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Literature summary extracted from

  • Kawahara, N.; Asano, Y.
    Mutagenesis of an Asn156 residue in a surface region of S-selective hydroxynitrile lyase from Baliospermum montanum enhances catalytic efficiency and enantioselectivity (2015), ChemBioChem, 16, 1891-1895 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.1.2.47 expression of wild-zyüe and mutant enzymes in Escherichia coli Baliospermum montanum

Protein Variants

EC Number Protein Variants Comment Organism
4.1.2.47 H103C the mutated enzyme shows low enantioselectivity and specific activity for (S)-mandelonitrile synthesis. (S)-Mandelonitrile enantiomeric excess is 60% Baliospermum montanum
4.1.2.47 H103C/N156D hydroxynitrile lyase of mutant enzyme H103C/N156D is approximately four times higher than that for mutant enzyme H103C. (S)-Mandelonitrile enantiomeric excess is 32% Baliospermum montanum
4.1.2.47 H103C/N156G the specific activity of the H103C/N156G mutant for (S)-mandelonitrile production is raised to 154 U/mg (wild-type hydroxynitrile lyase: 52 U/mg). The enantiomeric excess is increased to 93% (wild-type: 55%). Km-value for (R)-mandelonitrile and kcat for (S)-mandelonitrile increase by the mutation at Asn156, thus contributing to the increase in enantiomeric excess Baliospermum montanum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.1.2.47 0.28
-
 (R)-mandelonitrile pH 4.23, 25°C, mutant enzyme H103C Baliospermum montanum
4.1.2.47 0.34
-
 (S)-mandelonitrile pH 4.23, 25°C, mutant enzyme H103C Baliospermum montanum
4.1.2.47 0.47
-
 (S)-mandelonitrile pH 4.23, 25°C, mutant enzyme H103C/N156D Baliospermum montanum
4.1.2.47 0.49
-
 (S)-mandelonitrile pH 4.23, 25°C, mutant enzyme H103C/N156G Baliospermum montanum
4.1.2.47 0.61
-
 (R)-mandelonitrile pH 4.23, 25°C, mutant enzyme H103C/N156D Baliospermum montanum
4.1.2.47 1.49
-
 (R)-mandelonitrile pH 4.23, 25°C, mutant enzyme H103C/N156G Baliospermum montanum

Organism

EC Number Organism UniProt Comment Textmining
4.1.2.47 Baliospermum montanum D1MX73
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.1.2.47
-
 Baliospermum montanum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.1.2.47 (R)-mandelonitrile low activity Baliospermum montanum cyanide + benzaldehyde
-
 r
4.1.2.47 (S)-mandelonitrile
-
 Baliospermum montanum cyanide + benzaldehyde
-
 r
4.1.2.47 cyanide + benzaldehyde
-
 Baliospermum montanum (S)-mandelonitrile
-
 r

Synonyms

EC Number Synonyms Comment Organism
4.1.2.47 S-selective hydroxynitrile lyase
-
 Baliospermum montanum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.1.2.47 7.98
-
 (R)-mandelonitrile pH 4.23, 25°C, mutant enzyme H103C Baliospermum montanum
4.1.2.47 9.27
-
 (R)-mandelonitrile pH 4.23, 25°C, mutant enzyme H103C/N156D Baliospermum montanum
4.1.2.47 12.7
-
 (R)-mandelonitrile pH 4.23, 25°C, mutant enzyme H103C/N156G Baliospermum montanum
4.1.2.47 52.9
-
 (S)-mandelonitrile pH 4.23, 25°C, mutant enzyme H103C Baliospermum montanum
4.1.2.47 88.1
-
 (S)-mandelonitrile pH 4.23, 25°C, mutant enzyme H103C/N156D Baliospermum montanum
4.1.2.47 122
-
 (S)-mandelonitrile pH 4.23, 25°C, mutant enzyme H103C/N156G Baliospermum montanum

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
4.1.2.47 8.81
-
 (R)-mandelonitrile pH 4.23, 25°C, mutant enzyme H103C/N156G Baliospermum montanum
4.1.2.47 15.4
-
 (R)-mandelonitrile pH 4.23, 25°C, mutant enzyme H103C/N156D Baliospermum montanum
4.1.2.47 29.1
-
 (R)-mandelonitrile pH 4.23, 25°C, mutant enzyme H103C Baliospermum montanum
4.1.2.47 157
-
 (S)-mandelonitrile pH 4.23, 25°C, mutant enzyme H103C Baliospermum montanum
4.1.2.47 191
-
 (S)-mandelonitrile pH 4.23, 25°C, mutant enzyme H103C/N156D Baliospermum montanum
4.1.2.47 251
-
 (S)-mandelonitrile pH 4.23, 25°C, mutant enzyme H103C/N156G Baliospermum montanum