Journal of Applied Microbiology, Vol.119, No.1, 88-98, 2015
Unveiling the optimal parameters for cellulolytic characteristics of Talaromyces verruculosus SGMNPf3 and its secretory enzymes
AimsElucidation of different physico-chemical parameters and the secretory enzymes released by Talaromyces verruculosus SGMNPf3 during cellulosic biomass degradation. Methods and ResultsWe determined the optimal pH, temperature and time course parameters for the efficient degradation of different natural and commercial cellulosic substrates by T.verruculosus SGMNPf3, previously isolated from a forest soil. The optimal growth of the fungus and production of its cellulases were obtained when the culture condition was maintained at pH 33 and temperature 30 degrees C. Activity of the crude cellulases was maximum at 60 degrees C. Activity of cellulase enzymes produced on natural cellulose substrates was higher than that on commercial cellulose substrates. A continuous increase in cellulase activity at different time points indicated no apparent end product inhibition. This might be attributed to the high individual cellulases, notably -glucosidase (3161molg(-1)) production. Zymogram of extracellular crude proteins showed two dominant extracellular protein bands of molecular weight 723 and 614kDa, indicating their cellulolytic nature. MALDI-TOF and LC-MS/MS analysis of the 2DE spots also identified several enzymes including -glucosidase involved in the process of cellulose degradation. ConclusionsBased on its optimal parameters for cellulolytic activities, we suggest that the fungus is acido-mesophilic. There was apparently no end-product inhibition of the cellulase activity and this is attributed to the ability of the fungus to produce sufficient -glucosidase. The dominant proteins secreted by the fungus were confirmed to be cellulases. Significance and Impact of the StudyThe high individual cellulase activities, better cellulase production on natural substrates and apparent absence of end-product inhibition are characteristics of T.verruculosus SGMNPf3 for use in harvesting naturally endowed energy in cellulosic biomass.
Keywords:2DE-MALDI-TOF;MS;-glucosidase;cellulolytic micro-organisms;LC-MS;MS;SDS-PAGE;Talaromyces verruculosus;Zymography