Naringin is a flavanone bitter glycoside which is found in flowers and fruit of the grapefruit, which on further hyrdolysis yields naringenin and glucose. As naringin is responsible for the bitterness of the fruit, to neutralize the bitterness by eco-freindly method, microbial method is performed by using naringinase enzyme. In the presesnt study naringinase producing strain Apergills flavus KLA-80 which was previously isolated from soil sample of Kalyan-Karnataka region, India, and deposited in NFCCI with an accesion number of OQ152018 which was maintained in the lab conditions was used for optimization studies.Further to increase the activity of enzyme one factor at a time approach was carried out where the activity was found to be increased from 559 to 720U/mL in a temp of 350C, pH 4.5, carbon source 1.25% and nitrogen source of 0.15% at an agitation speed of 120rpm repectively. Eventually the production of naringinase from Aspergillus niger is optimized by Response Surface Methodology i.e., by CCD (Central Composite Design) method considering four factors at a time (pH,Agitation speed, Galactose, Peptone) as the variables. Based on the statstical method of optimization it is found that the enzyme activity was increased upto 20.41%.