Rapid autographic method for detection of enzymatic browning inhibitors based on enzyme immobilization

This work describes a TLC-coupled bioautographic assay suitable for the separation and detection of apple polyphenol oxidase (PPO) inhibitors from natural extracts. PPO was immobilised in agar containing L-DOPA as substrate and 3-methyl-2-benzothiazolinone hydrazone hydrochloride (MBTH) to enhance colour development. The inhibition was detected as white spots on reddish background. Minimum amount of PPO inhibitors detected was 0.0125 μg of 4-hexylresorcinol, 0.025 μg of ascorbic acid, 0.5 μg of cysteine and 1 μg of kojic acid. The assay was compatible with normal and reverse phase TLC systems and allows detecting compounds that directly had action on the enzyme as well as agents that could convert quinones back to their reduced form. The chromatographic run evidenced the different nature of enzymatic browning inhibitory compounds from garlic and onion extracts. Using natural enzymes will provide a fast and cheap alternative for target specific exploration of natural enzymatic inhibitors.

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Main Authors: Micheloni, Oscar B., Farroni, Abel Eduardo, Garcia, Paula, Furlan, Ricardo Luis Eugenio
Format: info:ar-repo/semantics/artículo biblioteca
Language:eng
Published: 2018-07
Subjects:Tecnología de Alimentos, Food Technology, Polifenoles, Pardeamiento Enzimático, Extractos Vegetales, Polyphenols, Enzymic Browning, Plant Extracts,
Online Access:http://hdl.handle.net/20.500.12123/2935
https://www.sciencedirect.com/science/article/pii/S0308814618311646
https://doi.org/10.1016/j.foodchem.2018.07.025
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spelling oai:localhost:20.500.12123-29352024-03-11T10:11:05Z Rapid autographic method for detection of enzymatic browning inhibitors based on enzyme immobilization Micheloni, Oscar B. Farroni, Abel Eduardo Garcia, Paula Furlan, Ricardo Luis Eugenio Tecnología de Alimentos Food Technology Polifenoles Pardeamiento Enzimático Extractos Vegetales Polyphenols Enzymic Browning Plant Extracts This work describes a TLC-coupled bioautographic assay suitable for the separation and detection of apple polyphenol oxidase (PPO) inhibitors from natural extracts. PPO was immobilised in agar containing L-DOPA as substrate and 3-methyl-2-benzothiazolinone hydrazone hydrochloride (MBTH) to enhance colour development. The inhibition was detected as white spots on reddish background. Minimum amount of PPO inhibitors detected was 0.0125 μg of 4-hexylresorcinol, 0.025 μg of ascorbic acid, 0.5 μg of cysteine and 1 μg of kojic acid. The assay was compatible with normal and reverse phase TLC systems and allows detecting compounds that directly had action on the enzyme as well as agents that could convert quinones back to their reduced form. The chromatographic run evidenced the different nature of enzymatic browning inhibitory compounds from garlic and onion extracts. Using natural enzymes will provide a fast and cheap alternative for target specific exploration of natural enzymatic inhibitors. EEA Pergamino Fil: Micheloni, Oscar B. Universidad Nacional del Noroeste de la Provincia de Buenos Aires (UNNOBA). Departamento de Ciencias Básicas; Argentina. Fil: Farroni, Abel E. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Pergamino. Laboratorio de Calidad de Alimento, Suelos y Agua; Argentina. Fil: García, Paula. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina. Fil: Furlan, Ricardo L.E.. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina. 2018-07-31T18:27:30Z 2018-07-31T18:27:30Z 2018-07 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/2935 https://www.sciencedirect.com/science/article/pii/S0308814618311646 0308-8146 https://doi.org/10.1016/j.foodchem.2018.07.025 eng info:eu-repo/semantics/restrictedAccess application/pdf Food chemistry. 269 : 638-643. (2018)
institution INTA AR
collection DSpace
country Argentina
countrycode AR
component Bibliográfico
access En linea
databasecode dig-inta-ar
tag biblioteca
region America del Sur
libraryname Biblioteca Central del INTA Argentina
language eng
topic Tecnología de Alimentos
Food Technology
Polifenoles
Pardeamiento Enzimático
Extractos Vegetales
Polyphenols
Enzymic Browning
Plant Extracts
Tecnología de Alimentos
Food Technology
Polifenoles
Pardeamiento Enzimático
Extractos Vegetales
Polyphenols
Enzymic Browning
Plant Extracts
spellingShingle Tecnología de Alimentos
Food Technology
Polifenoles
Pardeamiento Enzimático
Extractos Vegetales
Polyphenols
Enzymic Browning
Plant Extracts
Tecnología de Alimentos
Food Technology
Polifenoles
Pardeamiento Enzimático
Extractos Vegetales
Polyphenols
Enzymic Browning
Plant Extracts
Micheloni, Oscar B.
Farroni, Abel Eduardo
Garcia, Paula
Furlan, Ricardo Luis Eugenio
Rapid autographic method for detection of enzymatic browning inhibitors based on enzyme immobilization
description This work describes a TLC-coupled bioautographic assay suitable for the separation and detection of apple polyphenol oxidase (PPO) inhibitors from natural extracts. PPO was immobilised in agar containing L-DOPA as substrate and 3-methyl-2-benzothiazolinone hydrazone hydrochloride (MBTH) to enhance colour development. The inhibition was detected as white spots on reddish background. Minimum amount of PPO inhibitors detected was 0.0125 μg of 4-hexylresorcinol, 0.025 μg of ascorbic acid, 0.5 μg of cysteine and 1 μg of kojic acid. The assay was compatible with normal and reverse phase TLC systems and allows detecting compounds that directly had action on the enzyme as well as agents that could convert quinones back to their reduced form. The chromatographic run evidenced the different nature of enzymatic browning inhibitory compounds from garlic and onion extracts. Using natural enzymes will provide a fast and cheap alternative for target specific exploration of natural enzymatic inhibitors.
format info:ar-repo/semantics/artículo
topic_facet Tecnología de Alimentos
Food Technology
Polifenoles
Pardeamiento Enzimático
Extractos Vegetales
Polyphenols
Enzymic Browning
Plant Extracts
author Micheloni, Oscar B.
Farroni, Abel Eduardo
Garcia, Paula
Furlan, Ricardo Luis Eugenio
author_facet Micheloni, Oscar B.
Farroni, Abel Eduardo
Garcia, Paula
Furlan, Ricardo Luis Eugenio
author_sort Micheloni, Oscar B.
title Rapid autographic method for detection of enzymatic browning inhibitors based on enzyme immobilization
title_short Rapid autographic method for detection of enzymatic browning inhibitors based on enzyme immobilization
title_full Rapid autographic method for detection of enzymatic browning inhibitors based on enzyme immobilization
title_fullStr Rapid autographic method for detection of enzymatic browning inhibitors based on enzyme immobilization
title_full_unstemmed Rapid autographic method for detection of enzymatic browning inhibitors based on enzyme immobilization
title_sort rapid autographic method for detection of enzymatic browning inhibitors based on enzyme immobilization
publishDate 2018-07
url http://hdl.handle.net/20.500.12123/2935
https://www.sciencedirect.com/science/article/pii/S0308814618311646
https://doi.org/10.1016/j.foodchem.2018.07.025
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