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Chitinase activity

TN Takuya Nagata
SS Shoko Shinya
TO Takayuki Ohnuma
TF Tamo Fukamizo
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Chitinase activity was determined colorimetrically using glycol chitin, which was synthesized by the method reported by Yamada and Imoto43, as a substrate. Ten microliters of the enzyme solution was added to 500 μl of 0.2% (w/v) glycol chitin solution in 0.1 M sodium acetate buffer, pH 5.0. After incubation of the reaction mixture at 37 °C for 15 min, the reducing sugars were determined with ferri-ferrocyanide reagent using the method of Imoto and Yagishita44. An increase in reducing sugars was regarded as chitinase activity. One unit (U) of enzyme activity was defined as the amount of enzyme (mg) releasing 1 μmole of GlcNAc per min at 37 °C.

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