Immobilization of tyrosinase in chitosan film for an optical detection of phenol

Jaafar Abdullah, Musa Ahmad, Nadarajah Karuppiah, Yook Heng Lee, Hamidah Sidek

Research output: Contribution to journalArticle

72 Citations (Scopus)

Abstract

An optical biosensor based on the immobilized tyrosinase enzyme in a chitosan film is described for the detection of phenol. The film was prepared by depositing a chitosan solution containing tyrosinase onto a microscope glass slide via spin coating technique. The quinone produced from the enzymatic oxidation of phenol was reacted with 3-methyl-2-benzothiazolinone hydrazone (MBTH) to produce a maroon color adduct. The color intensity of the adduct was found to increase proportionally with the increase of the substrate concentrations after 15 min exposure to the substrate. The optimum enzyme loading and chitosan concentrations were found to be 0.9 mg and 2% (w/v), respectively. The biosensor also demonstrated optimum activity at pH 6-7, optimum temperature of 45 °C and a linear response in the phenol concentration range of 2.5-70.0 μM. The response characteristics of the biosensor towards different phenolic compounds were determined. It was found that the biosensor gave a linear response in the concentration range of 2.5-50.0 μM for 4-chlorophenol, 2.5-100.0 μM for m-cresol and 12.5-400.0 μM for p-cresol. The limit of detection and the apparent Michaels-Menten constant (K m) of the biosensor were 0.9, 1.0, 1.0, 3.0 μM and 0.038, 0.037, 0.056, 0.201 mM for 4-chlorophenol, phenol, m-cresol and p-cresol, respectively. It showed good stability for at least 2 months.

Original languageEnglish
Pages (from-to)604-609
Number of pages6
JournalSensors and Actuators, B: Chemical
Volume114
Issue number2
DOIs
Publication statusPublished - 26 Apr 2006

Fingerprint

Monophenol Monooxygenase
Chitosan
cresols
Phenol
immobilization
bioinstrumentation
Biosensors
phenols
Phenols
adducts
enzymes
Enzymes
Color
color
hydrazones
Coating techniques
Spin coating
quinones
Substrates
chutes

Keywords

  • Chitosan
  • MBTH
  • Optical biosensor
  • Phenol
  • Tyrosinase

ASJC Scopus subject areas

  • Analytical Chemistry
  • Electrochemistry
  • Electrical and Electronic Engineering

Cite this

Immobilization of tyrosinase in chitosan film for an optical detection of phenol. / Abdullah, Jaafar; Ahmad, Musa; Karuppiah, Nadarajah; Lee, Yook Heng; Sidek, Hamidah.

In: Sensors and Actuators, B: Chemical, Vol. 114, No. 2, 26.04.2006, p. 604-609.

Research output: Contribution to journalArticle

Abdullah, Jaafar ; Ahmad, Musa ; Karuppiah, Nadarajah ; Lee, Yook Heng ; Sidek, Hamidah. / Immobilization of tyrosinase in chitosan film for an optical detection of phenol. In: Sensors and Actuators, B: Chemical. 2006 ; Vol. 114, No. 2. pp. 604-609.
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