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Literature summary for 3.2.1.23 extracted from

  • Placier, G.; Watzlawick, H.; Rabiller, C.; Mattes, R.
    Evolved beta-galactosidases from Geobacillus stearothermophilus with improved transgalactosylation yield for galacto-oligosaccharide production (2009), Appl. Environ. Microbiol., 75, 6312-6321.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli JM109 cells Geobacillus stearothermophilus

Protein Variants

Protein Variants Comment Organism
R109K the mutant shows decreased specific activity towards 4-nitrophenyl-beta-D-galactopyranoside compared to the wild type enzyme, the apparent Km values toward the natural substrates lactose and lactosucrose are strongly increased Geobacillus stearothermophilus
R109V the mutant shows increased specific activity towards 4-nitrophenyl-beta-D-galactopyranoside compared to the wild type enzyme, the apparent Km values toward the natural substrates lactose and lactosucrose are strongly increased Geobacillus stearothermophilus
R109W the mutant shows increased specific activity towards 4-nitrophenyl-beta-D-galactopyranoside compared to the wild type enzyme, the apparent Km values toward the natural substrates lactose and lactosucrose are strongly increased Geobacillus stearothermophilus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.78
-
lactose wild type enzyme, at 37°C Geobacillus stearothermophilus
5
-
lactosucrose wild type enzyme, at 37°C Geobacillus stearothermophilus
84
-
lactose mutant enzyme R109K, at 37°C Geobacillus stearothermophilus
100
-
lactose mutant enzyme R109V, at 37°C Geobacillus stearothermophilus
114
-
lactose mutant enzyme R109W, at 37°C Geobacillus stearothermophilus
136
-
lactosucrose mutant enzyme R109K, at 37°C Geobacillus stearothermophilus
142
-
lactosucrose mutant enzyme R109V, at 37°C Geobacillus stearothermophilus
160
-
lactosucrose mutant enzyme R109W, at 37°C Geobacillus stearothermophilus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
78000
-
SDS-PAGE Geobacillus stearothermophilus

Organism

Organism UniProt Comment Textmining
Geobacillus stearothermophilus
-
-
-
Geobacillus stearothermophilus KVE39
-
-
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.19
-
wild type enzyme, using 20 mM 4-nitrophenyl-beta-D-galactopyranoside as substrate, at 37°C Geobacillus stearothermophilus
0.22
-
mutant enzyme R109K, using 2.65 mM 4-nitrophenyl-beta-D-galactopyranoside as substrate, at 37°C Geobacillus stearothermophilus
0.47
-
mutant enzyme R109K, using 20 mM 4-nitrophenyl-beta-D-galactopyranoside as substrate, at 37°C Geobacillus stearothermophilus
0.5
-
wild type enzyme, using 2.65 mM 4-nitrophenyl-beta-D-galactopyranoside as substrate, at 37°C Geobacillus stearothermophilus
0.6
-
mutant enzyme R109W, using 2.65 mM 4-nitrophenyl-beta-D-galactopyranoside as substrate, at 37°C Geobacillus stearothermophilus
1.52
-
mutant enzyme R109V, using 2.65 mM 4-nitrophenyl-beta-D-galactopyranoside as substrate, at 37°C Geobacillus stearothermophilus
1.79
-
mutant enzyme R109W, using 20 mM 4-nitrophenyl-beta-D-galactopyranoside as substrate, at 37°C Geobacillus stearothermophilus
4.9
-
mutant enzyme R109V, using 20 mM 4-nitrophenyl-beta-D-galactopyranoside as substrate, at 37°C Geobacillus stearothermophilus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl beta-D-galactopyranoside + H2O
-
Geobacillus stearothermophilus 4-nitrophenol + beta-D-galactose
-
?
4-nitrophenyl beta-D-galactopyranoside + H2O
-
Geobacillus stearothermophilus KVE39 4-nitrophenol + beta-D-galactose
-
?
lactose + H2O
-
Geobacillus stearothermophilus D-galactose + D-glucose
-
?
lactose + H2O
-
Geobacillus stearothermophilus KVE39 D-galactose + D-glucose
-
?
lactosucrose + H2O
-
Geobacillus stearothermophilus D-galactose + ?
-
?
lactosucrose + H2O
-
Geobacillus stearothermophilus KVE39 D-galactose + ?
-
?
additional information during the transglycosylation reaction with lactose of the evolved beta-galactosidases, a major trisaccharide, beta-D-galactopyranosyl-(1-3)-beta-D-galactopyranosyl-(1-4)-D-glucopyranoside (3'-galactosyl-lactose) is formed. At the lactose concentration of 18% (w/v), this trisaccharide is obtained in yields of 11.5% (w/w) with BgaB mutant enzyme R109K, 21% (w/w) with BgaB mutant enzyme R109V, 23% (w/w) with BgaB mutant enzyme R109W, and 2% (w/w) with the wild type BgaB enzyme Geobacillus stearothermophilus ?
-
?
additional information during the transglycosylation reaction with lactose of the evolved beta-galactosidases, a major trisaccharide, beta-D-galactopyranosyl-(1-3)-beta-D-galactopyranosyl-(1-4)-D-glucopyranoside (3'-galactosyl-lactose) is formed. At the lactose concentration of 18% (w/v), this trisaccharide is obtained in yields of 11.5% (w/w) with BgaB mutant enzyme R109K, 21% (w/w) with BgaB mutant enzyme R109V, 23% (w/w) with BgaB mutant enzyme R109W, and 2% (w/w) with the wild type BgaB enzyme Geobacillus stearothermophilus KVE39 ?
-
?

Synonyms

Synonyms Comment Organism
BgaB
-
Geobacillus stearothermophilus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
65
-
-
Geobacillus stearothermophilus