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Literature summary extracted from

  • Asano, Y.; Dadashipour, M.; Yamazaki, M.; Doi, N.; Komeda, H.
    Functional expression of a plant hydroxynitrile lyase in Escherichia coli by directed evolution: creation and characterization of highly in vivo soluble mutants (2011), Protein Eng. Des. Sel., 24, 607-616.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.1.2.47 expressed in Escherichia coli JM109 and BL21 (DE3) cells Manihot esculenta

Protein Variants

EC Number Protein Variants Comment Organism
4.1.2.47 H103C the mutant displays 9.3fold increase in total specific activity in the cell-free extract compared with the wild type Manihot esculenta
4.1.2.47 H103I the mutant displays 8.1fold increase in total specific activity in the cell-free extract compared with the wild type Manihot esculenta
4.1.2.47 H103K inactive Manihot esculenta
4.1.2.47 H103L the mutant displays 11.1fold increase in total specific activity in the cell-free extract compared with the wild type Manihot esculenta
4.1.2.47 H103M the mutant displays 9.07fold increase in total specific activity in the cell-free extract compared with the wild type Manihot esculenta
4.1.2.47 H103P inactive Manihot esculenta
4.1.2.47 H103Q the mutant displays 4.06fold increase in total specific activity in the cell-free extract compared with the wild type Manihot esculenta
4.1.2.47 H103R inactive Manihot esculenta
4.1.2.47 H103S the mutant displays 2.9fold increase in total specific activity in the cell-free extract compared with the wild type Manihot esculenta
4.1.2.47 H103T the mutant displays 3.4fold increase in total specific activity in the cell-free extract compared with the wild type Manihot esculenta
4.1.2.47 H103W inactive Manihot esculenta
4.1.2.47 H103Y inactive with (S)-mandelonitrile as substrate Manihot esculenta
4.1.2.47 K176P the mutant displays 2.02fold increase in total specific activity in the cell-free extract compared with the wild type Manihot esculenta
4.1.2.47 K176P/K199P/K224P the mutant displays 6.97fold increase in total specific activity in the cell-free extract compared with the wild type Manihot esculenta
4.1.2.47 K176P/K224P the mutant displays 5.05fold increase in total specific activity in the cell-free extract compared with the wild type Manihot esculenta
4.1.2.47 K199P the mutant displays 1.38fold increase in total specific activity in the cell-free extract compared with the wild type Manihot esculenta
4.1.2.47 K199P/K224P the mutant displays 4.25fold increase in total specific activity in the cell-free extract compared with the wild type Manihot esculenta
4.1.2.47 K224P the mutant displays 2.53fold increase in total specific activity in the cell-free extract compared with the wild type Manihot esculenta

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.1.2.47 1.86
-
 (S)-mandelonitrile mutant enzyme H103M, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication Manihot esculenta
4.1.2.47 2 3 benzaldehyde wild type enzyme, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication Manihot esculenta
4.1.2.47 2.5 5 (S)-mandelonitrile mutant enzyme H103L, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication Manihot esculenta
4.1.2.47 5.17
-
 (S)-mandelonitrile wild type enzyme, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication Manihot esculenta
4.1.2.47 5.54
-
 (S)-mandelonitrile mutant enzyme K176P/K199P/K224P, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication Manihot esculenta
4.1.2.47 6.98
-
 (S)-mandelonitrile mutant enzyme K176P, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication Manihot esculenta
4.1.2.47 12.3
-
 benzaldehyde mutant enzyme H103Y, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication Manihot esculenta
4.1.2.47 13.4
-
 benzaldehyde mutant enzyme H103L, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication Manihot esculenta
4.1.2.47 21.6
-
 benzaldehyde mutant enzyme K176P, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication Manihot esculenta
4.1.2.47 27
-
 benzaldehyde mutant enzyme K176P/K199P/K224P, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication Manihot esculenta
4.1.2.47 27.9
-
 benzaldehyde mutant enzyme H103M, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication Manihot esculenta

Organism

EC Number Organism UniProt Comment Textmining
4.1.2.47 Manihot esculenta P52705
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.1.2.47 (S)-mandelonitrile
-
 Manihot esculenta cyanide + benzaldehyde
-
 r
4.1.2.47 cyanide + benzaldehyde
-
 Manihot esculenta (S)-mandelonitrile
-
 r

Synonyms

EC Number Synonyms Comment Organism
4.1.2.47 HNL
-
 Manihot esculenta
4.1.2.47 S-hydroxynitrile lyase
-
 Manihot esculenta

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.1.2.47 21.5
-
 (S)-mandelonitrile mutant enzyme H103L, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication Manihot esculenta
4.1.2.47 23.9
-
 (S)-mandelonitrile mutant enzyme H103M, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication Manihot esculenta
4.1.2.47 34.5
-
 (S)-mandelonitrile wild type enzyme, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication Manihot esculenta
4.1.2.47 39.7
-
 (S)-mandelonitrile mutant enzyme K176P/K199P/K224P, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication Manihot esculenta
4.1.2.47 41.8
-
 (S)-mandelonitrile mutant enzyme K176P, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication Manihot esculenta
4.1.2.47 47.4
-
 benzaldehyde mutant enzyme H103Y, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication Manihot esculenta
4.1.2.47 83.2
-
 benzaldehyde mutant enzyme K176P, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication Manihot esculenta
4.1.2.47 90.4
-
 benzaldehyde mutant enzyme H103L, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication Manihot esculenta
4.1.2.47 93.5
-
 benzaldehyde mutant enzyme K176P/K199P/K224P, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication Manihot esculenta
4.1.2.47 96.1
-
 benzaldehyde mutant enzyme H103M, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication Manihot esculenta
4.1.2.47 96.6
-
 benzaldehyde wild type enzyme, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication Manihot esculenta

Cofactor

EC Number Cofactor Comment Organism Structure
4.1.2.47 FAD
-
 Manihot esculenta

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 3.44
-
 benzaldehyde mutant enzyme H103M, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication Manihot esculenta
4.1.2.47 3.46
-
 benzaldehyde mutant enzyme K176P/K199P/K224P, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication Manihot esculenta
4.1.2.47 3.85
-
 benzaldehyde mutant enzyme H103Y, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication Manihot esculenta
4.1.2.47 3.85
-
 benzaldehyde mutant enzyme K176P, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication Manihot esculenta
4.1.2.47 4.2
-
 benzaldehyde wild type enzyme, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication Manihot esculenta
4.1.2.47 6
-
 (S)-mandelonitrile mutant enzyme K176P, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication Manihot esculenta
4.1.2.47 6.67
-
 (S)-mandelonitrile wild type enzyme, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication Manihot esculenta
4.1.2.47 6.75
-
 benzaldehyde mutant enzyme H103L, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication Manihot esculenta
4.1.2.47 7.2
-
 (S)-mandelonitrile mutant enzyme K176P/K199P/K224P, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication Manihot esculenta
4.1.2.47 8.43
-
 (S)-mandelonitrile mutant enzyme H103L, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication Manihot esculenta
4.1.2.47 12.8
-
 (S)-mandelonitrile mutant enzyme H103M, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication Manihot esculenta