Determination of Michaelis-Menten and inhibitor constants by an open-closed flow injection approach (Application to the alkaline phosphatase/theophylline system)

Autor:	Juan Manuel Fernández-Romero, M. Sánchez-Cabezudo, M. D. Luque de Castro
Rok vydání:	1995
Předmět:	Chromatography Immobilized enzyme biology Substrate (chemistry) Phosphate Michaelis–Menten kinetics Analytical Chemistry chemistry.chemical_compound chemistry Biocatalysis Enzyme inhibitor medicine biology.protein Alkaline phosphatase Theophylline medicine.drug
Zdroj:	Talanta. 42:1103-1110
ISSN:	0039-9140
DOI:	10.1016/0039-9140(95)01532-g
Popis:	An automatic method for the estimation of Michaelis-Menten and inhibitor constants in biochemical systems was developed using an unsegmented open-closed flow manifold, and the Lineweaver-Burk and Dixon plot methods. The methods were applied to both the determination of Km of the enzymatic reaction catalyzed by bovine alkaline phosphatase using 4-methylumbelliferone phosphate as substrate, and the determination of the Ki constant from theophylline as an inhibitor of this biochemical system. The data treatment and comparative results are presented. The values obtained agree with those provided by a conventional method with the biocatalyst in solution.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_________::b9a3840f001f56877b73653e00abbabb https://doi.org/10.1016/0039-9140(95)01532-g Zobrazit plný text záznamu Full Text from ScienceDirect