HYUC_RHIML
ID HYUC_RHIML Reviewed; 416 AA.
AC Q6DTN4;
DT 10-MAY-2017, integrated into UniProtKB/Swiss-Prot.
DT 16-NOV-2011, sequence version 1.
DT 03-AUG-2022, entry version 51.
DE RecName: Full=N-carbamoyl-L-amino-acid hydrolase {ECO:0000303|PubMed:16254442};
DE EC=3.5.1.87 {ECO:0000269|PubMed:16254442};
DE AltName: Full=Hydantoin utilization protein C {ECO:0000250|UniProtKB:Q01264};
DE AltName: Full=L-N-carbamoylase {ECO:0000303|PubMed:16254442};
GN Name=hyuC {ECO:0000303|PubMed:16254442};
OS Rhizobium meliloti (Ensifer meliloti) (Sinorhizobium meliloti).
OC Bacteria; Proteobacteria; Alphaproteobacteria; Hyphomicrobiales;
OC Rhizobiaceae; Sinorhizobium/Ensifer group; Sinorhizobium.
OX NCBI_TaxID=382;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, ACTIVITY REGULATION, SUBSTRATE SPECIFICITY,
RP SUBUNIT, AND COFACTOR.
RC STRAIN=CECT 4114;
RX PubMed=16254442; DOI=10.1159/000088142;
RA Martinez-Rodriguez S., Clemente-Jimenez J.M., Rodriguez-Vico F.,
RA Las Heras-Vazquez F.J.;
RT "Molecular cloning and biochemical characterization of L-N-carbamoylase
RT from Sinorhizobium meliloti CECT4114.";
RL J. Mol. Microbiol. Biotechnol. 9:16-25(2005).
CC -!- FUNCTION: May be involved in the asymmetric conversion of racemic 5-
CC substituted hydantoins to the corresponding L-amino acids. Catalyzes
CC specifically the conversion of N-carbamoyl-L-amino acids to free L-
CC amino acids in an irreversible reaction. N-carbamoyl-L-methionine is
CC the best substrate. HyuC of R.meliloti is the first L-N-carbamoylase
CC that hydrolyzes N-carbamoyl-L-tryptophan as well as N-carbamoyl-L-amino
CC acids with aliphatic substituents. {ECO:0000269|PubMed:16254442}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=an N-carbamoyl-L-alpha-amino acid + 2 H(+) + H2O = an L-alpha-
CC amino acid + CO2 + NH4(+); Xref=Rhea:RHEA:17581, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:16526, ChEBI:CHEBI:28938,
CC ChEBI:CHEBI:58865, ChEBI:CHEBI:59869; EC=3.5.1.87;
CC Evidence={ECO:0000269|PubMed:16254442};
CC -!- COFACTOR:
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC Evidence={ECO:0000269|PubMed:16254442};
CC Name=Ni(2+); Xref=ChEBI:CHEBI:49786;
CC Evidence={ECO:0000269|PubMed:16254442};
CC Name=Co(2+); Xref=ChEBI:CHEBI:48828;
CC Evidence={ECO:0000269|PubMed:16254442};
CC Name=Fe(2+); Xref=ChEBI:CHEBI:29033;
CC Evidence={ECO:0000269|PubMed:16254442};
CC Note=Binds 2 divalent metal cations per subunit.
CC {ECO:0000250|UniProtKB:Q8VXY9};
CC -!- ACTIVITY REGULATION: Ni(2+), Mn(2+), Co(2+) and Fe(2+) ions greatly
CC increase hydrolase activity. Strongly inhibited by Hg(2+), Cu(2+),
CC Zn(2+), Pb(2+) and Fe(3+) ions, and slightly inhibited by Na(+) and
CC K(+) ions. Beta-mercaptoethanol and 5,5'-dithiobis-(2-nitrobenzoic
CC acid)(DTNB) cause 34% and 42% inhibition, respectively.
CC {ECO:0000269|PubMed:16254442}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.65 mM for N-carbamoyl-L-tryptophan (at 40 degrees Celsius)
CC {ECO:0000269|PubMed:16254442};
CC KM=0.69 mM for N-carbamoyl-L-methionine (at 40 degrees Celsius)
CC {ECO:0000269|PubMed:16254442};
CC KM=0.94 mM for N-carbamoyl-L-alanine (at 40 degrees Celsius)
CC {ECO:0000269|PubMed:16254442};
CC KM=2.61 mM for N-carbamoyl-L-phenylalanine (at 40 degrees Celsius)
CC {ECO:0000269|PubMed:16254442};
CC KM=4.80 mM for N-carbamoyl-L-tyrosine (at 40 degrees Celsius)
CC {ECO:0000269|PubMed:16254442};
CC KM=4.91 mM for N-carbamoyl-L-cysteine (at 40 degrees Celsius)
CC {ECO:0000269|PubMed:16254442};
CC KM=5.5 mM for N-acetyl-L-methionine (at 40 degrees Celsius)
CC {ECO:0000269|PubMed:16254442};
CC KM=12.9 mM for N-formyl-L-methionine (at 40 degrees Celsius)
CC {ECO:0000269|PubMed:16254442};
CC KM=34.47 mM for N-carbamoyl-L-valine (at 40 degrees Celsius)
CC {ECO:0000269|PubMed:16254442};
CC KM=51.23 mM for N-carbamoyl-L-glutamic acid (at 40 degrees Celsius)
CC {ECO:0000269|PubMed:16254442};
CC Note=kcat is 91.66 sec(-1) for formyl-L-methionine as substrate (at
CC 40 degrees Celsius). kcat is 66.84 sec(-1) for N-acetyl-L-methionine
CC as substrate (at 40 degrees Celsius). kcat is 16.9 sec(-1) for N-
CC carbamoyl-L-valine as substrate (at 40 degrees Celsius). kcat is
CC 14.46 sec(-1) for N-carbamoyl-L-methionine as substrate (at 40
CC degrees Celsius). kcat is 13.55 sec(-1) for N-carbamoyl-L-cysteine as
CC substrate (at 40 degrees Celsius). kcat is 4.26 sec(-1) for N-
CC carbamoyl-L-phenylalanine as substrate (at 40 degrees Celsius). kcat
CC is 2.59 sec(-1) for N-carbamoyl-L-alanine as substrate (at 40 degrees
CC Celsius). kcat is 1.52 sec(-1) for N-carbamoyl-L-glutamic acid as
CC substrate (at 40 degrees Celsius). kcat is 0.30 sec(-1) for N-
CC carbamoyl-L-tyrosine as substrate (at 40 degrees Celsius). kcat is
CC 0.15 sec(-1) for N-carbamoyl-L-tryptophan as substrate (at 40 degrees
CC Celsius). {ECO:0000269|PubMed:16254442};
CC pH dependence:
CC Optimum pH is 8. {ECO:0000269|PubMed:16254442};
CC Temperature dependence:
CC Optimum temperature is 60 degrees Celsius.
CC {ECO:0000269|PubMed:16254442};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:16254442}.
CC -!- SIMILARITY: Belongs to the peptidase M20 family. {ECO:0000305}.
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DR EMBL; AY646850; AAT66633.1; -; Genomic_DNA.
DR RefSeq; WP_010969946.1; NZ_WISY01000224.1.
DR AlphaFoldDB; Q6DTN4; -.
DR SMR; Q6DTN4; -.
DR STRING; 382.DU99_13665; -.
DR GeneID; 61603912; -.
DR PATRIC; fig|382.52.peg.2781; -.
DR OMA; IWPHGRW; -.
DR GO; GO:0016813; F:hydrolase activity, acting on carbon-nitrogen (but not peptide) bonds, in linear amidines; IEA:InterPro.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0050538; F:N-carbamoyl-L-amino-acid hydrolase activity; IEA:UniProtKB-EC.
DR GO; GO:0008652; P:cellular amino acid biosynthetic process; IEA:UniProtKB-KW.
DR InterPro; IPR010158; Amidase_Cbmase.
DR InterPro; IPR036264; Bact_exopeptidase_dim_dom.
DR InterPro; IPR002933; Peptidase_M20.
DR InterPro; IPR011650; Peptidase_M20_dimer.
DR PANTHER; PTHR32494; PTHR32494; 1.
DR Pfam; PF07687; M20_dimer; 1.
DR Pfam; PF01546; Peptidase_M20; 1.
DR PIRSF; PIRSF001235; Amidase_carbamoylase; 1.
DR SUPFAM; SSF55031; SSF55031; 1.
DR TIGRFAMs; TIGR01879; hydantase; 1.
PE 1: Evidence at protein level;
KW Amino-acid biosynthesis; Hydrolase; Manganese; Metal-binding.
FT CHAIN 1..416
FT /note="N-carbamoyl-L-amino-acid hydrolase"
FT /id="PRO_0000439931"
FT REGION 132..133
FT /note="Substrate"
FT /evidence="ECO:0000250|UniProtKB:Q8VXY9"
FT REGION 194..197
FT /note="Substrate"
FT /evidence="ECO:0000250|UniProtKB:Q8VXY9"
FT REGION 216..330
FT /note="Involved in dimerization"
FT /evidence="ECO:0000250|UniProtKB:Q8VXY9"
FT REGION 361..362
FT /note="Substrate"
FT /evidence="ECO:0000250|UniProtKB:Q8VXY9"
FT BINDING 87
FT /ligand="a divalent metal cation"
FT /ligand_id="ChEBI:CHEBI:60240"
FT /ligand_label="1"
FT /evidence="ECO:0000250|UniProtKB:Q8VXY9"
FT BINDING 98
FT /ligand="a divalent metal cation"
FT /ligand_id="ChEBI:CHEBI:60240"
FT /ligand_label="1"
FT /evidence="ECO:0000250|UniProtKB:Q8VXY9"
FT BINDING 98
FT /ligand="a divalent metal cation"
FT /ligand_id="ChEBI:CHEBI:60240"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:Q8VXY9"
FT BINDING 133
FT /ligand="a divalent metal cation"
FT /ligand_id="ChEBI:CHEBI:60240"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:Q8VXY9"
FT BINDING 194
FT /ligand="a divalent metal cation"
FT /ligand_id="ChEBI:CHEBI:60240"
FT /ligand_label="1"
FT /evidence="ECO:0000250|UniProtKB:Q8VXY9"
FT BINDING 230
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q8VXY9"
FT BINDING 279
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q8VXY9"
FT BINDING 292
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q8VXY9"
FT BINDING 386
FT /ligand="a divalent metal cation"
FT /ligand_id="ChEBI:CHEBI:60240"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:Q8VXY9"
FT BINDING 386
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q8VXY9"
FT SITE 239
FT /note="Necessary for dimerization"
FT /evidence="ECO:0000250|UniProtKB:Q53389"
SQ SEQUENCE 416 AA; 44857 MW; 85BA5F11BD855B97 CRC64;
MAAPGENRRV NADRLWDSLM EMAKIGPGVA GGNNRQTLTD ADGEGRRLFQ SWCEEAGLSM
GVDKMGTMFL TRPGTDPDAL PVHIGSHLDT QPTGGKFDGV LGVLSGLEAV RTMNDLGIKT
KHPIVVTNWT NEEGARFAPA MLASGVFAGV HTLEYAYARK DPEGKSFGDE LKRIGWLGDE
EVGARKMHAY FEYHIEQGPI LEAENKQIGV VTHCQGLWWL EFTLTGREAH TGSTPMDMRV
NAGLAMARIL EMVQTVAMEN QPGAVGGVGQ MFFSPNSRNV LPGKVVFTVD IRSPDQAKLD
GMRARIEAEA PKICERLGVG CSIEAVGHFD PVTFDPKLVE TVRGAAEKLG YSHMNLVSGA
GHDACWAAKV APTTMIMCPC VGGLSHNEAE DISREWAAAG ADVLFHAVLE TAEIVE
