POLYPHENOL OXIDASE AND PEROXIDASE ENZYME ASSAYS IN SWEET POTATO CULTIVARS HARVESTED AT DIFFERENT TIMES
Autor(a) principal: | |
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Data de Publicação: | 2019 |
Outros Autores: | , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Revista Caatinga |
Texto Completo: | https://periodicos.ufersa.edu.br/caatinga/article/view/8070 |
Resumo: | Enzyme assays are based on methodologies described in the literature. However, the enzyme kinetics must be adjusted to obtain more reliable results. This study aimed to adjust assays by testing different polyphenol oxidase (PPO) and peroxidase (POD) extract amounts and reaction times in sweet potato cultivars harvested at different times. Sweet potato cultivars Paraná, Mãe de Família, and ESAM1 were harvested at 120, 150, and 180 days after planting and minimally processed. A 0.25 g sample was used to determine PPO and POD activities immediately after minimal processing at each harvest. Extraction was performed in 1500 μL phosphate buffer (0.2 M, pH 6.0). The PPO assay was performed by adding 10–50 μL extract, 1490–1450 μL phosphate buffer (0.2 M, pH 6.0), and 1500 μL catechol (0.2 M). The POD assay was carried out by adding 10–50 μL extract to a reaction medium containing 1790–1750 μL phosphate buffer (0.2 M, pH 6), 100 μl guaiacol (5 g L-1), and 100 μL hydrogen peroxide (0.8 g L-1). In both cases, the evaluated reaction times were 1, 2, and 3 min. In the three cultivars, PPO and POD activities increased with the volume of extract and reaction time at all harvest times. The enzyme extract volume of 10 μL for 2 min promoted a continuous increase in PPO and POD enzyme activities in all studied cultivars and at all reaction times. |
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POLYPHENOL OXIDASE AND PEROXIDASE ENZYME ASSAYS IN SWEET POTATO CULTIVARS HARVESTED AT DIFFERENT TIMESENSAIOS ENZIMÁTICOS DA POLIFENOLOXIDASE E PEROXIDASE EM CULTIVARES DE BATATA DOCE COLHIDAS EM DIFERENTES ÉPOCASIpomoea batatas L. (Lam.). Standardization. Minimum processing.Ipomoea batatas L. (Lam.). Padronização. Processamento mínimo.Enzyme assays are based on methodologies described in the literature. However, the enzyme kinetics must be adjusted to obtain more reliable results. This study aimed to adjust assays by testing different polyphenol oxidase (PPO) and peroxidase (POD) extract amounts and reaction times in sweet potato cultivars harvested at different times. Sweet potato cultivars Paraná, Mãe de Família, and ESAM1 were harvested at 120, 150, and 180 days after planting and minimally processed. A 0.25 g sample was used to determine PPO and POD activities immediately after minimal processing at each harvest. Extraction was performed in 1500 μL phosphate buffer (0.2 M, pH 6.0). The PPO assay was performed by adding 10–50 μL extract, 1490–1450 μL phosphate buffer (0.2 M, pH 6.0), and 1500 μL catechol (0.2 M). The POD assay was carried out by adding 10–50 μL extract to a reaction medium containing 1790–1750 μL phosphate buffer (0.2 M, pH 6), 100 μl guaiacol (5 g L-1), and 100 μL hydrogen peroxide (0.8 g L-1). In both cases, the evaluated reaction times were 1, 2, and 3 min. In the three cultivars, PPO and POD activities increased with the volume of extract and reaction time at all harvest times. The enzyme extract volume of 10 μL for 2 min promoted a continuous increase in PPO and POD enzyme activities in all studied cultivars and at all reaction times.Ensaios enzimáticos são baseados em metodologias descritas na literatura. Entretanto, a cinética enzimática deve ser ajustada para resultados mais confiáveis. Objetivou-se ajustar os ensaios testando diferentes volumes de extrato de polifenoloxidase (PPO) e peroxidase (POD) e tempos de reação em cultivares de batata-doce colhidas em diferentes épocas. As cultivares Paraná, Mãe de Família e ESAM1 foram colhidas aos 120, 150 e 180 dias e minimamente processadas. Amostra de 0,25 g foi usada para determinar as atividades de PPO e POD imediatamente após o processamento mínimo em cada colheita. A extração foi realizada em 1500 μL de tampão fosfato (0,2 M, pH 6,0). O ensaio para PPO foi realizado pela adição de 10 a 50 μL de extrato, 1490 a 1450 μL de tampão fosfato (0,2 M, pH 6,0) e 1500 μL de catecol (0,2 M). O ensaio para a POD foi realizado adicionando 10–50 μL de extrato a um meio de reação contendo 1790 a 1750 μL de tampão fosfato (0,2 M, pH 6), 100 μl de guaiacol (5 g L-1) e 100 μL de peróxido de hidrogênio (0,8 g L-1). Em ambos os casos, os tempos de reação avaliados foram 1, 2 e 3 min. Nas três cultivares, as atividades da PPO e POD aumentaram com o volume de extrato e tempo de reação em todas as épocas de colheita. Alíquota de 10 μL de extrato por 2 minutos promoveu um incremento contínuo na atividade da PPO e POD em todas as cultivares e tempos de reação estudados.Universidade Federal Rural do Semi-Árido2019-05-22info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://periodicos.ufersa.edu.br/caatinga/article/view/807010.1590/1983-21252019v32n226rcREVISTA CAATINGA; Vol. 32 No. 2 (2019); 537-542Revista Caatinga; v. 32 n. 2 (2019); 537-5421983-21250100-316Xreponame:Revista Caatingainstname:Universidade Federal Rural do Semi-Árido (UFERSA)instacron:UFERSAenghttps://periodicos.ufersa.edu.br/caatinga/article/view/8070/9969Copyright (c) 2019 Revista Caatingainfo:eu-repo/semantics/openAccessAlmeida, Samara Lopes deMorais, Maria Aparecida dos SantosAlbuquerque, José Ricardo Tavares deBarros Júnior, Aurélio PaesSimões, Adriano do NascimentoFonseca, Kelem Silva2023-07-20T17:19:50Zoai:ojs.periodicos.ufersa.edu.br:article/8070Revistahttps://periodicos.ufersa.edu.br/index.php/caatinga/indexPUBhttps://periodicos.ufersa.edu.br/index.php/caatinga/oaipatricio@ufersa.edu.br|| caatinga@ufersa.edu.br1983-21250100-316Xopendoar:2024-04-29T09:46:38.091493Revista Caatinga - Universidade Federal Rural do Semi-Árido (UFERSA)true |
dc.title.none.fl_str_mv |
POLYPHENOL OXIDASE AND PEROXIDASE ENZYME ASSAYS IN SWEET POTATO CULTIVARS HARVESTED AT DIFFERENT TIMES ENSAIOS ENZIMÁTICOS DA POLIFENOLOXIDASE E PEROXIDASE EM CULTIVARES DE BATATA DOCE COLHIDAS EM DIFERENTES ÉPOCAS |
title |
POLYPHENOL OXIDASE AND PEROXIDASE ENZYME ASSAYS IN SWEET POTATO CULTIVARS HARVESTED AT DIFFERENT TIMES |
spellingShingle |
POLYPHENOL OXIDASE AND PEROXIDASE ENZYME ASSAYS IN SWEET POTATO CULTIVARS HARVESTED AT DIFFERENT TIMES Almeida, Samara Lopes de Ipomoea batatas L. (Lam.). Standardization. Minimum processing. Ipomoea batatas L. (Lam.). Padronização. Processamento mínimo. |
title_short |
POLYPHENOL OXIDASE AND PEROXIDASE ENZYME ASSAYS IN SWEET POTATO CULTIVARS HARVESTED AT DIFFERENT TIMES |
title_full |
POLYPHENOL OXIDASE AND PEROXIDASE ENZYME ASSAYS IN SWEET POTATO CULTIVARS HARVESTED AT DIFFERENT TIMES |
title_fullStr |
POLYPHENOL OXIDASE AND PEROXIDASE ENZYME ASSAYS IN SWEET POTATO CULTIVARS HARVESTED AT DIFFERENT TIMES |
title_full_unstemmed |
POLYPHENOL OXIDASE AND PEROXIDASE ENZYME ASSAYS IN SWEET POTATO CULTIVARS HARVESTED AT DIFFERENT TIMES |
title_sort |
POLYPHENOL OXIDASE AND PEROXIDASE ENZYME ASSAYS IN SWEET POTATO CULTIVARS HARVESTED AT DIFFERENT TIMES |
author |
Almeida, Samara Lopes de |
author_facet |
Almeida, Samara Lopes de Morais, Maria Aparecida dos Santos Albuquerque, José Ricardo Tavares de Barros Júnior, Aurélio Paes Simões, Adriano do Nascimento Fonseca, Kelem Silva |
author_role |
author |
author2 |
Morais, Maria Aparecida dos Santos Albuquerque, José Ricardo Tavares de Barros Júnior, Aurélio Paes Simões, Adriano do Nascimento Fonseca, Kelem Silva |
author2_role |
author author author author author |
dc.contributor.author.fl_str_mv |
Almeida, Samara Lopes de Morais, Maria Aparecida dos Santos Albuquerque, José Ricardo Tavares de Barros Júnior, Aurélio Paes Simões, Adriano do Nascimento Fonseca, Kelem Silva |
dc.subject.por.fl_str_mv |
Ipomoea batatas L. (Lam.). Standardization. Minimum processing. Ipomoea batatas L. (Lam.). Padronização. Processamento mínimo. |
topic |
Ipomoea batatas L. (Lam.). Standardization. Minimum processing. Ipomoea batatas L. (Lam.). Padronização. Processamento mínimo. |
description |
Enzyme assays are based on methodologies described in the literature. However, the enzyme kinetics must be adjusted to obtain more reliable results. This study aimed to adjust assays by testing different polyphenol oxidase (PPO) and peroxidase (POD) extract amounts and reaction times in sweet potato cultivars harvested at different times. Sweet potato cultivars Paraná, Mãe de Família, and ESAM1 were harvested at 120, 150, and 180 days after planting and minimally processed. A 0.25 g sample was used to determine PPO and POD activities immediately after minimal processing at each harvest. Extraction was performed in 1500 μL phosphate buffer (0.2 M, pH 6.0). The PPO assay was performed by adding 10–50 μL extract, 1490–1450 μL phosphate buffer (0.2 M, pH 6.0), and 1500 μL catechol (0.2 M). The POD assay was carried out by adding 10–50 μL extract to a reaction medium containing 1790–1750 μL phosphate buffer (0.2 M, pH 6), 100 μl guaiacol (5 g L-1), and 100 μL hydrogen peroxide (0.8 g L-1). In both cases, the evaluated reaction times were 1, 2, and 3 min. In the three cultivars, PPO and POD activities increased with the volume of extract and reaction time at all harvest times. The enzyme extract volume of 10 μL for 2 min promoted a continuous increase in PPO and POD enzyme activities in all studied cultivars and at all reaction times. |
publishDate |
2019 |
dc.date.none.fl_str_mv |
2019-05-22 |
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://periodicos.ufersa.edu.br/caatinga/article/view/8070 10.1590/1983-21252019v32n226rc |
url |
https://periodicos.ufersa.edu.br/caatinga/article/view/8070 |
identifier_str_mv |
10.1590/1983-21252019v32n226rc |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
https://periodicos.ufersa.edu.br/caatinga/article/view/8070/9969 |
dc.rights.driver.fl_str_mv |
Copyright (c) 2019 Revista Caatinga info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Copyright (c) 2019 Revista Caatinga |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal Rural do Semi-Árido |
publisher.none.fl_str_mv |
Universidade Federal Rural do Semi-Árido |
dc.source.none.fl_str_mv |
REVISTA CAATINGA; Vol. 32 No. 2 (2019); 537-542 Revista Caatinga; v. 32 n. 2 (2019); 537-542 1983-2125 0100-316X reponame:Revista Caatinga instname:Universidade Federal Rural do Semi-Árido (UFERSA) instacron:UFERSA |
instname_str |
Universidade Federal Rural do Semi-Árido (UFERSA) |
instacron_str |
UFERSA |
institution |
UFERSA |
reponame_str |
Revista Caatinga |
collection |
Revista Caatinga |
repository.name.fl_str_mv |
Revista Caatinga - Universidade Federal Rural do Semi-Árido (UFERSA) |
repository.mail.fl_str_mv |
patricio@ufersa.edu.br|| caatinga@ufersa.edu.br |
_version_ |
1797674027673714688 |