Stereochemical course of hydrolytic reaction catalyzed by alpha-galactosidase from cold adaptable marine bacterium of genus Pseudoalteromonas
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, trans...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2014-10-01
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| Series: | Frontiers in Chemistry |
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| Online Access: | http://journal.frontiersin.org/Journal/10.3389/fchem.2014.00089/full |
| _version_ | 1819076121010896896 |
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| author | Irina Yu Bakunina Larissa A Balabanova Vasiliy A Golotin Lyubov V Slepchenko Lyubov V Slepchenko Vladimir V Isakov Valeriy A Rasskazov |
| author_facet | Irina Yu Bakunina Larissa A Balabanova Vasiliy A Golotin Lyubov V Slepchenko Lyubov V Slepchenko Vladimir V Isakov Valeriy A Rasskazov |
| author_sort | Irina Yu Bakunina |
| collection | DOAJ |
| description | 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 E. 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 1H 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. |
| first_indexed | 2024-12-21T18:36:16Z |
| format | Article |
| id | doaj.art-e28bb854262e45bfa49d73895ad8ed09 |
| institution | Directory Open Access Journal |
| issn | 2296-2646 |
| language | English |
| last_indexed | 2024-12-21T18:36:16Z |
| publishDate | 2014-10-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Chemistry |
| spelling | doaj.art-e28bb854262e45bfa49d73895ad8ed092022-12-21T18:54:08ZengFrontiers Media S.A.Frontiers in Chemistry2296-26462014-10-01210.3389/fchem.2014.00089105710Stereochemical course of hydrolytic reaction catalyzed by alpha-galactosidase from cold adaptable marine bacterium of genus PseudoalteromonasIrina Yu Bakunina0Larissa A Balabanova1Vasiliy A Golotin2Lyubov V Slepchenko3Lyubov V Slepchenko4Vladimir V Isakov5Valeriy A Rasskazov6G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch, Russian Academy of SciencesG.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far Easte..G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch, Russian Academy of SciencesG.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far Easte..G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch, Russian Academy of SciencesG.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch, Russian Academy of SciencesG.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far Easte..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 E. 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 1H 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.http://journal.frontiersin.org/Journal/10.3389/fchem.2014.00089/fullstereochemistryα-galactosidasemarine bacterium Pseudoalteromonas sp.1H NMR spectrahydrolysis of glycosidesα-anomer |
| spellingShingle | Irina Yu Bakunina Larissa A Balabanova Vasiliy A Golotin Lyubov V Slepchenko Lyubov V Slepchenko Vladimir V Isakov Valeriy A Rasskazov Stereochemical course of hydrolytic reaction catalyzed by alpha-galactosidase from cold adaptable marine bacterium of genus Pseudoalteromonas Frontiers in Chemistry stereochemistry α-galactosidase marine bacterium Pseudoalteromonas sp. 1H NMR spectra hydrolysis of glycosides α-anomer |
| title | Stereochemical course of hydrolytic reaction catalyzed by alpha-galactosidase from cold adaptable marine bacterium of genus Pseudoalteromonas |
| title_full | Stereochemical course of hydrolytic reaction catalyzed by alpha-galactosidase from cold adaptable marine bacterium of genus Pseudoalteromonas |
| title_fullStr | Stereochemical course of hydrolytic reaction catalyzed by alpha-galactosidase from cold adaptable marine bacterium of genus Pseudoalteromonas |
| title_full_unstemmed | Stereochemical course of hydrolytic reaction catalyzed by alpha-galactosidase from cold adaptable marine bacterium of genus Pseudoalteromonas |
| title_short | Stereochemical course of hydrolytic reaction catalyzed by alpha-galactosidase from cold adaptable marine bacterium of genus Pseudoalteromonas |
| title_sort | stereochemical course of hydrolytic reaction catalyzed by alpha galactosidase from cold adaptable marine bacterium of genus pseudoalteromonas |
| topic | stereochemistry α-galactosidase marine bacterium Pseudoalteromonas sp. 1H NMR spectra hydrolysis of glycosides α-anomer |
| url | http://journal.frontiersin.org/Journal/10.3389/fchem.2014.00089/full |
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