Chitosan has been more popular in recent years due to its good bio-compatibility, low toxicity and rarely no allergic reaction to human body. Chitosan also has other bioactivity like reduce blood pressure, lower cholesterol, increase immunity, and its antibacterial property can be used in many clinical applications. For the best resource of Chitin/Chitosan we extracted from the seafood wastes (squid pens, shrimp or crab shells),which is abundant of polysaccharides, and glucosamine that we can use enzyme hydrolysis to achieve it. In traditional way, using a lot of chemical to receive chitosan and its oligomer, but in nowadays, we use the natural way, chitosanase bacteria strain TKU039 identified as Bacillus sp. from the soil in New Taipei City Linkou mountain. TKU039 was culture in medium with the squid pen powder (SPP) and shrimp powder (SP).The SPP medium was found most suitable with 1.0% SPP as a sole carbon/nitrogen source optimized under cultured at 37°C with 150 rpm for 5 days in 150 mL of medium containing 1% SPP, 0.1% K2HPO4 and 0.05 % MgSO4·7H2O.