Pectins are polysaccharides, ubiquitous in the plant kingdom and constitute the important major component of middle lamellae of plant cell walls. Pectin lyases are widely used in the food industry in the production of juice, fruit drinks and wines. In present study, pectin lyase producing fungi was isolated from fruit, vegetable, and dead organic waste soil. Totally 25 fungal organisms were isolated and identified. Screening of pectin lyase producing fungi were isolated using pectin screening agar medium. Out of 25 organisms, only two fungi namely Aspergillus niger and Penicillium citrinum were able to produce pectin lyase activity. The pH, temperature, carbon sources, nitrogen sources and trace elements were also optimized. Enzyme assay was determined by Thiobarbitutric method (TBA). There are a lot of industrial processes can be applied to improve the quality and the yield of the final product.