Selective Synthesis of Galactooligosaccharides Containing β(1→3) Linkages with β-Galactosidase from Bifidobacterium bifidum (Saphera)
[EN] The transglycosylation activity of a novel commercial β-galactosidase from Bifidobacterium bifidum (Saphera) was evaluated. The optimal conditions for the operation of this enzyme, measured with o-nitrophenyl-β-d-galactopyranoside, were 40 °C and pH around 6.0. Although at low lactose concentra...
| Autores: | , , , , , , , |
|---|---|
| Formato: | artículo |
| Estado: | Versión aceptada para publicación |
| Fecha de publicación: | 2020 |
| País: | España |
| Recursos: | Consejo Superior de Investigaciones Científicas (CSIC) |
| Repositorio: | DIGITAL.CSIC. Repositorio Institucional del CSIC |
| OAI Identifier: | oai:digital.csic.es:10261/216922 |
| Acesso em linha: | http://hdl.handle.net/10261/216922 |
| Access Level: | acceso abierto |
| Palavra-chave: | Galactooligosaccharides Prebiotics Transglycosylation β-Galactosidase Bifidobacteria Glycosidases. |
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Selective Synthesis of Galactooligosaccharides Containing β(1→3) Linkages with β-Galactosidase from Bifidobacterium bifidum (Saphera)Füreder, VeraRodríguez-Colinas, BárbaraCervantes, Fadia V.Fernández Arrojo, LucíaPoveda, AnaJiménez-Barbero, JesúsBallesteros Olmo, AntonioPlou Gasca, Francisco JoséGalactooligosaccharidesPrebioticsTransglycosylationβ-GalactosidaseBifidobacteriaGlycosidases.[EN] The transglycosylation activity of a novel commercial β-galactosidase from Bifidobacterium bifidum (Saphera) was evaluated. The optimal conditions for the operation of this enzyme, measured with o-nitrophenyl-β-d-galactopyranoside, were 40 °C and pH around 6.0. Although at low lactose concentrations the property of this enzyme was basically hydrolytic, an increase of lactose concentration to 400 g/L resulted in a significant formation (107.2 g/L, 27% yield) of prebiotic galactooligosaccharides (GOS). The maximum amount of GOS was obtained at a lactose conversion of approximately 90%, which contrasts with other β-galactosidases, for which the highest GOS yield is achieved at 40-50% lactose conversion. Using high-performance anion-exchange chromatography with pulsed amperometric detection, semipreparative high-performance liquid chromatography-hydrophilic interaction liquid chromatography, mass spectrometry, and 1D and 2D NMR, we determined the structure of most of the GOS synthesized by this enzyme. The main identified products were Gal-β(1→3)-Gal-β(1→4)-Glc (3′-O-β-galactosyl-lactose), Gal-β(1→6)-Glc (allolactose), Gal-β(1→3)-Glc (3-galactosyl-glucose), Gal-β(1→3)-Gal (3-galactobiose), and the tetrasaccharide Gal-β(1→3)-Gal-β(1→3)-Gal-β(1→4)-Glc. In general, B. bifidum β-galactosidase showed a tendency to form β(1→3) linkages followed by β(1→6) and more scarcely β(1→4).This work was supported by a grant from the Spanish Ministry of Economy and Competitiveness (BIO2016-76601-C3-1). The group at CIC bioGUNE thanks Agencia Estatal de Investigación (Spain) for the Severo Ochoa Excellence Accreditation (SEV-2016-0644).American Chemical SocietyMinisterio de Economía y Competitividad (España)Agencia Estatal de Investigación (España)Poveda, Ana [0000-0001-5060-2307]Jiménez-Barbero, Jesús [0000-0001-5421-8513]Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]2020202020202020info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Postprintinfo:eu-repo/semantics/acceptedVersionhttp://hdl.handle.net/10261/216922reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2016-76601-C3-1http://dx.doi.org/10.1021/acs.jafc.0c00997Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/2169222026-05-22T06:33:51Z |
| dc.title.none.fl_str_mv |
Selective Synthesis of Galactooligosaccharides Containing β(1→3) Linkages with β-Galactosidase from Bifidobacterium bifidum (Saphera) |
| title |
Selective Synthesis of Galactooligosaccharides Containing β(1→3) Linkages with β-Galactosidase from Bifidobacterium bifidum (Saphera) |
| spellingShingle |
Selective Synthesis of Galactooligosaccharides Containing β(1→3) Linkages with β-Galactosidase from Bifidobacterium bifidum (Saphera) Füreder, Vera Galactooligosaccharides Prebiotics Transglycosylation β-Galactosidase Bifidobacteria Glycosidases. |
| title_short |
Selective Synthesis of Galactooligosaccharides Containing β(1→3) Linkages with β-Galactosidase from Bifidobacterium bifidum (Saphera) |
| title_full |
Selective Synthesis of Galactooligosaccharides Containing β(1→3) Linkages with β-Galactosidase from Bifidobacterium bifidum (Saphera) |
| title_fullStr |
Selective Synthesis of Galactooligosaccharides Containing β(1→3) Linkages with β-Galactosidase from Bifidobacterium bifidum (Saphera) |
| title_full_unstemmed |
Selective Synthesis of Galactooligosaccharides Containing β(1→3) Linkages with β-Galactosidase from Bifidobacterium bifidum (Saphera) |
| title_sort |
Selective Synthesis of Galactooligosaccharides Containing β(1→3) Linkages with β-Galactosidase from Bifidobacterium bifidum (Saphera) |
| dc.creator.none.fl_str_mv |
Füreder, Vera Rodríguez-Colinas, Bárbara Cervantes, Fadia V. Fernández Arrojo, Lucía Poveda, Ana Jiménez-Barbero, Jesús Ballesteros Olmo, Antonio Plou Gasca, Francisco José |
| author |
Füreder, Vera |
| author_facet |
Füreder, Vera Rodríguez-Colinas, Bárbara Cervantes, Fadia V. Fernández Arrojo, Lucía Poveda, Ana Jiménez-Barbero, Jesús Ballesteros Olmo, Antonio Plou Gasca, Francisco José |
| author_role |
author |
| author2 |
Rodríguez-Colinas, Bárbara Cervantes, Fadia V. Fernández Arrojo, Lucía Poveda, Ana Jiménez-Barbero, Jesús Ballesteros Olmo, Antonio Plou Gasca, Francisco José |
| author2_role |
author author author author author author author |
| dc.contributor.none.fl_str_mv |
Ministerio de Economía y Competitividad (España) Agencia Estatal de Investigación (España) Poveda, Ana [0000-0001-5060-2307] Jiménez-Barbero, Jesús [0000-0001-5421-8513] Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72] |
| dc.subject.none.fl_str_mv |
Galactooligosaccharides Prebiotics Transglycosylation β-Galactosidase Bifidobacteria Glycosidases. |
| topic |
Galactooligosaccharides Prebiotics Transglycosylation β-Galactosidase Bifidobacteria Glycosidases. |
| description |
[EN] The transglycosylation activity of a novel commercial β-galactosidase from Bifidobacterium bifidum (Saphera) was evaluated. The optimal conditions for the operation of this enzyme, measured with o-nitrophenyl-β-d-galactopyranoside, were 40 °C and pH around 6.0. Although at low lactose concentrations the property of this enzyme was basically hydrolytic, an increase of lactose concentration to 400 g/L resulted in a significant formation (107.2 g/L, 27% yield) of prebiotic galactooligosaccharides (GOS). The maximum amount of GOS was obtained at a lactose conversion of approximately 90%, which contrasts with other β-galactosidases, for which the highest GOS yield is achieved at 40-50% lactose conversion. Using high-performance anion-exchange chromatography with pulsed amperometric detection, semipreparative high-performance liquid chromatography-hydrophilic interaction liquid chromatography, mass spectrometry, and 1D and 2D NMR, we determined the structure of most of the GOS synthesized by this enzyme. The main identified products were Gal-β(1→3)-Gal-β(1→4)-Glc (3′-O-β-galactosyl-lactose), Gal-β(1→6)-Glc (allolactose), Gal-β(1→3)-Glc (3-galactosyl-glucose), Gal-β(1→3)-Gal (3-galactobiose), and the tetrasaccharide Gal-β(1→3)-Gal-β(1→3)-Gal-β(1→4)-Glc. In general, B. bifidum β-galactosidase showed a tendency to form β(1→3) linkages followed by β(1→6) and more scarcely β(1→4). |
| publishDate |
2020 |
| dc.date.none.fl_str_mv |
2020 2020 2020 2020 |
| dc.type.none.fl_str_mv |
info:eu-repo/semantics/article http://purl.org/coar/resource_type/c_6501 Postprint info:eu-repo/semantics/acceptedVersion |
| format |
article |
| status_str |
acceptedVersion |
| dc.identifier.none.fl_str_mv |
http://hdl.handle.net/10261/216922 |
| url |
http://hdl.handle.net/10261/216922 |
| dc.language.none.fl_str_mv |
Inglés |
| language_invalid_str_mv |
Inglés |
| dc.relation.none.fl_str_mv |
#PLACEHOLDER_PARENT_METADATA_VALUE# info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2016-76601-C3-1 http://dx.doi.org/10.1021/acs.jafc.0c00997 Sí |
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info:eu-repo/semantics/openAccess |
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openAccess |
| dc.publisher.none.fl_str_mv |
American Chemical Society |
| publisher.none.fl_str_mv |
American Chemical Society |
| dc.source.none.fl_str_mv |
reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC instname:Consejo Superior de Investigaciones Científicas (CSIC) |
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Consejo Superior de Investigaciones Científicas (CSIC) |
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DIGITAL.CSIC. Repositorio Institucional del CSIC |
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DIGITAL.CSIC. Repositorio Institucional del CSIC |
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1869418267946254336 |
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15,812455 |