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

Abstract

The nematophagous – fungi Paecilomyces sp. are currently 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). This paper discusses the purification and determination of 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 exchanges chromatography and SDS - PAGE concentration of 12.5%; chitinase molecules weigh nearly 50kDa, having a specific activity of 133.3 U/mg, 2.1-fold higher than that of supernatant. Furthermore, tested with the nematode Meloidogyne sp., the ability to kill nematodes of Paecilomyces sp. P1 reached 58% efficiency in 96h. These results are a scientific basis for the application of Paecilomyces sp. P1 in the production of nematode insecticides.

Keywords

Paecilomyces sp. P1, chitinase, purify, biocontrol, Meloidogyne sp.

References

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