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Literature summary extracted from

  • Ma, Y.; Xu, W.; Zhang, J.; Zhang, S.; Hong, K.; Deng, Z.; Sun, Y.
    Nitrilase superfamily aryl acylamidase from the halotolerant mangrove Streptomyces sp. 211726 (2014), Appl. Microbiol. Biotechnol., 98, 8583-8590.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.5.1.4 additional information no effect by EDTA, 8-hydroxyquinoline, and 2-mercaptoethanol Streptomyces sp.
3.5.1.4 PMSF slight activation at 1 mM Streptomyces sp.
3.5.1.13 additional information no effect by EDTA, 8-hydroxyquinoline, and 2-mercaptoethanol Streptomyces sp.
3.5.1.13 PMSF slight activation at 1 mM Streptomyces sp.

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.1.4 gene azl13, DNA and amino acid sequence determination and analysis, sequence comparison, recombinant expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3) Streptomyces sp.
3.5.1.13 gene azl13, DNA and amino acid sequence determination and analysis, sequence comparison, recombinant expression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Streptomyces sp.

Protein Variants

EC Number Protein Variants Comment Organism
3.5.1.4 C141A site-directed mutagenesis, inactive mutant Streptomyces sp.
3.5.1.4 E41A site-directed mutagenesis, inactive mutant Streptomyces sp.
3.5.1.4 K107A site-directed mutagenesis, inactive mutant Streptomyces sp.
3.5.1.13 C141A site-directed mutagenesis, inactive mutant Streptomyces sp.
3.5.1.13 E41A site-directed mutagenesis, inactive mutant Streptomyces sp.
3.5.1.13 K107A site-directed mutagenesis, inactive mutant Streptomyces sp.

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.5.1.4 Ag+ complete inhibition at 1 mM Streptomyces sp.
3.5.1.4 Cd2+ complete inhibition at 1 mM Streptomyces sp.
3.5.1.4 Cu2+ complete inhibition at 1 mM Streptomyces sp.
3.5.1.4 Hg2+ complete inhibition at 1 mM Streptomyces sp.
3.5.1.4 additional information no effect by EDTA, 8-hydroxyquinoline, and 2-mercaptoethanol Streptomyces sp.
3.5.1.13 Ag+ complete inhibition at 1 mM Streptomyces sp.
3.5.1.13 Cd2+ complete inhibition at 1 mM Streptomyces sp.
3.5.1.13 Cu2+ complete inhibition at 1 mM Streptomyces sp.
3.5.1.13 Hg2+ complete inhibition at 1 mM Streptomyces sp.
3.5.1.13 additional information no effect by EDTA, 8-hydroxyquinoline, and 2-mercaptoethanol Streptomyces sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.1.4 2.3
-
 4-Guanidinobutyramide pH 8.0, 35°C, recombinant enzyme Streptomyces sp.
3.5.1.4 39.4
-
 acetamide pH 8.0, 35°C, recombinant enzyme Streptomyces sp.
3.5.1.4 45.1
-
 Propionamide pH 8.0, 35°C, recombinant enzyme Streptomyces sp.
3.5.1.4 135.6
-
 nicotinamide pH 8.0, 35°C, recombinant enzyme Streptomyces sp.

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.5.1.4 additional information no requirement for metal ions Streptomyces sp.
3.5.1.13 additional information no requirement for metal ions Streptomyces sp.

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.5.1.4 31000
-
 x * 31000, recombinant His-tagged enzyme, SDS-PAGE Streptomyces sp.
3.5.1.13 31000
-
 x * 31000, recombinant His-tagged enzyme, SDS-PAGE Streptomyces sp.

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.5.1.4 4-guanidinobutyramide + H2O Streptomyces sp. probably the natural substrate ?
-
 ?
3.5.1.13 additional information Streptomyces sp. 4-guanidinobutyramide probably is the natural substrate ?
-
 ?
3.5.1.13 additional information Streptomyces sp. 211726 4-guanidinobutyramide probably is the natural substrate ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.5.1.4 Streptomyces sp. A0A088BHP3 gene azl13
-
3.5.1.4 Streptomyces sp. 211726 A0A088BHP3 gene azl13
-
3.5.1.13 Streptomyces sp. A0A088BHP3 gene azl13
-
3.5.1.13 Streptomyces sp. 211726 A0A088BHP3 gene azl13
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.1.4 recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Streptomyces sp.
3.5.1.13 recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Streptomyces sp.

Source Tissue

EC Number Source Tissue Comment Organism Textmining
3.5.1.4 additional information halotolerant organism from mangrove swamps Streptomyces sp.
-
3.5.1.13 additional information halotolerant organism from mangrove swamps Streptomyces sp.
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.5.1.4 0.55
-
 recombinant purified His-tagged enzyme, pH 8.0, 35°C, substrate 3',4'-dichloropropionanilide Streptomyces sp.
3.5.1.4 0.69
-
 recombinant purified His-tagged enzyme, pH 8.0, 35°C, substrate 2',6'-dimethylacetanilide Streptomyces sp.
3.5.1.4 0.87
-
 recombinant purified His-tagged enzyme, pH 8.0, 35°C, substrate 4'-methylacetanilide Streptomyces sp.
3.5.1.13 0.55
-
 recombinant purified His-tagged enzyme, pH 8.0, 35°C, substrate 3',4'-dichloropropionanilide Streptomyces sp.
3.5.1.13 0.69
-
 recombinant purified His-tagged enzyme, pH 8.0, 35°C, substrate 2',6'-dimethylacetanilide Streptomyces sp.
3.5.1.13 0.87
-
 recombinant purified His-tagged enzyme, pH 8.0, 35°C, substrate p-acetotoluidide Streptomyces sp.

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.1.4 2',6'-dimethylacetanilide + H2O
-
 Streptomyces sp. 2,6-dimethylaniline + acetate
-
 ?
3.5.1.4 3',4'-dichloropropionanilide + H2O
-
 Streptomyces sp. 3,4-dichloroaniline + propanoate
-
 ?
3.5.1.4 4'-methylacetanilide + H2O
-
 Streptomyces sp. 4-methylaniline + acetate
-
 ?
3.5.1.4 4-guanidinobutyramide + H2O best substrate Streptomyces sp. ?
-
 ?
3.5.1.4 4-guanidinobutyramide + H2O probably the natural substrate Streptomyces sp. ?
-
 ?
3.5.1.4 acetamide + H2O
-
 Streptomyces sp. acetate + NH3
-
 ?
3.5.1.4 benzamide + H2O
-
 Streptomyces sp. benzoate + NH3
-
 ?
3.5.1.4 isobutyramide + H2O
-
 Streptomyces sp. isobutyrate + NH3
-
 ?
3.5.1.4 isonicotinamide + H2O
-
 Streptomyces sp. isonicotinate + NH3
-
 ?
3.5.1.4 additional information substrate specificity, overview. The recombinant enzyme displays an unusually wide substrate spectrum. It is moderately active on short-chain aliphatic amides and weakly active hydrolyzing aromatic and heterocyclic amides. Recombinant His6-tagged enzyme Azl13 also catalyzes acyl transfer to hydroxylamine from acetamide or the herbicide propanil. The enzyme also shows aryl acylamide hydrolase activity, EC 3.5.1.13, and hydroxylamine acyl transferase activity, EC 2.3.1.56. No activity with L-glutamine and L-asparagine Streptomyces sp. ?
-
 ?
3.5.1.4 nicotinamide + H2O reaction of EC 3.5.1.19, nicotinamidase Streptomyces sp. nicotinate + NH3
-
 ?
3.5.1.4 propionamide + H2O
-
 Streptomyces sp. propionate + NH3
-
 ?
3.5.1.4 pyrazinamide + H2O
-
 Streptomyces sp. pyrazinate + NH3
-
 ?
3.5.1.13 2',6'-dimethylacetanilide + H2O
-
 Streptomyces sp. ?
-
 ?
3.5.1.13 2',6'-dimethylacetanilide + H2O
-
 Streptomyces sp. 211726 ?
-
 ?
3.5.1.13 3',4'-dichloropropionanilide + H2O i.e. propanil Streptomyces sp. 3,4-dichloroaniline + propionate
-
 ?
3.5.1.13 3',4'-dichloropropionanilide + H2O i.e. propanil Streptomyces sp. 211726 3,4-dichloroaniline + propionate
-
 ?
3.5.1.13 additional information 4-guanidinobutyramide probably is the natural substrate Streptomyces sp. ?
-
 ?
3.5.1.13 additional information substrate specificity, overview. The recombinant enzyme displays an unusually wide substrate spectrum. It is moderately active on short-chain aliphatic amides and weakly active hydrolyzing aromatic and heterocyclic amides. Recombinant His6-tagged enzyme Azl13 also catalyzes acyl transfer to hydroxylamine from acetamide or the herbicide propanil. The enzyme also shows amidase, EC 3.5.1.4, and hydroxylamine acyl transferase activity, EC 2.3.1.56. No activity with benzonitrile. phenylacetonitrile, Flutolanil, L-glutamine, and L-asparagine Streptomyces sp. ?
-
 ?
3.5.1.13 additional information 4-guanidinobutyramide probably is the natural substrate Streptomyces sp. 211726 ?
-
 ?
3.5.1.13 additional information substrate specificity, overview. The recombinant enzyme displays an unusually wide substrate spectrum. It is moderately active on short-chain aliphatic amides and weakly active hydrolyzing aromatic and heterocyclic amides. Recombinant His6-tagged enzyme Azl13 also catalyzes acyl transfer to hydroxylamine from acetamide or the herbicide propanil. The enzyme also shows amidase, EC 3.5.1.4, and hydroxylamine acyl transferase activity, EC 2.3.1.56. No activity with benzonitrile. phenylacetonitrile, Flutolanil, L-glutamine, and L-asparagine Streptomyces sp. 211726 ?
-
 ?
3.5.1.13 p-acetotoluidide + H2O
-
 Streptomyces sp. ?
-
 ?
3.5.1.13 p-acetotoluidide + H2O
-
 Streptomyces sp. 211726 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.5.1.4 ? x * 31000, recombinant His-tagged enzyme, SDS-PAGE Streptomyces sp.
3.5.1.13 ? x * 31000, recombinant His-tagged enzyme, SDS-PAGE Streptomyces sp.

Synonyms

EC Number Synonyms Comment Organism
3.5.1.4 4-guanidinobutyramide hydrolase
-
 Streptomyces sp.
3.5.1.4 Azl13
-
 Streptomyces sp.
3.5.1.13 aryl acylamidase
-
 Streptomyces sp.
3.5.1.13 Azl13
-
 Streptomyces sp.

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.5.1.4 40
-
-
 Streptomyces sp.
3.5.1.13 40
-
-
 Streptomyces sp.

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.5.1.4 15 60 activity range Streptomyces sp.
3.5.1.13 15 60 activity range Streptomyces sp.

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.5.1.4 40
-
 purified His6-tagged enzyme, pH 8.0, stable at for 1 h Streptomyces sp.
3.5.1.4 45 50 purified His6-tagged enzyme, pH 8.0, 1 h, 50% activity remaining Streptomyces sp.
3.5.1.4 60
-
 purified His6-tagged enzyme, pH 8.0, 1 h, inactivation Streptomyces sp.
3.5.1.13 40
-
 purified His6-tagged enzyme, pH 8.0, stable at for 1 h Streptomyces sp.
3.5.1.13 45 50 purified His6-tagged enzyme, pH 8.0, 1 h, 50% activity remaining Streptomyces sp.
3.5.1.13 60
-
 purified His6-tagged enzyme, pH 8.0, 1 h, inactivation Streptomyces sp.

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.5.1.4 5.6
-
 nicotinamide pH 8.0, 35°C, recombinant enzyme Streptomyces sp.
3.5.1.4 28.6
-
 Propionamide pH 8.0, 35°C, recombinant enzyme Streptomyces sp.
3.5.1.4 29.9
-
 acetamide pH 8.0, 35°C, recombinant enzyme Streptomyces sp.
3.5.1.4 55.6
-
 4-Guanidinobutyramide pH 8.0, 35°C, recombinant enzyme Streptomyces sp.

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5.1.4 8
-
-
 Streptomyces sp.
3.5.1.13 8
-
-
 Streptomyces sp.

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.5.1.4 5.5 10 activity range Streptomyces sp.
3.5.1.13 5.5 10 activity range Streptomyces sp.

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
3.5.1.4 4.5 10 purified His6-tagged enzyme, over 50% activity remaining within this range after 1 h Streptomyces sp.
3.5.1.13 4.5 10 purified His6-tagged enzyme, over 50% activity remaining within this range after 1 h Streptomyces sp.

General Information

EC Number General Information Comment Organism
3.5.1.4 evolution the enzyme belongs to the nitrilase superfamily, subfamily in branch 13. The enzyme contains highly conserved E41-K107-C141 catalytic triad which is typical for nitrilase superfamily amidases Streptomyces sp.
3.5.1.13 evolution the enzyme belongs to the nitrilase superfamily, subfamily in branch 13. The enzyme contains highly conserved E41-K107-C141 catalytic triad which is typical for nitrilase superfamily amidases Streptomyces sp.

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.5.1.4 0.034
-
 Benzamide pH 8.0, 35°C, recombinant enzyme Streptomyces sp.
3.5.1.4 0.042
-
 nicotinamide pH 8.0, 35°C, recombinant enzyme Streptomyces sp.
3.5.1.4 0.64
-
 Propionamide pH 8.0, 35°C, recombinant enzyme Streptomyces sp.
3.5.1.4 0.76
-
 acetamide pH 8.0, 35°C, recombinant enzyme Streptomyces sp.
3.5.1.4 24.17
-
 4-Guanidinobutyramide pH 8.0, 35°C, recombinant enzyme Streptomyces sp.