NITR_PYRAB
ID NITR_PYRAB Reviewed; 262 AA.
AC Q9UYV8;
DT 11-JUN-2014, integrated into UniProtKB/Swiss-Prot.
DT 01-MAY-2000, sequence version 1.
DT 03-AUG-2022, entry version 109.
DE RecName: Full=Nitrilase;
DE EC=3.5.5.1;
DE AltName: Full=PaNit;
GN OrderedLocusNames=PYRAB13990; ORFNames=PAB1449;
OS Pyrococcus abyssi (strain GE5 / Orsay).
OC Archaea; Euryarchaeota; Thermococci; Thermococcales; Thermococcaceae;
OC Pyrococcus.
OX NCBI_TaxID=272844;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=GE5 / Orsay;
RX PubMed=12622808; DOI=10.1046/j.1365-2958.2003.03381.x;
RA Cohen G.N., Barbe V., Flament D., Galperin M., Heilig R., Lecompte O.,
RA Poch O., Prieur D., Querellou J., Ripp R., Thierry J.-C., Van der Oost J.,
RA Weissenbach J., Zivanovic Y., Forterre P.;
RT "An integrated analysis of the genome of the hyperthermophilic archaeon
RT Pyrococcus abyssi.";
RL Mol. Microbiol. 47:1495-1512(2003).
RN [2]
RP GENOME REANNOTATION.
RC STRAIN=GE5 / Orsay;
RX PubMed=22057919; DOI=10.1007/s00284-011-0035-x;
RA Gao J., Wang J.;
RT "Re-annotation of two hyperthermophilic archaea Pyrococcus abyssi GE5 and
RT Pyrococcus furiosus DSM 3638.";
RL Curr. Microbiol. 64:118-129(2012).
RN [3]
RP IDENTIFICATION, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL
RP PROPERTIES, SUBSTRATE SPECIFICITY, ACTIVITY REGULATION, AND SUBUNIT.
RX PubMed=16495079; DOI=10.1016/j.pep.2006.01.006;
RA Mueller P., Egorova K., Vorgias C.E., Boutou E., Trauthwein H., Verseck S.,
RA Antranikian G.;
RT "Cloning, overexpression, and characterization of a thermoactive nitrilase
RT from the hyperthermophilic archaeon Pyrococcus abyssi.";
RL Protein Expr. Purif. 47:672-681(2006).
RN [4]
RP X-RAY CRYSTALLOGRAPHY (1.57 ANGSTROMS) OF APOENZYME AND IN COMPLEXES WITH
RP ACETATE, SUBUNIT, DOCKING STUDIES, REACTION MECHANISM, AND ACTIVE SITES.
RC STRAIN=GE5 / Orsay;
RX PubMed=21095228; DOI=10.1016/j.jsb.2010.11.017;
RA Raczynska J.E., Vorgias C.E., Antranikian G., Rypniewski W.;
RT "Crystallographic analysis of a thermoactive nitrilase.";
RL J. Struct. Biol. 173:294-302(2011).
CC -!- FUNCTION: Nitrilase that hydrolyzes preferentially aliphatic nitriles
CC like malononitrile and fumaronitrile in vitro. These dinitriles are
CC converted to the corresponding monoacid mononitriles, showing the
CC enzyme is regioselective. Cannot hydrolyze compounds with a nitrile
CC group bound to an aromatic ring or amino acid. Its biological role is
CC unknown. {ECO:0000269|PubMed:16495079}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a nitrile + 2 H2O = a carboxylate + NH4(+);
CC Xref=Rhea:RHEA:21724, ChEBI:CHEBI:15377, ChEBI:CHEBI:18379,
CC ChEBI:CHEBI:28938, ChEBI:CHEBI:29067; EC=3.5.5.1;
CC Evidence={ECO:0000269|PubMed:16495079};
CC -!- ACTIVITY REGULATION: Enzymatic activity is inhibited in the presence of
CC acetone, methanol and metal ions such as Ag(2+) and Hg(2+). Is also
CC inhibited by various thiol reagents such as DTNB, p-
CC chloromercuribenzoate, p-hydroxymercuribenzoate, iodacetamide and
CC iodacetate. EDTA has no influence on activity.
CC {ECO:0000269|PubMed:16495079}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=3.47 mM for malononitrile {ECO:0000269|PubMed:16495079};
CC KM=9.48 mM for fumaronitrile {ECO:0000269|PubMed:16495079};
CC pH dependence:
CC Optimum pH is 7.4. Active within the pH range of 4.5-8.5.
CC {ECO:0000269|PubMed:16495079};
CC Temperature dependence:
CC Optimum temperature is 80 degrees Celsius. Active at a broad
CC temperature range (60-90 degrees Celsius). Highly thermostable with a
CC half-life of 25 hours at 70 degrees Celsius, 9 hours at 80 degrees
CC Celsius, and 6 hours at 90 degrees Celsius. Shows a Tm of 112.7
CC degrees Celsius. {ECO:0000269|PubMed:16495079};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:16495079,
CC ECO:0000269|PubMed:21095228}.
CC -!- SIMILARITY: Belongs to the carbon-nitrogen hydrolase superfamily.
CC {ECO:0000305}.
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DR EMBL; AJ248287; CAB50304.1; -; Genomic_DNA.
DR EMBL; HE613800; CCE70842.1; -; Genomic_DNA.
DR PIR; C75051; C75051.
DR RefSeq; WP_010868514.1; NC_000868.1.
DR PDB; 3IVZ; X-ray; 1.57 A; A/B=1-262.
DR PDB; 3IW3; X-ray; 1.80 A; A/B=1-262.
DR PDB; 3KI8; X-ray; 1.60 A; A/B=1-262.
DR PDB; 3KLC; X-ray; 1.76 A; A/B=1-262.
DR PDBsum; 3IVZ; -.
DR PDBsum; 3IW3; -.
DR PDBsum; 3KI8; -.
DR PDBsum; 3KLC; -.
DR AlphaFoldDB; Q9UYV8; -.
DR SMR; Q9UYV8; -.
DR STRING; 272844.PAB1449; -.
DR EnsemblBacteria; CAB50304; CAB50304; PAB1449.
DR GeneID; 1496788; -.
DR KEGG; pab:PAB1449; -.
DR PATRIC; fig|272844.11.peg.1486; -.
DR eggNOG; arCOG00062; Archaea.
DR HOGENOM; CLU_030130_3_1_2; -.
DR OMA; GNTYRES; -.
DR OrthoDB; 66517at2157; -.
DR PhylomeDB; Q9UYV8; -.
DR EvolutionaryTrace; Q9UYV8; -.
DR Proteomes; UP000000810; Chromosome.
DR Proteomes; UP000009139; Chromosome.
DR GO; GO:0080061; F:indole-3-acetonitrile nitrilase activity; IEA:UniProtKB-EC.
DR GO; GO:0006807; P:nitrogen compound metabolic process; IEA:InterPro.
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 3D-structure; Hydrolase.
FT CHAIN 1..262
FT /note="Nitrilase"
FT /id="PRO_0000429328"
FT DOMAIN 2..237
FT /note="CN hydrolase"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00054"
FT ACT_SITE 42
FT /note="Proton acceptor"
FT /evidence="ECO:0000305|PubMed:21095228"
FT ACT_SITE 113
FT /note="Proton donor"
FT /evidence="ECO:0000305|PubMed:21095228"
FT ACT_SITE 146
FT /note="Nucleophile"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00054,
FT ECO:0000269|PubMed:21095228"
FT BINDING 173..174
FT /ligand="substrate"
FT /evidence="ECO:0000255"
FT STRAND 3..8
FT /evidence="ECO:0007829|PDB:3IVZ"
FT HELIX 16..32
FT /evidence="ECO:0007829|PDB:3IVZ"
FT STRAND 36..39
FT /evidence="ECO:0007829|PDB:3IVZ"
FT TURN 42..46
FT /evidence="ECO:0007829|PDB:3IVZ"
FT HELIX 53..59
FT /evidence="ECO:0007829|PDB:3IVZ"
FT TURN 63..65
FT /evidence="ECO:0007829|PDB:3IVZ"
FT HELIX 67..79
FT /evidence="ECO:0007829|PDB:3IVZ"
FT STRAND 82..91
FT /evidence="ECO:0007829|PDB:3IVZ"
FT STRAND 94..103
FT /evidence="ECO:0007829|PDB:3IVZ"
FT STRAND 106..112
FT /evidence="ECO:0007829|PDB:3IVZ"
FT HELIX 118..122
FT /evidence="ECO:0007829|PDB:3IVZ"
FT STRAND 133..135
FT /evidence="ECO:0007829|PDB:3IVZ"
FT STRAND 140..143
FT /evidence="ECO:0007829|PDB:3IVZ"
FT HELIX 146..150
FT /evidence="ECO:0007829|PDB:3IVZ"
FT HELIX 152..160
FT /evidence="ECO:0007829|PDB:3IVZ"
FT STRAND 164..170
FT /evidence="ECO:0007829|PDB:3IVZ"
FT HELIX 177..188
FT /evidence="ECO:0007829|PDB:3IVZ"
FT STRAND 191..196
FT /evidence="ECO:0007829|PDB:3IVZ"
FT STRAND 199..201
FT /evidence="ECO:0007829|PDB:3IVZ"
FT STRAND 211..213
FT /evidence="ECO:0007829|PDB:3IVZ"
FT STRAND 219..222
FT /evidence="ECO:0007829|PDB:3IVZ"
FT STRAND 229..234
FT /evidence="ECO:0007829|PDB:3IVZ"
FT HELIX 237..241
FT /evidence="ECO:0007829|PDB:3IVZ"
FT STRAND 244..246
FT /evidence="ECO:0007829|PDB:3IVZ"
FT HELIX 251..254
FT /evidence="ECO:0007829|PDB:3IVZ"
FT HELIX 257..259
FT /evidence="ECO:0007829|PDB:3IVZ"
SQ SEQUENCE 262 AA; 29798 MW; C2EFA86AACF51D4F CRC64;
MVKVAYVQMN PQILEPDKNY SKAEKLIKEA SKQGAQLVVL PELFDTGYNF ETREEVFEIA
QKIPEGETTT FLMDVARDTG VYIVAGTAEK DGDVLYNSAV VVGPRGFIGK YRKIHLFYRE
KFFFEPGDLG FRVFDLGFMK VGVMICFDWF FPESARTLAL KGADVIAHPA NLVMPYAPRA
MPIRALENKV YTVTADRVGE ERGLKFIGKS LIASPKAEVL SMASETEEEV GVAEIDLYLV
RNKRINDLND IFKDRREEYY FR
