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Literature summary extracted from

  • Trummler, K.; Roos, J.; Schwaneberg, U.; Effenberger, F.; F๖rster, S.; Pfizenmaier, K.; Wajant, H.
    Expression of the Zn2+-containing hydroxynitrile lyase from flax (Linum usitatissimum) in Pichia pastoris - utilization of the recombinant enzyme for enzymatic analysis and site-directed mutagenesis (1998), Plant Sci., 139, 19-27.
No PubMed abstract available

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.1.2.46 cloning of a myc-His-tagged LuHNL-cDNA under control of the methanol-inducible AOX1 (alcohol oxidase) promotor of Pichia pastoris and introduction in the SMD1168 strain. Recombinant LuHNL is kinetically indistinguishable from the authentic flax enzyme Linum usitatissimum

Protein Variants

EC Number Protein Variants Comment Organism
4.1.2.46 G104A 5-10% of wild-type activity Linum usitatissimum
4.1.2.46 G95A complete destruction of enzymatic activity Linum usitatissimum

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
4.1.2.46 Zn2+ LuHNL has significant homologies to members of the Zn2+-containing alcohol dehydrogenases. In particular, residues responsible for coordination of Zn2+ ions or fulfilling structural or functional tasks in Zn2+-alcohol dehydrogenases are conserved. Contains about 2-4 mol zinc per mol of recombinant enzyme. Hydroxynitrile lyase from Linum usitatissimum and Zn2+-alcohol dehydrogenases have similar structural requirements with respect to maintaining a catalytically active structure. Residues essentially involved in catalysis of Zn2+-ADHs are also of functional importance in hydroxynitrile lyase from Linum usitatissimum Linum usitatissimum

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
4.1.2.46 40000
-
 2 * 40000, SDS-PAGE Linum usitatissimum
4.1.2.46 80000
-
 gel filtration Linum usitatissimum

Organism

EC Number Organism UniProt Comment Textmining
4.1.2.46 Linum usitatissimum P93243
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.1.2.46 cyanide + 2-pentanone 93% enantiomeric excess Linum usitatissimum (2R)-2-hydroxy-2-methylpentanenitrile
-
 ?
4.1.2.46 cyanide + acrolein 74% enantiomeric excess Linum usitatissimum (2R)-2-hydroxybut-3-enenitrile
-
 ?
4.1.2.46 cyanide + butan-2-one 95% enantiomeric excess Linum usitatissimum (2R)-2-hydroxy-2-methylbutanenitrile
-
 ?
4.1.2.46 cyanide + butyraldehyde 98% enantiomeric excess Linum usitatissimum (2R)-2-hydroxypentanenitrile
-
 ?
4.1.2.46 cyanide + crotonaldehyde 99% enantiomeric excess Linum usitatissimum (2R)-2-hydroxy-3-pentenenitrile
-
 ?
4.1.2.46 cyanide + hydroxypivaldehyde 73% enantiomeric excess Linum usitatissimum (2R)-2,4-dihydroxy-3,3-dimethylbutanenitrile
-
 ?
4.1.2.46 cyanide + isobutyraldehyde 93% enantiomeric excess Linum usitatissimum (2R)-2-hydroxy-4-methylpentanenitrile
-
 ?
4.1.2.46 cyanide + methacrolein 98% enantiomeric excess Linum usitatissimum (2R)-2-hydroxy-3-methylbut-3-enenitrile
-
 ?
4.1.2.46 cyanide + methyl vinyl ketone
-
 Linum usitatissimum (2R)-2-hydroxy-2-methylbut-3-enenitrile despite a short reaction time of 0.8 h, the conversion of methyl vinyl ketone results in a poor (38%) enantiomeric excess value. As in the same time there is almost no conversion without enzyme. This compound is one of the rare examples, where the enzyme exerts only a partial stereoselectivity for a defined substrate ?
4.1.2.46 cyanide + propionaldehyde 97% enantiomeric excess Linum usitatissimum (2R)-2-hydroxybutyronitrile
-
 ?
4.1.2.46 additional information the enzyme catalyzes the stereoselective synthesis of aliphatic (R)-cyanohydrins. Conversion of aromatic aldehydes (3-phenylpropionaldehyde or cinnamic aldehyde) and the aliphatic ketones is incomplete and gives poor enantiomeric excess-values, caused by the long reaction time Linum usitatissimum ?
-
 ?

Subunits

EC Number Subunits Comment Organism
4.1.2.46 dimer 2 * 40000, SDS-PAGE Linum usitatissimum

Synonyms

EC Number Synonyms Comment Organism
4.1.2.46 LuHNL
-
 Linum usitatissimum