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 |
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 |
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 |
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 |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
4.1.2.46 | Linum usitatissimum | P93243 | - |
- |
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 | ? | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
4.1.2.46 | dimer | 2 * 40000, SDS-PAGE | Linum usitatissimum |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
4.1.2.46 | LuHNL | - |
Linum usitatissimum |