Chitooligosaccharides has wide range of applications in medical and pharmacological fields due to water solubility and non toxicity properties. The present study was production of the chitosanase enzyme from marine Streptomyces massasporeus. The chitosanase producing actinomycetes isolates are isolated from marine wastes dumped soil, Out of 146 isolates,4 isolates were prominent of forming clear zone on the media.One among the isolate S15 produced highest chitosanase activity was identified as Streptomyces massasporeus KR873429 based on the molecular characterization. The chitosanase enzyme was optimized through RSM and the production of enzyme was increased to 1.913 U/ml.The crude enzymes were prepared and concentrated through (80% saturation) ammonium sulphate precipitatin followed by dialysis. The chitoligosaccarides hydrolysate was prepared with partially purified S15 chitosanase enzyme.The prepared COS mixture shown significant biocompatibility by MTT assay.