Modulating Glycoside Hydrolase Activity between Hydrolysis and Transfer Reactions Using an Evolutionary Approach

The proteins within the CAZy glycoside hydrolase family GH13 catalyze the hydrolysis of polysaccharides such as glycogen and starch. Many of these enzymes also perform transglycosylation in various degrees, ranging from secondary to predominant reactions. Identifying structural determinants associat...

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Autores principales:	Rodrigo A. Arreola-Barroso, Alexey Llopiz, Leticia Olvera, Gloria Saab-Rincón
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	transglycosidation hydrolysis contact-residues amylase glucanotransferase coevolution Organic chemistry QD241-441
Acceso en línea:	https://doaj.org/article/3d87d862e7c64ec6b489f79eba63c3ff
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spelling	oai:doaj.org-article:3d87d862e7c64ec6b489f79eba63c3ff2021-11-11T18:33:50ZModulating Glycoside Hydrolase Activity between Hydrolysis and Transfer Reactions Using an Evolutionary Approach10.3390/molecules262165861420-3049https://doaj.org/article/3d87d862e7c64ec6b489f79eba63c3ff2021-10-01T00:00:00Zhttps://www.mdpi.com/1420-3049/26/21/6586https://doaj.org/toc/1420-3049The proteins within the CAZy glycoside hydrolase family GH13 catalyze the hydrolysis of polysaccharides such as glycogen and starch. Many of these enzymes also perform transglycosylation in various degrees, ranging from secondary to predominant reactions. Identifying structural determinants associated with GH13 family reaction specificity is key to modifying and designing enzymes with increased specificity towards individual reactions for further applications in industrial, chemical, or biomedical fields. This work proposes a computational approach for decoding the determinant structural composition defining the reaction specificity. This method is based on the conservation of coevolving residues in spatial contacts associated with reaction specificity. To evaluate the algorithm, mutants of α-amylase (<i>TmAmyA</i>) and glucanotransferase (<i>TmGTase</i>) from <i>Thermotoga maritima</i> were constructed to modify the reaction specificity. The K98P/D99A/H222Q variant from <i>TmAmyA</i> doubled the transglycosydation/hydrolysis (T/H) ratio while the M279N variant from <i>TmGTase</i> increased the hydrolysis/transglycosidation ratio five-fold. Molecular dynamic simulations of the variants indicated changes in flexibility that can account for the modified T/H ratio. An essential contribution of the presented computational approach is its capacity to identify residues outside of the active center that affect the reaction specificity.Rodrigo A. Arreola-BarrosoAlexey LlopizLeticia OlveraGloria Saab-RincónMDPI AGarticletransglycosidationhydrolysiscontact-residuesamylaseglucanotransferasecoevolutionOrganic chemistryQD241-441ENMolecules, Vol 26, Iss 6586, p 6586 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	transglycosidation hydrolysis contact-residues amylase glucanotransferase coevolution Organic chemistry QD241-441
spellingShingle	transglycosidation hydrolysis contact-residues amylase glucanotransferase coevolution Organic chemistry QD241-441 Rodrigo A. Arreola-Barroso Alexey Llopiz Leticia Olvera Gloria Saab-Rincón Modulating Glycoside Hydrolase Activity between Hydrolysis and Transfer Reactions Using an Evolutionary Approach
description	The proteins within the CAZy glycoside hydrolase family GH13 catalyze the hydrolysis of polysaccharides such as glycogen and starch. Many of these enzymes also perform transglycosylation in various degrees, ranging from secondary to predominant reactions. Identifying structural determinants associated with GH13 family reaction specificity is key to modifying and designing enzymes with increased specificity towards individual reactions for further applications in industrial, chemical, or biomedical fields. This work proposes a computational approach for decoding the determinant structural composition defining the reaction specificity. This method is based on the conservation of coevolving residues in spatial contacts associated with reaction specificity. To evaluate the algorithm, mutants of α-amylase (<i>TmAmyA</i>) and glucanotransferase (<i>TmGTase</i>) from <i>Thermotoga maritima</i> were constructed to modify the reaction specificity. The K98P/D99A/H222Q variant from <i>TmAmyA</i> doubled the transglycosydation/hydrolysis (T/H) ratio while the M279N variant from <i>TmGTase</i> increased the hydrolysis/transglycosidation ratio five-fold. Molecular dynamic simulations of the variants indicated changes in flexibility that can account for the modified T/H ratio. An essential contribution of the presented computational approach is its capacity to identify residues outside of the active center that affect the reaction specificity.
format	article
author	Rodrigo A. Arreola-Barroso Alexey Llopiz Leticia Olvera Gloria Saab-Rincón
author_facet	Rodrigo A. Arreola-Barroso Alexey Llopiz Leticia Olvera Gloria Saab-Rincón
author_sort	Rodrigo A. Arreola-Barroso
title	Modulating Glycoside Hydrolase Activity between Hydrolysis and Transfer Reactions Using an Evolutionary Approach
title_short	Modulating Glycoside Hydrolase Activity between Hydrolysis and Transfer Reactions Using an Evolutionary Approach
title_full	Modulating Glycoside Hydrolase Activity between Hydrolysis and Transfer Reactions Using an Evolutionary Approach
title_fullStr	Modulating Glycoside Hydrolase Activity between Hydrolysis and Transfer Reactions Using an Evolutionary Approach
title_full_unstemmed	Modulating Glycoside Hydrolase Activity between Hydrolysis and Transfer Reactions Using an Evolutionary Approach
title_sort	modulating glycoside hydrolase activity between hydrolysis and transfer reactions using an evolutionary approach
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/3d87d862e7c64ec6b489f79eba63c3ff
work_keys_str_mv	AT rodrigoaarreolabarroso modulatingglycosidehydrolaseactivitybetweenhydrolysisandtransferreactionsusinganevolutionaryapproach AT alexeyllopiz modulatingglycosidehydrolaseactivitybetweenhydrolysisandtransferreactionsusinganevolutionaryapproach AT leticiaolvera modulatingglycosidehydrolaseactivitybetweenhydrolysisandtransferreactionsusinganevolutionaryapproach AT gloriasaabrincon modulatingglycosidehydrolaseactivitybetweenhydrolysisandtransferreactionsusinganevolutionaryapproach
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Modulating Glycoside Hydrolase Activity between Hydrolysis and Transfer Reactions Using an Evolutionary Approach

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