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Literature summary for 2.4.1.5 extracted from

  • Hellmuth, H.; Wittrock, S.; Kralj, S.; Dijkhuizen, L.; Hofer, B.; Seibel, J.
    Engineering the glucansucrase GTFR enzyme reaction and glycosidic bond specificity: toward tailor-made polymer and oligosaccharide products (2008), Biochemistry, 47, 6678-6684.
    View publication on PubMed
Search for more literature for 2.4.1.5

Cloned(Commentary)

Cloned (Comment) Organism
gene gtfR, expression of wild-type and mutant enzymes in Escherichia coli strain XL10-Gold Streptococcus oralis

Protein Variants

Protein Variants Comment Organism
additional information random mutagenesis of the most conserved motif around the transition state stabilizer in glucansucrase GTFR of Streptococcus oralis, yielding different variants with altered reaction specificity, generation of a mutant gtfR library, overview Streptococcus oralis
N1134S the mutation in GTFA results in a drastically changed specificity but no major changes in polymer versus oligosaccharide formation Streptococcus oralis
R624G the R624G mutations near the transition state stabilizer is involved in the phenotype which exhibits a drastic switch in regioselectivity from a dextran type with mainly alpha-1,6-glucosidic linkages to a mutant type polymer with predominantly alpha-1,3-glucosidic linkages Streptococcus oralis
R624G/V630I the mutant exhibits a drastic switch in regioselectivity from a dextran type with mainly alpha-1,6-glucosidic linkages to a mutant type polymer with predominantly alpha-1,3-glucosidic linkages, both mutations near the transition state stabilizer, R624G and V630I, are contributing to this alteration Streptococcus oralis
R624G/V630I/D717A the mutant exhibits a drastic switch in regioselectivity from a dextran type with mainly alpha-1,6-glucosidic linkages to a mutant type polymer with predominantly alpha-1,3-glucosidic linkages, both mutations near the transition state stabilizer, R624G and V630I, are contributing to this alteration Streptococcus oralis
S628D saturation mutagenesis, the mutation guides the reaction toward the synthesis of short chain oligosaccharides with a drastically increased yield of 47% isomaltose or 64% leucrose Streptococcus oralis
S628R saturation mutagenesis, the mutation guides the reaction toward the synthesis of short chain oligosaccharides with a drastically increased yield of 47% isomaltose or 64% leucrose Streptococcus oralis
V630I the V630I mutations near the transition state stabilizer is involved in the phenotype which exhibits a drastic switch in regioselectivity from a dextran type with mainly alpha-1,6-glucosidic linkages to a mutant type polymer with predominantly alpha-1,3-glucosidic linkages Streptococcus oralis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.4
-
 sucrose pH 7.0, 30°C, mutant R624G Streptococcus oralis
1.6
-
 sucrose pH 7.0, 30°C, mutant V630I Streptococcus oralis
2
-
 sucrose pH 7.0, 30°C, wild-type enzyme Streptococcus oralis
3.1
-
 sucrose pH 7.0, 30°C, mutant S628D Streptococcus oralis
3.5
-
 sucrose pH 7.0, 30°C, mutant R624G/V630I Streptococcus oralis
5
-
 sucrose pH 7.0, 30°C, mutant S628R Streptococcus oralis
9.8
-
 sucrose pH 7.0, 30°C, mutant R624G/V630I/D717A Streptococcus oralis

Organism

Organism UniProt Comment Textmining
Streptococcus oralis
-
 gene gtfR
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
sucrose + acceptor
-
 Streptococcus oralis ?
-
 ?

Synonyms

Synonyms Comment Organism
glucansucrase
-
 Streptococcus oralis
GTFR
-
 Streptococcus oralis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Streptococcus oralis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.6
-
 sucrose pH 7.0, 30°C, mutant S628R Streptococcus oralis
5.2
-
 sucrose pH 7.0, 30°C, mutant R624G Streptococcus oralis
14.5
-
 sucrose pH 7.0, 30°C, mutant R624G/V630I Streptococcus oralis
22.4
-
 sucrose pH 7.0, 30°C, wild-type enzyme Streptococcus oralis
25.6
-
 sucrose pH 7.0, 30°C, mutant R624G/V630I/D717A Streptococcus oralis
29.8
-
 sucrose pH 7.0, 30°C, mutant S628D Streptococcus oralis
32.6
-
 sucrose pH 7.0, 30°C, mutant V630I Streptococcus oralis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
 assay at Streptococcus oralis