Abstract
Chitinase gene from Trichoderma harzianum was cloned and hetrologously over expressed in M15 Escherichia coli. The recombinant protein of 42 kDa from E. coli was purified through Ni-NTA affinity column chromatography. The purified enzyme was active over broad range of pH (2.0 to 8.0) and temperature (10 to 60°C) with the peak activity at pH 5 (0.50 μg/ml) and 20°C with enzyme activity value (0.49 μg/ml). The purified protein fractions were tested for in vitro antifungal activity against different phytopathogens like Fusarium oxysporum f.sp. lycopersici, Sclerotioum rolfsii, Alternaria brassicae and Alternaria brassicicola. Purified endochitinase isolated from T. harzianum caused necrotic lesions, segmentation, branching and hyphal bursting at the concentration of 200 µg ml-1. Key words: Antifungal activity, Trichoderma harzianum, Fusarium oxysporum f.sp. lycopersici, Sclerotioum rolfsii, Alternaria brassicae and Alternaria brassicicola.
Highlights
Trichoderma spp. are among the most frequently isolated soil fungi, well known for their biocontrol ability against a wide range of plant pathogenic fungi (Howell, 2003; Sharma et al, 2011)
There are various modes of action associated with the ability of Trichoderma spp. to control plant pathogens but the best characterized and studied mechanism is mycoparasitism which involves a group of genes that encode for lytic enzyme chitinases
Expression of the active chitinase was optimized by inducing it with IPTG inducer at different time of induction
Summary
Trichoderma spp. are among the most frequently isolated soil fungi, well known for their biocontrol ability against a wide range of plant pathogenic fungi (Howell, 2003; Sharma et al, 2011). There are various modes of action associated with the ability of Trichoderma spp. to control plant pathogens but the best characterized and studied mechanism is mycoparasitism which involves a group of genes that encode for lytic enzyme chitinases 3.2.1.14), known to play important role in digesting chitin wall of the phytopathogenic fungi. Chitin, which is a significant component in the cell walls of large groups of fungi is made up of units of N-acetylglucosamine, linked together by 1,4-β-glycosidic bonds.
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