Activating Compound | Comment | Organism | Structure |
---|---|---|---|
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 | |
Mn2+ | 286.93% relative activity of non 8-hydroxyquinoline-5-sulfonic acid (HQSA) treated enzyme, 197% relative activity of HQSA pretreated enzyme | Geobacillus stearothermophilus | |
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 (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21 | Geobacillus stearothermophilus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
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 | |
5,5'-dithiobis(2-nitrobenzoic acid) | 2 mM, slight inhibition | Geobacillus stearothermophilus | |
8-Hydroxyquinoline-5-sulfonic acid | 10 mM, decreases the activity to 13% | Geobacillus stearothermophilus | |
Cs+ | 2 mM, slight inhibition | Geobacillus stearothermophilus | |
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 | |
EDTA | 10 mM, decreases the activity to 15% | Geobacillus stearothermophilus | |
Fe2+ | 2 mM, strong inhibition | Geobacillus stearothermophilus | |
Fe3+ | 2 mM, strong inhibition | Geobacillus stearothermophilus | |
Hg2+ | 2 mM, strong inhibition | Geobacillus stearothermophilus | |
Mg2+ | slightly inhibited activity of non 8-hydroxyquinoline-5-sulfonic acid (HQSA) treated enzyme and HQSA pretreated enzyme | Geobacillus stearothermophilus | |
Na+ | 2 mM, slight inhibition | Geobacillus stearothermophilus | |
Pb2+ | 2 mM, slight inhibition | Geobacillus stearothermophilus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.9 | - |
N-carbamoyl-L-methionine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
3.29 | - |
N-carbamoyl-L-valine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
4.19 | - |
N-carbamoyl-L-alanine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
5 | - |
N-formyl-L-alanine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
5.82 | - |
N-carbamoyl-L-phenylalanine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
6.75 | - |
N-formyl-L-valine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
9.77 | - |
N-formyl-L-methionine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
12.66 | - |
N-acetyl-L-methionine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
12.9 | - |
N-acetyl-L-phenylalanine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
12.99 | - |
N-carbamoyl-L-tyrosine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
13.82 | - |
N-formyl-L-phenylalanine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
13.92 | - |
N-acetyl-L-valine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
23.78 | - |
N-Formyl-L-tyrosine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
36.12 | - |
N-acetyl-L-alanine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
116 | - |
N-acetyl-L-tyrosine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
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 | |
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 | |
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 | |
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 | UniProt | Comment | Textmining |
---|---|---|---|
Geobacillus stearothermophilus | - |
CECT43, a thermophilic bacterium | - |
Purification (Comment) | Organism |
---|---|
- |
Geobacillus stearothermophilus |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
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 |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
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 |
33.34 | - |
activity of non 8-hydroxyquinoline-5-sulfonic acid (HQSA) treated enzyme, pH 7.5, 65°C | Geobacillus stearothermophilus |
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 |
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 |
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 |
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 |
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 |
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 |
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 | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
N-acetyl-L-alanine + H2O | - |
Geobacillus stearothermophilus | L-alanine + acetate | - |
? | |
N-acetyl-L-methionine + H2O | - |
Geobacillus stearothermophilus | L-methionine + acetate | - |
? | |
N-acetyl-L-phenylalanine + H2O | - |
Geobacillus stearothermophilus | L-phenylalanine + acetate | - |
? | |
N-acetyl-L-tyrosine + H2O | - |
Geobacillus stearothermophilus | L-tyrosine + acetate | - |
? | |
N-acetyl-L-valine + H2O | - |
Geobacillus stearothermophilus | L-valine + acetate | - |
? | |
N-carbamoyl-L-alanine + H2O | - |
Geobacillus stearothermophilus | L-alanine + CO2 + NH3 | - |
? | |
N-carbamoyl-L-methionine + H2O | - |
Geobacillus stearothermophilus | L-methionine + CO2 + NH3 | - |
? | |
N-carbamoyl-L-phenylalanine + H2O | - |
Geobacillus stearothermophilus | L-phenylalanine + CO2 + NH3 | - |
? | |
N-carbamoyl-L-tyrosine + H2O | - |
Geobacillus stearothermophilus | L-tyrosine + CO2 + NH3 | - |
? | |
N-carbamoyl-L-valine + H2O | - |
Geobacillus stearothermophilus | L-valine + CO2 + NH3 | - |
? | |
N-formyl-L-alanine + H2O | - |
Geobacillus stearothermophilus | L-alanine + CO2 | - |
? | |
N-formyl-L-methionine + H2O | - |
Geobacillus stearothermophilus | L-methionine + HCOOH | - |
? | |
N-formyl-L-phenylalanine + H2O | - |
Geobacillus stearothermophilus | L-phenylalanine + CO2 | - |
? | |
N-formyl-L-tyrosine + H2O | - |
Geobacillus stearothermophilus | L-tyrosine + CO2 | - |
? | |
N-formyl-L-valine + H2O | - |
Geobacillus stearothermophilus | L-valine + CO2 | - |
? |
Synonyms | Comment | Organism |
---|---|---|
BsLcar | - |
Geobacillus stearothermophilus |
N-carbamoyl-amino-acid amidohydrolase | - |
Geobacillus stearothermophilus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
65 | - |
maximal activity for hydrolysis of N-acetyl and N-formyl-amino acids and N-carbamoyl-amino acids | Geobacillus stearothermophilus |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
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 Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.01 | - |
N-acetyl-L-phenylalanine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
0.08 | - |
N-acetyl-L-tyrosine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
0.08 | - |
N-acetyl-L-valine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
0.16 | - |
N-carbamoyl-L-phenylalanine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
0.18 | - |
N-carbamoyl-L-tyrosine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
0.84 | - |
N-carbamoyl-L-valine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
1.46 | - |
N-Formyl-L-tyrosine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
2.26 | - |
N-acetyl-L-methionine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
5.32 | - |
N-carbamoyl-L-methionine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
12.12 | - |
N-formyl-L-phenylalanine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
27.26 | - |
N-formyl-L-valine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
27.86 | - |
N-acetyl-L-alanine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
48.96 | - |
N-carbamoyl-L-alanine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
293.5 | - |
N-formyl-L-alanine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
839.1 | - |
N-formyl-L-methionine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
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 Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.39 | - |
N-acetyl-L-phenylalanine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
0.69 | - |
N-acetyl-L-tyrosine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
5.96 | - |
N-acetyl-L-valine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
13.86 | - |
N-carbamoyl-L-tyrosine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
27.49 | - |
N-carbamoyl-L-phenylalanine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
61.4 | - |
N-Formyl-L-tyrosine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
178.5 | - |
N-acetyl-L-methionine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
255.3 | - |
N-carbamoyl-L-valine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
771.3 | - |
N-acetyl-L-alanine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
877 | - |
N-formyl-L-phenylalanine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
1834 | - |
N-carbamoyl-L-methionine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
4039 | - |
N-formyl-L-valine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
11680 | - |
N-carbamoyl-L-alanine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
58700 | - |
N-formyl-L-alanine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus | |
85880 | - |
N-formyl-L-methionine | pH 7.5, 65°C, preincubation with CoCl2 | Geobacillus stearothermophilus |