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Literature summary for 5.1.3.B12 extracted from

  • Kim, H.J.; Yeom, S.J.; Kim, K.; Rhee, S.; Kim, D.; Oh, D.K.
    Mutational analysis of the active site residues of a D-psicose 3-epimerase from Agrobacterium tumefaciens (2010), Biotechnol. Lett., 32, 261-268.
    View publication on PubMed

Protein Variants

Protein Variants Comment Organism
E156D mutant exhibits 63% of the wild-type activity Agrobacterium tumefaciens
R215K mutant exhibits 25% of the wild-type activity Agrobacterium tumefaciens
W112A mutant enzyme exhibits no detectable activity Agrobacterium tumefaciens
W112F mutant enzyme displays 19% of the wild type enzyme activity. kcat/Km for D-fructose is 100fold lower than wild-type value, kcat/Km fur D-psicose is 84fold lower than wild-type value Agrobacterium tumefaciens
W112H mutant enzyme displays 65% of the wild type enzyme activity Agrobacterium tumefaciens
W112Y mutant enzyme displays 54% of the wild type enzyme activity Agrobacterium tumefaciens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1 3 D-psicose pH 8.0, 45°C, wild-type enzyme Agrobacterium tumefaciens
33
-
D-fructose pH 8.0, 45°C, wild-type enzyme Agrobacterium tumefaciens
37
-
D-psicose pH 8.0, 45°C, mutant enzyme R215K Agrobacterium tumefaciens
48
-
D-psicose pH 8.0, 45°C, mutant enzyme E156D Agrobacterium tumefaciens
100
-
D-fructose pH 8.0, 45°C, mutant enzyme D183E Agrobacterium tumefaciens
100
-
D-psicose pH 8.0, 45°C, mutant enzyme W112F Agrobacterium tumefaciens
139
-
D-fructose pH 8.0, 45°C, mutant enzyme E156D Agrobacterium tumefaciens
270
-
D-fructose pH 8.0, 45°C, mutant enzyme W112F Agrobacterium tumefaciens
309
-
D-fructose pH 8.0, 45°C, mutant enzyme R215K Agrobacterium tumefaciens
427
-
D-fructose pH 8.0, 45°C, mutant enzyme H209A Agrobacterium tumefaciens

Metals/Ions

Metals/Ions Comment Organism Structure
Mn2+ the distances between the catalytic residues (Glu150 and Glu244) and Mn2+ are critical to the catalysis, and the negative charges of the metal-binding residues are important for interaction with metal ion Agrobacterium tumefaciens

Organism

Organism UniProt Comment Textmining
Agrobacterium tumefaciens A9CH28
-
-
Agrobacterium tumefaciens ATCC 33970 A9CH28
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-fructose
-
Agrobacterium tumefaciens D-psicose
-
r
D-fructose
-
Agrobacterium tumefaciens ATCC 33970 D-psicose
-
r

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
45
-
assay at Agrobacterium tumefaciens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.029
-
D-fructose pH 8.0, 45°C, mutant enzyme D183E Agrobacterium tumefaciens
4.9
-
D-fructose pH 8.0, 45°C, mutant enzyme R215K Agrobacterium tumefaciens
7.7
-
D-fructose pH 8.0, 45°C, mutant enzyme H209A Agrobacterium tumefaciens
17
-
D-fructose pH 8.0, 45°C, mutant enzyme E156D Agrobacterium tumefaciens
23
-
D-psicose pH 8.0, 45°C, mutant enzyme R215K Agrobacterium tumefaciens
26
-
D-fructose pH 8.0, 45°C, mutant enzyme W112F Agrobacterium tumefaciens
44
-
D-psicose pH 8.0, 45°C, mutant enzyme W112F Agrobacterium tumefaciens
46
-
D-psicose pH 8.0, 45°C, mutant enzyme E156D Agrobacterium tumefaciens
341
-
D-fructose pH 8.0, 45°C, wild-type enzyme Agrobacterium tumefaciens
478
-
D-psicose pH 8.0, 45°C, wild-type enzyme Agrobacterium tumefaciens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
assay at Agrobacterium tumefaciens

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.00029
-
D-fructose pH 8.0, 45°C, mutant enzyme D183E Agrobacterium tumefaciens
0.015
-
D-fructose pH 8.0, 45°C, mutant enzyme R215K Agrobacterium tumefaciens
0.018
-
D-fructose pH 8.0, 45°C, mutant enzyme H209A Agrobacterium tumefaciens
0.1
-
D-fructose pH 8.0, 45°C, mutant enzyme W112F Agrobacterium tumefaciens
0.12
-
D-fructose pH 8.0, 45°C, mutant enzyme E156D Agrobacterium tumefaciens
0.44
-
D-psicose pH 8.0, 45°C, mutant enzyme W112F Agrobacterium tumefaciens
0.62
-
D-psicose pH 8.0, 45°C, mutant enzyme R215K Agrobacterium tumefaciens
0.96
-
D-psicose pH 8.0, 45°C, mutant enzyme E156D Agrobacterium tumefaciens
10
-
D-fructose pH 8.0, 45°C, wild-type enzyme Agrobacterium tumefaciens
37
-
D-psicose pH 8.0, 45°C, wild-type enzyme Agrobacterium tumefaciens