Proteases are one of the important groups of enzymes having extensive applications in various industries. Although the protease enzymes were extracted from diverse group of organisms, microbial proteases are more advantageous and interesting, because of their wide range of stability. Hence, the aim of this study was isolation, biochemical characterization and enzyme kinetics for the production of alkaline serine protease from the isolated mid gut microbes of silk moth Bombyx mori larva. Results revealed the presence of gram positive, short tubular with endospores and other microbes in the gut of Bombyx mori larva. Various biochemical tests like citrate utilization test, gelatin liquefaction test, casein hydrolysis test, starch hydrolysis test and triple sugar test showed positive results for the presence of alkaline serine protease. The optimum culture conditions at which the purified serine protease enzyme showed highest activity were pH 9, temperature 35oC, 15 minutes of incubation and 0.8 % substrate concentration.