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Literature summary for 5.3.1.4 extracted from

  • Prabhu, P.; Jeya, M.; Lee, J.K.
    Probing the molecular determinant for the catalytic efficiency of L-arabinose isomerase from Bacillus licheniformis (2010), Appl. Environ. Microbiol., 76, 1653-1660.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 5.3.1.4

Cloned(Commentary)

Cloned (Comment) Organism
expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Bacillus licheniformis

Protein Variants

Protein Variants Comment Organism
I370A site-directed mutagenesis, the mutant catalytic activity is similar to the wild-type enzyme Bacillus licheniformis
L345A site-directed mutagenesis, the mutant catalytic activity is similar to the wild-type enzyme Bacillus licheniformis
M185A site-directed mutagenesis, the mutant catalytic activity is similar to the wild-type enzyme Bacillus licheniformis
M349A site-directed mutagenesis, the mutant catalytic activity is similar to the wild-type enzyme Bacillus licheniformis
T276A site-directed mutagenesis, the mutant catalytic activity is similar to the wild-type enzyme Bacillus licheniformis
W439A site-directed mutagenesis, the mutant catalytic activity is similar to the wild-type enzyme Bacillus licheniformis
Y333A site-directed mutagenesis, the catalytic site mutant shows 97.2% reduced activity compared to the wild-type enzyme Bacillus licheniformis
Y333D site-directed mutagenesis, the catalytic site mutant shows no activity Bacillus licheniformis
Y333E site-directed mutagenesis, the catalytic site mutant shows no activity Bacillus licheniformis
Y333I site-directed mutagenesis, the catalytic site mutant shows 72% reduced activity compared to the wild-type enzyme Bacillus licheniformis
Y333K site-directed mutagenesis, the catalytic site mutant shows no activity Bacillus licheniformis
Y333V site-directed mutagenesis, the catalytic site mutant shows 82% reduced activity compared to the wild-type enzyme Bacillus licheniformis
Y333X replacing Y333 with the aromatic amino acid Phe does not alter catalytic efficiency toward L-arabinose. In contrast, the activities of mutants containing a hydrophobic amino acid, Ala, Val, or Leu, decrease as the size of the hydrophobic side chain of the amino acid decreases. However, mutants containing hydrophilic and charged amino acids, such as Asp, Glu, and Lys, show almost no activity with L-arabinose Bacillus licheniformis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
8 16 L-arabinose pH 7.5, 50°C, recombinant mutant Y333V Bacillus licheniformis
369
-
 L-arabinose pH 7.5, 50°C, recombinant wild-type enzyme Bacillus licheniformis
492
-
 L-arabinose pH 7.5, 50°C, recombinant mutant Y333F Bacillus licheniformis
620
-
 L-arabinose pH 7.5, 50°C, recombinant mutant Y333W Bacillus licheniformis
750
-
 L-arabinose pH 7.5, 50°C, recombinant mutant Y333I Bacillus licheniformis

Metals/Ions

Metals/Ions Comment Organism Structure
Mn2+
-
 Bacillus licheniformis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-arabinose Bacillus licheniformis
-
 L-ribulose
-
 ?

Organism

Organism UniProt Comment Textmining
Bacillus licheniformis Q65J10
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Bacillus licheniformis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3
-
 purified recombinant mutant Y333A Bacillus licheniformis
18
-
 recombinant purified mutant Y333V Bacillus licheniformis
28
-
 recombinant purified mutant Y333I Bacillus licheniformis
105
-
 purified recombinant wild-type enzyme Bacillus licheniformis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-arabinose
-
 Bacillus licheniformis L-ribulose
-
 ?
L-arabinose the Bacillus licheniformis L-arabinose isomerase shows a high degree of substrate specificity for L-arabinose. The conserved amino acid Y333 in the substrate binding pocket of the wild-type L-AI is important for the catalytic efficiency, other putative catalytic residues are E306, E333, H350, and H450 Bacillus licheniformis L-ribulose
-
 ?

Subunits

Subunits Comment Organism
More three-dimensional homology models of the wild-type and mutant enzyme proteins from crystal structure, PDB ID 2ajtA, overview Bacillus licheniformis

Synonyms

Synonyms Comment Organism
L-AI
-
 Bacillus licheniformis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
50
-
 assay at Bacillus licheniformis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
38.9
-
 L-arabinose pH 7.5, 50°C, recombinant mutant Y333V Bacillus licheniformis
50.3
-
 L-arabinose pH 7.5, 50°C, recombinant mutant Y333I Bacillus licheniformis
148.8
-
 L-arabinose pH 7.5, 50°C, recombinant mutant Y333W Bacillus licheniformis
188.2
-
 L-arabinose pH 7.5, 50°C, recombinant mutant Y333F Bacillus licheniformis
207.5
-
 L-arabinose pH 7.5, 50°C, recombinant wild-type enzyme Bacillus licheniformis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Bacillus licheniformis

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.048
-
 L-arabinose pH 7.5, 50°C, recombinant mutant Y333V Bacillus licheniformis
0.067
-
 L-arabinose pH 7.5, 50°C, recombinant mutant Y333I Bacillus licheniformis
0.24
-
 L-arabinose pH 7.5, 50°C, recombinant mutant Y333W Bacillus licheniformis
0.38
-
 L-arabinose pH 7.5, 50°C, recombinant mutant Y333F Bacillus licheniformis
0.56
-
 L-arabinose pH 7.5, 50°C, recombinant wild-type enzyme Bacillus licheniformis