Automated Method, Based on Micro-Sequential Injection, for the Study of Enzyme Kinetics and Inhibition

DOI PDF Web Site Web Site Web Site 参考文献26件

この論文をさがす

抄録

A micro-reactor system with continual spectrophotometric detection has been operated in Sequential Injection lab-on-valve (SI-LOV) mode and applied to enzyme kinetics and inhibition studies, using acetylcholinesterase (AChE) and angiotensin-converting enzyme (ACE) as model systems. With the advantages of automation, real-time kinetic measurement, and thorough mixing, the SI-LOV micro-reactor system allows for the monitoring of initial reaction rates and determination of reactant concentrations in the reaction mixture, both of which are essential for the determination of kinetic constants for enzymes and inhibitors. Enzyme, substrate, and inhibitor are precisely metered by the syringe pump and delivered to a stirred micro-reactor, followed by a reference scan that establishes the baseline for the following reaction rate measurement. Michaelis constants (Km) for AChE and ACE were determined to be 0.16 mM and 0.30 mM, respectively, which are consistent with literature values. The type of inhibition (competitive, uncompetitive, or mixed), the dissociation constants for the inhibitors, and the inhibitor dose-response curves were also determined.

収録刊行物

  • Analytical Sciences

    Analytical Sciences 22 (1), 9-14, 2006

    社団法人 日本分析化学会

参考文献 (26)*注記

もっと見る

キーワード

詳細情報 詳細情報について

問題の指摘

ページトップへ