Isolation and purification of cuticle degrading extra cellular proteases from entomopathogenic fungal species of beauveria bassiana and metarhizium anisopliae.

Enhanced Pr1 and Pr2 activities were observed in entomopathogenic fungi Beauveria bassiana (UB9) and Metarhizium anisopliae (UM4) supplemented with casein. Highest activity was observed at 72 hours of incubation when compared to minimal medium (MM) and colloidal chitin amended media. The extra cellular proteases (Pr1 and Pr2) thus isolated were purified through dialysis and their activities were found to be increased by two-fold. Pr1 and Pr2 were subjected to SDS - PAGE analysis and activity gel electrophoresis. The molecular weight of protease produced by Beauveria bassiana (UB9) is 19 KD and that of Metarhizium anisopliae (UM4) is 21 KD.

Strain improvement of entomopathogenic fungal species beauveria bassiana and metarhizium anisopliae by protoplast fusion.

Protease hyper producing recombinant strains were produced by intergeneric protoplast fusion of entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae. β-glucuronidase and KCl were used as lysing enzyme and osmotic stabilizer. Along with inter-generic fusion, intra-strain and inter-strain fusions were also carried out using polyethylene glycol (PEG) as fusogen. When the fused protoplasts were regenerated on Czapekdox agar medium, they exhibited fast mycelial growth and abundant sporulation when compared to non-fusants. Pr1 and Pr2 specific activities were found to be increased by two- fold in recombinant strains than the nonfusants and the parental strains.