ISOLATION, SCREENING AND PARTIAL PURIFICATION OF CELLULASE FROM CELLULASE PRODUCING FUNGI

Shilpa Lokhande* and A. S. Pethe

ABSTRACT

The present investigation was undertaken to isolate and Screen the Cellulase Producing fungi. Microbial cultures were isolated from the soil sample collected from different villages of saline belt of Akola and Buldhana District, Maharashtra India. A Total of 146 isolates were isolated and identified based on Morphology and Biochemical characterization. Among all isolated organisms 72 fungal species were isolated, the one cellulolytic bacterial isolate shows maximum enzyme activity, and identified as Aspergillus niger. The best conditions like pH, carbon source, temperature and incubation period were also observed for cellulase producing organisms. Carboxy methyl cellulose [1.0% (w/v)] were found to be the best carbon source for the production of cellulase from Aspergillus niger and Optimum temperature and pH of the medium for the cellulase production were 40°C and 6, whereas it shows maximum enzyme activity after fifth day of incubation period. Further partial purification of cellulase was carried out by dialysis and ammonium sulphate precipitation and also to determine molecular weight by SDS-PAGE. The partial purification of the cellulase enzyme produced by Aspergillus niger had protein bands with the molecular weight of 45 kDa and FTIR was performed.

Keywords: Aspergillus niger, Cellulase, Partial purification.