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Detection of organophosphate pesticides is indispensable to improve the quality of the water and therefore required by the European Union law. Here, we propose a novel architecture of a pesticide biosensor where the enzyme and the electrode are placed into neighboring zones in the stripe form. This separation permits to use the optimal conditions for their immobilization resulting in undisrupted mechanical stability of the biosensor. The enzyme, acetylcholinesterase, generates thiocholine which is further oxidized at the carbon nanoparticles film electrode, exhibiting high electrocatalytic activity. The whole device was tested as a biosensor for malaoxon employing a wild or recombinant acetylcholinesterase. The latter allowed for incubation time decrease down to 10 minutes and lowered the detection limit to 0.25 nM. Moreover, the encapsulated enzyme activity was visualised by a scanning electrochemical …