INHA_PSEPU
ID INHA_PSEPU Reviewed; 228 AA.
AC Q8G9F9;
DT 05-SEP-2012, integrated into UniProtKB/Swiss-Prot.
DT 01-MAR-2003, sequence version 1.
DT 03-AUG-2022, entry version 41.
DE RecName: Full=Isonitrile hydratase;
DE EC=4.2.1.103;
DE AltName: Full=Cyclohexyl-isocyanide hydratase;
GN Name=inhA;
OS Pseudomonas putida (Arthrobacter siderocapsulatus).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Pseudomonadales;
OC Pseudomonadaceae; Pseudomonas.
OX NCBI_TaxID=303;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 2-79; 117-201 AND
RP 206-228, FUNCTION, CATALYTIC ACTIVITY, SUBUNIT, GENE NAME, AND MUTAGENESIS
RP OF GLU-79; GLU-81; CYS-101 AND THR-102.
RC STRAIN=N19-2;
RX PubMed=12244065; DOI=10.1074/jbc.m208571200;
RA Goda M., Hashimoto Y., Takase M., Herai S., Iwahara Y., Higashibata H.,
RA Kobayashi M.;
RT "Isonitrile hydratase from Pseudomonas putida N19-2. Cloning, sequencing,
RT gene expression, and identification of its active acid residue.";
RL J. Biol. Chem. 277:45860-45865(2002).
RN [2]
RP PROTEIN SEQUENCE OF 2-11, FUNCTION, CATALYTIC ACTIVITY, ACTIVITY
RP REGULATION, BIOPHYSICOCHEMICAL PROPERTIES, AND SUBUNIT.
RC STRAIN=N19-2;
RX PubMed=11306561; DOI=10.1074/jbc.m007856200;
RA Goda M., Hashimoto Y., Shimizu S., Kobayashi M.;
RT "Discovery of a novel enzyme, isonitrile hydratase, involved in nitrogen-
RT carbon triple bond cleavage.";
RL J. Biol. Chem. 276:23480-23485(2001).
CC -!- FUNCTION: Catalyzes the hydration of cyclohexyl isocyanide to N-
CC cyclohexylformamide. Acts on various isonitriles, but not on nitriles
CC or amides. Probably involved in detoxification.
CC {ECO:0000269|PubMed:11306561, ECO:0000269|PubMed:12244065}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H(+) + N-cyclohexylformamide = cyclohexyl isocyanide + H2O;
CC Xref=Rhea:RHEA:18197, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:17945, ChEBI:CHEBI:58334; EC=4.2.1.103;
CC Evidence={ECO:0000269|PubMed:11306561, ECO:0000269|PubMed:12244065};
CC -!- ACTIVITY REGULATION: Sensitive to thiol reagents and oxidizing
CC reagents, but is not influenced by chelators or reducing reagents.
CC {ECO:0000269|PubMed:11306561}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=16.2 mM for cyclohexyl isocyanide {ECO:0000269|PubMed:11306561};
CC Vmax=39.6 umol/min/mg enzyme {ECO:0000269|PubMed:11306561};
CC pH dependence:
CC Optimum pH is 6.0-6.5. {ECO:0000269|PubMed:11306561};
CC Temperature dependence:
CC Optimum temperature is 35 degrees Celsius. Activity is rapidly lost
CC above 40 degrees Celsius. {ECO:0000269|PubMed:11306561};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:11306561,
CC ECO:0000269|PubMed:12244065}.
CC -!- MISCELLANEOUS: Does not require metal ion for activity.
CC {ECO:0000305|PubMed:11306561}.
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DR EMBL; AB088117; BAC41251.1; -; Genomic_DNA.
DR AlphaFoldDB; Q8G9F9; -.
DR SMR; Q8G9F9; -.
DR KEGG; ag:BAC41251; -.
DR BioCyc; MetaCyc:MON-15823; -.
DR BRENDA; 4.2.1.103; 5092.
DR GO; GO:0050549; F:cyclohexyl-isocyanide hydratase activity; IDA:UniProtKB.
DR Gene3D; 3.40.50.880; -; 1.
DR InterPro; IPR029062; Class_I_gatase-like.
DR InterPro; IPR002818; DJ-1/PfpI.
DR Pfam; PF01965; DJ-1_PfpI; 1.
DR SUPFAM; SSF52317; SSF52317; 1.
PE 1: Evidence at protein level;
KW Direct protein sequencing; Lyase.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000269|PubMed:11306561,
FT ECO:0000269|PubMed:12244065"
FT CHAIN 2..228
FT /note="Isonitrile hydratase"
FT /id="PRO_0000418615"
FT ACT_SITE 101
FT /evidence="ECO:0000305"
FT MUTAGEN 79
FT /note="E->Q: Almost no change in activity."
FT /evidence="ECO:0000269|PubMed:12244065"
FT MUTAGEN 81
FT /note="E->Q: Almost no change in activity."
FT /evidence="ECO:0000269|PubMed:12244065"
FT MUTAGEN 101
FT /note="C->A: Lack of activity."
FT /evidence="ECO:0000269|PubMed:12244065"
FT MUTAGEN 102
FT /note="T->A: Strong decrease in activity."
FT /evidence="ECO:0000269|PubMed:12244065"
SQ SEQUENCE 228 AA; 24213 MW; F8A4722145A9CDF7 CRC64;
MALQIGFLLF PQVQQLDLTG PYDVLASLPD VQVHLVWKDL VPVTSSTGLQ LKPTTTFEDC
PVLDVICVPG GAGVGPLMED EQTLDFIRSQ AAQARYVTSV CTGSLVLGAA GLLQGKRATT
HWAYHDLLPT LGAIPVKDRV VRDGNLFTGG GITAGIDFAL TLAQELVGVD TAQLVQLQLE
YAPAPPFDSG SPDTAPSAVV DEARKRAAPS LKLRTEITER AAAKLNLR
