Enzyme coupled lactose sensors based on bienzyme membrane prepared by an avidin/biotin method

Tomonori Hoshi, Hidekazu Saiki, Katsushi Takeuchi

Research output: Contribution to journalArticlepeer-review

3 Citations (Scopus)


Lactose sensors were fabricated by an avidin/biotin method. Biotinylated glucose oxidase (GOx) and β-galactosidase (β-Gal) formed bienzyme membrane on a Pt electrode by a layer-by-layer deposition of avidin and the biotinylated enzymes. The modified electrode functioned satisfactorily as a lactose sensor, in which two enzymatic reactions were coupled. Ten layers of GOx were deposited on the electrode, and then β-Gal layers were deposited step by step on the GOx layer. The response current of the sensor to lactose increased with increasing the number of depositions of β-Gal; Imax values were 8.4 nA for 2 layers, and 81.3 nA for 12 layer of β-Gal. Three types of arrangement of the enzymes in the membrane were prepared; (5 × GOx)+(5 × β-Gal), (5 × β-Gal)+(5x GOx) and 5 × (GOx+β-Gal). The sensor modified with the alternate layers of GOx and β-Gal, 5x (GOx+β-Gal), exhibited a highest signal for lactose. The avidin/biotin method was shown to be useful to optimize composition of the bienzyme membrane.

Original languageEnglish
Pages (from-to)576-580
Number of pages5
JournalIEEJ Transactions on Sensors and Micromachines
Issue number11
Publication statusPublished - 1999


  • avidin
  • bienzyme membrane
  • biotin
  • lactose sensor

ASJC Scopus subject areas

  • Mechanical Engineering
  • Electrical and Electronic Engineering


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