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Literature summary extracted from

  • Pozo-Dengra, J.; Martinez-Gomez, A.I.; Martinez-Rodriguez, S.; Clemente-Jimenez, J.M.; Rodriguez-Vico, F.; Las Heras-Vazquez, F.J.
    Evaluation of substrate promiscuity of an L-carbamoyl amino acid amidohydrolase from Geobacillus stearothermophilus CECT43 (2010), Biotechnol. Prog., 26, 954-959.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.5.1.87 Co2+ 663.89% relative activity of non 8-hydroxyquinoline-5-sulfonic acid (HQSA) treated enzyme, 536.91% relative activity of HQSA pretreated enzyme, consequently, the enzyme is incubated with Co2+ before activity assays Geobacillus stearothermophilus
3.5.1.87 Mn2+ 286.93% relative activity of non 8-hydroxyquinoline-5-sulfonic acid (HQSA) treated enzyme, 197% relative activity of HQSA pretreated enzyme Geobacillus stearothermophilus
3.5.1.87 Ni2+ 341.94% relative activity of non 8-hydroxyquinoline-5-sulfonic acid (HQSA) treated enzyme, 333.96% relative activity of HQSA pretreated enzyme Geobacillus stearothermophilus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.1.87 expressed in Escherichia coli BL21 Geobacillus stearothermophilus

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.5.1.87 2-mercaptoethanol 10 mM causes 29% inhibition, suggesting that no cysteine residue is crucial for enzyme activity, although they might have a structural role Geobacillus stearothermophilus
3.5.1.87 5,5'-dithiobis(2-nitrobenzoic acid) 2 mM, slight inhibition Geobacillus stearothermophilus
3.5.1.87 8-Hydroxyquinoline-5-sulfonic acid 10 mM, decreases the activity to 13% Geobacillus stearothermophilus
3.5.1.87 Cs+ 2 mM, slight inhibition Geobacillus stearothermophilus
3.5.1.87 dithiothreitol 10 mM causes 41% inhibition, suggesting that no cysteine residue is crucial for enzyme activity, although they might have a structural role Geobacillus stearothermophilus
3.5.1.87 EDTA 10 mM, decreases the activity to 15% Geobacillus stearothermophilus
3.5.1.87 Fe2+ 2 mM, strong inhibition Geobacillus stearothermophilus
3.5.1.87 Fe3+ 2 mM, strong inhibition Geobacillus stearothermophilus
3.5.1.87 Hg2+ 2 mM, strong inhibition Geobacillus stearothermophilus
3.5.1.87 Mg2+ slightly inhibited activity of non 8-hydroxyquinoline-5-sulfonic acid (HQSA) treated enzyme and HQSA pretreated enzyme Geobacillus stearothermophilus
3.5.1.87 Na+ 2 mM, slight inhibition Geobacillus stearothermophilus
3.5.1.87 Pb2+ 2 mM, slight inhibition Geobacillus stearothermophilus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.1.87 2.9
-
 N-carbamoyl-L-methionine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 3.29
-
 N-carbamoyl-L-valine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 4.19
-
 N-carbamoyl-L-alanine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 5
-
 N-formyl-L-alanine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 5.82
-
 N-carbamoyl-L-phenylalanine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 6.75
-
 N-formyl-L-valine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 9.77
-
 N-formyl-L-methionine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 12.66
-
 N-acetyl-L-methionine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 12.9
-
 N-acetyl-L-phenylalanine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 12.99
-
 N-carbamoyl-L-tyrosine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 13.82
-
 N-formyl-L-phenylalanine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 13.92
-
 N-acetyl-L-valine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 23.78
-
 N-Formyl-L-tyrosine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 36.12
-
 N-acetyl-L-alanine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 116
-
 N-acetyl-L-tyrosine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.5.1.87 Ca2+ similar enzyme activity of non 8-hydroxyquinoline-5-sulfonic acid (HQSA) treated enzyme and HQSA pretreated enzyme compared to activity without addition of any effectors Geobacillus stearothermophilus
3.5.1.87 Cu2+ slightly activated activity of non 8-hydroxyquinoline-5-sulfonic acid (HQSA) treated enzyme and HQSA pretreated enzyme compared to activity without addition of any effectors Geobacillus stearothermophilus
3.5.1.87 additional information the enzymatic function is not affected by the sulfhydryl reagent iodoacetamide and is only slightly affected by 5,5'-dithiobis(2-nitrobenzoic acid) Geobacillus stearothermophilus
3.5.1.87 Zn2+ slightly activated activity of non 8-hydroxyquinoline-5-sulfonic acid (HQSA) treated enzyme, similar activity of 8-hydroxyquinoline-5-sulfonic acid pretreated enzyme compared to activity of non 8-hydroxyquinoline-5-sulfonic acid treated enzyme without addition of any effectors Geobacillus stearothermophilus

Organism

EC Number Organism UniProt Comment Textmining
3.5.1.87 Geobacillus stearothermophilus
-
 CECT43, a thermophilic bacterium
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.1.87
-
 Geobacillus stearothermophilus

Reaction

EC Number Reaction Comment Organism Reaction ID
3.5.1.87 an N-carbamoyl-L-2-amino acid (a 2-ureido carboxylate) + H2O = an L-2-amino acid + NH3 + CO2 recombinant BsLcar is able to hydrolyze N-acetyl and N-formyl-amino acids in addition to N-carbamoyl-amino acids, showing strict L-enantioselectivity for all the three. Additionally, the enzyme shows the fastest amidohydrolase activity on the N-formyl-amino acid followed by N-carbamoyl and N-acetyl ones Geobacillus stearothermophilus
a

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.5.1.87 additional information
-
 overview of effect of metal ions and other chemical agents on the enzyme activity of 8-hydroxyquinoline-5-sulfonic acid (HQSA) treated enzyme and the HQSA pretreated enzyme Geobacillus stearothermophilus
3.5.1.87 33.34
-
 activity of non 8-hydroxyquinoline-5-sulfonic acid (HQSA) treated enzyme, pH 7.5, 65°C Geobacillus stearothermophilus
3.5.1.87 40.2
-
 120% relative activity of 8-hydroxyquinoline-5-sulfonic acid (HQSA) pretreated enzyme compared to activity of non 8-hydroxyquinoline-5-sulfonic acid (HQSA) treated enzyme, pH 7.5, 65°C Geobacillus stearothermophilus
3.5.1.87 65.69
-
 Mn2+, 197% relative activity of 8-hydroxyquinoline-5-sulfonic acid (HQSA) pretreated enzyme compared to activity of non 8-hydroxyquinoline-5-sulfonic acid (HQSA) treated enzyme without addition of any effectors, pH 7.5, 65°C Geobacillus stearothermophilus
3.5.1.87 95.67
-
 Mn2+, 286.93% relative activity of non 8-hydroxyquinoline-5-sulfonic acid (HQSA) treated enzyme compared to activity without addition of any effectors, pH 7.5, 65°C Geobacillus stearothermophilus
3.5.1.87 111.3
-
 Ni2+, 333.96% relative activity of 8-hydroxyquinoline-5-sulfonic acid (HQSA) pretreated enzyme compared to activity of non 8-hydroxyquinoline-5-sulfonic acid (HQSA) treated enzyme without addition of any effectors, pH 7.5, 65°C Geobacillus stearothermophilus
3.5.1.87 114
-
 Ni2+, 341.94% relative activity of non 8-hydroxyquinoline-5-sulfonic acid (HQSA) treated enzyme compared to activity without addition of any effectors, pH 7.5, 65°C Geobacillus stearothermophilus
3.5.1.87 179
-
 Co2+, 536.91% relative activity of 8-hydroxyquinoline-5-sulfonic acid (HQSA) pretreated enzyme pretreated enzyme compared to activity of non 8-hydroxyquinoline-5-sulfonic acid (HQSA) treated enzyme without addition of any effectors, pH 7.5, 65°C Geobacillus stearothermophilus
3.5.1.87 221.4
-
 Co2+, 663.89% relative activity of non 8-hydroxyquinoline-5-sulfonic acid (HQSA) treated enzyme compared to activity without addition of any effectors, pH 7.5, 65°C Geobacillus stearothermophilus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.1.87 N-acetyl-L-alanine + H2O
-
 Geobacillus stearothermophilus L-alanine + acetate
-
 ?
3.5.1.87 N-acetyl-L-methionine + H2O
-
 Geobacillus stearothermophilus L-methionine + acetate
-
 ?
3.5.1.87 N-acetyl-L-phenylalanine + H2O
-
 Geobacillus stearothermophilus L-phenylalanine + acetate
-
 ?
3.5.1.87 N-acetyl-L-tyrosine + H2O
-
 Geobacillus stearothermophilus L-tyrosine + acetate
-
 ?
3.5.1.87 N-acetyl-L-valine + H2O
-
 Geobacillus stearothermophilus L-valine + acetate
-
 ?
3.5.1.87 N-carbamoyl-L-alanine + H2O
-
 Geobacillus stearothermophilus L-alanine + CO2 + NH3
-
 ?
3.5.1.87 N-carbamoyl-L-methionine + H2O
-
 Geobacillus stearothermophilus L-methionine + CO2 + NH3
-
 ?
3.5.1.87 N-carbamoyl-L-phenylalanine + H2O
-
 Geobacillus stearothermophilus L-phenylalanine + CO2 + NH3
-
 ?
3.5.1.87 N-carbamoyl-L-tyrosine + H2O
-
 Geobacillus stearothermophilus L-tyrosine + CO2 + NH3
-
 ?
3.5.1.87 N-carbamoyl-L-valine + H2O
-
 Geobacillus stearothermophilus L-valine + CO2 + NH3
-
 ?
3.5.1.87 N-formyl-L-alanine + H2O
-
 Geobacillus stearothermophilus L-alanine + CO2
-
 ?
3.5.1.87 N-formyl-L-methionine + H2O
-
 Geobacillus stearothermophilus L-methionine + HCOOH
-
 ?
3.5.1.87 N-formyl-L-phenylalanine + H2O
-
 Geobacillus stearothermophilus L-phenylalanine + CO2
-
 ?
3.5.1.87 N-formyl-L-tyrosine + H2O
-
 Geobacillus stearothermophilus L-tyrosine + CO2
-
 ?
3.5.1.87 N-formyl-L-valine + H2O
-
 Geobacillus stearothermophilus L-valine + CO2
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.5.1.87 BsLcar
-
 Geobacillus stearothermophilus
3.5.1.87 N-carbamoyl-amino-acid amidohydrolase
-
 Geobacillus stearothermophilus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.5.1.87 65
-
 maximal activity for hydrolysis of N-acetyl and N-formyl-amino acids and N-carbamoyl-amino acids Geobacillus stearothermophilus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.5.1.87 65 70 thermal stability studies performed on BsLcar show that maximum activity gradually decreases at temperatures over 65°C for 60 min, with 60% activity remaining after incubation at 70°C for 60 min Geobacillus stearothermophilus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.5.1.87 0.01
-
 N-acetyl-L-phenylalanine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 0.08
-
 N-acetyl-L-tyrosine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 0.08
-
 N-acetyl-L-valine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 0.16
-
 N-carbamoyl-L-phenylalanine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 0.18
-
 N-carbamoyl-L-tyrosine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 0.84
-
 N-carbamoyl-L-valine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 1.46
-
 N-Formyl-L-tyrosine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 2.26
-
 N-acetyl-L-methionine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 5.32
-
 N-carbamoyl-L-methionine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 12.12
-
 N-formyl-L-phenylalanine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 27.26
-
 N-formyl-L-valine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 27.86
-
 N-acetyl-L-alanine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 48.96
-
 N-carbamoyl-L-alanine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 293.5
-
 N-formyl-L-alanine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 839.1
-
 N-formyl-L-methionine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5.1.87 7.5
-
 maximal activity for hydrolysis of N-acetyl and N-formyl-amino acids and N-carbamoyl-amino acids, in sodium phosphate, 100 mM Geobacillus stearothermophilus

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.5.1.87 0.39
-
 N-acetyl-L-phenylalanine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 0.69
-
 N-acetyl-L-tyrosine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 5.96
-
 N-acetyl-L-valine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 13.86
-
 N-carbamoyl-L-tyrosine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 27.49
-
 N-carbamoyl-L-phenylalanine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 61.4
-
 N-Formyl-L-tyrosine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 178.5
-
 N-acetyl-L-methionine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 255.3
-
 N-carbamoyl-L-valine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 771.3
-
 N-acetyl-L-alanine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 877
-
 N-formyl-L-phenylalanine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 1834
-
 N-carbamoyl-L-methionine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 4039
-
 N-formyl-L-valine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 11680
-
 N-carbamoyl-L-alanine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 58700
-
 N-formyl-L-alanine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus
3.5.1.87 85880
-
 N-formyl-L-methionine pH 7.5, 65°C, preincubation with CoCl2 Geobacillus stearothermophilus