高感度電気化学的酵素活性測定法の研究

高灵敏度电化学酶活性测定方法研究

• 批准号: 05806016
• 负责人: 千田 貢
• 金额: $ 0.96万
• 依托单位: Fukui Prefectural University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05806016/
• 关键词: 高感度電気化学分析法; 酵素活性測定法; 4-アミノフェノール; ジアホラーゼ; NADH; グルコースオキシダーゼ; グルコース; ジクロロインドフェノール

本年度は特に本高感度法の基礎となる測定条件の詳細な検討を行った。(a)高感度達成の基礎となる酵素サイクリング反応には、酵素反応に特徴的な非直線性が伴う。この点を理論的、実験的に明らかにするため、4-アミノフェノール(PAP)/ジアフォラーゼ(DIA)/NADH系について、PAP及びNADHの濃度効果、特にその非直線性の表われる濃度範囲について理論的考察を加え、一方、実験的にも濃度の非直線性の表われることを確認し、またその濃度範囲が理論的考察とよく対応することを確かめた。分析条件として、NADHの濃度の変動による分析結果(電流側定値)の変動の極小となる条件下、PAPの濃度に対する電流測定値の直線的変化の条件を明らかにし、PAPの10^<-10>Mまでその検出が可能なこと、さらにアルカリホスフアターゼ及びガラクトシダーゼの酵素活性の測定についての予備実験の結果、たとえば前者について10^<-7>unitの測定が可能なことが明らかになった。(b)PAP以外に、その誘導体(4種)及び2-アミノフェノール体(2種)その他について、またDIAに代わる、PAP/グルコースオキシダーゼ/グルコース系についても検討を行なった。特に、PAPに代わる2,4-ジクロロインドフェノールについて、このものはそのその酸化体が比較的安定なことを利用して、上記非直線性の考察の基礎となる酵素反応式をあらためて実験的に確認し、本高感度法基礎をさらに確実なものにすることが出来た。得られた結果の範囲ではPAP/DIA/NADH系以上に著しく高感度が期待される系は見出されなかった。

This year, the high sensitivity method is used to determine the conditions of the test. (a) High sensitivity is the direct sexual companion of high sensitivity, high sensitivity and high sensitivity. This is a review of the theory, information, accuracy, accuracy and accuracy. A review of the theory in the range of degrees. Please make sure that you are in good condition. The results of the analysis of the conditions of analysis and the degree of NADH, the results of the dynamic analysis (current determination) under the condition of small temperature, the chemical conditions of the current measurement of the straight line under the condition of the temperature of PAP, PAPP 10 ^ & lt;-10&gt. The results of the measurement of enzyme activity, the results of the measurement of enzyme activity, the results of the measurement of enzyme activity, the results of the measurement of enzyme activity, the results of the measurement of enzyme activity, the results of the measurement of enzyme activity, the results of the measurement of enzyme activity, the results of the measurement of enzyme activity, the results of the measurement of enzyme activity, the results of the measurement of enzyme activity, the results of the measurement of enzyme activity, the results of the measurement of enzyme activity, the results of the measurement of enzyme activity, the results of the measurement of enzyme activity, the results of the measurement of enzyme activity, the results of the measurement of enzyme activity, the results of the measurement of enzyme activity, the results of the measurement of enzyme activity, the results of the measurement of enzyme activity, the results of the measurement of enzyme activity, the results of the measurement of enzyme activity, the results of the measurement of enzyme activity, the results of the measurement of enzyme activity, the results of the measurement of enzyme activity, the results of the measurement of enzyme activity, the results of the measurement of enzyme activity, the results of the measurement of enzyme activity, the (B) in addition to PAP, there are 4 kinds of drugs and 2 kinds of products. Other models, such as DIA, PAP/, and so on, are registered in other countries. In this paper, the authors studied the stability of the acidified body, the use of The results show that the PAP/DIA/NADH department has a high degree of sensitivity and expects that the system will produce a high level of sensitivity.

项目成果

M.Senda,S.Yamaguchi: "Proceedings of the 5th International Symposium on Redox Mechanisms and International Properties of Molecules of Biological Importance." The Electrochemical Society Inc., 406-414 (1993)

M.Senda，S.Yamaguchi：“第五届氧化还原机制和生物重要分子国际性质国际研讨会论文集”。

千田 貢・山口整毅: "p-Aminophenol/Diaphorase-NADH系における接触電流とそのアンペロメトリーの応用.II." ポーラログラフィー. 39. 36- (1993)

Mitsugu Senda 和 Seitake Yamaguchi：“对氨基苯酚/心肌黄酶-NADH 系统中的接触电流及其在电流分析法中的应用。II。39. 36- (1993)”