The enzyme chitin deacetylase was extracted from the fungus Mucor rouxii. The fungus was grown aerobically at pH 4.5, and a temperature of 28 +/- 2°C, using a shaker speed of 100 rpm. The highest enzymatic activity was obtained at 36 h of Mucor rouxii's growth. The protein content of Mucor rouxii was a function of incubation time and it increased in the course of the fungal growth up to 3 days. / The enzyme was partial purified by ammonium sulfate fractionation, followed by ion exchange chromatography using a Fast Protein Liquid Chromatography (FPLC) System. The specific activity in the 60--85% ammonium sulfate fraction showed higher specific activity (51.44 U/mg) and a 2.2-fold purification as compared with that of the crude extract (24.27 U/mg). The fraction after ion exchange chromatography step showed a specific activity of 233.65 U/mg, representing about 7.2-fold purification compared to the crude extract. This ion exchange fraction showed protein bands with molecular weights ranging from 45 to 75 kDa on SDS-PAGE (12%). / The enzyme showed optimal activity at a temperature of 50°C, and more than 90% of its activity was retained after 30 min incubation at this temperature. The optimal pH activity of chitin deacetylase from Mucor rouxii ranged from 5.5--6. The enzyme retained more than 80% of its activity in the pH range of 4.5--8.5 after 1 h incubation at room temperature, suggesting that this enzyme is quite stable in the slightly acidic to alkaline pH range. However, a lower pH (<4) and a higher pH (>9) caused the enzyme to lose more than 60% of its activity. / It was shown that the chitin deacetylase did not require the following metal ions (Co+2 and Mn+2) for activity. However, Mn+2 slightly increased the enzyme activity. The activity of the enzyme was also inhibited by EDTA.

The optimization of isolation and characterization of chitin deacetylase (CDa) from Mucor rouxii was the focus of this study. The crude extract in the active form was partially purified by ammonium sulfate fractionation, followed by column chromatography by ion exchange in the Fast Protein Liquid Chromatography (FPLC) system. / Using a 10 mM concentration, the order of effectiveness for the inhibitors in achieving 35--40% was CaCl2, CuSO4, MgCl 2 and EDTA. At a 25 mM concentration, propionic acid and sodium acetate inhibited the enzyme up to 40% & 50%, respectively. M. rouxii CDa activity was greater in the mycelial extract, and was capable of efficiently hydrolyzing the substrate glycol chitin. / Two major bands were observed after native PAGE of the partially purified enzyme using Mono Q column (FPLC). The estimated molecular weight of CDa bands by SDS-PAGE containing 0.1% glycol chitin was 21, 23, 26 and 44 kDa when compared to the migration of molecular weight markers. (Abstract shortened by UMI.)

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