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Literature summary for 3.5.1.101 extracted from

  • Komeda, H.; Harada, H.; Washika, S.; Sakamoto, T.; Ueda, M.; Asano, Y.
    S-stereoselective piperazine-2-tert-butylcarboxamide hydrolase from Pseudomonas azotoformans IAM 1603 is a novel L-amino acid amidase (2004), Eur. J. Biochem., 271, 1465-1475.
    View publication on PubMed
Search for more literature for 3.5.1.101

Application

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(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Pseudomonas azotoformans

Inhibitors

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 [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 [Da] Molecular Weight Maximum [Da] Comment Organism
32000
-
 gel filtration Pseudomonas azotoformans
34000
-
 1 * 34000, SDS-PAGE Pseudomonas azotoformans

Organism

Organism UniProt Comment Textmining
Pseudomonas azotoformans Q76KX0 IAM 1603
-
Pseudomonas azotoformans IAM 1603 Q76KX0 IAM 1603
-

Purification (Commentary)

Purification (Comment) Organism
-
 Pseudomonas azotoformans

Specific Activity [micromol/min/mg]

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

Storage Stability Organism
-20°C, stored without loss of activity for more than six months in the buffer containing 50% glycerol Pseudomonas azotoformans

Substrates and Products (Substrate)

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

Subunits Comment Organism
monomer 1 * 34000, SDS-PAGE Pseudomonas azotoformans

Synonyms

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 [°C] Temperature Optimum Maximum [°C] Comment Organism
45
-
-
 Pseudomonas azotoformans

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
45
-
 stable up to Pseudomonas azotoformans

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9
-
-
 Pseudomonas azotoformans