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Indolyl Septanoside Synthesis for In Vivo Screening of Bacterial Septanoside Hydrolases

Building-up and breaking-down of carbohydrates are processes common to all forms of life. Glycoside hydrolases are a broad class of enzymes that play a central role in the cleavage of glycosidic bonds, which is fundamental to carbohydrate degradation. The large majority of substrates are five- and s...

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Autores principales: Pote, Aditya R., Pascual, Sergi, Planas, Antoni, Peczuh, Mark W.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8123396/
https://www.ncbi.nlm.nih.gov/pubmed/33925857
http://dx.doi.org/10.3390/ijms22094497
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author Pote, Aditya R.
Pascual, Sergi
Planas, Antoni
Peczuh, Mark W.
author_facet Pote, Aditya R.
Pascual, Sergi
Planas, Antoni
Peczuh, Mark W.
author_sort Pote, Aditya R.
collection PubMed
description Building-up and breaking-down of carbohydrates are processes common to all forms of life. Glycoside hydrolases are a broad class of enzymes that play a central role in the cleavage of glycosidic bonds, which is fundamental to carbohydrate degradation. The large majority of substrates are five- and six-membered ring glycosides. Our interest in seven-membered ring septanose sugars has inspired the development of a way to search for septanoside hydrolase activity. Described here is a strategy for the discovery of septanoside hydrolases that uses synthetic indolyl septanosides as chromogenic substrates. Access to these tool compounds was enabled by a route where septanosyl halides act as glycosyl donors for the synthesis of the indolyl septanosides. The screening strategy leverages the known dimerization of 3-hydroxy-indoles to make colored dyes, as occurs when the β-galactosidase substrate X-Gal is hydrolyzed. Because screens in bacterial cells would enable searches in organisms that utilize heptoses or from metagenomics libraries, we also demonstrate that septanosides are capable of entering E. coli cells through the use of a BODIPY-labeled septanoside. The modularity of the indolyl septanoside synthesis should allow the screening of a variety of substrates that mimic natural structures via this general approach.
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spelling pubmed-81233962021-05-16 Indolyl Septanoside Synthesis for In Vivo Screening of Bacterial Septanoside Hydrolases Pote, Aditya R. Pascual, Sergi Planas, Antoni Peczuh, Mark W. Int J Mol Sci Article Building-up and breaking-down of carbohydrates are processes common to all forms of life. Glycoside hydrolases are a broad class of enzymes that play a central role in the cleavage of glycosidic bonds, which is fundamental to carbohydrate degradation. The large majority of substrates are five- and six-membered ring glycosides. Our interest in seven-membered ring septanose sugars has inspired the development of a way to search for septanoside hydrolase activity. Described here is a strategy for the discovery of septanoside hydrolases that uses synthetic indolyl septanosides as chromogenic substrates. Access to these tool compounds was enabled by a route where septanosyl halides act as glycosyl donors for the synthesis of the indolyl septanosides. The screening strategy leverages the known dimerization of 3-hydroxy-indoles to make colored dyes, as occurs when the β-galactosidase substrate X-Gal is hydrolyzed. Because screens in bacterial cells would enable searches in organisms that utilize heptoses or from metagenomics libraries, we also demonstrate that septanosides are capable of entering E. coli cells through the use of a BODIPY-labeled septanoside. The modularity of the indolyl septanoside synthesis should allow the screening of a variety of substrates that mimic natural structures via this general approach. MDPI 2021-04-26 /pmc/articles/PMC8123396/ /pubmed/33925857 http://dx.doi.org/10.3390/ijms22094497 Text en © 2021 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
Pote, Aditya R.
Pascual, Sergi
Planas, Antoni
Peczuh, Mark W.
Indolyl Septanoside Synthesis for In Vivo Screening of Bacterial Septanoside Hydrolases
title Indolyl Septanoside Synthesis for In Vivo Screening of Bacterial Septanoside Hydrolases
title_full Indolyl Septanoside Synthesis for In Vivo Screening of Bacterial Septanoside Hydrolases
title_fullStr Indolyl Septanoside Synthesis for In Vivo Screening of Bacterial Septanoside Hydrolases
title_full_unstemmed Indolyl Septanoside Synthesis for In Vivo Screening of Bacterial Septanoside Hydrolases
title_short Indolyl Septanoside Synthesis for In Vivo Screening of Bacterial Septanoside Hydrolases
title_sort indolyl septanoside synthesis for in vivo screening of bacterial septanoside hydrolases
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8123396/
https://www.ncbi.nlm.nih.gov/pubmed/33925857
http://dx.doi.org/10.3390/ijms22094497
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