ID   HNL_MANES               Reviewed;         258 AA.
AC   P52705;
DT   01-OCT-1996, integrated into UniProtKB/Swiss-Prot.
DT   23-JAN-2007, sequence version 3.
DT   24-JAN-2024, entry version 107.
DE   RecName: Full=(S)-hydroxynitrile lyase {ECO:0000303|Ref.2};
DE            Short=MeHNL {ECO:0000303|PubMed:11173464, ECO:0000303|PubMed:11316882, ECO:0000303|PubMed:8824213, ECO:0000303|Ref.2};
DE            EC=4.1.2.47 {ECO:0000269|Ref.2};
DE   AltName: Full=2-hydroxy-2-methylpropanenitrile lyase {ECO:0000303|PubMed:11316882};
DE   AltName: Full=Acetone cyanohydrin lyase {ECO:0000303|PubMed:11316882, ECO:0000303|PubMed:8203915};
DE   AltName: Full=Alpha-hydroxynitrile lyase {ECO:0000303|PubMed:8203915};
DE   AltName: Full=Hydroxynitrile lyase {ECO:0000303|PubMed:11173464, ECO:0000303|PubMed:11316882, ECO:0000303|PubMed:11742123, ECO:0000303|PubMed:8824213};
GN   Name=HNL {ECO:0000303|PubMed:11173464, ECO:0000303|PubMed:11316882,
GN   ECO:0000303|PubMed:8203915};
OS   Manihot esculenta (Cassava) (Jatropha manihot).
OC   Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC   Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae;
OC   rosids; fabids; Malpighiales; Euphorbiaceae; Crotonoideae; Manihoteae;
OC   Manihot.
OX   NCBI_TaxID=3983;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [MRNA], PROTEIN SEQUENCE OF 2-37 AND 169-192, FUNCTION,
RP   CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL PROPERTIES.
RC   TISSUE=Cotyledon;
RX   PubMed=8203915; DOI=10.1006/abbi.1994.1267;
RA   Hughes J., Carvalho F.J.P.D.C., Hughes M.A.;
RT   "Purification, characterization, and cloning of alpha-hydroxynitrile lyase
RT   from cassava (Manihot esculenta Crantz).";
RL   Arch. Biochem. Biophys. 311:496-502(1994).
RN   [2]
RP   FUNCTION, CATALYTIC ACTIVITY, AND BIOTECHNOLOGY.
RX   DOI=10.1002/anie.199604371;
RA   Foerster S., Roos J., Effenberger F., Wajant H., Sprauer A.;
RT   "The first recombinant hydroxynitrile lyase and its application in the
RT   synthesis of (S)-cyanohydrins.";
RL   Angew. Chem. Int. Ed. Engl. 35:437-439(1996).
RN   [3]
RP   FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, AND
RP   MUTAGENESIS OF SER-80; ASP-208 AND HIS-236.
RX   PubMed=8824213; DOI=10.1074/jbc.271.42.25830;
RA   Wajant H., Pfizenmaier K.;
RT   "Identification of potential active-site residues in the hydroxynitrile
RT   lyase from Manihot esculenta by site-directed mutagenesis.";
RL   J. Biol. Chem. 271:25830-25834(1996).
RN   [4] {ECO:0007744|PDB:1DWO, ECO:0007744|PDB:1DWP, ECO:0007744|PDB:1DWQ}
RP   X-RAY CRYSTALLOGRAPHY (2.20 ANGSTROMS) IN SUBSTRATE-FREE FORM AND IN
RP   COMPLEXES WITH ACETONE AND CHLOROACETONE, AND REACTION MECHANISM.
RX   PubMed=11173464; DOI=10.1107/s0907444900015766;
RA   Lauble H., Forster S., Miehlich B., Wajant H., Effenberger F.;
RT   "Structure of hydroxynitrile lyase from Manihot esculenta in complex with
RT   substrates acetone and chloroacetone: implications for the mechanism of
RT   cyanogenesis.";
RL   Acta Crystallogr. D 57:194-200(2001).
RN   [5] {ECO:0007744|PDB:1E89, ECO:0007744|PDB:1E8D}
RP   X-RAY CRYSTALLOGRAPHY (2.10 ANGSTROMS) OF INACTIVE MUTANT ALA-80 IN
RP   SUBSTRATE-FREE FORM AND IN COMPLEX WITH 2-HYDROXY-2-METHYLPROPANENITRILE,
RP   FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF
RP   THR-11 AND CYS-81, AND REACTION MECHANISM.
RX   PubMed=11316882; DOI=10.1110/ps.01301;
RA   Lauble H., Miehlich B., Foerster S., Wajant H., Effenberger F.;
RT   "Mechanistic aspects of cyanogenesis from active-site mutant Ser80Ala of
RT   hydroxynitrile lyase from Manihot esculenta in complex with acetone
RT   cyanohydrin.";
RL   Protein Sci. 10:1015-1022(2001).
RN   [6] {ECO:0007744|PDB:1EB8, ECO:0007744|PDB:1EB9}
RP   X-RAY CRYSTALLOGRAPHY (2.10 ANGSTROMS) OF MUTANT ALA-128 IN SUBSTRATE-FREE
RP   FORM AND IN COMPLEX WITH 4-HYDROXYBENZALDEHYDE, MUTAGENESIS OF TRP-128,
RP   FUNCTION, CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL PROPERTIES.
RX   PubMed=11742123; DOI=10.1110/ps.33702;
RA   Lauble H., Miehlich B., Foerster S., Kobler C., Wajant H., Effenberger F.;
RT   "Structure determinants of substrate specificity of hydroxynitrile lyase
RT   from Manihot esculenta.";
RL   Protein Sci. 11:65-71(2002).
CC   -!- FUNCTION: Involved in cyanogenesis, the release of HCN from cyanogenic
CC       glycosides in injured tissues; the release of toxic HCN is believed to
CC       play a central role in the defense mechanism of plants against
CC       herbivores and microbial attack (Probable). Decomposes a variety of
CC       cyanohydrins (alpha-hydroxynitriles) into HCN and the corresponding
CC       aldehydes or ketones; two natural substrates are 2-hydroxy-2-
CC       methylpropanenitrile (acetone cyanohydrin) and 2-hydroxy-2-
CC       methylbutanenitrile (2-butanone cyanohydrin), but in vitro can also act
CC       on 2-hydroxy-2-methylpentanenitrile (2-pentanone cyanohydrin) and
CC       mandelonitrile (PubMed:8203915, PubMed:8824213, PubMed:11316882,
CC       PubMed:11742123). Is also able to catalyze the reverse reaction in
CC       vitro, leading to the stereospecific synthesis of aliphatic, aromatic,
CC       and heterocyclic cyanohydrins, important intermediates in the
CC       production of various agrochemicals or pharmaceuticals (Ref.2).
CC       {ECO:0000269|PubMed:11316882, ECO:0000269|PubMed:11742123,
CC       ECO:0000269|PubMed:8203915, ECO:0000269|PubMed:8824213,
CC       ECO:0000269|Ref.2, ECO:0000305|PubMed:11316882,
CC       ECO:0000305|PubMed:8203915, ECO:0000305|PubMed:8824213}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=a monosubstituted aliphatic (S)-hydroxynitrile = an aldehyde +
CC         hydrogen cyanide; Xref=Rhea:RHEA:56588, ChEBI:CHEBI:17478,
CC         ChEBI:CHEBI:18407, ChEBI:CHEBI:140596; EC=4.1.2.47;
CC         Evidence={ECO:0000269|Ref.2};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=a disubstituted aliphatic (S)-hydroxynitrile = a ketone +
CC         hydrogen cyanide; Xref=Rhea:RHEA:56592, ChEBI:CHEBI:17087,
CC         ChEBI:CHEBI:18407, ChEBI:CHEBI:140597; EC=4.1.2.47;
CC         Evidence={ECO:0000269|Ref.2, ECO:0000305|PubMed:8203915};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=an aromatic (S)-hydroxynitrile = an aromatic aldehyde +
CC         hydrogen cyanide; Xref=Rhea:RHEA:54660, ChEBI:CHEBI:18407,
CC         ChEBI:CHEBI:33855, ChEBI:CHEBI:138306; EC=4.1.2.47;
CC         Evidence={ECO:0000269|Ref.2};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=2-hydroxy-2-methylpropanenitrile = acetone + hydrogen cyanide;
CC         Xref=Rhea:RHEA:11932, ChEBI:CHEBI:15347, ChEBI:CHEBI:15348,
CC         ChEBI:CHEBI:18407; Evidence={ECO:0000269|PubMed:11316882,
CC         ECO:0000269|PubMed:11742123, ECO:0000269|PubMed:8203915,
CC         ECO:0000269|PubMed:8824213};
CC       PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:11933;
CC         Evidence={ECO:0000305|PubMed:11316882, ECO:0000305|PubMed:8203915,
CC         ECO:0000305|PubMed:8824213};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=butan-2-one + hydrogen cyanide = 2-hydroxy-2-
CC         methylbutanenitrile; Xref=Rhea:RHEA:77467, ChEBI:CHEBI:18407,
CC         ChEBI:CHEBI:28398, ChEBI:CHEBI:60954;
CC         Evidence={ECO:0000269|PubMed:8203915, ECO:0000269|Ref.2};
CC       PhysiologicalDirection=right-to-left; Xref=Rhea:RHEA:77469;
CC         Evidence={ECO:0000305|PubMed:8203915};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=hydrogen cyanide + pentan-2-one = (2S)-2-hydroxy-2-
CC         methylpentanenitrile; Xref=Rhea:RHEA:77471, ChEBI:CHEBI:16472,
CC         ChEBI:CHEBI:18407, ChEBI:CHEBI:197344; Evidence={ECO:0000269|Ref.2,
CC         ECO:0000305|PubMed:8203915};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=hexan-2-one + hydrogen cyanide = (2S)-2-hydroxy-2-
CC         methylhexanenitrile; Xref=Rhea:RHEA:77479, ChEBI:CHEBI:18407,
CC         ChEBI:CHEBI:89206, ChEBI:CHEBI:197345; Evidence={ECO:0000269|Ref.2};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=heptan-2-one + hydrogen cyanide = (2S)-2-hydroxy-2-
CC         methylheptanenitrile; Xref=Rhea:RHEA:77483, ChEBI:CHEBI:5672,
CC         ChEBI:CHEBI:18407, ChEBI:CHEBI:197346; Evidence={ECO:0000269|Ref.2};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=4-methylpentan-2-one + hydrogen cyanide = (2S)-2-hydroxy-2,4-
CC         dimethylpentanenitrile; Xref=Rhea:RHEA:77487, ChEBI:CHEBI:18407,
CC         ChEBI:CHEBI:82344, ChEBI:CHEBI:197348; Evidence={ECO:0000269|Ref.2};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=3,3-dimethylbutan-2-one + hydrogen cyanide = (2S)-2-hydroxy-2-
CC         methyl-3,3-dimethylbutanenitrile; Xref=Rhea:RHEA:77491,
CC         ChEBI:CHEBI:18407, ChEBI:CHEBI:197349, ChEBI:CHEBI:197350;
CC         Evidence={ECO:0000269|Ref.2};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=acetophenone + hydrogen cyanide = (2S)-2-hydroxy-2-
CC         phenylpropanenitrile; Xref=Rhea:RHEA:77495, ChEBI:CHEBI:18407,
CC         ChEBI:CHEBI:27632, ChEBI:CHEBI:197351; Evidence={ECO:0000269|Ref.2};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=hydrogen cyanide + propanal = (2S)-2-hydroxybutanenitrile;
CC         Xref=Rhea:RHEA:77395, ChEBI:CHEBI:17153, ChEBI:CHEBI:18407,
CC         ChEBI:CHEBI:197352; Evidence={ECO:0000269|Ref.2};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=hydrogen cyanide + pentanal = (2S)-2-hydroxyhexanenitrile;
CC         Xref=Rhea:RHEA:77399, ChEBI:CHEBI:18407, ChEBI:CHEBI:84069,
CC         ChEBI:CHEBI:197353; Evidence={ECO:0000269|Ref.2};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=2-methylpropanal + hydrogen cyanide = (2S)-2-hydroxy-3-
CC         methylbutanenitrile; Xref=Rhea:RHEA:77403, ChEBI:CHEBI:18407,
CC         ChEBI:CHEBI:48943, ChEBI:CHEBI:197354; Evidence={ECO:0000269|Ref.2};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=2,2-dimethylpropanal + hydrogen cyanide = (2S)-2-hydroxy-3,3-
CC         dimethylbutanenitrile; Xref=Rhea:RHEA:77407, ChEBI:CHEBI:18407,
CC         ChEBI:CHEBI:141557, ChEBI:CHEBI:197355; Evidence={ECO:0000269|Ref.2};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=acrolein + hydrogen cyanide = (2S)-2-hydroxybut-3-enenitrile;
CC         Xref=Rhea:RHEA:77411, ChEBI:CHEBI:15368, ChEBI:CHEBI:18407,
CC         ChEBI:CHEBI:197356; Evidence={ECO:0000269|Ref.2};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=(2E)-but-2-enal + hydrogen cyanide = (2S,3E)-2-hydroxypent-3-
CC         enenitrile; Xref=Rhea:RHEA:77415, ChEBI:CHEBI:18407,
CC         ChEBI:CHEBI:41607, ChEBI:CHEBI:197357; Evidence={ECO:0000269|Ref.2};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=(E)-hex-2-enal + hydrogen cyanide = (2S,3E)-2-hydroxyhept-3-
CC         enenitrile; Xref=Rhea:RHEA:77419, ChEBI:CHEBI:18407,
CC         ChEBI:CHEBI:28913, ChEBI:CHEBI:197358; Evidence={ECO:0000269|Ref.2};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=cyclohexanecarbaldehyde + hydrogen cyanide = (2S)-2-
CC         cyclohexyl-2-hydroxyacetonitrile; Xref=Rhea:RHEA:77423,
CC         ChEBI:CHEBI:18407, ChEBI:CHEBI:197359, ChEBI:CHEBI:197360;
CC         Evidence={ECO:0000269|Ref.2};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=benzaldehyde + hydrogen cyanide = (S)-mandelonitrile;
CC         Xref=Rhea:RHEA:77427, ChEBI:CHEBI:17169, ChEBI:CHEBI:18407,
CC         ChEBI:CHEBI:36941; Evidence={ECO:0000269|Ref.2,
CC         ECO:0000305|PubMed:11742123};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=4-methoxybenzaldehyde + hydrogen cyanide = (2S)-2-hydroxy-2-
CC         (4-methoxyphenyl)acetonitrile; Xref=Rhea:RHEA:77447,
CC         ChEBI:CHEBI:18407, ChEBI:CHEBI:28235, ChEBI:CHEBI:197328;
CC         Evidence={ECO:0000269|Ref.2};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=hydrogen cyanide + piperonal = (2S)-2-(2H-1,3-benzodioxol-5-
CC         yl)-2-hydroxyacetonitrile; Xref=Rhea:RHEA:77451, ChEBI:CHEBI:8240,
CC         ChEBI:CHEBI:18407, ChEBI:CHEBI:197361; Evidence={ECO:0000269|Ref.2};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=formylthiophene + hydrogen cyanide = (2R)-2-hydroxy-2-
CC         (thiophen-2-yl)acetonitrile; Xref=Rhea:RHEA:77455, ChEBI:CHEBI:18407,
CC         ChEBI:CHEBI:87301, ChEBI:CHEBI:197332; Evidence={ECO:0000269|Ref.2};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=3-formylthiophene + hydrogen cyanide = (2S)-2-hydroxy-2-
CC         (thiophen-3-yl)acetonitrile; Xref=Rhea:RHEA:77459, ChEBI:CHEBI:18407,
CC         ChEBI:CHEBI:87611, ChEBI:CHEBI:197333; Evidence={ECO:0000269|Ref.2};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=furan-3-carbaldehyde + hydrogen cyanide = (2S)-2-(furan-3-yl)-
CC         2-hydroxyacetonitrile; Xref=Rhea:RHEA:77463, ChEBI:CHEBI:18407,
CC         ChEBI:CHEBI:87609, ChEBI:CHEBI:197362; Evidence={ECO:0000269|Ref.2};
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       Kinetic parameters:
CC         KM=105 mM for 2-hydroxy-2-methylpropanenitrile
CC         {ECO:0000269|PubMed:8203915};
CC         KM=101 mM for 2-hydroxy-2-methylpropanenitrile
CC         {ECO:0000269|PubMed:8824213};
CC         KM=174 mM for 2-hydroxy-2-methylpropanenitrile
CC         {ECO:0000269|PubMed:11316882};
CC         KM=67 mM for 2-hydroxy-2-methylpropanenitrile
CC         {ECO:0000269|PubMed:11742123};
CC         KM=30 mM for mandelonitrile {ECO:0000269|PubMed:11742123};
CC         Note=The large KM for 2-hydroxy-2-methylpropanenitrile is not
CC         inconsistent with the concentration of cyanoglucosides in cassava
CC         leaves. {ECO:0000269|PubMed:8203915};
CC       pH dependence:
CC         Exact pH optimum pH is difficult to determine due to the fact that
CC         acetone cyanohydrin is very unstable above pH 6.5 but HNL activity
CC         was found to rise from pH 3.5 to pH 5.4. The pH of the cell sap of
CC         damages cassava leaves was found to be 5.3.
CC         {ECO:0000269|PubMed:8203915};
CC   -!- SUBUNIT: Homotetramer. {ECO:0000269|PubMed:8824213,
CC       ECO:0000305|PubMed:11316882}.
CC   -!- BIOTECHNOLOGY: This enzyme can be used as a biocatalyst for
CC       stereoselective synthesis of a wide array of (S)-cyanohydrins by
CC       addition of HCN to aldehydes or ketones. Optically active cyanohydrins
CC       are interesting intermediates for the synthesis of alpha-hydroxy acids,
CC       alpha-hydroxy ketones, or beta-ethanolamines, all of which are
CC       important building blocks in organic synthesis. {ECO:0000269|Ref.2}.
CC   -!- SIMILARITY: Belongs to the AB hydrolase superfamily. Hydroxynitrile
CC       lyase family. {ECO:0000305}.
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DR   EMBL; Z29091; CAA82334.1; -; mRNA.
DR   PDB; 1DWO; X-ray; 2.20 A; A/B=2-258.
DR   PDB; 1DWP; X-ray; 2.20 A; A/B=2-258.
DR   PDB; 1DWQ; X-ray; 2.20 A; A/B=2-258.
DR   PDB; 1E89; X-ray; 2.10 A; A/B=2-258.
DR   PDB; 1E8D; X-ray; 2.20 A; A/B=2-258.
DR   PDB; 1EB8; X-ray; 2.10 A; A/B=2-258.
DR   PDB; 1EB9; X-ray; 2.10 A; A/B=2-258.
DR   PDB; 3RKS; X-ray; 2.50 A; A/B/C/D=1-258.
DR   PDB; 3X3H; X-ray; 2.88 A; A/B/C/D/E/F/G/H=1-258.
DR   PDB; 4YK7; X-ray; 2.60 A; A/B/C/D=1-258.
DR   PDBsum; 1DWO; -.
DR   PDBsum; 1DWP; -.
DR   PDBsum; 1DWQ; -.
DR   PDBsum; 1E89; -.
DR   PDBsum; 1E8D; -.
DR   PDBsum; 1EB8; -.
DR   PDBsum; 1EB9; -.
DR   PDBsum; 3RKS; -.
DR   PDBsum; 3X3H; -.
DR   PDBsum; 4YK7; -.
DR   AlphaFoldDB; P52705; -.
DR   SMR; P52705; -.
DR   ESTHER; manes-hnl; Hydroxynitrile_lyase.
DR   EnsemblPlants; OAY33393; OAY33393; MANES_13G092100.
DR   Gramene; OAY33393; OAY33393; MANES_13G092100.
DR   KEGG; ag:CAA82334; -.
DR   OrthoDB; 31810at3646; -.
DR   BioCyc; MetaCyc:MONOMER-6902; -.
DR   BRENDA; 4.1.2.47; 3175.
DR   SABIO-RK; P52705; -.
DR   EvolutionaryTrace; P52705; -.
DR   GO; GO:0052891; F:aliphatic (S)-hydroxynitrile lyase activity; IEA:RHEA.
DR   GO; GO:0052892; F:aromatic (S)-hydroxynitrile lyase activity; IEA:RHEA.
DR   Gene3D; 3.40.50.1820; alpha/beta hydrolase; 1.
DR   InterPro; IPR029058; AB_hydrolase.
DR   InterPro; IPR000073; AB_hydrolase_1.
DR   InterPro; IPR045889; MES/HNL.
DR   PANTHER; PTHR10992:SF1078; AB HYDROLASE-1 DOMAIN-CONTAINING PROTEIN; 1.
DR   PANTHER; PTHR10992; METHYLESTERASE FAMILY MEMBER; 1.
DR   Pfam; PF00561; Abhydrolase_1; 1.
DR   SUPFAM; SSF53474; alpha/beta-Hydrolases; 1.
PE   1: Evidence at protein level;
KW   3D-structure; Direct protein sequencing; Lyase.
FT   INIT_MET        1
FT                   /note="Removed"
FT                   /evidence="ECO:0000269|PubMed:8203915"
FT   CHAIN           2..258
FT                   /note="(S)-hydroxynitrile lyase"
FT                   /id="PRO_0000084018"
FT   DOMAIN          5..242
FT                   /note="AB hydrolase-1"
FT                   /evidence="ECO:0000255"
FT   ACT_SITE        80
FT                   /note="Proton donor/acceptor"
FT                   /evidence="ECO:0000305|PubMed:11173464,
FT                   ECO:0000305|PubMed:11316882"
FT   ACT_SITE        236
FT                   /note="Proton donor/acceptor"
FT                   /evidence="ECO:0000305|PubMed:11173464,
FT                   ECO:0000305|PubMed:11316882"
FT   BINDING         11
FT                   /ligand="2-hydroxy-2-methylpropanenitrile"
FT                   /ligand_id="ChEBI:CHEBI:15348"
FT                   /evidence="ECO:0000269|PubMed:11316882,
FT                   ECO:0007744|PDB:1E8D"
FT   BINDING         11
FT                   /ligand="acetone"
FT                   /ligand_id="ChEBI:CHEBI:15347"
FT                   /evidence="ECO:0000269|PubMed:11173464,
FT                   ECO:0007744|PDB:1DWO"
FT   BINDING         80
FT                   /ligand="2-hydroxy-2-methylpropanenitrile"
FT                   /ligand_id="ChEBI:CHEBI:15348"
FT                   /evidence="ECO:0000250|UniProtKB:P52704"
FT   BINDING         80
FT                   /ligand="acetone"
FT                   /ligand_id="ChEBI:CHEBI:15347"
FT                   /evidence="ECO:0000269|PubMed:11173464,
FT                   ECO:0007744|PDB:1DWO"
FT   BINDING         81
FT                   /ligand="acetone"
FT                   /ligand_id="ChEBI:CHEBI:15347"
FT                   /evidence="ECO:0000269|PubMed:11173464,
FT                   ECO:0007744|PDB:1DWO"
FT   SITE            208
FT                   /note="Increases basicity of active site His"
FT                   /evidence="ECO:0000305|PubMed:11316882"
FT   MUTAGEN         11
FT                   /note="T->A: 95% decrease in catalytic activity."
FT                   /evidence="ECO:0000269|PubMed:11316882"
FT   MUTAGEN         80
FT                   /note="S->A: Loss of catalytic activity."
FT                   /evidence="ECO:0000269|PubMed:8824213"
FT   MUTAGEN         81
FT                   /note="C->A: No change in catalytic activity and substrate
FT                   affinity."
FT                   /evidence="ECO:0000269|PubMed:11316882"
FT   MUTAGEN         128
FT                   /note="W->A: Slightly reduced affinity and catalytic
FT                   activity toward acetone cyanohydrin but increased
FT                   efficiency on mandelonitrile. Acquired ability to use
FT                   4-hydroxymandelonitrile as substrate."
FT                   /evidence="ECO:0000269|PubMed:11742123"
FT   MUTAGEN         208
FT                   /note="D->A: 80% decrease in catalytic activity."
FT                   /evidence="ECO:0000269|PubMed:8824213"
FT   MUTAGEN         236
FT                   /note="H->A: Loss of catalytic activity."
FT                   /evidence="ECO:0000269|PubMed:8824213"
FT   STRAND          5..9
FT                   /evidence="ECO:0007829|PDB:1E89"
FT   HELIX           16..19
FT                   /evidence="ECO:0007829|PDB:1E89"
FT   HELIX           22..28
FT                   /evidence="ECO:0007829|PDB:1E89"
FT   STRAND          32..36
FT                   /evidence="ECO:0007829|PDB:1E89"
FT   HELIX           48..50
FT                   /evidence="ECO:0007829|PDB:1E89"
FT   HELIX           54..57
FT                   /evidence="ECO:0007829|PDB:1E89"
FT   HELIX           59..67
FT                   /evidence="ECO:0007829|PDB:1E89"
FT   STRAND          74..80
FT                   /evidence="ECO:0007829|PDB:1E89"
FT   HELIX           83..93
FT                   /evidence="ECO:0007829|PDB:1E89"
FT   HELIX           94..96
FT                   /evidence="ECO:0007829|PDB:1E89"
FT   STRAND          97..105
FT                   /evidence="ECO:0007829|PDB:1E89"
FT   STRAND          110..112
FT                   /evidence="ECO:0007829|PDB:1E89"
FT   HELIX           116..124
FT                   /evidence="ECO:0007829|PDB:1E89"
FT   STRAND          132..137
FT                   /evidence="ECO:0007829|PDB:1E89"
FT   STRAND          143..148
FT                   /evidence="ECO:0007829|PDB:1E89"
FT   HELIX           151..157
FT                   /evidence="ECO:0007829|PDB:1E89"
FT   HELIX           164..173
FT                   /evidence="ECO:0007829|PDB:1E89"
FT   HELIX           181..185
FT                   /evidence="ECO:0007829|PDB:1E89"
FT   TURN            192..194
FT                   /evidence="ECO:0007829|PDB:1E89"
FT   HELIX           195..197
FT                   /evidence="ECO:0007829|PDB:1E89"
FT   STRAND          200..204
FT                   /evidence="ECO:0007829|PDB:1E89"
FT   STRAND          209..211
FT                   /evidence="ECO:0007829|PDB:1E89"
FT   HELIX           213..222
FT                   /evidence="ECO:0007829|PDB:1E89"
FT   STRAND          226..230
FT                   /evidence="ECO:0007829|PDB:1E89"
FT   HELIX           238..241
FT                   /evidence="ECO:0007829|PDB:1E89"
FT   HELIX           243..257
FT                   /evidence="ECO:0007829|PDB:1E89"
SQ   SEQUENCE   258 AA;  29372 MW;  98ABA050AC8AF1C5 CRC64;
     MVTAHFVLIH TICHGAWIWH KLKPALERAG HKVTALDMAA SGIDPRQIEQ INSFDEYSEP
     LLTFLEKLPQ GEKVIIVGES CAGLNIAIAA DRYVDKIAAG VFHNSLLPDT VHSPSYTVEK
     LLESFPDWRD TEYFTFTNIT GETITTMKLG FVLLRENLFT KCTDGEYELA KMVMRKGSLF
     QNVLAQRPKF TEKGYGSIKK VYIWTDQDKI FLPDFQRWQI ANYKPDKVYQ VQGGDHKLQL
     TKTEEVAHIL QEVADAYA
//