Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 3.5.1.4 extracted from

  • Shen, W.; Chen, H.; Jia, K.; Ni, J.; Yan, X.; Li, S.
    Cloning and characterization of a novel amidase from Paracoccus sp. M-1, showing aryl acylamidase and acyl transferase activities (2012), Appl. Microbiol. Biotechnol., 94, 1007-1018.
    View publication on PubMed
Search for more literature for 3.5.1.4

Cloned(Commentary)

Cloned (Comment) Organism
gene pamh, genetic organization, 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). The enzyme is only successfully expressed using autoinduction media under low-temperature incubation at 25°C, otherwise, the expressed proteins forms inclusion bodies Paracoccus sp.

Protein Variants

Protein Variants Comment Organism
K84A site-directed mutagenesis, the mutant shows no detectable hydrolysis activity Paracoccus sp.
S159A site-directed mutagenesis, the mutant shows no detectable hydrolysis activity Paracoccus sp.
S183A site-directed mutagenesis, the mutant shows no detectable hydrolysis activity Paracoccus sp.

Inhibitors

Inhibitors Comment Organism Structure
1,10-phenanthroline 20-30% inhibition at 1 mM Paracoccus sp.
2-mercaptoethanol 52% inhibition at 1 mM Paracoccus sp.
4-chloromercurybenzoate
-
 Paracoccus sp.
Co2+ strong inhibition Paracoccus sp.
Cr2+
-
 Paracoccus sp.
Cu2+ strong inhibition Paracoccus sp.
EDTA 20-30% inhibition at 1 mM Paracoccus sp.
Fe2+
-
 Paracoccus sp.
Hg2+ complete inhibition Paracoccus sp.
iodoacetamide
-
 Paracoccus sp.
Ni2+
-
 Paracoccus sp.
PMSF strong inhibition Paracoccus sp.
SDS
-
 Paracoccus sp.
Triton X-100
-
 Paracoccus sp.
Tween-80
-
 Paracoccus sp.
Zn2+
-
 Paracoccus sp.

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.8
-
 Acrylamide pH 7.0, 35°C, recombinant enzyme Paracoccus sp.

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ activates at 1 mM Paracoccus sp.
Mg2+ activates at 1 mM Paracoccus sp.
Mn2+ activates at 1 mM Paracoccus sp.

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
52000
-
 x * 52000, about, sequence calculation Paracoccus sp.

Organism

Organism UniProt Comment Textmining
Paracoccus sp. G9FKH7 gene pamH
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His6-tagged wild-type enzyme 5.82fold from Escherichia coli strain BL21 (DE3) by ammonium sulfate fractionation, dialysis, anion exchange chromatography, gel filtration, and ultrafiltration Paracoccus sp.

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
14.33
-
 purified recombinant wild-type enzyme, pH 7.0, 35°C, substrate acrylamides Paracoccus sp.
39.82
-
 purified recombinant wild-type enzyme, pH 7.0, 35°C, substrate benzamide Paracoccus sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
acetamide + H2O
-
 Paracoccus sp. acetate + NH3
-
 ?
acrylamide + H2O
-
 Paracoccus sp. acrylate + NH3
-
 ?
benzamide + H2O
-
 Paracoccus sp. benzoate + NH3
-
 ?
additional information the PamH enzyme exhibits amidase activity, aryl acylamidase activity (EC 3.5.1.13), and acyl transferase activity. It shows excellent activity toward the majority of the aromatic and aliphatic amides, such as acetamide, propionamide, phenylacetamide, and benzamide. The aromatic amides, with substitutions of one or two carbons in the ring by a nitrogen, have a negative influence on amidase activity, leading to low specific activity values for pyrazinamide and nicotinamide. No activity is detectable on long-chain aliphatic amide hexanoamides. Amino acid amides are also hydrolyzed by the enzyme. The enzyme possesses urease activity, but N-methyl substituted is not hydrolyzed by the enzyme. The amidase shows low activity on asparagines (9%), L-glutamine (17%), and D-glutamine (13%) corresponding to benzamide (100%). The anilide substrate range of the enzyme is very narrow and cannot hydrolyze butachlor, acetochlor, 4-nitroacetanilide, p-chloroacetanilide, or other structurally analogous compounds Paracoccus sp. ?
-
 ?
nicotinamide + H2O low activity, reaction of EC 3.5.1.19 Paracoccus sp. nicotinate + NH3
-
 ?
phenylacetamide + H2O
-
 Paracoccus sp. phenylacetate + NH3
-
 ?
propionamide + H2O
-
 Paracoccus sp. propionate + NH3
-
 ?
pyrazinamide + H2O low activity Paracoccus sp. pyrazinate + NH3
-
 ?

Subunits

Subunits Comment Organism
? x * 52000, about, sequence calculation Paracoccus sp.

Synonyms

Synonyms Comment Organism
PamH
-
 Paracoccus sp.

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
45
-
 recombinant enzyme Paracoccus sp.

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
15 70 activity range, profile overview Paracoccus sp.

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
40 60 the purified recombinant wild-type enzyme is fairly stable up to 40°C, has 25% residual activity at 50°C after 1 h, and is completely inactivated at 60°C after 30 min Paracoccus sp.

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
 recombinant enzyme Paracoccus sp.

pH Range

pH Minimum pH Maximum Comment Organism
5.5 10 activity range, profile overview Paracoccus sp.

pH Stability

pH Stability pH Stability Maximum Comment Organism
4.5 10 purified recombinant enzyme, over 50% activity within this range at 35°C Paracoccus sp.

pI Value

Organism Comment pI Value Maximum pI Value
Paracoccus sp. isoelectric focusing
-
 5.13

General Information

General Information Comment Organism
evolution the enzyme belongs to the amidase signature enzyme family Paracoccus sp.
additional information the enzyme maintains a core alpha/beta/alpha structure and the G-(GAV)-S-(GS)2-GX-(GSAE)-(GSAVYCT)-X-(LIVMT)-(GSA)-X6-(GSAT)-X-(GA)-X-(DE)-X-(GA)-X-S-(LIVM)-R-X-P-(GSACTL) sequence motif Paracoccus sp.