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Kinetic analysis of an enzymatic hydrolysis of p-nitrophenyl acetate with carboxylesterase by pressure-assisted capillary electrophoresis/dynamic frontal analysis
https://tokushima-u.repo.nii.ac.jp/records/2008415
https://tokushima-u.repo.nii.ac.jp/records/2008415e082fef8-acbb-4f59-bee0-67c1028d8845
| 名前 / ファイル | ライセンス | アクション |
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am_12_48_5846.pdf (1.44 MB)
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| Item type | 文献 / Documents(1) | |||||||||||||||||||||||||||||
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| 公開日 | 2020-11-30 | |||||||||||||||||||||||||||||
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| アクセス権 | open access | |||||||||||||||||||||||||||||
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| 資源タイプ識別子 | http://purl.org/coar/resource_type/c_6501 | |||||||||||||||||||||||||||||
| 資源タイプ | journal article | |||||||||||||||||||||||||||||
| 出版社版DOI | ||||||||||||||||||||||||||||||
| 識別子タイプ | DOI | |||||||||||||||||||||||||||||
| 関連識別子 | https://doi.org/10.1039/D0AY01736A | |||||||||||||||||||||||||||||
| 言語 | ja | |||||||||||||||||||||||||||||
| 関連名称 | 10.1039/D0AY01736A | |||||||||||||||||||||||||||||
| 出版タイプ | ||||||||||||||||||||||||||||||
| 出版タイプ | AM | |||||||||||||||||||||||||||||
| 出版タイプResource | http://purl.org/coar/version/c_ab4af688f83e57aa | |||||||||||||||||||||||||||||
| タイトル | ||||||||||||||||||||||||||||||
| タイトル | Kinetic analysis of an enzymatic hydrolysis of p-nitrophenyl acetate with carboxylesterase by pressure-assisted capillary electrophoresis/dynamic frontal analysis | |||||||||||||||||||||||||||||
| 言語 | en | |||||||||||||||||||||||||||||
| 著者 |
ミネ, マサノリ
× ミネ, マサノリ
× マツモト, ナオヤ
× 水口, 仁志
WEKO
807
× 髙栁, 俊夫
WEKO
549
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| 内容記述タイプ | Abstract | |||||||||||||||||||||||||||||
| 内容記述 | An enzymatic hydrolysis of p‐nitrophenyl acetate with carboxyesterase was analyzed by capillary electrophoresis/dynamic frontal analysis (CE/DFA). A plateau signal was expected with the anionic product of p‐nitrophenol by the CE/DFA applying in‐capillary reaction and the continuous CE resolution of the product from the substrate zone. However, the plateau height was not sufficient, and/or the plateau signal fluctuated and drifted. Therefore, a pressure assist was utilized in the CE/DFA to detect the product zone fast and to average the fluctuated plateau signal by mixing in a laminar flow. The plateau signal became relatively flat and its height was developed by the pressure‐assisted capillary electrophoresis/dynamic frontal analysis (pCE/DFA). The plateau height was used for the Michaelis‐Menten analysis, and a Michaelis‐Menten constant was determined as KM = 0.83 mmol L−1. An enzyme inhibition was also examined with bis (p‐nitrophenyl) phosphate by adding it in the separation buffer. The height of the plateau signal decreased by the inhibition, and a 50% inhibitory concentration was determined as IC50 = 0.79 μmol L−1. The values of KM and IC50 obtained in this study agreed well with the reported values. Since the proposed pCE/DFA includes electrophoretic migration of the substrate zone in a capillary, it is also noticed that the deactivation of the enzyme by ethanol on the preparation of the substrate solution can be avoided, as well as the exclusion of the inhibition by the product. | |||||||||||||||||||||||||||||
| 言語 | en | |||||||||||||||||||||||||||||
| 書誌情報 |
en : Analytical Methods 巻 12, 号 48, p. 5846-5851, 発行日 2020-10-26 |
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| 収録物識別子タイプ | ISSN | |||||||||||||||||||||||||||||
| 収録物識別子 | 17599679 | |||||||||||||||||||||||||||||
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| 出版者 | The Royal Society of Chemistry | |||||||||||||||||||||||||||||
| 言語 | en | |||||||||||||||||||||||||||||
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| 識別子 | 372525 | |||||||||||||||||||||||||||||
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| 言語 | eng | |||||||||||||||||||||||||||||
