| Summary: | 碩士 === 淡江大學 === 生命科學研究所碩士班 === 97 === A protease- and chitosanase-producing strain, Serratia marcescens TKU011, was cultivated by using shrimp head shell powder (SHP) as the sole carbon/nitrogen source. In this study, the optimized condition for protease and chitosanase production using squid pen powder (SPP) as the carbon/nitrogen source was found to be when the culture was shaken at 25℃ for five days in 100 mL of medium (pH 7) containing 1% SPP, 0.1 % K2HPO4, 0.05% MgSO4．7H2O. In a prolonged autoclave (at 121℃ for 120 min), the protease and chitosanase activity were measured were better than others. The culture supernatants from various time of the medium in an autoclave by SDS-PAGE analysis was found that the enzyme concentration increased with an increase in heating time. The dry weight of the recovered squid pen decreased with an increase in cultivation. The peptides and oligosaccharides in the culture supernatants was determined. The antioxidative activity of the supernatant was derermind by DPPH radical-scavenging activity, reducing activity, total phenolic, Fe2+-Chelating activity. Two chitosanase( or chitinase) (C1,C2) were purified from the culture supernatant by chromatography procedures of DEAE-Sepharose and Phenyl Sepharose. The molecular mass of C1 and C2 determined by SDS-PAGE was approximately 49 kDa and 66 kDa, respectively. The substrate specificity and biochemical characterization of the enzymes were studied.
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