Some Minor Characteristics of Spectrophotometric Determination of Antioxidant System and Phenolic Metabolism Enzyme Activity in Wood Plant Tissues of Pinus sylvestris L.
Abstract: A comprehensive study of enzymes of the antioxidant system (AOS) and phenolic metabolism is an actual subject of biochemical research; changes in the activity of these enzymes can be used as a diagnostic sign. At the same time, practically little attention has been paid to describing the r...
Published in: | Protein & Peptide Letters |
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2022
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crbenthamsciepub:10.2174/0929866529666220414104747 2024-06-23T07:54:19+00:00 Some Minor Characteristics of Spectrophotometric Determination of Antioxidant System and Phenolic Metabolism Enzyme Activity in Wood Plant Tissues of Pinus sylvestris L. Ershova, Maria A. Nikerova, Kseniya M. Galibina, Natalia A. Sofronova, Irina N. Borodina, Marina N. Russian Science Foundation 2022 http://dx.doi.org/10.2174/0929866529666220414104747 https://www.eurekaselect.com/article/download?doi=10.2174/0929866529666220414104747 https://www.eurekaselect.com/203572/article en eng Bentham Science Publishers Ltd. Protein & Peptide Letters volume 29, issue 8, page 711-720 ISSN 0929-8665 journal-article 2022 crbenthamsciepub https://doi.org/10.2174/0929866529666220414104747 2024-06-13T04:10:33Z Abstract: A comprehensive study of enzymes of the antioxidant system (AOS) and phenolic metabolism is an actual subject of biochemical research; changes in the activity of these enzymes can be used as a diagnostic sign. At the same time, practically little attention has been paid to describing the regularities of these enzymatic reactions. The article presents the chemical kinetics study of reactions catalyzed by superoxide dismutase, catalase, peroxidase, polyphenol oxidase, and phenylalanine ammonia-lyase in Scots pine trunk tissues (Pinus sylvestris L.). The dependence of the enzyme reaction rate on the enzyme concentration and the substrate concentration is presented, and the pH-optimum for each reaction is established. Background: Determination of AOS enzyme activity and PAL activity in woody plants has many difficulties. The chemical composition of pine trunk tissues affects determining AOS enzyme activity and PAL activity. Spectrophotometric determination of AOS enzyme activity and PAL activity gives perfect results when considering all additional controls by taking into account minor characteristics. Objective: This study aimed at determining the AOS enzyme activity in 40-year-old Scots pine (Pinus sylvestris L.) plants growing in the Karelian (Russia) forest seed plantation. Method: Plant tissues were ground in liquid nitrogen to a uniform mass and homogenized at 4 °C in the buffer containing 50 mM HEPES (pH 7.5), 1 mM EDTA, 1 mM EGTA, 3 mM DTT, 5 mM MgCl2, and 0.5 mM PMSF. After 15-min of extraction, the homogenate was centrifuged at 12000 g for 10 min (MPW-351R centrifuge, Poland). The supernatant was purified on 20 cm3 columns with Sephadex G-250. Aliquots with the highest protein amount were collected. In tissues, the protein concentration was 10-50 μg/ml. Proteins in the extracts were quantified by a Bradford assay. The enzyme activity was determined spectrophotometrically on a SpectroStar Nano plate spectrophotometer (BMG Labtech, Germany). Results: Our study made it possible to modify the methods ... Article in Journal/Newspaper karelia* karelian Bentham Science Publishers Protein & Peptide Letters 29 |
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Bentham Science Publishers |
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crbenthamsciepub |
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English |
description |
Abstract: A comprehensive study of enzymes of the antioxidant system (AOS) and phenolic metabolism is an actual subject of biochemical research; changes in the activity of these enzymes can be used as a diagnostic sign. At the same time, practically little attention has been paid to describing the regularities of these enzymatic reactions. The article presents the chemical kinetics study of reactions catalyzed by superoxide dismutase, catalase, peroxidase, polyphenol oxidase, and phenylalanine ammonia-lyase in Scots pine trunk tissues (Pinus sylvestris L.). The dependence of the enzyme reaction rate on the enzyme concentration and the substrate concentration is presented, and the pH-optimum for each reaction is established. Background: Determination of AOS enzyme activity and PAL activity in woody plants has many difficulties. The chemical composition of pine trunk tissues affects determining AOS enzyme activity and PAL activity. Spectrophotometric determination of AOS enzyme activity and PAL activity gives perfect results when considering all additional controls by taking into account minor characteristics. Objective: This study aimed at determining the AOS enzyme activity in 40-year-old Scots pine (Pinus sylvestris L.) plants growing in the Karelian (Russia) forest seed plantation. Method: Plant tissues were ground in liquid nitrogen to a uniform mass and homogenized at 4 °C in the buffer containing 50 mM HEPES (pH 7.5), 1 mM EDTA, 1 mM EGTA, 3 mM DTT, 5 mM MgCl2, and 0.5 mM PMSF. After 15-min of extraction, the homogenate was centrifuged at 12000 g for 10 min (MPW-351R centrifuge, Poland). The supernatant was purified on 20 cm3 columns with Sephadex G-250. Aliquots with the highest protein amount were collected. In tissues, the protein concentration was 10-50 μg/ml. Proteins in the extracts were quantified by a Bradford assay. The enzyme activity was determined spectrophotometrically on a SpectroStar Nano plate spectrophotometer (BMG Labtech, Germany). Results: Our study made it possible to modify the methods ... |
author2 |
Russian Science Foundation |
format |
Article in Journal/Newspaper |
author |
Ershova, Maria A. Nikerova, Kseniya M. Galibina, Natalia A. Sofronova, Irina N. Borodina, Marina N. |
spellingShingle |
Ershova, Maria A. Nikerova, Kseniya M. Galibina, Natalia A. Sofronova, Irina N. Borodina, Marina N. Some Minor Characteristics of Spectrophotometric Determination of Antioxidant System and Phenolic Metabolism Enzyme Activity in Wood Plant Tissues of Pinus sylvestris L. |
author_facet |
Ershova, Maria A. Nikerova, Kseniya M. Galibina, Natalia A. Sofronova, Irina N. Borodina, Marina N. |
author_sort |
Ershova, Maria A. |
title |
Some Minor Characteristics of Spectrophotometric Determination of Antioxidant System and Phenolic Metabolism Enzyme Activity in Wood Plant Tissues of Pinus sylvestris L. |
title_short |
Some Minor Characteristics of Spectrophotometric Determination of Antioxidant System and Phenolic Metabolism Enzyme Activity in Wood Plant Tissues of Pinus sylvestris L. |
title_full |
Some Minor Characteristics of Spectrophotometric Determination of Antioxidant System and Phenolic Metabolism Enzyme Activity in Wood Plant Tissues of Pinus sylvestris L. |
title_fullStr |
Some Minor Characteristics of Spectrophotometric Determination of Antioxidant System and Phenolic Metabolism Enzyme Activity in Wood Plant Tissues of Pinus sylvestris L. |
title_full_unstemmed |
Some Minor Characteristics of Spectrophotometric Determination of Antioxidant System and Phenolic Metabolism Enzyme Activity in Wood Plant Tissues of Pinus sylvestris L. |
title_sort |
some minor characteristics of spectrophotometric determination of antioxidant system and phenolic metabolism enzyme activity in wood plant tissues of pinus sylvestris l. |
publisher |
Bentham Science Publishers Ltd. |
publishDate |
2022 |
url |
http://dx.doi.org/10.2174/0929866529666220414104747 https://www.eurekaselect.com/article/download?doi=10.2174/0929866529666220414104747 https://www.eurekaselect.com/203572/article |
genre |
karelia* karelian |
genre_facet |
karelia* karelian |
op_source |
Protein & Peptide Letters volume 29, issue 8, page 711-720 ISSN 0929-8665 |
op_doi |
https://doi.org/10.2174/0929866529666220414104747 |
container_title |
Protein & Peptide Letters |
container_volume |
29 |
_version_ |
1802646449520902144 |