Sequence diversity and catalytic properties of phytases
Autor(a) principal: | |
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Data de Publicação: | 2022 |
Outros Autores: | , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Research, Society and Development |
Texto Completo: | https://rsdjournal.org/index.php/rsd/article/view/32765 |
Resumo: | Phytic acid is an antinutritional factor in cereal feeds, and the use of phytases increases the bioavailability of nutrients bound to this molecule. However, the application of these enzymes depends on their thermal stability and activity at acidic pH. Therefore, in this study we created a database composed of 59 phytase sequences and analyzed the interactions that stabilize their structures in order to understand whether they contribute to the biochemical properties observed. The sequences were aligned and grouped at 30 % similarity, generating 5 clusters, which highlights the high variability among them. A comparative structural analysis of the cluster 3 phytases revealed conserved catalytic domains, as well as eight cysteine residues along the primary sequence, forming disulfide bonds for stabilizing the three-dimensional structure. However, the number of Van der Waals, ionic, and hydrogen interactions, and disulfide bonds was not determinant for the biochemical characteristics presented by these enzymes. The phytase KM873028, from cluster 3, was selected for characterization studies, but its expression in Pichia pastoris generated a protein with properties distinct from those derived from the same sequence expressed in a prokaryotic system. It is likely that the differences observed are associated with the location of the interactions in the structures, non-conserved amino acid residues found around the catalytic site, and post-translational modifications inherent to the expression systems. These possibilities highlight the relevance of strategic choices related to enzyme expression aiming at its production and industrial feasibility. |
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Sequence diversity and catalytic properties of phytasesDiversidad en secuencias y propiedades catalíticas en fitasasDiversidade de sequências e propriedades catalíticas de fitasesConservación de secuencias enzimáticasÁcido fíticoAlimentación animalEstabilidad enzimáticaEstructura enzimática.Conservação de sequências de enzimasÁcido fíticoAlimentação animalEstabilidade de enzimasEstrutura de enzimas.Enzyme sequence conservationPhytic acidAnimal feedEnzyme stabilityEnzyme structure. Phytic acid is an antinutritional factor in cereal feeds, and the use of phytases increases the bioavailability of nutrients bound to this molecule. However, the application of these enzymes depends on their thermal stability and activity at acidic pH. Therefore, in this study we created a database composed of 59 phytase sequences and analyzed the interactions that stabilize their structures in order to understand whether they contribute to the biochemical properties observed. The sequences were aligned and grouped at 30 % similarity, generating 5 clusters, which highlights the high variability among them. A comparative structural analysis of the cluster 3 phytases revealed conserved catalytic domains, as well as eight cysteine residues along the primary sequence, forming disulfide bonds for stabilizing the three-dimensional structure. However, the number of Van der Waals, ionic, and hydrogen interactions, and disulfide bonds was not determinant for the biochemical characteristics presented by these enzymes. The phytase KM873028, from cluster 3, was selected for characterization studies, but its expression in Pichia pastoris generated a protein with properties distinct from those derived from the same sequence expressed in a prokaryotic system. It is likely that the differences observed are associated with the location of the interactions in the structures, non-conserved amino acid residues found around the catalytic site, and post-translational modifications inherent to the expression systems. These possibilities highlight the relevance of strategic choices related to enzyme expression aiming at its production and industrial feasibility.El ácido fítico es un factor antinutricional en los cereales que componen las raciones, y el uso de fitasas aumenta la biodisponibilidad de los nutrientes en esta molécula. La aplicación de estas enzimas depende de su estabilidad térmica y actividad a pH ácido. Por lo tanto, creamos una base de datos con 59 secuencias de fitasa y analizamos las interacciones que estabilizan sus estructuras para comprender si contribuyen a las propiedades bioquímicas observadas. Las secuencias fueron agrupadas al 30% de similitud, generando 5 clusters, lo que demuestra la alta variabilidad. Un análisis estructural comparativo de las fitasas del grupo 3 reveló dominios catalíticos conservados, así como ocho residuos de cisteína a lo largo de la secuencia primaria, formando enlaces disulfuro para estabilizar la estrutura. Sin embargo, el número de interacciones de hidrógeno, Van der Waals, iónico y disulfuro no es determinante para las características bioquímicas que presentan estas enzimas. La fitasa KM873028, del grupo 3, fue seleccionada para estudios de caracterización, pero su expresión en Pichia pastoris generó una proteína con propiedades diferentes a las derivadas de la misma secuencia expresada en un sistema procariótico. Es probable que las diferencias observadas estén asociadas con la ubicación de las interacciones en las estructuras, los residuos de aminoácidos no conservados que se encuentran alrededor del sitio catalítico, y las modificaciones postraduccionales inherentes a los sistemas de expresión elegidos. Estas posibilidades muestran la relevancia de elecciones estratégicas relacionadas con la expresión de la enzima, con el objetivo de su producción y viabilidad industrial.O ácido fítico é um fator antinutricional de cereais que compõem rações, e o uso de fitases aumenta a biodisponibilidade de nutrientes ligados a esta molécula. No entanto, a aplicação dessas enzimas depende de sua estabilidade térmica e atividade em pH ácido. Por isso, nesse estudo criamos um banco de dados com 59 sequências de fitases e analisamos as interações que estabilizam suas estruturas a fim de compreender se estas contribuem para as propriedades bioquímicas observadas. As sequências foram alinhadas e agrupadas a 30% de similaridade, gerando 5 clusters, o que evidencia a alta variabilidade entre elas. Uma análise comparativa estrutural das fitases do cluster 3 revelou domínios catalíticos conservados, assim como oito resíduos de cisteína ao longo da sequência primária, formando pontes dissulfeto para estabilização da estrutura tridimensional proteica. Entretanto, o número de interações de hidrogênio, Van der Waals e iônicas e pontes dissulfeto não foi determinante para as características bioquímicas apresentadas por essas enzimas. A fitase KM873028, do cluster 3, foi selecionada para estudos de caracterização, mas a sua expressão em Pichia pastoris gerou uma proteína com propriedades distintas daquelas derivadas da mesma sequência expressa em sistema procarioto. É provável que as diferenças observadas estejam associadas à localização das interações nas estruturas, resíduos de aminoácidos não conservados encontrados ao redor do sítio catalítico, além de modificações pós-traducionais inerentes aos sistemas de expressão escolhidos. Essas possibilidades evidenciam a relevância das escolhas estratégicas relacionadas à expressão da enzima visando sua produção e viabilização industrial.Research, Society and Development2022-08-06info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://rsdjournal.org/index.php/rsd/article/view/3276510.33448/rsd-v11i10.32765Research, Society and Development; Vol. 11 No. 10; e427111032765Research, Society and Development; Vol. 11 Núm. 10; e427111032765Research, Society and Development; v. 11 n. 10; e4271110327652525-3409reponame:Research, Society and Developmentinstname:Universidade Federal de Itajubá (UNIFEI)instacron:UNIFEIenghttps://rsdjournal.org/index.php/rsd/article/view/32765/27940Copyright (c) 2022 Elizabeth Bárbara Epalanga Pires ; Marcelo Depólo Polêto; Pedro Marcus Pereira Vidigal; Matheus Ítalo Bonfim Aragão ; Tarley Araújo Barros ; Rafael Locatelli Salgado; Valéria Monteze Guimarães ; Monique Renon Ellerhttps://creativecommons.org/licenses/by/4.0info:eu-repo/semantics/openAccessPires , Elizabeth Bárbara EpalangaPolêto, Marcelo DepóloVidigal, Pedro Marcus PereiraAragão , Matheus Ítalo BonfimBarros , Tarley AraújoSalgado, Rafael LocatelliGuimarães , Valéria MontezeEller, Monique Renon2022-08-12T22:23:03Zoai:ojs.pkp.sfu.ca:article/32765Revistahttps://rsdjournal.org/index.php/rsd/indexPUBhttps://rsdjournal.org/index.php/rsd/oairsd.articles@gmail.com2525-34092525-3409opendoar:2024-01-17T09:48:38.951830Research, Society and Development - Universidade Federal de Itajubá (UNIFEI)false |
dc.title.none.fl_str_mv |
Sequence diversity and catalytic properties of phytases Diversidad en secuencias y propiedades catalíticas en fitasas Diversidade de sequências e propriedades catalíticas de fitases |
title |
Sequence diversity and catalytic properties of phytases |
spellingShingle |
Sequence diversity and catalytic properties of phytases Pires , Elizabeth Bárbara Epalanga Conservación de secuencias enzimáticas Ácido fítico Alimentación animal Estabilidad enzimática Estructura enzimática. Conservação de sequências de enzimas Ácido fítico Alimentação animal Estabilidade de enzimas Estrutura de enzimas. Enzyme sequence conservation Phytic acid Animal feed Enzyme stability Enzyme structure. |
title_short |
Sequence diversity and catalytic properties of phytases |
title_full |
Sequence diversity and catalytic properties of phytases |
title_fullStr |
Sequence diversity and catalytic properties of phytases |
title_full_unstemmed |
Sequence diversity and catalytic properties of phytases |
title_sort |
Sequence diversity and catalytic properties of phytases |
author |
Pires , Elizabeth Bárbara Epalanga |
author_facet |
Pires , Elizabeth Bárbara Epalanga Polêto, Marcelo Depólo Vidigal, Pedro Marcus Pereira Aragão , Matheus Ítalo Bonfim Barros , Tarley Araújo Salgado, Rafael Locatelli Guimarães , Valéria Monteze Eller, Monique Renon |
author_role |
author |
author2 |
Polêto, Marcelo Depólo Vidigal, Pedro Marcus Pereira Aragão , Matheus Ítalo Bonfim Barros , Tarley Araújo Salgado, Rafael Locatelli Guimarães , Valéria Monteze Eller, Monique Renon |
author2_role |
author author author author author author author |
dc.contributor.author.fl_str_mv |
Pires , Elizabeth Bárbara Epalanga Polêto, Marcelo Depólo Vidigal, Pedro Marcus Pereira Aragão , Matheus Ítalo Bonfim Barros , Tarley Araújo Salgado, Rafael Locatelli Guimarães , Valéria Monteze Eller, Monique Renon |
dc.subject.por.fl_str_mv |
Conservación de secuencias enzimáticas Ácido fítico Alimentación animal Estabilidad enzimática Estructura enzimática. Conservação de sequências de enzimas Ácido fítico Alimentação animal Estabilidade de enzimas Estrutura de enzimas. Enzyme sequence conservation Phytic acid Animal feed Enzyme stability Enzyme structure. |
topic |
Conservación de secuencias enzimáticas Ácido fítico Alimentación animal Estabilidad enzimática Estructura enzimática. Conservação de sequências de enzimas Ácido fítico Alimentação animal Estabilidade de enzimas Estrutura de enzimas. Enzyme sequence conservation Phytic acid Animal feed Enzyme stability Enzyme structure. |
description |
Phytic acid is an antinutritional factor in cereal feeds, and the use of phytases increases the bioavailability of nutrients bound to this molecule. However, the application of these enzymes depends on their thermal stability and activity at acidic pH. Therefore, in this study we created a database composed of 59 phytase sequences and analyzed the interactions that stabilize their structures in order to understand whether they contribute to the biochemical properties observed. The sequences were aligned and grouped at 30 % similarity, generating 5 clusters, which highlights the high variability among them. A comparative structural analysis of the cluster 3 phytases revealed conserved catalytic domains, as well as eight cysteine residues along the primary sequence, forming disulfide bonds for stabilizing the three-dimensional structure. However, the number of Van der Waals, ionic, and hydrogen interactions, and disulfide bonds was not determinant for the biochemical characteristics presented by these enzymes. The phytase KM873028, from cluster 3, was selected for characterization studies, but its expression in Pichia pastoris generated a protein with properties distinct from those derived from the same sequence expressed in a prokaryotic system. It is likely that the differences observed are associated with the location of the interactions in the structures, non-conserved amino acid residues found around the catalytic site, and post-translational modifications inherent to the expression systems. These possibilities highlight the relevance of strategic choices related to enzyme expression aiming at its production and industrial feasibility. |
publishDate |
2022 |
dc.date.none.fl_str_mv |
2022-08-06 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://rsdjournal.org/index.php/rsd/article/view/32765 10.33448/rsd-v11i10.32765 |
url |
https://rsdjournal.org/index.php/rsd/article/view/32765 |
identifier_str_mv |
10.33448/rsd-v11i10.32765 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
https://rsdjournal.org/index.php/rsd/article/view/32765/27940 |
dc.rights.driver.fl_str_mv |
https://creativecommons.org/licenses/by/4.0 info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
https://creativecommons.org/licenses/by/4.0 |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Research, Society and Development |
publisher.none.fl_str_mv |
Research, Society and Development |
dc.source.none.fl_str_mv |
Research, Society and Development; Vol. 11 No. 10; e427111032765 Research, Society and Development; Vol. 11 Núm. 10; e427111032765 Research, Society and Development; v. 11 n. 10; e427111032765 2525-3409 reponame:Research, Society and Development instname:Universidade Federal de Itajubá (UNIFEI) instacron:UNIFEI |
instname_str |
Universidade Federal de Itajubá (UNIFEI) |
instacron_str |
UNIFEI |
institution |
UNIFEI |
reponame_str |
Research, Society and Development |
collection |
Research, Society and Development |
repository.name.fl_str_mv |
Research, Society and Development - Universidade Federal de Itajubá (UNIFEI) |
repository.mail.fl_str_mv |
rsd.articles@gmail.com |
_version_ |
1797052821666791424 |