Improvement of Cellulomonas fimi endoglucanase CenA by multienzymatic display on a decameric structural scaffold

Autores
Iglesias Rando, Matías Rubén; Gorojovsky, Natalia; Zylberman, Vanesa; Goldbaum, Fernando Alberto; Craig, Patricio Oliver
Año de publicación
2023
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The development of multifunctional particles using polymeric scaffolds is an emergingtechnology for many nanobiotechnological applications. Here we present a system forthe production of multifunctional complexes, based on the high affinity non-covalentinteraction of cohesin and dockerin modules complementary fused to decamericBrucella abortus lumazine synthase (BLS) subunits, and selected target proteins,respectively. The cohesin-BLS scaffold was solubly expressed in high yield inEscherichia coli, and revealed a high thermostability. The production of multienzymaticparticles using this system was evaluated using the catalytic domain of Cellulomonasfimi endoglucanase CenA recombinantly fused to a dockerin module. Coupling of theenzyme to the scaffold was highly efficient and occurred with the expectedstoichiometry. The decavalent enzymatic complexes obtained, showed highercellulolytic activity and association to the substrate compared to equivalent amounts ofthe free enzyme. This phenomenon was dependent on the multiplicity and proximity ofthe enzymes coupled to the scaffold, and was attributed to an avidity effect in thepolyvalent enzyme interaction with the substrate. Our results highlight the usefulness ofthe scaffold presented in this work for the development of multifunctional particles, andthe improvement of lignocellulose degradation among other applications.
Fil: Iglesias Rando, Matías Rubén. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Gorojovsky, Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Zylberman, Vanesa. Inmunova; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Goldbaum, Fernando Alberto. Inmunova; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina
Fil: Craig, Patricio Oliver. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina
Materia
ARTIFICIAL CELLULOSOME
BLS
CELLULASE
MULTIENZYMATIC COMPLEX
SCAFFOLD
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/220413
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/220413
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Improvement of Cellulomonas fimi endoglucanase CenA by multienzymatic display on a decameric structural scaffoldIglesias Rando, Matías RubénGorojovsky, NataliaZylberman, VanesaGoldbaum, Fernando AlbertoCraig, Patricio OliverARTIFICIAL CELLULOSOMEBLSCELLULASEMULTIENZYMATIC COMPLEXSCAFFOLDhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The development of multifunctional particles using polymeric scaffolds is an emergingtechnology for many nanobiotechnological applications. Here we present a system forthe production of multifunctional complexes, based on the high affinity non-covalentinteraction of cohesin and dockerin modules complementary fused to decamericBrucella abortus lumazine synthase (BLS) subunits, and selected target proteins,respectively. The cohesin-BLS scaffold was solubly expressed in high yield inEscherichia coli, and revealed a high thermostability. The production of multienzymaticparticles using this system was evaluated using the catalytic domain of Cellulomonasfimi endoglucanase CenA recombinantly fused to a dockerin module. Coupling of theenzyme to the scaffold was highly efficient and occurred with the expectedstoichiometry. The decavalent enzymatic complexes obtained, showed highercellulolytic activity and association to the substrate compared to equivalent amounts ofthe free enzyme. This phenomenon was dependent on the multiplicity and proximity ofthe enzymes coupled to the scaffold, and was attributed to an avidity effect in thepolyvalent enzyme interaction with the substrate. Our results highlight the usefulness ofthe scaffold presented in this work for the development of multifunctional particles, andthe improvement of lignocellulose degradation among other applications.Fil: Iglesias Rando, Matías Rubén. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Gorojovsky, Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Zylberman, Vanesa. Inmunova; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Goldbaum, Fernando Alberto. Inmunova; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Craig, Patricio Oliver. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; ArgentinaSpringer2023-05info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/220413Iglesias Rando, Matías Rubén; Gorojovsky, Natalia; Zylberman, Vanesa; Goldbaum, Fernando Alberto; Craig, Patricio Oliver; Improvement of Cellulomonas fimi endoglucanase CenA by multienzymatic display on a decameric structural scaffold; Springer; Applied Microbiology and Biotechnology; 107; 13; 5-2023; 4261-42740175-7598CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://link.springer.com/article/10.1007/s00253-023-12581-6#citeasinfo:eu-repo/semantics/altIdentifier/doi/10.1007/s00253-023-12581-6info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T15:07:14Zoai:ri.conicet.gov.ar:11336/220413instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-10-15 15:07:14.502CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Improvement of Cellulomonas fimi endoglucanase CenA by multienzymatic display on a decameric structural scaffold
title Improvement of Cellulomonas fimi endoglucanase CenA by multienzymatic display on a decameric structural scaffold
spellingShingle Improvement of Cellulomonas fimi endoglucanase CenA by multienzymatic display on a decameric structural scaffold
Iglesias Rando, Matías Rubén
ARTIFICIAL CELLULOSOME
BLS
CELLULASE
MULTIENZYMATIC COMPLEX
SCAFFOLD
title_short Improvement of Cellulomonas fimi endoglucanase CenA by multienzymatic display on a decameric structural scaffold
title_full Improvement of Cellulomonas fimi endoglucanase CenA by multienzymatic display on a decameric structural scaffold
title_fullStr Improvement of Cellulomonas fimi endoglucanase CenA by multienzymatic display on a decameric structural scaffold
title_full_unstemmed Improvement of Cellulomonas fimi endoglucanase CenA by multienzymatic display on a decameric structural scaffold
title_sort Improvement of Cellulomonas fimi endoglucanase CenA by multienzymatic display on a decameric structural scaffold
dc.creator.none.fl_str_mv Iglesias Rando, Matías Rubén
Gorojovsky, Natalia
Zylberman, Vanesa
Goldbaum, Fernando Alberto
Craig, Patricio Oliver
author Iglesias Rando, Matías Rubén
author_facet Iglesias Rando, Matías Rubén
Gorojovsky, Natalia
Zylberman, Vanesa
Goldbaum, Fernando Alberto
Craig, Patricio Oliver
author_role author
author2 Gorojovsky, Natalia
Zylberman, Vanesa
Goldbaum, Fernando Alberto
Craig, Patricio Oliver
author2_role author
author
author
author
dc.subject.none.fl_str_mv ARTIFICIAL CELLULOSOME
BLS
CELLULASE
MULTIENZYMATIC COMPLEX
SCAFFOLD
topic ARTIFICIAL CELLULOSOME
BLS
CELLULASE
MULTIENZYMATIC COMPLEX
SCAFFOLD
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The development of multifunctional particles using polymeric scaffolds is an emergingtechnology for many nanobiotechnological applications. Here we present a system forthe production of multifunctional complexes, based on the high affinity non-covalentinteraction of cohesin and dockerin modules complementary fused to decamericBrucella abortus lumazine synthase (BLS) subunits, and selected target proteins,respectively. The cohesin-BLS scaffold was solubly expressed in high yield inEscherichia coli, and revealed a high thermostability. The production of multienzymaticparticles using this system was evaluated using the catalytic domain of Cellulomonasfimi endoglucanase CenA recombinantly fused to a dockerin module. Coupling of theenzyme to the scaffold was highly efficient and occurred with the expectedstoichiometry. The decavalent enzymatic complexes obtained, showed highercellulolytic activity and association to the substrate compared to equivalent amounts ofthe free enzyme. This phenomenon was dependent on the multiplicity and proximity ofthe enzymes coupled to the scaffold, and was attributed to an avidity effect in thepolyvalent enzyme interaction with the substrate. Our results highlight the usefulness ofthe scaffold presented in this work for the development of multifunctional particles, andthe improvement of lignocellulose degradation among other applications.
Fil: Iglesias Rando, Matías Rubén. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Gorojovsky, Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Zylberman, Vanesa. Inmunova; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Goldbaum, Fernando Alberto. Inmunova; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina
Fil: Craig, Patricio Oliver. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina
description The development of multifunctional particles using polymeric scaffolds is an emergingtechnology for many nanobiotechnological applications. Here we present a system forthe production of multifunctional complexes, based on the high affinity non-covalentinteraction of cohesin and dockerin modules complementary fused to decamericBrucella abortus lumazine synthase (BLS) subunits, and selected target proteins,respectively. The cohesin-BLS scaffold was solubly expressed in high yield inEscherichia coli, and revealed a high thermostability. The production of multienzymaticparticles using this system was evaluated using the catalytic domain of Cellulomonasfimi endoglucanase CenA recombinantly fused to a dockerin module. Coupling of theenzyme to the scaffold was highly efficient and occurred with the expectedstoichiometry. The decavalent enzymatic complexes obtained, showed highercellulolytic activity and association to the substrate compared to equivalent amounts ofthe free enzyme. This phenomenon was dependent on the multiplicity and proximity ofthe enzymes coupled to the scaffold, and was attributed to an avidity effect in thepolyvalent enzyme interaction with the substrate. Our results highlight the usefulness ofthe scaffold presented in this work for the development of multifunctional particles, andthe improvement of lignocellulose degradation among other applications.
publishDate 2023
dc.date.none.fl_str_mv 2023-05
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/220413
Iglesias Rando, Matías Rubén; Gorojovsky, Natalia; Zylberman, Vanesa; Goldbaum, Fernando Alberto; Craig, Patricio Oliver; Improvement of Cellulomonas fimi endoglucanase CenA by multienzymatic display on a decameric structural scaffold; Springer; Applied Microbiology and Biotechnology; 107; 13; 5-2023; 4261-4274
0175-7598
CONICET Digital
CONICET
url http://hdl.handle.net/11336/220413
identifier_str_mv Iglesias Rando, Matías Rubén; Gorojovsky, Natalia; Zylberman, Vanesa; Goldbaum, Fernando Alberto; Craig, Patricio Oliver; Improvement of Cellulomonas fimi endoglucanase CenA by multienzymatic display on a decameric structural scaffold; Springer; Applied Microbiology and Biotechnology; 107; 13; 5-2023; 4261-4274
0175-7598
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://link.springer.com/article/10.1007/s00253-023-12581-6#citeas
info:eu-repo/semantics/altIdentifier/doi/10.1007/s00253-023-12581-6
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Springer
publisher.none.fl_str_mv Springer
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.22299

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