Abstract: Objective: In order to overcome the defects of chemical hydrolysis approach to prepare glucosamine, an enzymatic hydrolysis method was developed. Methods: glucosamine was prepared by hydrolyzing chitosan, employing α-Amylase initially, and subsequently, glucoamylase. Results: The optimal hydrolyzing conditions were as follows: reaction time, 4 h; pH, 5.0; temperature, 50 °C; and, α-Amylase, 80 U/g for the initial reaction. Subsequently, glucoamylase was added in the presence of α-Amylase. The optimal reaction conditions were found to be: reaction time, 8 h; pH, 4.5; temperature, 55 °C; and, glucoamylase, 4000 U/g. The hydrolysates were subject to filtrating, concentrating to about 20% (w/w), precipitating with five volumes of ethanol, and drying at 60 °C for 2 h. The content and the yield of glucosamine in the dried precipitate were 91.3% (w/w) and 86.2% (w/w), respectively. Conclusions: The method developed in this study is a promising option in the preparation of glucosamine.

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