Enzymatic Synthesis of Galactosylated Serine/Threonine Derivatives by β-Galactosidase from Escherichia coli
The transgalactosylations of serine/threonine derivatives were investigated using β-galactosidase from Escherichia coli as biocatalyst. Using ortho-nitrophenyl-β-D-galactoside as donor, the highest bioconversion yield of transgalactosylated N-carboxy benzyl L-serine benzyl ester (23.2%) was achieved...
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MDPI AG
2015-06-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | http://www.mdpi.com/1422-0067/16/6/13714 |
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| author | Sooyoun Seo Joseph Rebehmed Alexandre G. de Brevern Salwa Karboune |
| author_facet | Sooyoun Seo Joseph Rebehmed Alexandre G. de Brevern Salwa Karboune |
| author_sort | Sooyoun Seo |
| collection | DOAJ |
| description | The transgalactosylations of serine/threonine derivatives were investigated using β-galactosidase from Escherichia coli as biocatalyst. Using ortho-nitrophenyl-β-D-galactoside as donor, the highest bioconversion yield of transgalactosylated N-carboxy benzyl L-serine benzyl ester (23.2%) was achieved in heptane:buffer medium (70:30), whereas with the lactose, the highest bioconversion yield (3.94%) was obtained in the buffer reaction system. The structures of most abundant galactosylated serine products were characterized by MS/MS. The molecular docking simulation revealed that the binding of serine/threonine derivatives to the enzyme’s active site was stronger (−4.6~−7.9 kcal/mol) than that of the natural acceptor, glucose, and mainly occurred through interactions with aromatic residues. For N-tert-butoxycarbonyl serine methyl ester (6.8%) and N-carboxybenzyl serine benzyl ester (3.4%), their binding affinities and the distances between their hydroxyl side chain and the 1′-OH group of galactose moiety were in good accordance with the quantified bioconversion yields. Despite its lower predicted bioconversion yield, the high experimental bioconversion yield obtained with N-carboxybenzyl serine methyl ester (23.2%) demonstrated the importance of the thermodynamically-driven nature of the transgalactosylation reaction. |
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| issn | 1422-0067 |
| language | English |
| last_indexed | 2024-04-13T18:10:56Z |
| publishDate | 2015-06-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-237e0e8188e64608953b1d7569b92aa12022-12-22T02:35:52ZengMDPI AGInternational Journal of Molecular Sciences1422-00672015-06-01166137141372810.3390/ijms160613714ijms160613714Enzymatic Synthesis of Galactosylated Serine/Threonine Derivatives by β-Galactosidase from Escherichia coliSooyoun Seo0Joseph Rebehmed1Alexandre G. de Brevern2Salwa Karboune3Department of Food Science and Agricultural Chemistry, McGill University, 21,111 Lakeshore, Ste Anne de Bellevue, QC H9X-3V9, CanadaINSERM UMR_S 1134, Dynamique des Structures et Interactions des Macromolécules Biologiques (DSIMB), University Paris Diderot, Sorbonne Paris Cité, INTS, Laboratory of excellence, GR-Ex, 6, rue Alexandre Cabanel, 75739 Paris Cedex 15, FranceINSERM UMR_S 1134, Dynamique des Structures et Interactions des Macromolécules Biologiques (DSIMB), University Paris Diderot, Sorbonne Paris Cité, INTS, Laboratory of excellence, GR-Ex, 6, rue Alexandre Cabanel, 75739 Paris Cedex 15, FranceDepartment of Food Science and Agricultural Chemistry, McGill University, 21,111 Lakeshore, Ste Anne de Bellevue, QC H9X-3V9, CanadaThe transgalactosylations of serine/threonine derivatives were investigated using β-galactosidase from Escherichia coli as biocatalyst. Using ortho-nitrophenyl-β-D-galactoside as donor, the highest bioconversion yield of transgalactosylated N-carboxy benzyl L-serine benzyl ester (23.2%) was achieved in heptane:buffer medium (70:30), whereas with the lactose, the highest bioconversion yield (3.94%) was obtained in the buffer reaction system. The structures of most abundant galactosylated serine products were characterized by MS/MS. The molecular docking simulation revealed that the binding of serine/threonine derivatives to the enzyme’s active site was stronger (−4.6~−7.9 kcal/mol) than that of the natural acceptor, glucose, and mainly occurred through interactions with aromatic residues. For N-tert-butoxycarbonyl serine methyl ester (6.8%) and N-carboxybenzyl serine benzyl ester (3.4%), their binding affinities and the distances between their hydroxyl side chain and the 1′-OH group of galactose moiety were in good accordance with the quantified bioconversion yields. Despite its lower predicted bioconversion yield, the high experimental bioconversion yield obtained with N-carboxybenzyl serine methyl ester (23.2%) demonstrated the importance of the thermodynamically-driven nature of the transgalactosylation reaction.http://www.mdpi.com/1422-0067/16/6/13714β-galactosidasetransgalactosylationserine/threonine derivativesmolecular dockingbinding affinitiesgalactosyl acceptor |
| spellingShingle | Sooyoun Seo Joseph Rebehmed Alexandre G. de Brevern Salwa Karboune Enzymatic Synthesis of Galactosylated Serine/Threonine Derivatives by β-Galactosidase from Escherichia coli International Journal of Molecular Sciences β-galactosidase transgalactosylation serine/threonine derivatives molecular docking binding affinities galactosyl acceptor |
| title | Enzymatic Synthesis of Galactosylated Serine/Threonine Derivatives by β-Galactosidase from Escherichia coli |
| title_full | Enzymatic Synthesis of Galactosylated Serine/Threonine Derivatives by β-Galactosidase from Escherichia coli |
| title_fullStr | Enzymatic Synthesis of Galactosylated Serine/Threonine Derivatives by β-Galactosidase from Escherichia coli |
| title_full_unstemmed | Enzymatic Synthesis of Galactosylated Serine/Threonine Derivatives by β-Galactosidase from Escherichia coli |
| title_short | Enzymatic Synthesis of Galactosylated Serine/Threonine Derivatives by β-Galactosidase from Escherichia coli |
| title_sort | enzymatic synthesis of galactosylated serine threonine derivatives by β galactosidase from escherichia coli |
| topic | β-galactosidase transgalactosylation serine/threonine derivatives molecular docking binding affinities galactosyl acceptor |
| url | http://www.mdpi.com/1422-0067/16/6/13714 |
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