Organophosphorus pesticides, widely used as insecticides for crop protection, are classified as an extremely toxic class of chemical compounds by the World Health Organization (WHO). Organophosphorus pesticides show marked specificity for the enzyme acetylcholinesterase, and can cause irreversible harm to the nervous system. The excessive use of organophosphorus pesticides, specifically in developing countries, made WHO to impose a ban on certain organophosphorus pesticides being sold to the developing nations. WHO and the Food and Agriculture Organization of United Nations, in joint meetings on pesticide residues, have stressed the need to develop easy, rapid, and sensitive methods to detect the presence of organophosphorus pesticides in food and water. In this regard, a number of review articles have been published explaining a myriad of detection methods for organophosphorus pesticides. Among these, optical detection methods, using biological molecules as the recognition element, provide a number of advantages, such as, high sensitivity and selectivity, simple operation, fast response and cheap instrumentation, and can really prove to be an effective alternative to the time-consuming traditional methods in the current scenario. Therefore, in this review article, we have discussed the recent advances in the field of biosensors employing the optical detection of organophosphorus pesticides. Specifically, various biosensors developed, using enzymes acetylcholinesterase and organophosphate hydrolase, have been categorized on the basis of the material used for their fabrication. A systematic discussion of the working principles, analytical parameters, and advantages and disadvantages of the various biosensors, has been attempted, along with a future perspective, on the challenges and possible improvements in the field of optical detection of organophosphorus pesticides.