The inhibitory effect of nicotine on the activity of choline oxidase immobilized on nylon net has been investigated. Nicotine was found to inhibit choline oxidase in a manner similar to its effect on acetylcholinesterase. A nicotine - sensitive probe was realised via nicotine inhibition of choline oxidase. No appreciable difference in this biosensor performance and the performance of nicotine probe incorporating acetylcholinesterase coimmobilized with choline oxidase was observed.