Literature summary for 3.2.1.26 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.

Cloned(Commentary)

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

Crystallization (Commentary)

Crystallization (Comment)	Organism
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

Crystallization (Comment)

Organism

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

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

Organism

Organism	UniProt	Comment	Textmining
Thermotoga maritima	O33833	-	-

Purification (Commentary)

Purification (Comment)	Organism
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

Reaction	Comment	Organism	Reaction ID
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)

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

Subunits

Subunits	Comment	Organism
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

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

Temperature Optimum [°C]

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

pH Optimum

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