Application | Comment | Organism |
---|---|---|
synthesis | Escherichia coli cells overexpressing the laaA gene have been demonstrated to be applicable to the S-stereoselective hydrolysis of (R,S)-piperazine-2-tert-butylcarboxamide to produce (S)-piperazine-2-carboxylic acid with high optical purity. Enantiomerically pure piperazine-2-carboxylic acid and its tert-butylcarboxamide derivative are important chiral building blocks for some pharmacologically active compounds such as N-methyl-D-aspartate antagonist for glutamate receptor, cardioprotective nucleoside transport blocker and HIV protease inhibitor | Pseudomonas azotoformans |
Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Pseudomonas azotoformans |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Ag+ | 1 mM, complete inhibition | Pseudomonas azotoformans | |
Cd2+ | 1 mM, complete inhibition | Pseudomonas azotoformans | |
CoCl2 | 1 mM, 52% inhibition | Pseudomonas azotoformans | |
Hg2+ | 1 mM, complete inhibition | Pseudomonas azotoformans | |
iodoacetate | 1 mM, 40% inhibition | Pseudomonas azotoformans | |
NEM | 1 mM, 24% inhibition | Pseudomonas azotoformans | |
NiCl2 | 1 mM, 70% inhibition | Pseudomonas azotoformans | |
p-chloromercuribenzoate | 1 mM, 67% inhibition | Pseudomonas azotoformans | |
PbCl2 | 1 mM, 73% inhibition | Pseudomonas azotoformans | |
phenylhydrazine | 1 mM, complete inhibition | Pseudomonas azotoformans | |
Zn2+ | 1 mM, complete inhibition | Pseudomonas azotoformans |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.58 | - |
L-proline-p-nitroanilide | 30°C, pH 7.0 | Pseudomonas azotoformans |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
32000 | - |
gel filtration | Pseudomonas azotoformans |
34000 | - |
1 * 34000, SDS-PAGE | Pseudomonas azotoformans |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas azotoformans | Q76KX0 | IAM 1603 | - |
Pseudomonas azotoformans IAM 1603 | Q76KX0 | IAM 1603 | - |
Purification (Comment) | Organism |
---|---|
- |
Pseudomonas azotoformans |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.000894 | - |
S-stereoselective amidase from Pseudomonas azotoformans IAM 1603 | Pseudomonas azotoformans |
192 | - |
LaaA from Escharichia coli JM109 harboring pSTB20 | Pseudomonas azotoformans |
Storage Stability | Organism |
---|---|
-20°C, stored without loss of activity for more than six months in the buffer containing 50% glycerol | Pseudomonas azotoformans |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(R,S)-piperazine-2-tert-butylcarboxamide + H2O | % of the activity with L-proline amide | Pseudomonas azotoformans | (S)-piperazine-2-tert-butylcarboxylic acid + NH3 | - |
? | |
(R,S)-piperazine-2-tert-butylcarboxamide + H2O | % of the activity with L-proline amide | Pseudomonas azotoformans IAM 1603 | (S)-piperazine-2-tert-butylcarboxylic acid + NH3 | - |
? | |
(S)-piperazine-2-carboxamide + H2O | 3.7% of the activity with L-proline amide | Pseudomonas azotoformans | (S)-piperazine-2-carboxylic acid + NH3 | - |
? | |
(S)-piperazine-2-carboxamide + H2O | 3.7% of the activity with L-proline amide | Pseudomonas azotoformans IAM 1603 | (S)-piperazine-2-carboxylic acid + NH3 | - |
? | |
(S)-piperazine-2-tert-butylcarboxamide + H2O | 0.2% of the activity with L-proline amide | Pseudomonas azotoformans | (S)-piperazine-2-carboxylic acid + tert-butylamine | - |
? | |
(S)-piperidine-2-carboxamide + H2O | 32% of the activity with L-proline amide | Pseudomonas azotoformans | (S)-piperidine-2-carboxylic acid + NH3 | - |
? | |
L-alanine amide + H2O | 10.6% of the activity with L-proline amide | Pseudomonas azotoformans | L-alanine + NH3 | - |
? | |
L-isoleucine amide + H2O | 0.17% of the activity with L-proline amide | Pseudomonas azotoformans | L-isoleucine + NH3 | - |
? | |
L-leucine amide + H2O | 0.46% of the activity with L-proline amide | Pseudomonas azotoformans | L-leucine + NH3 | - |
? | |
L-leucine amide + H2O | 0.46% of the activity with L-proline amide | Pseudomonas azotoformans IAM 1603 | L-leucine + NH3 | - |
? | |
L-methionine amide + H2O | 4.2% of the activity with L-proline amide | Pseudomonas azotoformans | L-methionine + NH3 | - |
? | |
L-phenylalanine amide + H2O | 0.97% of the activity with L-proline amide | Pseudomonas azotoformans | L-phenylalanine + NH3 | - |
? | |
L-phenylalanine amide + H2O | 0.97% of the activity with L-proline amide | Pseudomonas azotoformans IAM 1603 | L-phenylalanine + NH3 | - |
? | |
L-proline amide + H2O | - |
Pseudomonas azotoformans | L-proline + NH3 | - |
? | |
L-proline-p-nitroanilide + H2O | 40.9% of the activity with L-proline amide | Pseudomonas azotoformans | ? + NH3 | - |
? | |
L-serine amide + H2O | 0.43% of the activity with L-proline amide | Pseudomonas azotoformans | L-serine + NH3 | - |
? | |
L-threonine amide + H2O | 0.12% of the activity with L-proline amide | Pseudomonas azotoformans | L-threonine + NH3 | - |
? | |
L-tryptophan amide + H2O | 0.2% of the activity with L-proline amide | Pseudomonas azotoformans | L-tryptophan + NH3 | - |
? | |
L-tyrosine amide + H2O | 0.086% of the activity with L-proline amide | Pseudomonas azotoformans | L-tyrosine + NH3 | - |
? | |
additional information | the following compounds are not substrates for the amidase: L-arginine amide, L-asparagine amide, L-isoasparagine, L-glutaminamide, L-isoglutamine, glycine amide, L-histidine amide, L-lysine amide, L-valine amide, D-proline amide, L-alanyl-L-alanine, L-alanylglycine, glycylglycine, L-prolyl-L-alanine and L-prolylglycine. LaaA can not act on the peptide substrates such as L-prolyl-L-alanine, L-prolylglycine, L-alanyl-L-alanine, L-alanylglycine and glycylglycine | Pseudomonas azotoformans | ? | - |
? | |
additional information | the following compounds are not substrates for the amidase: L-arginine amide, L-asparagine amide, L-isoasparagine, L-glutaminamide, L-isoglutamine, glycine amide, L-histidine amide, L-lysine amide, L-valine amide, D-proline amide, L-alanyl-L-alanine, L-alanylglycine, glycylglycine, L-prolyl-L-alanine and L-prolylglycine. LaaA can not act on the peptide substrates such as L-prolyl-L-alanine, L-prolylglycine, L-alanyl-L-alanine, L-alanylglycine and glycylglycine | Pseudomonas azotoformans IAM 1603 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
monomer | 1 * 34000, SDS-PAGE | Pseudomonas azotoformans |
Synonyms | Comment | Organism |
---|---|---|
L-amino acid amidase | - |
Pseudomonas azotoformans |
LaaA | - |
Pseudomonas azotoformans |
S-stereoselective piperazine-2-tert-butylcarboxamide hydrolase | - |
Pseudomonas azotoformans |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
45 | - |
- |
Pseudomonas azotoformans |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
45 | - |
stable up to | Pseudomonas azotoformans |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
9 | - |
- |
Pseudomonas azotoformans |