Novosphingobium capsulatum

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.4.11.2 45
-
 20 min, purified enzyme, loss of 40% activity Novosphingobium capsulatum
3.4.11.2 50
-
 20 min, purified enzyme, inactivation Novosphingobium capsulatum
3.4.11.22 45
-
 20 min, loss of 40% activity Novosphingobium capsulatum
3.4.11.22 50
-
 20 min, complete loss of activity Novosphingobium capsulatum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.4.11.2 860
-
 L-Ala-2-naphthylamide pH 7.5, 43°C Novosphingobium capsulatum
3.4.11.2 7600
-
 L-Ala-4-nitroanilide pH 7.5, 43°C Novosphingobium capsulatum
3.4.11.22 0.52
-
 Ala-beta-naphthylamide pH 7.5, 21°C, native enzyme Novosphingobium capsulatum
3.4.11.22 126.7
-
 Ala-4-nitroanilide pH 7.5, 21°C, native enzyme Novosphingobium capsulatum
3.4.11.22 138
-
 Ala-4-nitroanilide pH 7.5, 21°C, native enzyme Novosphingobium capsulatum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.4.11.2 7.2 7.4
-
 Novosphingobium capsulatum
3.4.11.22 7.2 7.4
-
 Novosphingobium capsulatum

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
3.4.11.2 additional information
-
 additional information
-
 Novosphingobium capsulatum
3.4.11.22 3
-
 phosphate pH 7.5, 21°C Novosphingobium capsulatum
3.4.11.22 4.2
-
 ferrocyanide pH 7.5, 21°C Novosphingobium capsulatum
3.4.11.22 11
-
 iodate pH 7.5, 21°C Novosphingobium capsulatum