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Literature summary for 3.2.1.1 extracted from

  • Mizuno, M.; Tonozuka, T.; Uechi, A.; Ohtaki, A.; Ichikawa, K.; Kamitori, S.; Nishikawa, A.; Sakano, Y.
    The crystal structure of Thermoactinomyces vulgaris R-47 alpha-amylase II (TVA II) complexed with transglycosylated product (2004), Eur. J. Biochem., 271, 2530-2538.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expresion of mutant enzymes in Escherichai coli strain MV1184 Thermoactinomyces vulgaris

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant TVAII mutant enzyme D325N complexed with alpha-panosylpanose, hanging drop vapour diffusion method, 0.0015 ml of 20 mg/ml protein in 5 m Tris-HCl, pH 7.5, is mixed with 0.0015 ml of reservoir solution containing 1% w/v PEG 6000, 5 mM CaCl2, and 40 mM MES-NaOH, pH 6.1, soaking of crystals in cryoprotection solution containing 20% v/v 2-methyl-2,4-pentanediol, 20% w/v PEG 6000, and 2.5 mM CaCl2, X-ray diffraction structure determination and analysis at 2.2 A resolution, modelling Thermoactinomyces vulgaris

Protein Variants

Protein Variants Comment Organism
D325N site-directed mutagenesis, inactive mutant Thermoactinomyces vulgaris
Y374A site-directed mutagenesis, the mutant shows reduced activity, and altered substrate specificity and kinetics compared to the wild-type enzyme Thermoactinomyces vulgaris

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information kinetic study Thermoactinomyces vulgaris
0.19
-
starch mutant Y374A Thermoactinomyces vulgaris
0.23
-
starch wild-type enzyme Thermoactinomyces vulgaris
0.57
-
pullulan mutant Y374A Thermoactinomyces vulgaris
1.4
-
pullulan wild-type enzyme Thermoactinomyces vulgaris

Organism

Organism UniProt Comment Textmining
Thermoactinomyces vulgaris
-
-
-
Thermoactinomyces vulgaris R-47
-
-
-

Reaction

Reaction Comment Organism Reaction ID
(alpha-D-glucopyranosyl-(1-4))n-alpha-D-glucopyranose + H2O = (alpha-D-glucopyranosyl-(1-4))n-m-alpha-D-glucopyranose + (alpha-D-glucopyranosyl-(1-4))m-alpha-D-glucopyranose TVAII is a bifunctional enzyme showing alpha-amylase as well as cyclodextrin-hydrolyzing activity, the enzyme hydrolyzes alpha-1,4-glucosidic linkages and alpha-1,6-glucosidic linkages, active site structure and substrate binding structure, Trp356 is involved in substrate binding, and Tyr374 is involved in substrate orientation for catalysis Thermoactinomyces vulgaris

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
-
Thermoactinomyces vulgaris

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
cyclodextrin + H2O cyclomaltodextrinase activity, hydrolysis of alpha-1,4-glucosidic linkages Thermoactinomyces vulgaris panose + maltoheptaose
-
?
isopanose + H2O cyclomaltodextrinase activity, hydrolysis of alpha-1,6-glucosidic linkages Thermoactinomyces vulgaris ?
-
?
additional information substrate specificity of the bifunctional enzyme, the enzyme hydrolyzes alpha-1,4-glucosidic linkages and alpha-1,6-glucosidic linkages, and performs transglycosylation reactions, overview Thermoactinomyces vulgaris ?
-
?
pullulan + H2O cyclomaltodextrinase activity, hydrolysis of alpha-1,4-glucosidic linkages Thermoactinomyces vulgaris panose + ?
-
?
starch + H2O alpha-amylase activity, preferred substrate, hydrolysis of alpha-1,4-glucosidic linkages Thermoactinomyces vulgaris maltooligosaccharides
-
?

Synonyms

Synonyms Comment Organism
alpha-amylase II
-
Thermoactinomyces vulgaris
More cf. EC 3.2.1.54, cyclomaltodextrinase Thermoactinomyces vulgaris
TVA II
-
Thermoactinomyces vulgaris

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.8
-
pullulan mutant Y374A Thermoactinomyces vulgaris
9.4
-
starch mutant Y374A Thermoactinomyces vulgaris
120
-
starch wild-type enzyme Thermoactinomyces vulgaris
140
-
pullulan wild-type enzyme Thermoactinomyces vulgaris