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Literature summary for 2.4.1.64 extracted from

  • Van der Borght, J.; Soetaert, W.; Desmet, T.
    Engineering the acceptor specificity of trehalose phosphorylase for the production of trehalose analogs (2012), Biotechnol. Prog., 28, 1257-1262.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Thermoanaerobacter brockii

Protein Variants

Protein Variants Comment Organism
A431T the mutant shows strongly decreased catalytic efficiency for D-glucose and decreased catalytic efficiency for D-galactose compared to the wild type enzyme Thermoanaerobacter brockii
A440E the mutant shows increased catalytic efficiency for D-glucose and wild type catalytic efficiency for D-galactose compared to the wild type enzyme Thermoanaerobacter brockii
A440K the mutant shows increased catalytic efficiencies for D-glucose and D-galactose compared to the wild type enzyme Thermoanaerobacter brockii
A440M the mutant shows increased catalytic efficiencies for D-glucose and D-galactose compared to the wild type enzyme Thermoanaerobacter brockii
A440V the mutant shows increased catalytic efficiencies for D-glucose and D-galactose compared to the wild type enzyme Thermoanaerobacter brockii
A440V/N657Y the mutant shows increased catalytic efficiencies for D-glucose and D-galactose compared to the wild type enzyme Thermoanaerobacter brockii
A440V/R448S the mutant shows increased catalytic efficiency for D-glucose and strongly increased catalytic efficiency for D-galactose compared to the wild type enzyme Thermoanaerobacter brockii
N657D the mutant shows increased catalytic efficiencies for D-glucose and D-galactose compared to the wild type enzyme Thermoanaerobacter brockii
N657G the mutant shows increased catalytic efficiencies for D-glucose and D-galactose compared to the wild type enzyme Thermoanaerobacter brockii
N657I the mutant shows approximate 2fold increase in affinity for D-galactose Thermoanaerobacter brockii
N657L the mutant shows increased catalytic efficiencies for D-glucose and D-galactose compared to the wild type enzyme Thermoanaerobacter brockii
N657R the mutant shows increased catalytic efficiencies for D-glucose and D-galactose compared to the wild type enzyme Thermoanaerobacter brockii
N657Y the mutant shows approximate 2fold increase in affinity for D-galactose Thermoanaerobacter brockii
P588H the mutant shows increased catalytic efficiencies for D-glucose and D-galactose compared to the wild type enzyme Thermoanaerobacter brockii
R448D the mutant shows increased catalytic efficiencies for D-glucose and D-galactose compared to the wild type enzyme Thermoanaerobacter brockii
R448F the mutant shows increased catalytic efficiencies for D-glucose and D-galactose compared to the wild type enzyme Thermoanaerobacter brockii
R448N the mutant shows increased catalytic efficiencies for D-glucose and D-galactose compared to the wild type enzyme Thermoanaerobacter brockii
R448S the mutant shows increased catalytic efficiencies for D-glucose and D-galactose compared to the wild type enzyme Thermoanaerobacter brockii
R448S/N657Y the mutant shows about wild type catalytic efficiencies for D-glucose and D-galactose Thermoanaerobacter brockii
R448V the mutant shows increased catalytic efficiencies for D-glucose and D-galactose compared to the wild type enzyme Thermoanaerobacter brockii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.6
-
D-glucose mutant enzyme A440V/N657Y, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
0.6
-
D-glucose mutant enzyme R448S/N657Y, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
0.7
-
D-glucose mutant enzyme A440V/R448S, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
1.2
-
D-glucose mutant enzyme R448D, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
1.2
-
D-glucose mutant enzyme R448F, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
1.3
-
D-glucose mutant enzyme N657R, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
1.5
-
D-glucose mutant enzyme P588H, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
1.5
-
D-glucose mutant enzyme R448N, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
1.5
-
D-glucose mutant enzyme R448S, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
1.8
-
D-glucose mutant enzyme N657D, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
1.9
-
D-glucose mutant enzyme A440V, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
1.9
-
D-glucose mutant enzyme R448V, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
2.1
-
D-glucose mutant enzyme A440M, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
2.1
-
D-glucose mutant enzyme N657I, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
2.2
-
D-glucose mutant enzyme N657L, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
2.3
-
D-glucose mutant enzyme A440E, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
2.3
-
D-glucose mutant enzyme A440K, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
2.5
-
D-glucose mutant enzyme N657G, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
3
-
D-glucose mutant enzyme N657Y, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
3 3.5 D-galactose mutant enzyme R448S, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
3.2
-
D-glucose wild type enzyme, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
30.1
-
D-galactose mutant enzyme A440V/R448S, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
35.1
-
D-galactose mutant enzyme A440V/N657Y, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
36.8
-
D-glucose mutant enzyme A431T, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
38
-
D-galactose mutant enzyme R448S/N657Y, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
43.7
-
D-galactose mutant enzyme R448V, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
43.8
-
D-galactose mutant enzyme A440M, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
45.2
-
D-galactose mutant enzyme R448F, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
45.7
-
D-galactose mutant enzyme A440V, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
45.8
-
D-galactose mutant enzyme N657Y, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
47.5
-
D-galactose mutant enzyme R448D, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
53.5
-
D-galactose mutant enzyme N657I, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
53.6
-
D-galactose mutant enzyme R448N, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
56.8
-
D-galactose mutant enzyme A440K, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
58.1
-
D-galactose mutant enzyme N657G, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
61.1
-
D-galactose mutant enzyme N657L, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
61.9
-
D-galactose mutant enzyme N657D, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
62.5
-
D-galactose mutant enzyme N657R, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
68.2
-
D-galactose mutant enzyme A440E, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
96
-
D-galactose mutant enzyme P588H, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
98
-
D-galactose wild type enzyme, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
447.6
-
D-galactose mutant enzyme A431T, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii

Organism

Organism UniProt Comment Textmining
Thermoanaerobacter brockii Q8L164 previously known as Thermoanaerobium brockii
-

Purification (Commentary)

Purification (Comment) Organism
-
Thermoanaerobacter brockii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
beta-D-glucose 1-phosphate + D-galactose low activity. The Km value for D-galactose is about 30fold higher than that for D-glucose Thermoanaerobacter brockii ? + phosphate
-
r
D-glucose + beta-D-glucose 1-phosphate best substrate combination Thermoanaerobacter brockii alpha,alpha-trehalose + phosphate
-
r
additional information the wild type enzyme shows low activity with D-fructose and intermediate activity with D-mannose Thermoanaerobacter brockii ?
-
?

Synonyms

Synonyms Comment Organism
trehalose phosphorylase
-
Thermoanaerobacter brockii

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
60
-
the half-life of the wild type enzyme remains constant for about 13 h at 60°C Thermoanaerobacter brockii

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
7
-
D-glucose mutant enzyme R448S/N657Y, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
14
-
D-galactose mutant enzyme R448S/N657Y, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
16.1
-
D-glucose mutant enzyme A440V/N657Y, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
17
-
D-galactose mutant enzyme A440M, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
18.6
-
D-glucose mutant enzyme R448D, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
21.1
-
D-galactose mutant enzyme R448D, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
21.7
-
D-glucose mutant enzyme R448F, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
22.6
-
D-galactose mutant enzyme R448F, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
23
-
D-galactose mutant enzyme A440E, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
23.3
-
D-galactose mutant enzyme N657Y, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
23.6
-
D-galactose mutant enzyme A440V/N657Y, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
24
-
D-galactose mutant enzyme N657G, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
25
-
D-glucose mutant enzyme A440V/R448S, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
26.2
-
D-galactose mutant enzyme R448V, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
26.3
-
D-glucose mutant enzyme R448S, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
27.4
-
D-glucose mutant enzyme A440M, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
27.7
-
D-galactose mutant enzyme A440K, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
30.6
-
D-glucose mutant enzyme R448N, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
31.4
-
D-glucose mutant enzyme N657R, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
31.9
-
D-galactose mutant enzyme R448S, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
32.1
-
D-glucose mutant enzyme A431T, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
32.4
-
D-galactose mutant enzyme R448N, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
32.7
-
D-glucose mutant enzyme A440E, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
32.9
-
D-galactose mutant enzyme A440V, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
32.9
-
D-galactose wild type enzyme, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
33.3
-
D-galactose mutant enzyme N657L, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
33.9
-
D-glucose wild type enzyme, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
35.5
-
D-galactose mutant enzyme N657D, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
36.1
-
D-glucose mutant enzyme R448V, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
38.3
-
D-galactose mutant enzyme N657R, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
38.9
-
D-galactose mutant enzyme A440V/R448S, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
41.2
-
D-glucose mutant enzyme A440K, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
42
-
D-glucose mutant enzyme A440V, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
42.2
-
D-galactose mutant enzyme N657I, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
42.7
-
D-glucose mutant enzyme N657G, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
43.3
-
D-glucose mutant enzyme N657L, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
43.5
-
D-glucose mutant enzyme P588H, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
46.9
-
D-glucose mutant enzyme N657I, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
51.2
-
D-glucose mutant enzyme N657D, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
53.9
-
D-glucose mutant enzyme N657Y, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
54.3
-
D-galactose mutant enzyme P588H, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
66.5
-
D-galactose mutant enzyme A431T, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.149
-
D-galactose mutant enzyme A431T, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
0.336
-
D-galactose wild type enzyme, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
0.337
-
D-galactose mutant enzyme A440E, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
0.368
-
D-galactose mutant enzyme R448S/N657Y, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
0.388
-
D-galactose mutant enzyme A440M, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
0.413
-
D-galactose mutant enzyme N657G, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
0.444
-
D-galactose mutant enzyme R448D, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
0.488
-
D-galactose mutant enzyme A440K, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
0.5
-
D-galactose mutant enzyme R448F, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
0.509
-
D-galactose mutant enzyme N657Y, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
0.545
-
D-galactose mutant enzyme N657L, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
0.566
-
D-galactose mutant enzyme P588H, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
0.574
-
D-galactose mutant enzyme N657D, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
0.6
-
D-galactose mutant enzyme R448V, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
0.604
-
D-galactose mutant enzyme R448N, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
0.613
-
D-galactose mutant enzyme N657R, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
0.672
-
D-galactose mutant enzyme A440V/N657Y, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
0.72
-
D-galactose mutant enzyme A440V, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
0.789
-
D-galactose mutant enzyme N657I, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
0.872
-
D-glucose mutant enzyme A431T, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
0.952
-
D-galactose mutant enzyme R448S, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
1.29
-
D-galactose mutant enzyme A440V/R448S, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
11
-
D-glucose wild type enzyme, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
12
-
D-glucose mutant enzyme R448S/N657Y, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
13
-
D-glucose mutant enzyme A440M, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
14
-
D-glucose mutant enzyme A440E, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
16
-
D-glucose mutant enzyme R448D, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
17
-
D-glucose mutant enzyme N657G, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
18
-
D-glucose mutant enzyme A440K, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
18
-
D-glucose mutant enzyme N657Y, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
18
-
D-glucose mutant enzyme R448F, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
18
-
D-glucose mutant enzyme R448S, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
19
-
D-glucose mutant enzyme R448V, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
20
-
D-glucose mutant enzyme N657L, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
20
-
D-glucose mutant enzyme R448N, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
22
-
D-glucose mutant enzyme A440V, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
22
-
D-glucose mutant enzyme N657I, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
24
-
D-glucose mutant enzyme N657R, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
27
-
D-glucose mutant enzyme A440V/N657Y, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
28
-
D-glucose mutant enzyme N657D, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
29
-
D-glucose mutant enzyme P588H, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii
36
-
D-glucose mutant enzyme A440V/R448S, in 50 mM MES buffer at pH 6.0 and 60°C Thermoanaerobacter brockii