ABSTRACT
A highly sensitive ethanol biosensor based on Acetobacter aceti biofilm, which contains pyrroloquinolinequinone-alcohol dehydrogenase enzyme for halal detection in food and beverages, has been developed. In thisstudy, A. aceti cells were purified by determining the conditions of growth time, incubation temperature, andpH to suppress the yeast growth. Selection of optimum conditions for biofilm formation was also carried out toobtain a stable sensor operation. The optimum biofilm formation was 14 days. The performance of biosensor wasinvestigated by the cyclic voltammetry technique. Linear range, sensitivity, limit of detection (LOD), and limitof quantification (LOQ) were found to be 5 × 10−5–3 × 10−4 %, 43,076 µA %−1, 2.32 × 10−5, and 7.03 × 10−5 %,respectively. The repeatability of biosensor was tested in 1 × 10−5 − 1% ethanol concentration and the relativestandard deviation was 1.08% (for n = 7). The stability studies have shown that the cyclic voltammetry responseof biosensor to ethanol leave the act of 98.99% at the 7 week. Thus, the developed biosensor is promising to beused for simple analysis of halal products and has been tested on real sample.