Partial purification of chitinase from Trichoderma atroviride

ABSTRACTChitinases [E.C.3.2.1.14] are enzymes which hydrolyze chitin, a linear homopolymer of β-1,4-linked N-asetylglucosamine. Chitin is a major component of most fungal and algae cellwalls, insects and arthropods exoskeletons, and shells of marine invertebrates.Chitin and chitinolytic enzymes are gaining importance for their biotechnologicalapplications. Particularly, chitinases are used in agriculture to control plant pathogens.Chitinases and chitooligomers produced by enzymatic hydrolysis of highly activepreparations at reasonable cost. Therefore, the understanding of biochemistry and genetics ofchitinolytic enzymes, their phylogenetic relationships and methods of estimation will makethem more useful in a variety of processes in near furute.Chitin degradation and chitinases are important in a wide variety of biological andbiotechnological processes, ranging from the exploitation and environmental clean-up ofchitinous wastes, to human therapy, plant defense systems and biological control. For thisreason, being a mold, frequently used in plant biocontrol Trichoderma atroviride producingchitinase was examined. T. atroviride culture was maintained on potato-dextrose agar.Chitinase induction was carried out on liquid medium which had been proposed by Dana et al.It was determined that the maximum chitinase activity obtained with T. atroviride when theliquid medium which was contained 0.1% colloidal chitin, the method of replacement wasapplied, and the cultivation was carried out at 30 ºC for 48 hours with rotary shaker. Then,amonium sulfate, ethanol, acetone and trichloroacetic acid were examined upon the chitinaseactivity.Keywords: Chitinase, Trichoderma atroviride, enzyme activity, purification.2