AHNL_LINUS
ID AHNL_LINUS Reviewed; 422 AA.
AC P93243; O22574;
DT 25-JAN-2012, integrated into UniProtKB/Swiss-Prot.
DT 01-MAY-1997, sequence version 1.
DT 03-AUG-2022, entry version 88.
DE RecName: Full=Aliphatic (R)-hydroxynitrile lyase;
DE Short=LuHNL;
DE EC=4.1.2.46;
OS Linum usitatissimum (Flax) (Linum humile).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae;
OC rosids; fabids; Malpighiales; Linaceae; Linum.
OX NCBI_TaxID=4006;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], CHARACTERIZATION, AND BIOPHYSICOCHEMICAL
RP PROPERTIES.
RX PubMed=9030531; DOI=10.1074/jbc.272.8.4770;
RA Trummler K., Wajant H.;
RT "Molecular cloning of acetone cyanohydrin lyase from flax (Linum
RT usitatissimum). Definition of a novel class of hydroxynitrile lyases.";
RL J. Biol. Chem. 272:4770-4774(1997).
RN [2]
RP NUCLEOTIDE SEQUENCE [MRNA], AND PROTEIN SEQUENCE OF 15-47.
RA Breithaupt H., Pohl M., Boenigk W., Heim P., Schimz K.-L., Kula M.-R.;
RT "Cloning and expression of (R)-hydroxynitrile lyase from Linum
RT usitatissimum (flax).";
RL J. Mol. Catal., B Enzym. 6:315-332(1999).
RN [3]
RP PROTEIN SEQUENCE OF 9-47, AND CATALYTIC ACTIVITY.
RX PubMed=8387315;
RA Albrecht J., Jansen I., Kula M.R.;
RT "Improved purification of an (R)-oxynitrilase from Linum usitatissimum
RT (flax) and investigation of the substrate range.";
RL Biotechnol. Appl. Biochem. 17:191-203(1993).
RN [4]
RP CHARACTERIZATION, BIOPHYSICOCHEMICAL PROPERTIES, AND SUBUNIT.
RX PubMed=3377504; DOI=10.1016/0003-9861(88)90634-0;
RA Xu L.L., Singh B.K., Conn E.E.;
RT "Purification and characterization of acetone cyanohydrin lyase from Linum
RT usitatissimum.";
RL Arch. Biochem. Biophys. 263:256-263(1988).
RN [5]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, AND MUTAGENESIS OF CYS-63; THR-65;
RP GLY-84; HIS-85; GLY-95; GLY-104; CYS-118; CYS-129 AND CYS-199.
RX DOI=10.1016/S0168-9452(98)00173-3;
RA Trummler K., Roos J., Schwaneberg U., Effenberger F., Foerster S.,
RA Pfizenmaier K., Wajant H.;
RT "Expression of the Zn(2+)-containing hydroxynitrile lyase from flax (Linum
RT usitatissimum) in Pichia pastoris - utilization of the recombinant enzyme
RT for enzymatic analysis and site-directed mutagenesis.";
RL Plant Sci. 139:19-27(1998).
CC -!- FUNCTION: Involved in the catabolism of cyanogenic glycosides.
CC Naturally occurring substrates are the aliphatic acetone cyanohydrin
CC and butan-2-one cyanohydrin, which are the aglycones of the cyanogenic
CC glycosides linamarin, lotaustralin, linustatin and neolinustatin. Can
CC use various aliphatic ketones and aldehydes as substrates, but not
CC aromatic ketones. {ECO:0000269|Ref.5}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(2R)-2-hydroxy-2-methylbutanenitrile = butan-2-one + hydrogen
CC cyanide; Xref=Rhea:RHEA:28170, ChEBI:CHEBI:18407, ChEBI:CHEBI:28398,
CC ChEBI:CHEBI:60908; EC=4.1.2.46; Evidence={ECO:0000269|PubMed:8387315,
CC ECO:0000269|Ref.5};
CC -!- COFACTOR:
CC Name=Zn(2+); Xref=ChEBI:CHEBI:29105; Evidence={ECO:0000269|Ref.5};
CC Note=Binds 2 Zn(2+) ions per subunit. {ECO:0000269|Ref.5};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=1.9 mM for acetone cyanohydrin {ECO:0000269|PubMed:3377504,
CC ECO:0000269|PubMed:9030531};
CC Vmax=71 umol/min/mg enzyme with acetone cyanohydrin as substrate
CC {ECO:0000269|PubMed:3377504, ECO:0000269|PubMed:9030531};
CC pH dependence:
CC Optimum pH is 5.5. {ECO:0000269|PubMed:3377504,
CC ECO:0000269|PubMed:9030531};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:3377504}.
CC -!- MISCELLANEOUS: In contrast to the enzyme from rosaceae, this enzyme is
CC not glycosylated and does not contain FAD. Not inhibited by reagents
CC interfering with Zn(2+) coordination.
CC -!- SIMILARITY: Belongs to the zinc-containing alcohol dehydrogenase
CC family. {ECO:0000305}.
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DR EMBL; Y09084; CAA70304.1; -; mRNA.
DR EMBL; AF024588; AAB81956.1; -; mRNA.
DR PIR; T07967; T07967.
DR PDB; 7VB3; X-ray; 1.48 A; A/B/C/D=1-422.
DR PDB; 7VB5; X-ray; 1.58 A; A/B/C/D=1-422.
DR PDB; 7VB6; X-ray; 1.74 A; A/B/C/D=1-422.
DR PDBsum; 7VB3; -.
DR PDBsum; 7VB5; -.
DR PDBsum; 7VB6; -.
DR AlphaFoldDB; P93243; -.
DR SMR; P93243; -.
DR KEGG; ag:CAA70304; -.
DR BioCyc; MetaCyc:MON-15903; -.
DR BRENDA; 4.1.2.46; 3037.
DR GO; GO:0052920; F:(2R)-2-hydroxy-2-methylbutanenitrile lyase activity; IEA:UniProtKB-EC.
DR GO; GO:0052921; F:acetone-cyanohydrin acetone-lyase (cyanide-forming) activity; IEA:UniProtKB-EC.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR InterPro; IPR013149; ADH-like_C.
DR InterPro; IPR013154; ADH_N.
DR InterPro; IPR011032; GroES-like_sf.
DR InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR Pfam; PF08240; ADH_N; 1.
DR Pfam; PF00107; ADH_zinc_N; 1.
DR SUPFAM; SSF50129; SSF50129; 1.
DR SUPFAM; SSF51735; SSF51735; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Direct protein sequencing; Lyase; Metal-binding; Zinc.
FT CHAIN 1..422
FT /note="Aliphatic (R)-hydroxynitrile lyase"
FT /id="PRO_0000415393"
FT BINDING 63
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="1"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000250"
FT BINDING 85
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="1"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000250"
FT BINDING 115
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="2"
FT /evidence="ECO:0000250"
FT BINDING 118
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="2"
FT /evidence="ECO:0000250"
FT BINDING 121
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="2"
FT /evidence="ECO:0000250"
FT BINDING 129
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="2"
FT /evidence="ECO:0000250"
FT BINDING 199
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="1"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000250"
FT MUTAGEN 63
FT /note="C->A: Loss of activity."
FT /evidence="ECO:0000269|Ref.5"
FT MUTAGEN 65
FT /note="T->A: Loss of activity."
FT /evidence="ECO:0000269|Ref.5"
FT MUTAGEN 84
FT /note="G->A: Loss of activity."
FT /evidence="ECO:0000269|Ref.5"
FT MUTAGEN 85
FT /note="H->A: Loss of activity."
FT /evidence="ECO:0000269|Ref.5"
FT MUTAGEN 95
FT /note="G->A: Loss of activity."
FT /evidence="ECO:0000269|Ref.5"
FT MUTAGEN 104
FT /note="G->A: 90% reduction of activity."
FT /evidence="ECO:0000269|Ref.5"
FT MUTAGEN 118
FT /note="C->A: Loss of activity."
FT /evidence="ECO:0000269|Ref.5"
FT MUTAGEN 129
FT /note="C->A: Loss of activity."
FT /evidence="ECO:0000269|Ref.5"
FT MUTAGEN 199
FT /note="C->A: Loss of activity."
FT /evidence="ECO:0000269|Ref.5"
FT CONFLICT 117
FT /note="V -> T (in Ref. 2; AAB81956)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 422 AA; 45783 MW; 178901103F1783FF CRC64;
MASLPVSFAK PDKNGVITCK AIMLKEAKLP GMSYADTVQI IDIQVDPPQN VELRVKMLCA
SVCRTDILTI EGFMAPTQFP KINGHEGVGI IESMGPDTKN FKVGDVIVAP TLGECQVCSS
CRSGRTNFCQ NYGANESALE PDGTSRFSYI DSDGKKKLLY YKLGCSTWTQ YMVVDSNYAT
KLNEIAPELP PPHGSILSCA FATGYGAVWL DAAVQEGDSV AIFGVGSVGI SAVIAAKELK
AKQIIVVDRN EYKLKMAMEL GATHCINSEK LPEGVTPSQA VRKLTPKEVG VDASIESSGY
DVFMNEAMKA AIHGKAKTVI TGEGIYENDR IFFDFKDFLF GGNVVGNVTG RVRIHSDFPG
LLRKAQEPVI RAGMDKILGY DAATMKCKYE VDIREGTPAL LKALEEVENV DCVKLVIKLN
DY
