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Literature summary extracted from

  • Kim, Y.W.; Chen, H.; Kim, J.H.; Withers, S.G.
    Catalytic properties of a mutant beta-galactosidase from Xanthomonas manihotis engineered to synthesize galactosyl-thio-beta-1,3 and -beta-1,4-glycosides (2006), FEBS Lett., 580, 4377-4381.
    View publication on PubMed

Application

EC Number Application Comment Organism
3.2.1.23 synthesis enzyme mutant E184A is a valuable catalyst for the synthesis of metabolically stable analogues of the important glycosidic linkages to the 3 and 4 positions of glucosides and galactosides Xanthomonas phaseoli pv. manihotis

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.23 expression of mutant enzymes in Escherichia coli Xanthomonas phaseoli pv. manihotis

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.23 E184A site-directed mutagenesis, the mutant functions as an efficient thioglycoligase, which synthesises thiogalactosides with linkages to the 3 and 4 positions of glucosides and galactosides in high yields, the mutant shows reduced galactohydrolase activity compared to the wild-type enzyme Xanthomonas phaseoli pv. manihotis
3.2.1.23 E184Q site-directed mutagenesis, the mutant does not function as a thioglycoligase, the mutant shows reduced galactohydrolase activity compared to the wild-type enzyme Xanthomonas phaseoli pv. manihotis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.23 additional information
-
 additional information kinetics of the thioglycoligase activity of mutant E184A, overview Xanthomonas phaseoli pv. manihotis
3.2.1.23 0.00008
-
 3,4-dinitrophenyl beta-D-galactopyranoside pH 7.0, 30°C, mutant E184A, glycohydrolase activity Xanthomonas phaseoli pv. manihotis
3.2.1.23 0.002
-
 4-nitrophenyl-beta-D-galactopyranoside pH 7.0, 30°C, mutant E184A, glycohydrolase activity Xanthomonas phaseoli pv. manihotis
3.2.1.23 0.05
-
 4-nitrophenyl-beta-D-galactopyranoside pH 7.0, 30°C, wild-type enzyme, glycohydrolase activity Xanthomonas phaseoli pv. manihotis

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.23 Xanthomonas phaseoli pv. manihotis
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.23 recombinant mutant enzymes from Escherichia coli by ammonium sulfate fractionation and hydrophobic interaction chromatography Xanthomonas phaseoli pv. manihotis

Reaction

EC Number Reaction Comment Organism Reaction ID
3.2.1.23 beta-D-galactopyranosyl-(1-4)-beta-D-galactopyranosyl-(1-6)-beta-D-galactopyranosyl-(1-3)-beta-D-galactopyranose + 3 H2O = 4 beta-D-galactopyranose Glu184 acts as acid/base catalyst Xanthomonas phaseoli pv. manihotis
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.23 3,4-dinitrophenyl beta-D-galactopyranoside + H2O
-
 Xanthomonas phaseoli pv. manihotis 3,4-dinitrophenol + beta-D-galactose
-
 ?
3.2.1.23 4-nitrophenyl beta-D-galactopyranoside + H2O
-
 Xanthomonas phaseoli pv. manihotis 4-nitrophenol + beta-D-galactose
-
 ?
3.2.1.23 additional information the mutant E184A enzyme also acts as thioglycoligase, reaction mechanism, overview Xanthomonas phaseoli pv. manihotis ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.2.1.23 BgaX
-
 Xanthomonas phaseoli pv. manihotis
3.2.1.23 More the enzyme belongs to the glycosyl hydrolase family 35 Xanthomonas phaseoli pv. manihotis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.23 25
-
 thioglycoligase assay at Xanthomonas phaseoli pv. manihotis
3.2.1.23 30
-
 glycohydrolase assay at Xanthomonas phaseoli pv. manihotis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.23 2.1
-
 4-nitrophenyl-beta-D-galactopyranoside pH 7.0, 30°C, mutant E184A, glycohydrolase activity Xanthomonas phaseoli pv. manihotis
3.2.1.23 2.6
-
 3,4-dinitrophenyl beta-D-galactopyranoside pH 7.0, 30°C, mutant E184A, glycohydrolase activity Xanthomonas phaseoli pv. manihotis
3.2.1.23 2140
-
 4-nitrophenyl-beta-D-galactopyranoside pH 7.0, 30°C, wild-type enzyme, glycohydrolase activity Xanthomonas phaseoli pv. manihotis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.23 7
-
 assay at Xanthomonas phaseoli pv. manihotis