LAAA_PSEAZ
ID LAAA_PSEAZ Reviewed; 310 AA.
AC Q76KX0;
DT 02-NOV-2010, integrated into UniProtKB/Swiss-Prot.
DT 05-JUL-2004, sequence version 1.
DT 03-AUG-2022, entry version 66.
DE RecName: Full=L-amino acid amidase {ECO:0000303|PubMed:15066172};
DE EC=3.5.1.101 {ECO:0000269|PubMed:15066172};
GN Name=laaA {ECO:0000312|EMBL:BAD15092.1};
OS Pseudomonas azotoformans.
OC Bacteria; Proteobacteria; Gammaproteobacteria; Pseudomonadales;
OC Pseudomonadaceae; Pseudomonas.
OX NCBI_TaxID=47878;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 1-10 AND 53-62,
RP FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION, BIOPHYSICOCHEMICAL
RP PROPERTIES, AND SUBUNIT.
RC STRAIN=DSM 1862 / CIP 106744 / IAM 1603 / JCM 2777 / NCBRC 12693
RC {ECO:0000312|EMBL:BAD15092.1};
RX PubMed=15066172; DOI=10.1111/j.1432-1033.2004.04056.x;
RA Komeda H., Harada H., Washika S., Sakamoto T., Ueda M., Asano Y.;
RT "S-stereoselective piperazine-2-tert-butylcarboxamide hydrolase from
RT Pseudomonas azotoformans IAM 1603 is a novel L-amino acid amidase.";
RL Eur. J. Biochem. 271:1465-1475(2004).
CC -!- FUNCTION: Hydrolyzes L-prolinamide, L-proline-p-nitroanilide, L-
CC alaninamide, L-methioninamide, piperidine-2-carboxamide and piperazine-
CC 2-carboxamide. Has a much lower activity towards piperazine-2-tert-
CC butylcarboxamide. Does not hydrolyze dipeptides and D-prolinamide.
CC {ECO:0000269|PubMed:15066172}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(S)-piperazine-2-carboxamide + H2O = (S)-piperazine-2-
CC carboxylate + NH4(+); Xref=Rhea:RHEA:26550, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:28938, ChEBI:CHEBI:58919, ChEBI:CHEBI:58920;
CC EC=3.5.1.101; Evidence={ECO:0000269|PubMed:15066172};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + L-prolinamide = L-proline + NH4(+);
CC Xref=Rhea:RHEA:26510, ChEBI:CHEBI:15377, ChEBI:CHEBI:28938,
CC ChEBI:CHEBI:58495, ChEBI:CHEBI:60039; EC=3.5.1.101;
CC Evidence={ECO:0000269|PubMed:15066172};
CC -!- ACTIVITY REGULATION: Completely inhibited by ZnSO(4), ZnCl(2), AgNO(3),
CC CdCl(2) and HgCl(2). Partially inhibited by PbCl(2), NiCl(2) and
CC CoCl(2). Unaffected by LiBr, H(2)BO(3), NaCl, MgSO(4), AlCl(3), KCl,
CC CaCl(2), CrCl(3), MnCl(2), FeSO(4), Fe(NH(4))(2)(SO(4))(2), CuSO(4),
CC RbCl, Na(2)MoO(4), (NH(4))(6)Mo(7)O(24), SnCl(2), CsCl and BaCl(2).
CC Completely inhibited by phenylhydrazine, but not by the other carbonyl
CC reagents hydroxylamine, hydrazine, D,L-penicillamine and D-cycloserine.
CC Unaffected by the chelating agents o-phenanthroline, 8-
CC hydroxyquinoline, enthylenediaminetetraacetic acid and alpha,alpha'-
CC dipyridyl. Partially inhibited by the thiol reagents p-
CC chloromercuribenzoate, iodoacetate and N-ethylmaleimide. Not affected
CC by the serine protease inhibitor phenylmethanesulfonyl fluoride, the
CC serine/cysteine protease inhibitor leupeptine or the aspartic protease
CC inhibitor pepstatin. {ECO:0000269|PubMed:15066172}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.58 mM for L-proline-p-nitroanilide
CC {ECO:0000269|PubMed:15066172};
CC Vmax=80.9 umol/min/mg enzyme with L-proline-p-nitroanilide as
CC substrate {ECO:0000269|PubMed:15066172};
CC pH dependence:
CC Optimum pH is 9.0. Stable from pH 6.0 to 9.5.
CC {ECO:0000269|PubMed:15066172};
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 5 minutes incubation at 55
CC degrees Celsius, and only retains 25% of activity after 5 minutes
CC incubation at 50 degrees Celsius. {ECO:0000269|PubMed:15066172};
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:15066172}.
CC -!- SIMILARITY: Belongs to the peptidase S33 family. {ECO:0000305}.
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DR EMBL; AB087498; BAD15092.1; -; Genomic_DNA.
DR RefSeq; WP_071496689.1; NZ_UYXP01000006.1.
DR AlphaFoldDB; Q76KX0; -.
DR SMR; Q76KX0; -.
DR ESTHER; psefs-laaa; Proline_iminopeptidase.
DR GeneID; 57378433; -.
DR KEGG; ag:BAD15092; -.
DR BioCyc; MetaCyc:MON-15016; -.
DR BRENDA; 3.5.1.101; 10312.
DR GO; GO:0008233; F:peptidase activity; IEA:InterPro.
DR GO; GO:0006508; P:proteolysis; IEA:InterPro.
DR Gene3D; 3.40.50.1820; -; 1.
DR InterPro; IPR029058; AB_hydrolase.
DR InterPro; IPR000073; AB_hydrolase_1.
DR InterPro; IPR002410; Peptidase_S33.
DR InterPro; IPR005945; Pro_imino_pep.
DR Pfam; PF00561; Abhydrolase_1; 1.
DR PIRSF; PIRSF005539; Pept_S33_TRI_F1; 1.
DR PRINTS; PR00793; PROAMNOPTASE.
DR SUPFAM; SSF53474; SSF53474; 1.
DR TIGRFAMs; TIGR01250; pro_imino_pep_2; 1.
PE 1: Evidence at protein level;
KW Direct protein sequencing; Hydrolase.
FT CHAIN 1..310
FT /note="L-amino acid amidase"
FT /id="PRO_0000400095"
FT DOMAIN 33..284
FT /note="AB hydrolase-1"
FT /evidence="ECO:0000255"
FT ACT_SITE 111
FT /note="Nucleophile"
FT /evidence="ECO:0000250"
FT ACT_SITE 251
FT /evidence="ECO:0000250"
FT ACT_SITE 278
FT /note="Proton donor"
FT /evidence="ECO:0000250"
SQ SEQUENCE 310 AA; 34514 MW; 79D8C36C6AC7A63D CRC64;
MEFIEKIREG YAAFGAYQTW YRVTGDLSSG RTPLVVIHGG PGCTHDYVDA FKDVAASGHA
VIHYDQLGNG RSTHLPDKDP SFWTVGLFLE ELNNLLDHLQ ISDNYAILGQ SWGGMLGSEH
AILQPKGLRA FIPANSPTCM RTWVSEANRL RKLLPEGVHE TLLKHETAGT YQDPEYLAAS
RVFYDHHVCR VIPWPEEVAR TFAAVDADPT VYHAMSGPTE FHVIGSLKDW KSTGRLSAIN
VPTLVISGRH DEATPLVVKP FLDEIADVRW ALFEDSSHMP HVEERQACMG TVVKFLDEVC
SAKYKVLKAS
