| IED ID | IndEnz0002001441 |
| Enzyme Type ID | protease001441 |
| Protein Name |
R -stereoselective amidase EC 3.5.1.100 |
| Gene Name | ramA |
| Organism | Pseudomonas sp. |
| Taxonomic Lineage | cellular organisms Bacteria Proteobacteria Gammaproteobacteria Pseudomonadales Pseudomonadaceae Pseudomonas unclassified Pseudomonas Pseudomonas sp. |
| Enzyme Sequence | MKIELVQLAGRDGDTAYNLSRTLNAIATCAGDTDLLVFPETYLSGFVGGAQLAQVAEPLHGTTLQTLLQAVRQRDVAVVLGFAEVHQGRFYNSSVLVTPEGIALQYRKTHLWPSERSDFSPGDRFTTVLWRGVRVGLLICYDIELPETSRALAQLGAEVVIVTNGNMDPYGPVHRTAIMARAQENQLFAVMVNRVGAGDDGLVFAGGSMAVDPFGRVLFEAGRDEVRHVVELDLDQLKAARRDYDYLKDRRLMLSGEQTEHPDGRRELLIGASQ |
| Enzyme Length | 274 |
| Uniprot Accession Number | Q75SP7 |
| Absorption | |
| Active Site | ACT_SITE 40; /note=Proton acceptor; /evidence=ECO:0000255|PROSITE-ProRule:PRU00054; ACT_SITE 108; /note=Proton donor; /evidence=ECO:0000255|PROSITE-ProRule:PRU00054; ACT_SITE 140; /note=Nucleophile; /evidence=ECO:0000255|PROSITE-ProRule:PRU00054 |
| Activity Regulation | ACTIVITY REGULATION: Completely inhibited by p-chloromercuribenzoate, N-ethylmaleimide, MnSO(4), MnCl(2), CoCl(2), NiCl(2), CuSO(4), CuCl(2), ZnSO(4), ZnCl(2), AgNO(3), CdCl(2), HgCl(2) and PbCl(2). Partially inhibited by FeCl(3) and Fe(NH(4))(2)(SO(4))(2). Slightly enhanced by dithiothreitol. Unaffected by LiBr, H(2)BO(3), NaCl, MgSO(4), MgCl(2), AlCl(3), KCl, CaCl(2), CrCl(3), RbCl, Na(2)MoO(4), (NH(4))(6)Mo(7)O(24), CsCl and BaCl(2). Unaffected by the chelating agents o-phenanthroline, 8-hydroxyquinoline, enthylenediaminetetraacetic acid and alpha,alpha'-dipyridyl. Not inhibited by the carbonyl reagents hydroxylamine, phenylhydrazine, hydrazine, D,L-penicillamine and D-cycloserine. Not affected by the serine protease inhibitor phenylmethanesulfonyl fluoride, the serine/cysteine protease inhibitor leupeptine or the aspartic protease inhibitor pepstatin. {ECO:0000269|PubMed:15066183}. |
| Binding Site | |
| Calcium Binding | |
| catalytic Activity | CATALYTIC ACTIVITY: Reaction=(R)-piperazine-2-carboxamide + H2O = (R)-piperazine-2-carboxylate + NH4(+); Xref=Rhea:RHEA:26542, ChEBI:CHEBI:15377, ChEBI:CHEBI:28938, ChEBI:CHEBI:58916, ChEBI:CHEBI:58917; EC=3.5.1.100; Evidence={ECO:0000269|PubMed:15066183}; CATALYTIC ACTIVITY: Reaction=beta-alaninamide + H2O = beta-alanine + NH4(+); Xref=Rhea:RHEA:26546, ChEBI:CHEBI:15377, ChEBI:CHEBI:28938, ChEBI:CHEBI:57966, ChEBI:CHEBI:58918; EC=3.5.1.100; Evidence={ECO:0000269|PubMed:15066183}; |
| DNA Binding | |
| EC Number | 3.5.1.100 |
| Enzyme Function | FUNCTION: Hydrolyzes (R)-piperazine-2-carboxamide and (R)-piperazine-2-tert-butylcarboxamide with strict R-stereoselectivity. Also active towards beta-alaninamide, piperidine-3-carboxmide, D-glutaminamide and slightly active towards L-glutaminamide and piperidine-4-carboxamide. {ECO:0000269|PubMed:15066183}. |
| Temperature Dependency | BIOPHYSICOCHEMICAL PROPERTIES: Temperature dependence: Optimum temperature is 45 degrees Celsius, activity decreases rapidly above 45 degrees Celsius possibly due to instability at higher temperatures. Inactivated following 10 minutes incubation at 55 degrees Celsius, and only retains 2.6% of activity after 10 minutes incubation at 50 degrees Celsius. {ECO:0000269|PubMed:15066183}; |
| PH Dependency | BIOPHYSICOCHEMICAL PROPERTIES: pH dependence: Optimum pH is 8.0. Stable from pH 6.0 to 9.0. {ECO:0000269|PubMed:15066183}; |
| Pathway | |
| nucleotide Binding | |
| Features | Active site (3); Chain (1); Domain (1); Sequence conflict (1) |
| Keywords | Direct protein sequencing;Hydrolase |
| Interact With | |
| Induction | |
| Subcellular Location | |
| Modified Residue | |
| Post Translational Modification | |
| Signal Peptide | |
| Structure 3D | |
| Cross Reference PDB | - |
| Mapped Pubmed ID | - |
| Motif | |
| Gene Encoded By | |
| Mass | 30,128 |
| Kinetics | |
| Metal Binding | |
| Rhea ID | RHEA:26542; RHEA:26546 |
| Cross Reference Brenda | 3.5.1.100; |