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Literature summary for 2.4.1.4 extracted from

  • Skov, L.K.; Pizzut-Serin, S.; Remaud-Simeon, M.; Ernst, H.A.; Gajhede, M.; Mirza, O.
    The structure of amylosucrase from Deinococcus radiodurans has an unusual open active-site topology (2013), Acta Crystallogr. Sect. F, 69, 973-978.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression in Escherichia coli strain BL21 Deinococcus radiodurans

Crystallization (Commentary)

Crystallization (Comment) Organism
purified wild-type and mutant ligand-free enzyme, hanging drop vapour diffusion method, mixing of 0.0025 ml of 3.3 mg/ml protein in 50 mM HEPES, pH 7.0, 150 mM NaCl, 1 mM EDTA, and 1 mM DTT, with 0.0025 ml of reservoir solution containing 1 M sodium acetate, 0.1 M imidazole, 0.1 M LiCl, pH 6.5, 4°C, X-ray diffraction structure determination and analysis at 3.15 A resolution, modeling Deinococcus radiodurans

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
sucrose + [(1->4)-alpha-D-glucosyl]n Deinococcus radiodurans amylosucrases catalyze the formation of an alpha-1,4-glucosidic linkage by transferring a glucosyl unit from sucrose onto an acceptor alpha-1,4-glucan D-fructose + [(1->4)-alpha-D-glucosyl]n+1
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?

Organism

Organism UniProt Comment Textmining
Deinococcus radiodurans
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Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme from Escherichia coli strain BL21 to homogeneity using GST-affinity chromatography followed by dialysis and subsequent tag removal via PreScission protease and reverse affinity chromatography Deinococcus radiodurans

Reaction

Reaction Comment Organism Reaction ID
sucrose + [(1->4)-alpha-D-glucosyl]n = D-fructose + [(1->4)-alpha-D-glucosyl]n+1 the elongation mechanism involves enzyme conformational changes to allow the entry of sucrose to the active site Deinococcus radiodurans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
sucrose + [(1->4)-alpha-D-glucosyl]n amylosucrases catalyze the formation of an alpha-1,4-glucosidic linkage by transferring a glucosyl unit from sucrose onto an acceptor alpha-1,4-glucan Deinococcus radiodurans D-fructose + [(1->4)-alpha-D-glucosyl]n+1
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?

Subunits

Subunits Comment Organism
More the putative enzyme interface is primarily mediated by two regions from domain N, residues Thr22, Leu25, Arg26, Arg29 and Tyr30 and residues Glu73, Leu76 and Leu77, which are involved in hydrophobic interactions with the corresponding residues from the second protomer Deinococcus radiodurans

Synonyms

Synonyms Comment Organism
DRAS
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Deinococcus radiodurans

General Information

General Information Comment Organism
additional information ligand binding causes extensive changes in active-site topology Deinococcus radiodurans