Structure–Function Insights into the Fungal Endo-Chitinase Chit33 Depict its Mechanism on Chitinous Material

15 pags., 6 figs., 2 tabs.

Detalles Bibliográficos
Autores: Jiménez-Ortega, Elena, Kidibule, Peter E, Fernández Lobato, María, Sanz-Aparicio, J.
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2022
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/283600
Acceso en línea:http://hdl.handle.net/10261/283600
Access Level:acceso abierto
Palabra clave:Chitinase
Crystal structure
Chito-oligosaccharides
NAG6 binding mode
Hydrolytic activity
Specificity
id ES_b3589e32584ed6a6dfd627e3f65fbaef
oai_identifier_str oai:digital.csic.es:10261/283600
network_acronym_str ES
network_name_str España
repository_id_str
spelling Structure–Function Insights into the Fungal Endo-Chitinase Chit33 Depict its Mechanism on Chitinous MaterialJiménez-Ortega, ElenaKidibule, Peter EFernández Lobato, MaríaSanz-Aparicio, J.ChitinaseCrystal structureChito-oligosaccharidesNAG6 binding modeHydrolytic activitySpecificity15 pags., 6 figs., 2 tabs.Chitin is the most widespread amino renewable carbohydrate polymer in nature and the second most abundant polysaccharide. Therefore, chitin and chitinolytic enzymes are becoming more importance for biotechnological applications in food, health and agricultural fields, the design of effective enzymes being a paramount issue. We report the crystal structure of the plant-type endo-chitinase Chit33 from Trichoderma harzianum and its D165A/E167A-Chit33-(NAG) complex, which showed an extended catalytic cleft with six binding subsites lined with many polar interactions. The major trait of Chit33 is the location of the non-conserved Asp117 and Arg274 acting as a clamp, fixing the distorted conformation of the sugar at subsite –1 and the bent shape of the substrate, which occupies the full catalytic groove. Relevant residues were selected for mutagenesis experiments, the variants being biochemically characterized through their hydrolytic activity against colloidal chitin and other polymeric substrates with different molecular weights and deacetylation percentages. The mutant S118Y stands out, showing a superior performance in all the substrates tested, as well as detectable transglycosylation capacity, with this variant providing a promising platform for generation of novel Chit33 variants with adjusted performance by further design of rational mutants’. The putative role of Tyr in binding was extrapolated from molecular dynamics simulation.This work was supported by grants from the Spanish Ministry of Economy and Competitiveness through grants PID2019-105838RB-C33/-C32, Fundacioón Ramón Areces (XIX Call of Research Grants in Life and Material Sciences, and EU EMFF-Blue Economy-2018 (Fish4Fish-863697). We are grateful to the staff of the Synchrotron Radiation Sources at Alba (Barcelona, Spain) for providing access and for technical assistance with BL13-XALOC beamline and to the Fundacioón Ramón Areces for an institutional grant to the Centre of Molecular Biology Severo Ochoa.Molecular Diversity Preservation InternationalMinisterio de Economía y Competitividad (España)Fundación Ramón ArecesEuropean Maritime and Fisheries FundALBA SynchrotronCSIC-UAM - Centro de Biología Molecular Severo Ochoa (CBM)Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]2022202220222022info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/283600reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PID2019-105838RB-C33info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PID2019-105838RB-C32http://dx.doi.org/10.3390/ijms23147599Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/2836002026-05-22T06:33:51Z
dc.title.none.fl_str_mv Structure–Function Insights into the Fungal Endo-Chitinase Chit33 Depict its Mechanism on Chitinous Material
title Structure–Function Insights into the Fungal Endo-Chitinase Chit33 Depict its Mechanism on Chitinous Material
spellingShingle Structure–Function Insights into the Fungal Endo-Chitinase Chit33 Depict its Mechanism on Chitinous Material
Jiménez-Ortega, Elena
Chitinase
Crystal structure
Chito-oligosaccharides
NAG6 binding mode
Hydrolytic activity
Specificity
title_short Structure–Function Insights into the Fungal Endo-Chitinase Chit33 Depict its Mechanism on Chitinous Material
title_full Structure–Function Insights into the Fungal Endo-Chitinase Chit33 Depict its Mechanism on Chitinous Material
title_fullStr Structure–Function Insights into the Fungal Endo-Chitinase Chit33 Depict its Mechanism on Chitinous Material
title_full_unstemmed Structure–Function Insights into the Fungal Endo-Chitinase Chit33 Depict its Mechanism on Chitinous Material
title_sort Structure–Function Insights into the Fungal Endo-Chitinase Chit33 Depict its Mechanism on Chitinous Material
dc.creator.none.fl_str_mv Jiménez-Ortega, Elena
Kidibule, Peter E
Fernández Lobato, María
Sanz-Aparicio, J.
author Jiménez-Ortega, Elena
author_facet Jiménez-Ortega, Elena
Kidibule, Peter E
Fernández Lobato, María
Sanz-Aparicio, J.
author_role author
author2 Kidibule, Peter E
Fernández Lobato, María
Sanz-Aparicio, J.
author2_role author
author
author
dc.contributor.none.fl_str_mv Ministerio de Economía y Competitividad (España)
Fundación Ramón Areces
European Maritime and Fisheries Fund
ALBA Synchrotron
CSIC-UAM - Centro de Biología Molecular Severo Ochoa (CBM)
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Chitinase
Crystal structure
Chito-oligosaccharides
NAG6 binding mode
Hydrolytic activity
Specificity
topic Chitinase
Crystal structure
Chito-oligosaccharides
NAG6 binding mode
Hydrolytic activity
Specificity
description 15 pags., 6 figs., 2 tabs.
publishDate 2022
dc.date.none.fl_str_mv 2022
2022
2022
2022
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/283600
url http://hdl.handle.net/10261/283600
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
#PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PID2019-105838RB-C33
info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PID2019-105838RB-C32
http://dx.doi.org/10.3390/ijms23147599
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Molecular Diversity Preservation International
publisher.none.fl_str_mv Molecular Diversity Preservation International
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869417169778900992
score 15.811543

Similares en LA Referencia