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Stereochemical Course of Hydrolytic Reaction Catalyzed by Alpha-Galactosidase From Cold Adaptable Marine Bacterium of Genus Pseudoalteromonas

Stereochemical Course of Hydrolytic Reaction Catalyzed by Alpha-Galactosidase From Cold Adaptable Marine Bacterium of Genus Pseudoalteromonas

Irina Y Bakunina, Larissa A Balabanova, Vasiliy A Golotin, Lyubov V Slepchenko, Vladimir V Isakov, Valeriy A Rasskazov

Front Chem. 2014 Oct 13;2:89.

PMID: 25353020

Abstract:

The recombinant α-galactosidase of the marine bacterium (α-PsGal) was synthesized with the use of the plasmid 40Gal, consisting of plasmid pET-40b (+) (Novagen) and the gene corresponding to the open reading frame of the mature α-galactosidase of marine bacterium Pseudoalteromonas sp. KMM 701, transformed into the Escherichia coli Rosetta(DE3) cells. In order to understand the mechanism of action, the stereochemistry of hydrolysis of 4-nitrophenyl α-D-galactopyranoside (4-NPGP) by α-PsGal was measured by (1)H NMR spectroscopy. The kinetics of formation of α- and β-anomer of galactose showed that α-anomer initially formed and accumulated, and then an appreciable amount of β-anomer appeared as a result of mutarotation. The data clearly show that the enzymatic hydrolysis of 4-NPGP proceeds with the retention of anomeric configuration, probably, due to a double displacement mechanism of reaction.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP7493950	4-Nitrophenyl α-D-galactopyranoside		7493-95-0	Price

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