Optimization of different factors for efficient protoplast release from entomopathogenic fungus<Emphasis Type="Italic">Metarhizium anisopliae</Emphasis>

Optimization of different factors for efficient protoplast release fromMetarhizium anisopliae was investigated. Factors like culture media, age of the mycelium, incubation time, different enzymatic combinations and osmotic stabilizer were studied. Mycelium harvested at 40^th h in Sabouraud Dextrose broth showed the best protoplast yield over the other media and age of mycelium tested. An incubation time of 3 h and Lysing enzyme at a concentration of 10 mg/ml was the best among the different enzymatic concentrations and combinations tested and yielded a protoplast release of 7.3×10⁸ protoplasts/ml. The most suitable osmotic stabilizer for efficient protoplast release was 0.7M KCl.