The current study was aimed at L-asparaginase production and optimization using one of the promising bacterial isolates from soil. The isolate was grown on different media compositions, varied parameters like growth temperature, media pH etc and checked for L-asparaginase enzyme production. The enzyme was analyzed and the maximum enzyme activity was observed to be 2.54IU/ml, which was obtained from media composition having 2% glucose as carbon content, incubation temperature of 350C at a media pH of 7.4 and an incubation time of 24hrs at 150 rpm on a rotary shaker incubator.