Production and Characterization of Alkaline Chitinase and Protease From an Alkaliphilic Actinomycete

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Addis Ababa University


Chitinase and protease are enzymes produced by a number of organisms and have wide industrial applications. This study was aimed at production and characterization of alkaline chitinase and protease from alkaliphilic microorganisms and evaluation of their possible applications. Alkaliphilic actinomycetes isolated from alkaline soda lakes were screened for the production of chitinase and protease. Out of 122 isolates tested 54 % produced appreciable chitinase activity indicating the presence of diverse chitinolytic microorganisms in these habitats. Based on the level of chitinase and protease production, isolate designated as A1 was selected for further study. When grown in colloidal chitin, the isolate A1 showed peak production of chitinase after 72 h while it was 120 h for protease production. The maximum amount of chitinase obtained at the peak production time was 22.6 U/ml for and 110.4 U/ml for protease. Both of the enzymes showed wide temperature and pH optima where the peak pH for chitinase activity was 8.5 while it was 10.5 for protease. The optimum temperatures for chitinase and protease activity were 45 °C and 55 °C respectively. Chitinase was stable over the wide range of pH and maintained more than 80% of its original activity after incubation in different buffers of pH 6.5-11 for one hour at room temperature. Protease A1 was stable in a wide range of pH and temperature. It has kept over 75% of its original activity in the pH range of 9-11 after one hour at 50°C. Protease does not require CaCl2 for its stability. It showed good stability in the presence and absence of CaCl2 where its stability was maintained after 60 min of incubation at 45°C. Both protease and chitinase can be used in wide applications of biotechnology and various industries



Alkaline, Alkaliphilic, Protease, Chitinase, Enzyme