Analysis of the structure-function relationship of alpha amylase complexed with polyacrylic acid

Autores
Porfiri, María Cecilia; Melnichuk, Natasha; Braia, Mauricio Javier; Brinatti, César; Loh, Watson; Romanini, Diana
Año de publicación
2020
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Alpha-amylase is frequently used in technologies that require its immobilization, stabilization or encapsulation. Polyacrylic acid is a very suitable polymer for these purposes because it can bind to enzymes and then be released under certain conditions without altering the functional capacity of enzymes. The consequences produced by polyacrylic acid on alpha-amylase structure and function have been investigated through various techniques. Calorimetric measurements allowed examining the nature of the binding reaction, stoichiometry and affinity, while spectroscopic techniques provided additional information about functional and structural perturbations of the enzyme. Isothermal titration calorimetry (ITC) revealed a mixed interaction and a binding model with a large number of molecules of protein per molecule of polyacrylic acid. One the one hand circular dichroism (CD) spectroscopy showed that alpha-amylase loses its secondary structure in the presence of increasing concentrations of polyacrylic acid, while it is stabilized by the polyelectrolyte at low pH. On the other hand, fluorescence spectra revealed that the three-dimensional enzyme structure was not affected in the microenvironment of tryptophan residues. Differential scanning calorimetry (DSC) thermograms showed that only one domain of alpha-amylase is affected in its conformational stability by the polymer. The unfolding process proved to be partially reversible. Finally, the enzyme retained more than 90 % of its catalytic activity even in excess of the polymer.
Fil: Porfiri, María Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Investigación en Funcionalidad y Tecnología de Alimentos; Argentina
Fil: Melnichuk, Natasha. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; Argentina
Fil: Braia, Mauricio Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; Argentina
Fil: Brinatti, César. Universidade Estadual Do Campinas. Instituto de Química.; Brasil
Fil: Loh, Watson. Universidade Estadual Do Campinas. Instituto de Química.; Brasil
Fil: Romanini, Diana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; Argentina
Materia
ALPHA-AMYLASE
CALORIMETRY
POLYACRYLIC ACID
SPECTROSCOPIC ANALYSIS
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/168579
Acceder
id CONICETDig_0d9c9d457ebc6e15bec3d9ffd84ecdff
oai_identifier_str oai:ri.conicet.gov.ar:11336/168579
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Analysis of the structure-function relationship of alpha amylase complexed with polyacrylic acidPorfiri, María CeciliaMelnichuk, NatashaBraia, Mauricio JavierBrinatti, CésarLoh, WatsonRomanini, DianaALPHA-AMYLASECALORIMETRYPOLYACRYLIC ACIDSPECTROSCOPIC ANALYSIShttps://purl.org/becyt/ford/1.4https://purl.org/becyt/ford/1Alpha-amylase is frequently used in technologies that require its immobilization, stabilization or encapsulation. Polyacrylic acid is a very suitable polymer for these purposes because it can bind to enzymes and then be released under certain conditions without altering the functional capacity of enzymes. The consequences produced by polyacrylic acid on alpha-amylase structure and function have been investigated through various techniques. Calorimetric measurements allowed examining the nature of the binding reaction, stoichiometry and affinity, while spectroscopic techniques provided additional information about functional and structural perturbations of the enzyme. Isothermal titration calorimetry (ITC) revealed a mixed interaction and a binding model with a large number of molecules of protein per molecule of polyacrylic acid. One the one hand circular dichroism (CD) spectroscopy showed that alpha-amylase loses its secondary structure in the presence of increasing concentrations of polyacrylic acid, while it is stabilized by the polyelectrolyte at low pH. On the other hand, fluorescence spectra revealed that the three-dimensional enzyme structure was not affected in the microenvironment of tryptophan residues. Differential scanning calorimetry (DSC) thermograms showed that only one domain of alpha-amylase is affected in its conformational stability by the polymer. The unfolding process proved to be partially reversible. Finally, the enzyme retained more than 90 % of its catalytic activity even in excess of the polymer.Fil: Porfiri, María Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Investigación en Funcionalidad y Tecnología de Alimentos; ArgentinaFil: Melnichuk, Natasha. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Braia, Mauricio Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Brinatti, César. Universidade Estadual Do Campinas. Instituto de Química.; BrasilFil: Loh, Watson. Universidade Estadual Do Campinas. Instituto de Química.; BrasilFil: Romanini, Diana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaElsevier Science2020-04info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/168579Porfiri, María Cecilia; Melnichuk, Natasha; Braia, Mauricio Javier; Brinatti, César; Loh, Watson; et al.; Analysis of the structure-function relationship of alpha amylase complexed with polyacrylic acid; Elsevier Science; Colloids and Surfaces B: Biointerfaces; 188; 4-2020; 1-310927-7765CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://linkinghub.elsevier.com/retrieve/pii/S0927776520300175info:eu-repo/semantics/altIdentifier/doi/10.1016/j.colsurfb.2020.110787info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:16:36Zoai:ri.conicet.gov.ar:11336/168579instacron: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:34982025-09-29 10:16:36.626CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Analysis of the structure-function relationship of alpha amylase complexed with polyacrylic acid
title Analysis of the structure-function relationship of alpha amylase complexed with polyacrylic acid
spellingShingle Analysis of the structure-function relationship of alpha amylase complexed with polyacrylic acid
Porfiri, María Cecilia
ALPHA-AMYLASE
CALORIMETRY
POLYACRYLIC ACID
SPECTROSCOPIC ANALYSIS
title_short Analysis of the structure-function relationship of alpha amylase complexed with polyacrylic acid
title_full Analysis of the structure-function relationship of alpha amylase complexed with polyacrylic acid
title_fullStr Analysis of the structure-function relationship of alpha amylase complexed with polyacrylic acid
title_full_unstemmed Analysis of the structure-function relationship of alpha amylase complexed with polyacrylic acid
title_sort Analysis of the structure-function relationship of alpha amylase complexed with polyacrylic acid
dc.creator.none.fl_str_mv Porfiri, María Cecilia
Melnichuk, Natasha
Braia, Mauricio Javier
Brinatti, César
Loh, Watson
Romanini, Diana
author Porfiri, María Cecilia
author_facet Porfiri, María Cecilia
Melnichuk, Natasha
Braia, Mauricio Javier
Brinatti, César
Loh, Watson
Romanini, Diana
author_role author
author2 Melnichuk, Natasha
Braia, Mauricio Javier
Brinatti, César
Loh, Watson
Romanini, Diana
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv ALPHA-AMYLASE
CALORIMETRY
POLYACRYLIC ACID
SPECTROSCOPIC ANALYSIS
topic ALPHA-AMYLASE
CALORIMETRY
POLYACRYLIC ACID
SPECTROSCOPIC ANALYSIS
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.4
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Alpha-amylase is frequently used in technologies that require its immobilization, stabilization or encapsulation. Polyacrylic acid is a very suitable polymer for these purposes because it can bind to enzymes and then be released under certain conditions without altering the functional capacity of enzymes. The consequences produced by polyacrylic acid on alpha-amylase structure and function have been investigated through various techniques. Calorimetric measurements allowed examining the nature of the binding reaction, stoichiometry and affinity, while spectroscopic techniques provided additional information about functional and structural perturbations of the enzyme. Isothermal titration calorimetry (ITC) revealed a mixed interaction and a binding model with a large number of molecules of protein per molecule of polyacrylic acid. One the one hand circular dichroism (CD) spectroscopy showed that alpha-amylase loses its secondary structure in the presence of increasing concentrations of polyacrylic acid, while it is stabilized by the polyelectrolyte at low pH. On the other hand, fluorescence spectra revealed that the three-dimensional enzyme structure was not affected in the microenvironment of tryptophan residues. Differential scanning calorimetry (DSC) thermograms showed that only one domain of alpha-amylase is affected in its conformational stability by the polymer. The unfolding process proved to be partially reversible. Finally, the enzyme retained more than 90 % of its catalytic activity even in excess of the polymer.
Fil: Porfiri, María Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Investigación en Funcionalidad y Tecnología de Alimentos; Argentina
Fil: Melnichuk, Natasha. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; Argentina
Fil: Braia, Mauricio Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; Argentina
Fil: Brinatti, César. Universidade Estadual Do Campinas. Instituto de Química.; Brasil
Fil: Loh, Watson. Universidade Estadual Do Campinas. Instituto de Química.; Brasil
Fil: Romanini, Diana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; Argentina
description Alpha-amylase is frequently used in technologies that require its immobilization, stabilization or encapsulation. Polyacrylic acid is a very suitable polymer for these purposes because it can bind to enzymes and then be released under certain conditions without altering the functional capacity of enzymes. The consequences produced by polyacrylic acid on alpha-amylase structure and function have been investigated through various techniques. Calorimetric measurements allowed examining the nature of the binding reaction, stoichiometry and affinity, while spectroscopic techniques provided additional information about functional and structural perturbations of the enzyme. Isothermal titration calorimetry (ITC) revealed a mixed interaction and a binding model with a large number of molecules of protein per molecule of polyacrylic acid. One the one hand circular dichroism (CD) spectroscopy showed that alpha-amylase loses its secondary structure in the presence of increasing concentrations of polyacrylic acid, while it is stabilized by the polyelectrolyte at low pH. On the other hand, fluorescence spectra revealed that the three-dimensional enzyme structure was not affected in the microenvironment of tryptophan residues. Differential scanning calorimetry (DSC) thermograms showed that only one domain of alpha-amylase is affected in its conformational stability by the polymer. The unfolding process proved to be partially reversible. Finally, the enzyme retained more than 90 % of its catalytic activity even in excess of the polymer.
publishDate 2020
dc.date.none.fl_str_mv 2020-04
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/11336/168579
Porfiri, María Cecilia; Melnichuk, Natasha; Braia, Mauricio Javier; Brinatti, César; Loh, Watson; et al.; Analysis of the structure-function relationship of alpha amylase complexed with polyacrylic acid; Elsevier Science; Colloids and Surfaces B: Biointerfaces; 188; 4-2020; 1-31
0927-7765
CONICET Digital
CONICET
url http://hdl.handle.net/11336/168579
identifier_str_mv Porfiri, María Cecilia; Melnichuk, Natasha; Braia, Mauricio Javier; Brinatti, César; Loh, Watson; et al.; Analysis of the structure-function relationship of alpha amylase complexed with polyacrylic acid; Elsevier Science; Colloids and Surfaces B: Biointerfaces; 188; 4-2020; 1-31
0927-7765
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://linkinghub.elsevier.com/retrieve/pii/S0927776520300175
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.colsurfb.2020.110787
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier Science
publisher.none.fl_str_mv Elsevier Science
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
_version_ 1844614112029442048
score 13.070432

Publicaciones similares