ID   CHT_ASPNG               Reviewed;         356 AA.
AC   A9QXE0;
DT   04-MAR-2015, integrated into UniProtKB/Swiss-Prot.
DT   05-FEB-2008, sequence version 1.
DT   03-AUG-2022, entry version 45.
DE   RecName: Full=Cyanide hydratase {ECO:0000255|HAMAP-Rule:MF_03224, ECO:0000303|PubMed:23475593};
DE            Short=CHT {ECO:0000255|HAMAP-Rule:MF_03224};
DE            EC=4.2.1.66 {ECO:0000255|HAMAP-Rule:MF_03224, ECO:0000269|PubMed:23475593};
DE   AltName: Full=Cyanide-degrading nitrilase {ECO:0000255|HAMAP-Rule:MF_03224};
DE   AltName: Full=Formamide hydrolyase {ECO:0000255|HAMAP-Rule:MF_03224};
DE   AltName: Full=NitAn1 {ECO:0000303|PubMed:23475593};
OS   Aspergillus niger.
OC   Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Eurotiomycetes;
OC   Eurotiomycetidae; Eurotiales; Aspergillaceae; Aspergillus;
OC   Aspergillus subgen. Circumdati.
OX   NCBI_TaxID=5061;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [MRNA].
RC   STRAIN=K10 / CCF 3411;
RA   Kaplan O., Martinkova L., Plihal O., Bezouska K., Kavan D.;
RL   Submitted (NOV-2007) to the EMBL/GenBank/DDBJ databases.
RN   [2]
RP   FUNCTION, AND CATALYTIC ACTIVITY.
RC   STRAIN=K10 / CCF 3411;
RX   PubMed=17061133; DOI=10.1007/s00253-006-0503-6;
RA   Kaplan O., Vejvoda V., Plihal O., Pompach P., Kavan D., Bojarova P.,
RA   Bezouska K., Mackova M., Cantarella M., Jirku V., Kren V., Martinkova L.;
RT   "Purification and characterization of a nitrilase from Aspergillus niger
RT   K10.";
RL   Appl. Microbiol. Biotechnol. 73:567-575(2006).
RN   [3]
RP   ERRATUM OF PUBMED:17061133, AND IDENTIFICATION BY MASS SPECTROMETRY.
RX   PubMed=23455586; DOI=10.1007/s00253-013-4743-y;
RA   Kaplan O., Vejvoda V., Plihal O., Pompach P., Kavan D., Bojarova P.,
RA   Bezouska K., Mackova M., Cantarella M., Jirku V., Kren V., Martinkova L.;
RL   Appl. Microbiol. Biotechnol. 97:3745-3746(2013).
RN   [4]
RP   RETRACTED PAPER.
RX   PubMed=21210990; DOI=10.1186/1472-6750-11-2;
RA   Kaplan O., Bezouska K., Plihal O., Ettrich R., Kulik N., Vanek O.,
RA   Kavan D., Benada O., Malandra A., Sveda O., Vesela A.B., Rinagelova A.,
RA   Slamova K., Cantarella M., Felsberg J., Duskova J., Dohnalek J., Kotik M.,
RA   Kren V., Martinkova L.;
RT   "Heterologous expression, purification and characterization of nitrilase
RT   from Aspergillus niger K10.";
RL   BMC Biotechnol. 11:2-2(2011).
RN   [5]
RP   RETRACTION NOTICE OF PUBMED:21210990.
RX   PubMed=23870008; DOI=10.1186/1472-6750-13-57;
RA   Kaplan O., Bezouska K., Plihal O., Ettrich R., Kulik N., Vanek O.,
RA   Kavan D., Benada O., Malandra A., Sveda O., Vesela A.B., Rinagelova A.,
RA   Slamova K., Cantarella M., Felsberg J., Duskova J., Dohnalek J., Kotik M.,
RA   Kren V., Martinkova L.;
RL   BMC Biotechnol. 13:57-57(2013).
RN   [6]
RP   FUNCTION.
RX   PubMed=21892598; DOI=10.1007/s00253-011-3525-7;
RA   Petrickova A., Vesela A.B., Kaplan O., Kubac D., Uhnakova B., Malandra A.,
RA   Felsberg J., Rinagelova A., Weyrauch P., Kren V., Bezouska K.,
RA   Martinkova L.;
RT   "Purification and characterization of heterologously expressed nitrilases
RT   from filamentous fungi.";
RL   Appl. Microbiol. Biotechnol. 93:1553-1561(2012).
RN   [7]
RP   ERRATUM OF PUBMED:21892598.
RX   DOI=10.1007/s00253-013-5204-3;
RA   Petrickova A., Vesela A.B., Kaplan O., Kubac D., Uhnakova B., Malandra A.,
RA   Felsberg J., Rinagelova A., Weyrauch P., Kren V., Bezouska K.,
RA   Martinkova L.;
RL   Appl. Microbiol. Biotechnol. 97:9263-9264(2013).
RN   [8]
RP   FUNCTION, AND CATALYTIC ACTIVITY.
RC   STRAIN=K10 / CCF 3411;
RX   PubMed=23475593; DOI=10.1007/s12033-013-9656-6;
RA   Kaplan O., Vesela A.B., Petrickova A., Pasquarelli F., Picmanova M.,
RA   Rinagelova A., Bhalla T.C., Patek M., Martinkova L.;
RT   "A comparative study of nitrilases identified by genome mining.";
RL   Mol. Biotechnol. 54:996-1003(2013).
RN   [9]
RP   FUNCTION, CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL PROPERTIES.
RC   STRAIN=K10 / CCF 3411;
RX   DOI=10.1016/j.procbio.2013.12.008;
RA   Rinagelova A., Kaplan O., Vesela A.B., Chmatal M., Krenkova A., Plihal O.,
RA   Pasquarelli F., Cantarella M., Martinkova L.;
RT   "Cyanide hydratase from Aspergillus niger K10: Overproduction in
RT   Escherichia coli, purification, characterization and use in continuous
RT   cyanide degradation.";
RL   Process Biochem. 49:445-450(2014).
CC   -!- FUNCTION: Catalyzes the hydration of cyanide to formamide. Degradation
CC       of cyanide may be important for plant pathogenic fungi in infection of
CC       cyanogenic plants (PubMed:17061133, PubMed:23475593, Ref.9). Can also
CC       transform some nitriles like 2-cyanopyridine and fumaronitrile (Ref.9).
CC       {ECO:0000255|HAMAP-Rule:MF_03224, ECO:0000269|PubMed:17061133,
CC       ECO:0000269|PubMed:23475593, ECO:0000269|Ref.9}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=formamide = H2O + hydrogen cyanide; Xref=Rhea:RHEA:21720,
CC         ChEBI:CHEBI:15377, ChEBI:CHEBI:16397, ChEBI:CHEBI:18407; EC=4.2.1.66;
CC         Evidence={ECO:0000255|HAMAP-Rule:MF_03224,
CC         ECO:0000269|PubMed:17061133, ECO:0000269|PubMed:23475593,
CC         ECO:0000269|Ref.9};
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       Kinetic parameters:
CC         KM=109 mM for cyanide {ECO:0000269|Ref.9};
CC         KM=14.7 mM for fumaronitrile {ECO:0000269|Ref.9};
CC         KM=3.7 mM for 2-cyanopyridine {ECO:0000269|Ref.9};
CC         Vmax=6.8 mmol/min/mg enzyme with cyanide as substrate
CC         {ECO:0000269|Ref.9};
CC         Vmax=18.8 mmol/min/mg enzyme with fumaronitrile as substrate
CC         {ECO:0000269|Ref.9};
CC         Vmax=10.3 mmol/min/mg enzyme with 2-cyanopyridine as substrate
CC         {ECO:0000269|Ref.9};
CC       pH dependence:
CC         Optimum pH is 8-9. Active from pH 5.5 to pH 10. {ECO:0000269|Ref.9};
CC       Temperature dependence:
CC         Optimum temperature is 45 degrees Celsius. {ECO:0000269|Ref.9};
CC   -!- SUBUNIT: Oligomer of dimers, forming left-handed helical fibers.
CC       {ECO:0000255|HAMAP-Rule:MF_03224}.
CC   -!- INDUCTION: By cyanide. {ECO:0000255|HAMAP-Rule:MF_03224}.
CC   -!- SIMILARITY: Belongs to the carbon-nitrogen hydrolase superfamily.
CC       Nitrilase family. {ECO:0000255|HAMAP-Rule:MF_03224, ECO:0000305}.
CC   -!- CAUTION: It was initially shown that the substrate specificities of the
CC       enzyme natively expressed in Aspergillus niger and the recombinant
CC       enzyme expressed in E.coli were different (PubMed:17061133), and it was
CC       hypothesized that the difference may be due to a misfolding or a post-
CC       translational modification (PubMed:21210990). However, both papers were
CC       corrected (PubMed:23455586) or retracted (PubMed:23870008), because it
CC       was shown that the 2 enzymes analyzed were different in terms of their
CC       primary structure (Ref.7). {ECO:0000305|PubMed:23455586,
CC       ECO:0000305|PubMed:23870008, ECO:0000305|Ref.7}.
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DR   EMBL; EU282000; ABX75546.1; -; mRNA.
DR   AlphaFoldDB; A9QXE0; -.
DR   SMR; A9QXE0; -.
DR   STRING; 5061.CADANGAP00001395; -.
DR   VEuPathDB; FungiDB:An01g14550; -.
DR   VEuPathDB; FungiDB:ASPNIDRAFT2_1118133; -.
DR   VEuPathDB; FungiDB:ATCC64974_11800; -.
DR   VEuPathDB; FungiDB:M747DRAFT_70451; -.
DR   eggNOG; KOG0805; Eukaryota.
DR   BRENDA; 3.5.5.1; 518.
DR   BRENDA; 3.5.5.5; 518.
DR   BRENDA; 4.2.1.66; 518.
DR   GO; GO:0030196; F:cyanide hydratase activity; IEA:UniProtKB-UniRule.
DR   GO; GO:0000257; F:nitrilase activity; IEA:UniProt.
DR   GO; GO:0019500; P:cyanide catabolic process; IEA:UniProtKB-UniRule.
DR   CDD; cd07564; nitrilases_CHs; 1.
DR   Gene3D; 3.60.110.10; -; 1.
DR   HAMAP; MF_03224; CN_hydrolase; 1.
DR   InterPro; IPR003010; C-N_Hydrolase.
DR   InterPro; IPR036526; C-N_Hydrolase_sf.
DR   InterPro; IPR037544; CN_hydrolase.
DR   InterPro; IPR000132; Nitrilase/CN_hydratase_CS.
DR   InterPro; IPR044149; Nitrilases_CHs.
DR   PANTHER; PTHR46044; PTHR46044; 1.
DR   Pfam; PF00795; CN_hydrolase; 1.
DR   SUPFAM; SSF56317; SSF56317; 1.
DR   PROSITE; PS50263; CN_HYDROLASE; 1.
DR   PROSITE; PS00920; NITRIL_CHT_1; 1.
PE   1: Evidence at protein level;
KW   Hydrolase; Lyase.
FT   CHAIN           1..356
FT                   /note="Cyanide hydratase"
FT                   /id="PRO_0000432182"
FT   DOMAIN          8..287
FT                   /note="CN hydrolase"
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU00054"
FT   ACT_SITE        48
FT                   /note="Proton acceptor"
FT                   /evidence="ECO:0000255|HAMAP-Rule:MF_03224"
FT   ACT_SITE        130
FT                   /evidence="ECO:0000255|HAMAP-Rule:MF_03224"
FT   ACT_SITE        165
FT                   /note="Nucleophile"
FT                   /evidence="ECO:0000255|HAMAP-Rule:MF_03224"
SQ   SEQUENCE   356 AA;  40022 MW;  0F0EBCBAD80706B4 CRC64;
     MAPVLKKYKA AAVNAEPGWF NLEESVRRTI HWIDEAGKAG CKFIAFPELW IPGYPYWMWK
     VNYQESLPLL KKYRENSLPS DSDEMRRIRN AARANKIYVS LGYSEVDLAS LYTTQVMISP
     SGDILNHRRK IRATHVERLV FGDGTGDTTE SVIQTDIGRV GHLNCWENMN PFMKAYAASL
     GEQVHVAAWP LYPGKETLKY PDPFTNVAEA NADLVTPAYA IETGTYTLAP WQTITAEGIK
     LNTPPGKDLE DPHIYNGHGR IFGPDGQNLV PHPDKDFEGL LFVDIDLDEC HLSKSLADFG
     GHYMRPDLIR LLVDTNRKDL VVREDRVNGG VEYTRTVDRV GLSTPLDIAN TVDSEN