Purification and Characterization of Chitinase from the Nematode – Fungus Paecilomyces sp. P1

The nematophagous – fungi Paecilomyces sp. is curently developed as a biocontrol agent against plant parasitic nematodes (Khan et al., 2003; Yang et al., 2007). Biological control agents can infiltrate certain nematode sites and destroy them by producing some enzymes including chitinase (Khadijeh et al., 2017). The purpose of this study was to purify, determine the chitinase activity from Paecilomyces sp. P1. With Lugol reagent, chitinase of this strain was characterized by diffusion on agar plate. Chitinase specific activity was determined by measuring the release of reducing saccharides from colloidal chitin by the N-acetyl-glucosamine-dinitrosalicylate method at 540 nm. By using the saturated (NH4)2SO4 precipitation at 65% concentration, DEAE A-50 ion exchange chromatography and SDS - PAGE concentration 12.5%, chitinase molecules weigh nearly 50kDa, having a specific activity of 133,3 U/mg, 2,1-fold higher than that of supernatant. 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