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Engineered Glycosidases for the Synthesis of Analogs of Human Milk Oligosaccharides

Enzymatic synthesis is an elegant biocompatible approach to complex compounds such as human milk oligosaccharides (HMOs). These compounds are vital for healthy neonatal development with a positive impact on the immune system. Although HMOs may be prepared by glycosyltransferases, this pathway is oft...

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Autores principales: Nekvasilová, Pavlína, Hovorková, Michaela, Mészáros, Zuzana, Petrásková, Lucie, Pelantová, Helena, Křen, Vladimír, Slámová, Kristýna, Bojarová, Pavla
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
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Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9027921/
https://www.ncbi.nlm.nih.gov/pubmed/35456924
http://dx.doi.org/10.3390/ijms23084106
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author Nekvasilová, Pavlína
Hovorková, Michaela
Mészáros, Zuzana
Petrásková, Lucie
Pelantová, Helena
Křen, Vladimír
Slámová, Kristýna
Bojarová, Pavla
author_facet Nekvasilová, Pavlína
Hovorková, Michaela
Mészáros, Zuzana
Petrásková, Lucie
Pelantová, Helena
Křen, Vladimír
Slámová, Kristýna
Bojarová, Pavla
author_sort Nekvasilová, Pavlína
collection PubMed
description Enzymatic synthesis is an elegant biocompatible approach to complex compounds such as human milk oligosaccharides (HMOs). These compounds are vital for healthy neonatal development with a positive impact on the immune system. Although HMOs may be prepared by glycosyltransferases, this pathway is often complicated by the high price of sugar nucleotides, stringent substrate specificity, and low enzyme stability. Engineered glycosidases (EC 3.2.1) represent a good synthetic alternative, especially if variations in the substrate structure are desired. Site-directed mutagenesis can improve the synthetic process with higher yields and/or increased reaction selectivity. So far, the synthesis of human milk oligosaccharides by glycosidases has mostly been limited to analytical reactions with mass spectrometry detection. The present work reveals the potential of a library of engineered glycosidases in the preparative synthesis of three tetrasaccharides derived from lacto-N-tetraose (Galβ4GlcNAcβ3Galβ4Glc), employing sequential cascade reactions catalyzed by β3-N-acetylhexosaminidase BbhI from Bifidobacterium bifidum, β4-galactosidase BgaD-B from Bacillus circulans, β4-N-acetylgalactosaminidase from Talaromyces flavus, and β3-galactosynthase BgaC from B. circulans. The reaction products were isolated and structurally characterized. This work expands the insight into the multi-step catalysis by glycosidases and shows the path to modified derivatives of complex carbohydrates that cannot be prepared by standard glycosyltransferase methods.
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spelling pubmed-90279212022-04-23 Engineered Glycosidases for the Synthesis of Analogs of Human Milk Oligosaccharides Nekvasilová, Pavlína Hovorková, Michaela Mészáros, Zuzana Petrásková, Lucie Pelantová, Helena Křen, Vladimír Slámová, Kristýna Bojarová, Pavla Int J Mol Sci Article Enzymatic synthesis is an elegant biocompatible approach to complex compounds such as human milk oligosaccharides (HMOs). These compounds are vital for healthy neonatal development with a positive impact on the immune system. Although HMOs may be prepared by glycosyltransferases, this pathway is often complicated by the high price of sugar nucleotides, stringent substrate specificity, and low enzyme stability. Engineered glycosidases (EC 3.2.1) represent a good synthetic alternative, especially if variations in the substrate structure are desired. Site-directed mutagenesis can improve the synthetic process with higher yields and/or increased reaction selectivity. So far, the synthesis of human milk oligosaccharides by glycosidases has mostly been limited to analytical reactions with mass spectrometry detection. The present work reveals the potential of a library of engineered glycosidases in the preparative synthesis of three tetrasaccharides derived from lacto-N-tetraose (Galβ4GlcNAcβ3Galβ4Glc), employing sequential cascade reactions catalyzed by β3-N-acetylhexosaminidase BbhI from Bifidobacterium bifidum, β4-galactosidase BgaD-B from Bacillus circulans, β4-N-acetylgalactosaminidase from Talaromyces flavus, and β3-galactosynthase BgaC from B. circulans. The reaction products were isolated and structurally characterized. This work expands the insight into the multi-step catalysis by glycosidases and shows the path to modified derivatives of complex carbohydrates that cannot be prepared by standard glycosyltransferase methods. MDPI 2022-04-07 /pmc/articles/PMC9027921/ /pubmed/35456924 http://dx.doi.org/10.3390/ijms23084106 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Nekvasilová, Pavlína
Hovorková, Michaela
Mészáros, Zuzana
Petrásková, Lucie
Pelantová, Helena
Křen, Vladimír
Slámová, Kristýna
Bojarová, Pavla
Engineered Glycosidases for the Synthesis of Analogs of Human Milk Oligosaccharides
title Engineered Glycosidases for the Synthesis of Analogs of Human Milk Oligosaccharides
title_full Engineered Glycosidases for the Synthesis of Analogs of Human Milk Oligosaccharides
title_fullStr Engineered Glycosidases for the Synthesis of Analogs of Human Milk Oligosaccharides
title_full_unstemmed Engineered Glycosidases for the Synthesis of Analogs of Human Milk Oligosaccharides
title_short Engineered Glycosidases for the Synthesis of Analogs of Human Milk Oligosaccharides
title_sort engineered glycosidases for the synthesis of analogs of human milk oligosaccharides
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9027921/
https://www.ncbi.nlm.nih.gov/pubmed/35456924
http://dx.doi.org/10.3390/ijms23084106
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