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Literature summary for 3.2.1.22 extracted from

  • Bobrov, K.; Borisova, A.; Eneyskaya, E.; Ivanen, D.; Shabalin, K.; Kulminskaya, A.; Rychkov, G.
    Improvement of the efficiency of transglycosylation catalyzed by alpha-galactosidase from Thermotoga maritima by protein engineering (2013), Biochemistry, 78, 1112-1123 .
    View publication on PubMed
Search for more literature for 3.2.1.22

Protein Variants

Protein Variants Comment Organism
F328A transglycosylation is markedly increased compared to wild-type enzyme. kcat/Km value is more than twofold lower than for the wild type enzyme. 16-fold higher yield of products with the (alpha1,2)-bond as compared to products produced by the wild type enzyme Thermotoga maritima
G385L transglycosylation is markedly increased compared to wild-type enzyme. Mutation G385L results in a twofold decrease in the specific activity of the enzyme during its purification. The kinetic parameters are not determined for the mutant G385L due to its instability Thermotoga maritima
L195C kcat/Km value is more than twofold lower than for the wild type enzyme Thermotoga maritima
P402D transglycosylation is markedly increased compared to wild-type enzyme. kcat/Km value is more than twofold lower than for the wild type enzyme. 4-fold higher yield of products with the (alpha1,2)-bond as compared to products produced by the wild type enzyme Thermotoga maritima
P402S no significant changes in the kinetic parameters of hydrolysis Thermotoga maritima
W85Y no significant changes in the kinetic parameters of hydrolysis Thermotoga maritima

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.1
-
 4-nitrophenyl alpha-D-galactopyranoside 37°C, pH 5.0, mutant enzyme 328A Thermotoga maritima
0.1
-
 4-nitrophenyl alpha-D-galactopyranoside 37°C, pH 5.0, mutant enzyme F194K Thermotoga maritima
0.11
-
 4-nitrophenyl alpha-D-galactopyranoside 37°C, pH 5.0, wild-type enzyme Thermotoga maritima
0.11
-
 4-nitrophenyl alpha-D-galactopyranoside 37°C, pH 5.0, mutant enzyme G385L Thermotoga maritima
0.11
-
 4-nitrophenyl alpha-D-galactopyranoside 37°C, pH 5.0, mutant enzyme P402S Thermotoga maritima
0.32
-
 4-nitrophenyl alpha-D-galactopyranoside 37°C, pH 5.0, mutant enzyme L195C Thermotoga maritima
0.43
-
 4-nitrophenyl alpha-D-galactopyranoside 37°C, pH 5.0, mutant enzyme P402D Thermotoga maritima

Organism

Organism UniProt Comment Textmining
Thermotoga maritima G4FEF4
-
-
Thermotoga maritima DSM 3109 G4FEF4
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl alpha-D-galactopyranoside + H2O hydrolysis is accompanied by transglycosylation resulting in production of a mixture of (alpha1,2)-, (alpha1,3)-, and (alpha1,6)-4-nitrophenyl-digalactosides Thermotoga maritima 4-nitrophenol + D-galactopyranose
-
 ?
4-nitrophenyl alpha-D-galactopyranoside + H2O hydrolysis is accompanied by transglycosylation resulting in production of a mixture of (alpha1,2)-, (alpha1,3)-, and (alpha1,6)-4-nitrophenyl-digalactosides Thermotoga maritima DSM 3109 4-nitrophenol + D-galactopyranose
-
 ?

Subunits

Subunits Comment Organism
? x * 60000, wild type enzyme and its mutants W85Y, F194K, L195C, F328A, P402D, P402S, and G385L, SDS-PAGE Thermotoga maritima

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1
-
 4-nitrophenyl alpha-D-galactopyranoside 37°C, pH 5.0, mutant enzyme 328A Thermotoga maritima
8
-
 4-nitrophenyl alpha-D-galactopyranoside 37°C, pH 5.0, wild-type enzyme Thermotoga maritima
8.15
-
 4-nitrophenyl alpha-D-galactopyranoside 37°C, pH 5.0, mutant enzyme G385L Thermotoga maritima
9.8
-
 4-nitrophenyl alpha-D-galactopyranoside 37°C, pH 5.0, mutant enzyme P402S Thermotoga maritima
10.73
-
 4-nitrophenyl alpha-D-galactopyranoside 37°C, pH 5.0, mutant enzyme P402D Thermotoga maritima
12.2
-
 4-nitrophenyl alpha-D-galactopyranoside 37°C, pH 5.0, mutant enzyme L195C Thermotoga maritima
20.4
-
 4-nitrophenyl alpha-D-galactopyranoside 37°C, pH 5.0, mutant enzyme F194K Thermotoga maritima

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
11.57
-
 4-nitrophenyl alpha-D-galactopyranoside 37°C, pH 5.0, mutant enzyme 328A Thermotoga maritima
24.95
-
 4-nitrophenyl alpha-D-galactopyranoside 37°C, pH 5.0, mutant enzyme P402D Thermotoga maritima
38.76
-
 4-nitrophenyl alpha-D-galactopyranoside 37°C, pH 5.0, mutant enzyme L195C Thermotoga maritima
72.07
-
 4-nitrophenyl alpha-D-galactopyranoside 37°C, pH 5.0, wild-type enzyme Thermotoga maritima
74.09
-
 4-nitrophenyl alpha-D-galactopyranoside 37°C, pH 5.0, mutant enzyme G385L Thermotoga maritima
88.28
-
 4-nitrophenyl alpha-D-galactopyranoside 37°C, pH 5.0, mutant enzyme P402S Thermotoga maritima
198.06
-
 4-nitrophenyl alpha-D-galactopyranoside 37°C, pH 5.0, mutant enzyme F194K Thermotoga maritima