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Partial purification and characterization of cellulases from digestive tracts of the African giant snail (Achatina achatina)

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Crude cellulase was extracted from the digestive tracts of 30 mature Achatina achatina and subjected to a 3-step purification process of ammonium sulfate precipitation, dialysis, and gel filtration. | Turkish Journal of Biology http://journals.tubitak.gov.tr/biology/ Research Article Turk J Biol (2013) 37: 199-205 © TÜBİTAK doi:10.3906/biy-1205-11 Partial purification and characterization of cellulases from digestive tracts of the African giant snail (Achatina achatina) Paul Chijioke OZIOKO, Sabinus Oscar Onyebuchi EZE*, Ferdinand Chiemeka CHILAKA Department of Biochemistry, University of Nigeria, Nsukka, Enugu State, Nigeria Received: 08.05.2012 Accepted: 27.09.2012 Published Online: 25.03.2013 Printed: 25.04.2013 Abstract: Crude cellulase was extracted from the digestive tracts of 30 mature Achatina achatina and subjected to a 3-step purification process of ammonium sulfate precipitation, dialysis, and gel filtration. This purification procedure gave 3 prominent enzyme activity peaks that coincided with protein peaks and were designated A, B, and C, corresponding to high endoglucanase, β-glucosidase, and total cellulase activities, respectively. Temperature optima of 50 °C were recorded for β-glucosidase and total cellulase while 45 °C was recorded for endoglucanase. Total cellulase, glucosidase, and endoglucanase showed maximum activities at pH values of 5.5, 4.5, and 7.5, respectively. Kinetic studies show that total cellulase has a Vmax and Km of 2427.18 µmol/min and 15.12 mg cellulose and endoglucanase has values of 955.11 µmol/min and 2.39 mg Na-CMC, while the values of β-glucosidase are 946.97 µmol/min and 4.3 mM cellobiose, respectively. This study shows that cellulases from digestive tracts of A. achatina could be utilized for degradation of cellulose-containing materials because of their high thermostability and acid/alkali stability, which reflect the potential commercial significance of the enzyme. Key words: Achatina achatina, cellulases, purifications, characterization, sodium carboxymethyl cellulose (Na-CMC), cellobiose 1. Introduction Cellulase (EC 3.2.1.4) refers to a class of hydrolase enzymes produced by fungi, bacteria, protozoans, .

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