ID   DCAS_ENSAD              Reviewed;         304 AA.
AC   Q5S260;
DT   16-MAR-2016, integrated into UniProtKB/Swiss-Prot.
DT   21-DEC-2004, sequence version 1.
DT   25-MAY-2022, entry version 44.
DE   RecName: Full=N-carbamoyl-D-amino acid hydrolase {ECO:0000305|PubMed:16546310};
DE            EC=3.5.1.77 {ECO:0000269|PubMed:16546310};
DE   AltName: Full=D-carbamoylase {ECO:0000303|PubMed:16546310};
OS   Ensifer adhaerens (Sinorhizobium morelense).
OC   Bacteria; Proteobacteria; Alphaproteobacteria; Hyphomicrobiales;
OC   Rhizobiaceae; Sinorhizobium/Ensifer group; Ensifer.
OX   NCBI_TaxID=106592 {ECO:0000312|EMBL:AAV53595.1};
RN   [1] {ECO:0000312|EMBL:AAV53595.1}
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 2-11, FUNCTION,
RP   CATALYTIC ACTIVITY, ACTIVITY REGULATION, BIOPHYSICOCHEMICAL PROPERTIES,
RP   SUBSTRATE SPECIFICITY, SUBUNIT, AND BIOTECHNOLOGY.
RC   STRAIN=S-5 {ECO:0000303|PubMed:16546310};
RX   PubMed=16546310; DOI=10.1016/j.biochi.2005.05.013;
RA   Wu S., Liu Y., Zhao G., Wang J., Sun W.;
RT   "Thermostable D-carbamoylase from Sinorhizobium morelens S-5: purification,
RT   characterization and gene expression in Escherichia coli.";
RL   Biochimie 88:237-244(2006).
CC   -!- FUNCTION: Catalyzes the hydrolysis of N-carbamoyl-D-amino acids to the
CC       corresponding D-amino acids. Hydrolyzes aromatic and aliphatic N-
CC       carbamoyl-D-amino acids in vitro. Effectively hydrolyzes N-carbamoyl-D-
CC       p-hydroxyphenylglycine and N-carbamoyl-DL-p-hydroxyphenylglycine, and
CC       to a lesser extent N-carbamoyl-D-methionine. No activity for N-
CC       carbamoyl-L-amino acids, N-carbamoyl-beta-alanine or (RS)-alpha-ethyl-
CC       N-carbamoylphenylglycine in vitro. {ECO:0000269|PubMed:16546310}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=an N-carbamoyl-D-amino acid + 2 H(+) + H2O = a D-alpha-amino
CC         acid + CO2 + NH4(+); Xref=Rhea:RHEA:11000, ChEBI:CHEBI:15377,
CC         ChEBI:CHEBI:15378, ChEBI:CHEBI:16526, ChEBI:CHEBI:28938,
CC         ChEBI:CHEBI:59871, ChEBI:CHEBI:85602; EC=3.5.1.77;
CC         Evidence={ECO:0000269|PubMed:16546310};
CC   -!- ACTIVITY REGULATION: The activity decreases with increasing
CC       concentration of H(2)O(2). Has 68% and 43% of activity remaining upon
CC       treatment with 0.1 and 0.2 mM H(2)O(2) for 30 minutes, respectively.
CC       Inhibited significantly by 2 mM Zn(2+), Cu(2+) and Ag(+), moderately by
CC       Co(2+), Mn(2+), Sn(2+) and Mg(2+), and only slightly by Ba(2+).
CC       Slightly activated by Fe(2+) and Ca(2+). No effect on activity by metal
CC       chelators EDTA and 8-hydroxyquinoline at 2 mM or by dithiothreitol, 2-
CC       mercaptoethanol or phenylmethanesulfonyl fluoride.
CC       {ECO:0000269|PubMed:16546310}.
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       Kinetic parameters:
CC         KM=3.76 mM for N-carbamoyl-D-p-hydroxyphenylglycine
CC         {ECO:0000269|PubMed:16546310};
CC         Vmax=383 umol/min/mg enzyme {ECO:0000269|PubMed:16546310};
CC       pH dependence:
CC         Optimum pH is 7.0. Stable at pH 6.5-8.2.
CC         {ECO:0000269|PubMed:16546310};
CC       Temperature dependence:
CC         Optimum temperature is 60 degrees Celsius. Very thermostable as still
CC         has 98% of its activity after incubation for 30 minutes at 50 degrees
CC         Celsius. {ECO:0000269|PubMed:16546310};
CC   -!- SUBUNIT: Homotetramer. {ECO:0000269|PubMed:16546310}.
CC   -!- BIOTECHNOLOGY: Has potential for industrial application in the
CC       enzymatic production of D-amino acids, which are very valuable
CC       intermediates for the production of semisynthetic antibiotics, peptide
CC       hormones, and pesticides. {ECO:0000303|PubMed:16546310}.
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DR   EMBL; AY787759; AAV53595.1; -; Genomic_DNA.
DR   AlphaFoldDB; Q5S260; -.
DR   SMR; Q5S260; -.
DR   KEGG; ag:AAV53595; -.
DR   BRENDA; 3.5.1.77; 8538.
DR   GO; GO:0047417; F:N-carbamoyl-D-amino acid hydrolase activity; IDA:UniProtKB.
DR   GO; GO:0006807; P:nitrogen compound metabolic process; IEA:InterPro.
DR   GO; GO:0051289; P:protein homotetramerization; IDA:UniProtKB.
DR   Gene3D; 3.60.110.10; -; 1.
DR   InterPro; IPR003010; C-N_Hydrolase.
DR   InterPro; IPR036526; C-N_Hydrolase_sf.
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..304
FT                   /note="N-carbamoyl-D-amino acid hydrolase"
FT                   /id="PRO_0000435673"
FT   DOMAIN          5..276
FT                   /note="CN hydrolase"
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU00054"
FT   ACT_SITE        47
FT                   /evidence="ECO:0000250|UniProtKB:Q44185"
FT   ACT_SITE        127
FT                   /evidence="ECO:0000250|UniProtKB:Q44185"
FT   ACT_SITE        172
FT                   /evidence="ECO:0000250|UniProtKB:Q44185"
SQ   SEQUENCE   304 AA;  34366 MW;  55B042B6A339D46C CRC64;
     MTRQMILAVG QQGPIARAET REQVVVRLLY MLTKAASRGA NFIVFPELAF TTFFPRWHFT
     DEAELDSFYE TEMPGPVVRP LFEKAAELGI GFNLGYAELV VEGGVKRRFN TSILVDKPGK
     IVGKYRKIHL PGHKEYEAYR PFQHLEKRYF EPGDLGFPVY DVDAAKMGMF ICNDRRWPEA
     WRVMGLRGAE IICGGYNTPT HNPPVPQHDH LTSFHHLLSM QAGSYQNGAW SAAAGKVGME
     ENCMLLGHSC IVAPTGEIVA LTTTLEDEVI TAAVCLDRCR ELREHIFNFK QHRQPQHYGL
     IAEL