Hyper production of carboxy methyl cellulase by Thermomyces dupontii utilizing physical and chemical mutagenesis

Utilization of cellulases as substitute of chemical process gained huge momentum in the field of biotechnology  Now there is dire need to find out un explore reveres of fungi possessing greater potential for efficient cellulase production.  This boosted isolation of novel thermo tolerant fungal strains capable of producing the targeted product. In this investigation 70 thermophilic cellulolytic fungal strains were isolated. All the strains were screened via submerged fermentation. The strain showing highest CMCase activity was identified by conventional method i.e.  based on morphology and microscopic features and confirmed by 18S rDNAgene sequencing, using specific ITS primers. The modified CTAB method was used for rapid extraction of DNA from thermo tolerant strain.  The selected strain subsequently subjected to sequencing and phylogenetic analysis. The result indicates the selected strain was found to be T. dupontii. For strain improvement the T. dupontii was subjected to random mutagenesis by using (UV) irradiation and EMS treatment. Out of 40 screened mutant T. dupontii EMS 15 had 2.4 fold more yield (21.8 U/ml/min) as compared to parental strain. The Five fermentation media were also screened. The Medium3 gave higher titer of cellulase activity (29.3 U/ml/min) and found to be the best medium.

Keywords: Molecular identification, Fungi, CMCase, Mutation.