RSAM_PSESP
ID RSAM_PSESP Reviewed; 274 AA.
AC Q75SP7;
DT 02-NOV-2010, integrated into UniProtKB/Swiss-Prot.
DT 05-JUL-2004, sequence version 1.
DT 03-AUG-2022, entry version 57.
DE RecName: Full=(R)-stereoselective amidase {ECO:0000303|PubMed:15066183, ECO:0000312|EMBL:BAD15093.1};
DE EC=3.5.1.100 {ECO:0000269|PubMed:15066183};
GN Name=ramA {ECO:0000312|EMBL:BAD15093.1};
OS Pseudomonas sp.
OC Bacteria; Proteobacteria; Gammaproteobacteria; Pseudomonadales;
OC Pseudomonadaceae; Pseudomonas.
OX NCBI_TaxID=306;
RN [1] {ECO:0000305, ECO:0000312|EMBL:BAD15093.1}
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 1-14, FUNCTION,
RP CATALYTIC ACTIVITY, ACTIVITY REGULATION, BIOPHYSICOCHEMICAL PROPERTIES, AND
RP SUBUNIT.
RC STRAIN=MCI3434 {ECO:0000312|EMBL:BAD15093.1};
RX PubMed=15066183; DOI=10.1111/j.1432-1033.2004.04069.x;
RA Komeda H., Harada H., Washika S., Sakamoto T., Ueda M., Asano Y.;
RT "A novel R-stereoselective amidase from Pseudomonas sp. MCI3434 acting on
RT piperazine-2-tert-butylcarboxamide.";
RL Eur. J. Biochem. 271:1580-1590(2004).
RN [2] {ECO:0000312|EMBL:BAE02667.1}
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=MCI3434 {ECO:0000312|EMBL:BAE02667.1};
RX PubMed=15955066; DOI=10.1111/j.1742-4658.2005.04721.x;
RA Komeda H., Asano Y.;
RT "A DmpA-homologous protein from Pseudomonas sp. is a dipeptidase specific
RT for beta-alanyl dipeptides.";
RL FEBS J. 272:3075-3084(2005).
CC -!- FUNCTION: Hydrolyzes (R)-piperazine-2-carboxamide and (R)-piperazine-2-
CC tert-butylcarboxamide with strict R-stereoselectivity. Also active
CC towards beta-alaninamide, piperidine-3-carboxmide, D-glutaminamide and
CC slightly active towards L-glutaminamide and piperidine-4-carboxamide.
CC {ECO:0000269|PubMed:15066183}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(R)-piperazine-2-carboxamide + H2O = (R)-piperazine-2-
CC carboxylate + NH4(+); Xref=Rhea:RHEA:26542, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:28938, ChEBI:CHEBI:58916, ChEBI:CHEBI:58917;
CC EC=3.5.1.100; Evidence={ECO:0000269|PubMed:15066183};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=beta-alaninamide + H2O = beta-alanine + NH4(+);
CC Xref=Rhea:RHEA:26546, ChEBI:CHEBI:15377, ChEBI:CHEBI:28938,
CC ChEBI:CHEBI:57966, ChEBI:CHEBI:58918; EC=3.5.1.100;
CC Evidence={ECO:0000269|PubMed:15066183};
CC -!- ACTIVITY REGULATION: Completely inhibited by p-chloromercuribenzoate,
CC N-ethylmaleimide, MnSO(4), MnCl(2), CoCl(2), NiCl(2), CuSO(4), CuCl(2),
CC ZnSO(4), ZnCl(2), AgNO(3), CdCl(2), HgCl(2) and PbCl(2). Partially
CC inhibited by FeCl(3) and Fe(NH(4))(2)(SO(4))(2). Slightly enhanced by
CC dithiothreitol. Unaffected by LiBr, H(2)BO(3), NaCl, MgSO(4), MgCl(2),
CC AlCl(3), KCl, CaCl(2), CrCl(3), RbCl, Na(2)MoO(4),
CC (NH(4))(6)Mo(7)O(24), CsCl and BaCl(2). Unaffected by the chelating
CC agents o-phenanthroline, 8-hydroxyquinoline,
CC enthylenediaminetetraacetic acid and alpha,alpha'-dipyridyl. Not
CC inhibited by the carbonyl reagents hydroxylamine, phenylhydrazine,
CC hydrazine, D,L-penicillamine and D-cycloserine. Not affected by the
CC serine protease inhibitor phenylmethanesulfonyl fluoride, the
CC serine/cysteine protease inhibitor leupeptine or the aspartic protease
CC inhibitor pepstatin. {ECO:0000269|PubMed:15066183}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC pH dependence:
CC Optimum pH is 8.0. Stable from pH 6.0 to 9.0.
CC {ECO:0000269|PubMed:15066183};
CC Temperature dependence:
CC Optimum temperature is 45 degrees Celsius, activity decreases rapidly
CC above 45 degrees Celsius possibly due to instability at higher
CC temperatures. Inactivated following 10 minutes incubation at 55
CC degrees Celsius, and only retains 2.6% of activity after 10 minutes
CC incubation at 50 degrees Celsius. {ECO:0000269|PubMed:15066183};
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:15066183}.
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DR EMBL; AB154368; BAD15093.1; -; Genomic_DNA.
DR EMBL; AB158573; BAE02667.1; -; Genomic_DNA.
DR AlphaFoldDB; Q75SP7; -.
DR SMR; Q75SP7; -.
DR KEGG; ag:BAD15093; -.
DR BioCyc; MetaCyc:MON-15017; -.
DR BRENDA; 3.5.1.100; 5085.
DR GO; GO:0016787; F:hydrolase activity; IEA:UniProtKB-KW.
DR GO; GO:0006807; P:nitrogen compound metabolic process; IEA:InterPro.
DR CDD; cd07576; R-amidase_like; 1.
DR Gene3D; 3.60.110.10; -; 1.
DR InterPro; IPR003010; C-N_Hydrolase.
DR InterPro; IPR036526; C-N_Hydrolase_sf.
DR InterPro; IPR044083; RamA-like.
DR Pfam; PF00795; CN_hydrolase; 1.
DR SUPFAM; SSF56317; SSF56317; 1.
DR PROSITE; PS50263; CN_HYDROLASE; 1.
PE 1: Evidence at protein level;
KW Direct protein sequencing; Hydrolase.
FT CHAIN 1..274
FT /note="(R)-stereoselective amidase"
FT /id="PRO_0000400092"
FT DOMAIN 1..234
FT /note="CN hydrolase"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00054"
FT ACT_SITE 40
FT /note="Proton acceptor"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00054"
FT ACT_SITE 108
FT /note="Proton donor"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00054"
FT ACT_SITE 140
FT /note="Nucleophile"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00054"
FT CONFLICT 2
FT /note="K -> A (in Ref. 1; AA sequence)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 274 AA; 30128 MW; 1C1B027C88046BD3 CRC64;
MKIELVQLAG RDGDTAYNLS RTLNAIATCA GDTDLLVFPE TYLSGFVGGA QLAQVAEPLH
GTTLQTLLQA VRQRDVAVVL GFAEVHQGRF YNSSVLVTPE GIALQYRKTH LWPSERSDFS
PGDRFTTVLW RGVRVGLLIC YDIELPETSR ALAQLGAEVV IVTNGNMDPY GPVHRTAIMA
RAQENQLFAV MVNRVGAGDD GLVFAGGSMA VDPFGRVLFE AGRDEVRHVV ELDLDQLKAA
RRDYDYLKDR RLMLSGEQTE HPDGRRELLI GASQ
