著作: 峯 大典/[水口 仁志]/[髙栁 俊夫]/Kinetic analyses of two-steps oxidation from L-tyrosine to L-dopaquinone with tyrosinase by capillary electrophoresis/dynamic frontal analysis/[Analytical Biochemistry: Methods in the Biological Sciences]
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種別 | 必須 | 学術論文(審査論文) | |||
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言語 | 必須 | 英語 | |||
招待 | 推奨 | ||||
審査 | 推奨 | Peer Review | |||
カテゴリ | 推奨 | ||||
共著種別 | 推奨 | 単独著作(徳島大学内の単一の研究グループ(研究室等)内の研究 (単著も含む)) | |||
学究種別 | 推奨 | ||||
組織 | 推奨 | ||||
著者 | 必須 | ||||
題名 | 必須 |
(英) Kinetic analyses of two-steps oxidation from L-tyrosine to L-dopaquinone with tyrosinase by capillary electrophoresis/dynamic frontal analysis |
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副題 | 任意 | ||||
要約 | 任意 |
(英) Tyrosinase catalyzes the oxidation of l-tyrosine in two stages to produce l-dopa and l-dopaquinone stepwise, and l-dopaquinone is subsequently converted to dopachrome. Most of the conventional analyses subjected only one-step reaction from l-tyrosine to l-dopa or from l-dopa to l-dopaquinone. In this study, kinetic analyses of two-steps oxidation of l-tyrosine with tyrosinase were made by capillary electrophoresis/dynamic frontal analysis (CE/DFA). When l-dopa was introduced into a capillary as a sample plug in a CE/DFA format, the enzymatic oxidation continuously occurred during the electrophoresis, and the product l-dopaquinone was subsequently converted to dopachrome which was detected as a plateau signal. A Michaelis-Menten constant of the second-step kinetic reaction, K, was determined as 0.45 ± 0.03 mmol L. In the analysis of the first-step kinetic reaction from l-tyrosine to l-dopa, l-dopa was not resolved by CE/DFA because both l-tyrosine and l-dopa are electrically neutral. The l-dopa formed and co-migrated at the l-tyrosine zone was calibrated beforehand with the final product of dopachrome detected as a plateau signal. Constantly formed l-dopa was successfully detected as a plateau signal of dopachrome, and a Michaelis-Menten constant of K was also determined as 0.061 ± 0.009 mmol L by the CE/DFA. CE/DFA is applicable to two-steps enzymatic reactions. |
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キーワード | 推奨 |
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発行所 | 推奨 | Elsevier Science | |||
誌名 | 必須 |
Analytical Biochemistry: Methods in the Biological Sciences([Elsevier Science])
(pISSN: 0003-2697, eISSN: 1096-0309)
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巻 | 必須 | 655 | |||
号 | 必須 | --- | |||
頁 | 必須 | 114856 114856 | |||
都市 | 任意 | ||||
年月日 | 必須 | 2022年 10月 15日 | |||
URL | 任意 | ||||
DOI | 任意 | 10.1016/j.ab.2022.114856 (→Scopusで検索) | |||
PMID | 任意 | 35964734 (→Scopusで検索) | |||
CRID | 任意 | ||||
WOS | 任意 | ||||
Scopus | 任意 | ||||
評価値 | 任意 | ||||
被引用数 | 任意 | ||||
指導教員 | 推奨 | ||||
備考 | 任意 |
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