Immobilized-enzyme electrode for nicotinamide adenine dinucleotide (reduced form) (NADH) sensing and application to the kinetic studies of NADH dependent dehydrogenases

Hsien Chang Chang, Akinori Ueno, Hiroshi Yamada, Tomokazu Matsue, Isamu Uchida

Research output: Contribution to journalArticlepeer-review

26 Citations (Scopus)

Abstract

Amperometric determination of nicotinamide adenine dinucleotide (reduced form) (NADH) at an immobilized-diaphorase (Dp) electrode is described. The measurement was conducted using ferrocenylmethanol as a mediator in a stirred solution at 0.20 V versus a saturated calomel electrode. A linear relationship between the steady-state current and the concentration of NADH was found over the range 0.005-0.125 mmol dm-3. The immobilized-Dp electrode showed outstanding stability and the current response reached a steady state within 2-3 seconds upon addition of NADH. The proposed electrode was used to follow the reactions of pig heart lactate dehydrogenase and horse liver alcohol dehydrogenase. The kinetic investigation using the immobilized-Dp electrode gave the kinetic parameters (Michaelis constants, Km values, and maximum velocities, Vm values), which were in satisfactory agreement with those determined by a conventional spectrophotometric method.

Original languageEnglish
Pages (from-to)793-796
Number of pages4
JournalThe Analyst
Volume116
Issue number8
DOIs
Publication statusPublished - 1991

Keywords

  • Diaphorase
  • Immobilized-enzyme electrode
  • Nicotinamide adenine dinucleotide sensor

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Environmental Chemistry
  • Spectroscopy
  • Electrochemistry

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