A Dual-Enzyme Electrode for the Determination of Flavin Adenine Dinucleotide in the Presence of Riboflavin and Flavin Mononucleotide
作者:
Abdul Rashid,
Mohd. Yatim,
ThomasL. Riechel,
期刊:
Analytical Letters
(Taylor Available online 1984)
卷期:
Volume 17,
issue 9
页码: 835-855
ISSN:0003-2719
年代: 1984
DOI:10.1080/00032718408081484
出版商: Taylor & Francis Group
关键词: Enzyme Electrode;Flavin Adenine Dinucleotide;Riboflavin;Flavin Mononucleotide
数据来源: Taylor
摘要:
An electrode method has been developed for the determination of flavin adenine dinucleotide (FAD) using a potentiometric gas sensor and commercially available enzyme preparations. The construction of the FAD-sensitive electrode is based on immobilizing alkaline phosphatase (E.C. 3.1.3.1) and adenosine deaminase (E.C. 3.5.4.4) on the sensing tip of an ammonia gas sensor. Alkaline phosphatase enzymatically catalyzes the hydrolysis of FAD to adenosine which is subsequently converted to ammonia by adenosine deaminase. The response of the dual-enzyme electrode is linear between 8 × 10−5Mand 4 × 10−3MFAD with a slope of 43 mV/decade concentration at pH 8.5 and 37[ddot]C. The optimum buffer system is 0.5Mdiethanolamine, 1 × 10−3MTris-HCl and 1 × 10−3MMgCl2. Electrodes constructed with enzymes immobilized by cross-linking with glutaraldehyde and bovine serum albumin showed longer life times than electrodes using enzymes entrapped by a dialysis membrane. The electrode is highly selective over riboflavin and flavin mononucleotide, but it does respond to other nucleotides.
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