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Literature summary extracted from

  • Fan, C.; Xu, W.; Zhang, T.; Zhou, L.; Jiang, B.; Mu, W.
    Engineering of Alicyclobacillus hesperidum L-arabinose isomerase for improved catalytic activity and reduced pH optimum using random and site-directed mutagenesis (2015), Appl. Biochem. Biotechnol., 177, 1480-1492 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
5.3.1.4 expression in Escherichia coli Alicyclobacillus hesperidum

Protein Variants

EC Number Protein Variants Comment Organism
5.3.1.4 D478A mutant retains more than 80 % of the maximum relative activity of the wild-type at 75°C Alicyclobacillus hesperidum
5.3.1.4 D478K mutant displays a decreased optimum pH value Alicyclobacillus hesperidum
5.3.1.4 D478N mutant shows increased activity for D-galactose isomerization and retains more than 80 % of the maximum relative activity of the wild-type at 75°C. Mutant displays a decreased optimum pH value Alicyclobacillus hesperidum
5.3.1.4 D478Q mutant shows increased activity for D-galactose isomerization and retains more than 80 % of the maximum relative activity of the wild-type at 75°C and displays a decreased optimum pH value Alicyclobacillus hesperidum
5.3.1.4 D478R mutant displays a decreased optimum pH value Alicyclobacillus hesperidum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
5.3.1.4 34.2
-
 L-arabinose mutant D478A, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 41.4
-
 D-galactose mutant D478Q, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 54.7
-
 D-galactose wild-type, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 55
-
 L-arabinose mutant D478K, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 56
-
 L-arabinose mutant D478R, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 56.3
-
 L-arabinose mutant D478N, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 60.7
-
 D-galactose mutant D478R, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 66
-
 D-galactose mutant D478N, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 75.4
-
 D-galactose mutant D478K, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 83.2
-
 D-galactose mutant D478A, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 94.2
-
 L-arabinose mutant D478Q, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 105.2
-
 L-arabinose wild-type, pH 7.0, 65°C Alicyclobacillus hesperidum

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
5.3.1.4 Co2+ 3fold increase in activity, assay in presence of 0.5 mM Alicyclobacillus hesperidum
5.3.1.4 Mn2+ 3fold increase in activity, assay in presence of 1 mM Alicyclobacillus hesperidum

Organism

EC Number Organism UniProt Comment Textmining
5.3.1.4 Alicyclobacillus hesperidum J9E728
-
-
5.3.1.4 Alicyclobacillus hesperidum URH17-3-68 J9E728
-
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
5.3.1.4 0.33
-
 substrate D-galactose, mutant D478R, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 0.52
-
 substrate D-galactose, wild-type, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 0.73
-
 substrate D-galactose, mutant D478A, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 0.75
-
 substrate D-galactose, mutant D478N, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 0.76
-
 substrate D-galactose, mutant D478K, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 0.81
-
 substrate D-galactose, mutant D478Q, pH 7.0, 65°C Alicyclobacillus hesperidum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5.3.1.4 D-galactose
-
 Alicyclobacillus hesperidum D-tagatose
-
 ?
5.3.1.4 D-galactose
-
 Alicyclobacillus hesperidum URH17-3-68 D-tagatose
-
 ?
5.3.1.4 L-arabinose
-
 Alicyclobacillus hesperidum L-ribulose
-
 ?
5.3.1.4 L-arabinose
-
 Alicyclobacillus hesperidum URH17-3-68 L-ribulose
-
 ?

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
5.3.1.4 70
-
 wild-type Alicyclobacillus hesperidum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
5.3.1.4 0.98
-
 D-galactose mutant D478Q, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 1.13
-
 D-galactose wild-type, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 1.26
-
 D-galactose mutant D478R, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 1.36
-
 D-galactose mutant D478K, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 1.58
-
 D-galactose mutant D478A, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 1.65
-
 D-galactose mutant D478N, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 71.8
-
 L-arabinose wild-type, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 72.9
-
 L-arabinose mutant D478R, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 85
-
 L-arabinose mutant D478A, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 99.6
-
 L-arabinose mutant D478K, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 105
-
 L-arabinose mutant D478Q, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 110
-
 L-arabinose mutant D478N, pH 7.0, 65°C Alicyclobacillus hesperidum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
5.3.1.4 6 6.5 mutants D478R, D478N Alicyclobacillus hesperidum
5.3.1.4 7
-
 wild-type and mutant D478A Alicyclobacillus hesperidum

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
5.3.1.4 0.018
-
 D-galactose mutant D478K, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 0.019
-
 D-galactose mutant D478A, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 0.02
-
 D-galactose wild-type, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 0.021
-
 D-galactose mutant D478R, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 0.024
-
 D-galactose mutant D478Q, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 0.025
-
 D-galactose mutant D478N, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 0.68
-
 L-arabinose wild-type, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 1.12
-
 L-arabinose mutant D478Q, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 1.3
-
 L-arabinose mutant D478R, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 1.81
-
 L-arabinose mutant D478K, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 1.95
-
 L-arabinose mutant D478N, pH 7.0, 65°C Alicyclobacillus hesperidum
5.3.1.4 2.48
-
 L-arabinose mutant D478A, pH 7.0, 65°C Alicyclobacillus hesperidum