Naringin is a flavanone found in citrus fruits and vegetables which are responsible for the asperity, due to which citrus fruit are unable to consume and from which we are unable to get the valuable nutrition in our body. To overcome this asperity in citrus fruits, present investigation is carried out, which provides a detailed report of isolation and screening of naringinase producing micro-organisms from different samples collected from Kalyan -Karnataka region, India. A sum of around 50 strains were isolated from 20 different samples which showed the positive result for naringinase activity, among 50 strains obtained 42 strains are fungi and remaining 8 were bacteria which showed good to moderate results in preliminary screening by 1% FeCl3 method, further 5 fungi isolates which showed a good naringinase activity in submerged fermentation were assayed by Spectrophotometric method. As a result, KLA-80 showed a maximum result with an activity of 559 U/ml. Further from the identification of the isolate KLA-80 by BLAST analysis found to be having similarity with Aspergillus flavus and the accession number of OQ152018 is obtained.