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Literature summary for 3.5.1.14 extracted from

  • Koreishi, M.; Nakatani, Y.; Ooi, M.; Imanaka, H.; Imamura, K.; Nakanishi, K.
    Purification, characterization, molecular cloning, and expression of a new aminoacylase from Streptomyces mobaraensis that can hydrolyze N-(middle/long)-chain-fatty-acyl-L-amino acids as well as N-short-chain-acyl-L-amino acids (2009), Biosci. Biotechnol. Biochem., 73, 1940-1947.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
DNA and amino acid sequence determination and analysis, sequence comparisons, subcloning in Escherichia coli strain DH5alpha, expression in Streptomyces lividans strain TK24 resulting in 42fold higher activity compared to the native wild-type Streptomyces mobaraensis strain Streptomyces mobaraensis
expressed in Streptomyces lividans strain TK24 Streptomyces mobaraensis

Inhibitors

Inhibitors Comment Organism Structure
1,10-phenanthroline 10% residual activity at 1 mM; 90% inhibition at 1 mM Streptomyces mobaraensis
2-mercaptoethanol 20% inhibition at 1 mM; 81% residual activity at 1 mM Streptomyces mobaraensis
Ca2+ 86% residual activity at 0.005 mM Streptomyces mobaraensis
Co2+ 82% residual activity at 0.005 mM; slight inhibition at 0.5 mM Streptomyces mobaraensis
Cu2+ 93% residual activity at 0.005 mM; slight inhibition at 0.5 mM Streptomyces mobaraensis
Fe2+ 86% residual activity at 0.005 mM; 90% inhibition at 0.5 mM Streptomyces mobaraensis
iodoacetamide 15% inhibition at 1 mM; 85% residual activity at 1 mM Streptomyces mobaraensis
Mg2+ 96% residual activity at 0.005 mM Streptomyces mobaraensis
Mn2+ 88% residual activity at 0.005 mM; slight inhibition at 0.5 mM Streptomyces mobaraensis
additional information the activity of the recombinant enzyme is not influenced by dithiothreitol, reduced glutathione, EDTA, p-chloromercuribenzoic acid, and L-Cys Streptomyces mobaraensis
Ni2+ slight inhibition at 0.5 mM Streptomyces mobaraensis
Zn2+ activates 57% at 500 nM, 20% at 0.005 mM, but inhibits 61% at 2.5 mM; slight inhibition at 0.5 mM; the relative remaining activity is decreased to 39% by incubation with 2.5 mM Zn2+ Streptomyces mobaraensis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
11.3
-
N-acetyl-L-Met recombinant enzyme, at 37°C, pH not specified in the publication Streptomyces mobaraensis

Metals/Ions

Metals/Ions Comment Organism Structure
additional information the enzyme activity is not affected by Ca2+ or Mg2+ at 0.5 mM Streptomyces mobaraensis
Ni2+ 110% relative activity at 0.005 mM Streptomyces mobaraensis
Zn2+ 120% relative activity at 0.0005 mM Streptomyces mobaraensis
Zn2+ activates 57% at 500 nM, 20% at 0.005 mM, but inhibits 61% at 2.5 mM Streptomyces mobaraensis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
50000
-
SDS-PAGE Streptomyces mobaraensis
55000
-
-
Streptomyces mobaraensis
55000
-
blue native PAGE Streptomyces mobaraensis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Streptomyces mobaraensis the enzyme shows high hydrolytic activity towards various N-acetyl-L-amino acids and N-(middle/long)-chain-fatty-acyl-L-amino acids with a preference for the acyl derivatives of L-Met, L-Ala, L-Cys, etc ?
-
?
N-acetyl-L-alanine + H2O Streptomyces mobaraensis
-
acetate + L-alanine
-
?
N-acetyl-L-cysteine + H2O Streptomyces mobaraensis
-
acetate + L-cysteine
-
?
N-acetyl-L-methionine + H2O Streptomyces mobaraensis
-
acetate + L-methionine
-
?

Organism

Organism UniProt Comment Textmining
Streptomyces mobaraensis C9K2Z6 strain NBRC (IFO) 13819
-
Streptomyces mobaraensis C9K2Z6 strain NBRC 13819
-

Purification (Commentary)

Purification (Comment) Organism
ammonium sulfate precipitation, octyl-Sepharose column chromatography, HiTrap chelating column chromatography, and DEAE Sephadex A-50 ion exchange chromatography Streptomyces mobaraensis
Streptomyces lividans strain TK24, native enzyme 494fold from by ammonium sulfate fractionation, anion exchange chromatography, and hydrophobic interaction chromatography Streptomyces mobaraensis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
substrate specificities of recombinant and native enzyme, overview Streptomyces mobaraensis
0.89
-
native wild-type enzyme, culture supernatant, substrate N-acetyl-L-methionine, pH 7.5, 37°C Streptomyces mobaraensis
0.89
-
recombinant wild-type enzyme, culture supernatant, substrate N-acetyl-L-methionine, pH 7.5, 37°C Streptomyces mobaraensis
4
-
using N-lauroyl-L-Arg as substrate, at 37°C, pH not specified in the publication Streptomyces mobaraensis
6
-
using N-lauroyl-L-Asn as substrate, at 37°C, pH not specified in the publication Streptomyces mobaraensis
6
-
using N-lauroyl-L-His as substrate, at 37°C, pH not specified in the publication Streptomyces mobaraensis
8
-
using N-acetyl-Gly as substrate, at 37°C, pH not specified in the publication Streptomyces mobaraensis
8
-
using N-acetyl-L-Asp as substrate, at 37°C, pH not specified in the publication Streptomyces mobaraensis
14
-
using N-acetyl-L-Gln as substrate, at 37°C, pH not specified in the publication Streptomyces mobaraensis
14
-
using N-lauroyl-L-Lys as substrate, at 37°C, pH not specified in the publication Streptomyces mobaraensis
17
-
using N-lauroyl-L-Cys as substrate, at 37°C, pH not specified in the publication Streptomyces mobaraensis
19
-
using N-acetyl-L-Phe as substrate, at 37°C, pH not specified in the publication Streptomyces mobaraensis
24
-
using N-acetyl-L-Val as substrate, at 37°C, pH not specified in the publication Streptomyces mobaraensis
31
-
using N-acetyl-L-Asn as substrate, at 37°C, pH not specified in the publication Streptomyces mobaraensis
34
-
using N-lauroyl-Gly as substrate, at 37°C, pH not specified in the publication Streptomyces mobaraensis
36
-
using N-lauroyl-L-Ser as substrate, at 37°C, pH not specified in the publication Streptomyces mobaraensis
59
-
using N-acetyl-L-His as substrate, at 37°C, pH not specified in the publication Streptomyces mobaraensis
60
-
using N-acetyl-L-Leu as substrate, at 37°C, pH not specified in the publication Streptomyces mobaraensis
66
-
using N-acetyl-L-Arg as substrate, at 37°C, pH not specified in the publication Streptomyces mobaraensis
72
-
using 3 mM N-lauroyl-L-Met as substrate, at 37°C, pH not specified in the publication Streptomyces mobaraensis
89
-
using 3 mM N-decanoyl-L-Met as substrate, at 37°C, pH not specified in the publication Streptomyces mobaraensis
96
-
purified recombinant wild-type enzyme, substrate N-lauryl-L-alanine, pH 7.5, 37°C Streptomyces mobaraensis
96
-
using N-lauroyl-L-Ala as substrate, at 37°C, pH not specified in the publication Streptomyces mobaraensis
105
-
using 3 mM N-palmitoyl-L-Met as substrate, at 37°C, pH not specified in the publication Streptomyces mobaraensis
106
-
using 3 mM N-myristoyl-L-Met as substrate, at 37°C, pH not specified in the publication Streptomyces mobaraensis
169
-
using 3 mM N-butyryl-L-Met as substrate, at 37°C, pH not specified in the publication Streptomyces mobaraensis
173
-
using 3 mM N-acetyl-L-Met as substrate, at 37°C, pH not specified in the publication Streptomyces mobaraensis
175
-
using 3 mM N-hexanoyl-L-Met as substrate, at 37°C, pH not specified in the publication Streptomyces mobaraensis
194
-
using N-acetyl-L-Ala as substrate, at 37°C, pH not specified in the publication Streptomyces mobaraensis
251
-
using 3 mM N-octanoyl-L-Met as substrate, at 37°C, pH not specified in the publication Streptomyces mobaraensis
412
-
purified recombinant wild-type enzyme, substrate N-acetyl-L-cysteine, pH 7.5, 37°C Streptomyces mobaraensis
412
-
using N-acetyl-L-Cys as substrate, at 37°C, pH not specified in the publication Streptomyces mobaraensis
440
-
purified native wild-type enzyme, substrate N-acetyl-L-methionine, pH 7.5, 37°C Streptomyces mobaraensis
530
-
purified recombinant wild-type enzyme, substrate N-acetyl-L-methionine, pH 7.5, 37°C Streptomyces mobaraensis
530
-
using 15 mM N-acetyl-L-Met as substrate, at 37°C, pH not specified in the publication Streptomyces mobaraensis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information the enzyme shows high hydrolytic activity towards various N-acetyl-L-amino acids and N-(middle/long)-chain-fatty-acyl-L-amino acids with a preference for the acyl derivatives of L-Met, L-Ala, L-Cys, etc Streptomyces mobaraensis ?
-
?
additional information no activity towards N-acetyl-L-Lys, N-acetyl-L-Glu, N-acetyl-L-Pro, N-acetyl-L-Trp, N-acetyl-L-Tyr, N-lauroyl-L-Ile, N-lauroyl-L-Leu, N-lauroyl-L-Phe, N-lauroyl-L-Pro, N-lauroyl-L-Trp, N-lauroyl-L-Tyr, and N-lauroyl-L-Val Streptomyces mobaraensis ?
-
?
additional information the enzyme hydrolyzes N-(middle/long)-chain-fatty-acyl-L-amino acids as well as N-short-chain-acyl-L-amino acids. Poor or no activity with N-acetyl-amino acids containing L-Lys, L-Asp, L-Cys, L-Pro, L-Trp, and L-Tyr, as well as with N-leuroyl-amino acids containing L-Arg, L-His, L-Asn, L-Ile, L-Leu, L-Phe, L-Pro, L-Trp, L-Tyr, and L-Val, substrate specificity, overview Streptomyces mobaraensis ?
-
?
N-Acetyl-Gly + H2O
-
Streptomyces mobaraensis acetate + Gly
-
?
N-acetyl-L-Ala + H2O
-
Streptomyces mobaraensis acetate + L-Ala
-
?
N-acetyl-L-alanine + H2O
-
Streptomyces mobaraensis acetate + L-alanine
-
?
N-acetyl-L-Arg + H2O
-
Streptomyces mobaraensis acetate + L-Arg
-
?
N-acetyl-L-arginine + H2O
-
Streptomyces mobaraensis acetate + L-arginine
-
?
N-acetyl-L-Asn + H2O
-
Streptomyces mobaraensis acetate + L-Asn
-
?
N-acetyl-L-Asp + H2O
-
Streptomyces mobaraensis acetate + L-Asp
-
?
N-acetyl-L-asparagine + H2O
-
Streptomyces mobaraensis acetate + L-asparagine
-
?
N-acetyl-L-Cys + H2O
-
Streptomyces mobaraensis acetate + L-Cys
-
?
N-acetyl-L-cysteine + H2O
-
Streptomyces mobaraensis acetate + L-cysteine
-
?
N-acetyl-L-Gln + H2O
-
Streptomyces mobaraensis acetate + L-Gln
-
?
N-acetyl-L-glutamine + H2O
-
Streptomyces mobaraensis acetate + L-glutamine
-
?
N-acetyl-L-histidine + H2O
-
Streptomyces mobaraensis acetate + L-histidine
-
?
N-acetyl-L-Leu + H2O
-
Streptomyces mobaraensis acetate + L-Leu
-
?
N-acetyl-L-leucine + H2O
-
Streptomyces mobaraensis acetate + L-leucine
-
?
N-acetyl-L-Met + H2O highest activity Streptomyces mobaraensis acetate + L-Met
-
?
N-acetyl-L-methionine + H2O
-
Streptomyces mobaraensis acetate + L-methionine
-
?
N-acetyl-L-methionine + H2O preferred substrate Streptomyces mobaraensis acetate + L-methionine
-
?
N-acetyl-L-Phe + H2O
-
Streptomyces mobaraensis acetate + L-Phe
-
?
N-acetyl-L-phenylalanine + H2O
-
Streptomyces mobaraensis acetate + L-phenylalanine
-
?
N-acetyl-L-Val + H2O
-
Streptomyces mobaraensis acetate + L-Val
-
?
N-acetyl-L-valine + H2O
-
Streptomyces mobaraensis acetate + L-valine
-
?
N-butyryl-L-Met + H2O
-
Streptomyces mobaraensis butanoate + L-Met
-
?
N-butyryl-L-methionine + H2O
-
Streptomyces mobaraensis butyrate + L-methionine
-
?
N-decanoyl-L-Met + H2O
-
Streptomyces mobaraensis decanoate + L-Met
-
?
N-decanoyl-L-methionine + H2O
-
Streptomyces mobaraensis decanoate + L-methionine
-
?
N-hexanoyl-L-Met + H2O
-
Streptomyces mobaraensis hexanoate + L-Met
-
?
N-hexanoyl-L-methionine + H2O
-
Streptomyces mobaraensis hexanoate + L-methionine
-
?
N-lauroyl-Gly + H2O
-
Streptomyces mobaraensis laurate + Gly
-
?
N-lauroyl-L-Ala + H2O
-
Streptomyces mobaraensis laurate + L-Ala
-
?
N-lauroyl-L-Arg + H2O
-
Streptomyces mobaraensis laurate + L-Arg
-
?
N-lauroyl-L-Asn + H2O
-
Streptomyces mobaraensis laurate + L-Asn
-
?
N-lauroyl-L-Cys + H2O
-
Streptomyces mobaraensis laurate + L-Cys
-
?
N-lauroyl-L-His + H2O
-
Streptomyces mobaraensis laurate + L-His
-
?
N-lauroyl-L-Lys + H2O
-
Streptomyces mobaraensis laurate + L-Lys
-
?
N-lauroyl-L-Met + H2O
-
Streptomyces mobaraensis laurate + L-Met
-
?
N-lauroyl-L-Ser + H2O
-
Streptomyces mobaraensis laurate + L-Ser
-
?
N-lauryl-glycine + H2O
-
Streptomyces mobaraensis laurate + glycine
-
?
N-lauryl-L-alanine + H2O
-
Streptomyces mobaraensis laurate + L-alanine
-
?
N-lauryl-L-cysteine + H2O
-
Streptomyces mobaraensis laurate + L-cysteine
-
?
N-lauryl-L-lysine + H2O
-
Streptomyces mobaraensis laurate + L-lysine
-
?
N-lauryl-L-methionine + H2O
-
Streptomyces mobaraensis laurate + L-methionine
-
?
N-lauryl-L-serine + H2O
-
Streptomyces mobaraensis laurate + L-serine
-
?
N-myristoyl-L-Met + H2O
-
Streptomyces mobaraensis myristate + L-Met
-
?
N-myristoyl-L-methionine + H2O
-
Streptomyces mobaraensis myristate + L-methionine
-
?
N-octanoyl-L-Met + H2O
-
Streptomyces mobaraensis octanoate + L-Met
-
?
N-octanoyl-L-methionine + H2O
-
Streptomyces mobaraensis octanoate + L-methionine
-
?
N-palmitoyl-L-Met + H2O
-
Streptomyces mobaraensis palmitate + L-Met
-
?
N-palmitoyl-L-methionine + H2O
-
Streptomyces mobaraensis palmitate + L-methionine
-
?

Subunits

Subunits Comment Organism
monomer 1 * 55000 Streptomyces mobaraensis
monomer 1 * 55000, purified wild type enzyme, SDS-PAGE Streptomyces mobaraensis

Synonyms

Synonyms Comment Organism
More the enzyme belongs to the peptidase M20 family Streptomyces mobaraensis
N-acyl-L-amino-acid amidohydrolase
-
Streptomyces mobaraensis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
50
-
-
Streptomyces mobaraensis
50
-
at pH 7.5 Streptomyces mobaraensis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
-
Streptomyces mobaraensis