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Literature summary extracted from

  • Alberto, F.; Jordi, E.; Henrissat, B.; Czjzek, M.
    Crystal structure of inactivated Thermotoga maritima invertase in complex with the trisaccharide substrate raffinose (2006), Biochem. J., 395, 457-462.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.26 expression of His-tagged wild-type enzyme and mutants E190A and E190D in Escherichia coli strain BL21(DE3) Thermotoga maritima

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.2.1.26 purified recombinant inactivated mutant enzyme E190D in complex with substrate raffinose, hanging drop vapour diffusion method, 11 mg/ml protein in 15-17% PEG 1000, 150 mM Li2SO4, and 100 mM sodium citrate, pH 4.2, soaking of crystals in 5 mM substrate solution, flash freezing at -173°C, X-ray diffraction structure determination and analysis at a.87 A resolution, structure modelling, no successful crystal structure analysis with crystals of wild-type enzyme and mutant E190A built by the same method Thermotoga maritima

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.26 E190A site-directed mutagenesis, inactivated mutant Thermotoga maritima
3.2.1.26 E190D site-directed mutagenesis, inactivated mutant Thermotoga maritima

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.26 Thermotoga maritima O33833
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.26 recombinant His-tagged wild-type enzyme and mutants E190A and E190D from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration Thermotoga maritima

Reaction

EC Number Reaction Comment Organism Reaction ID
3.2.1.26 raffinose + H2O = beta-D-fructofuranose + 6-O alpha-D-galactopyranosyl-alpha-D-glucopyranose double-displacement reaction mechanism, cleavage of beta-fructopyranosidic linkages involving a nucleophilic aspartate and a catalytic glutamic acid acting as general acid/base, structure of the active site substrate binding pocket possessing three binding subsites Thermotoga maritima
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.26 inulin + H2O
-
 Thermotoga maritima D-fructose + ?
-
 ?
3.2.1.26 raffinose + H2O
-
 Thermotoga maritima alpha-D-galactosyl-1,6-alpha-D-glucose + D-fructose
-
 ?
3.2.1.26 sucrose + H2O best substrate Thermotoga maritima D-glucose + D-fructose
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.26 More the enzyme is bimodular with a five bladed beta-propeller catalytic domain linked to a beta-sandwich, three-dimensional structure, overview Thermotoga maritima

Synonyms

EC Number Synonyms Comment Organism
3.2.1.26 beta-fructosidase
-
 Thermotoga maritima
3.2.1.26 invertase
-
 Thermotoga maritima
3.2.1.26 More the enzyme belongs to glycoside hydrolase family GH-32 Thermotoga maritima

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.26 37
-
 assay at Thermotoga maritima

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.26 5.5
-
 assay at Thermotoga maritima