1Nuclear Science and Technology Research Institute (NSTRI), Atomic Energy Organization of IRAN (AEOI), Karaj, Iran.
2Department of Horticulture and Plant Protection, Shahrood University of Technology, Shahrood, Iran.
This study was undertaken to ﬁnd out the optimum physicochemical parameters of fermentation, i.e. pH, incubation temperature and incubation time for the cellulase enzyme production of Trichoderma harzianum. The extracellular protein content was estimated by the dye binding method of Bradford. Endo-glucanase (EG), exoglucanase (or Cellobiohydrolase; CBH), β-glucosidase and total cellulase activity were investigated. The molecular weight of cellulase enzymes was studied using SDS-PAGE. To identify the predominant catalytic components in optimum conditions of enzyme production, cellulases were separated by an adapted two-dimensional electrophoresis technique. Estimated optimum conditions for cellulase enzyme were found as: pH 6.5, incubation temperature 28°C and incubation time 72 h. The SDS-PAGE profiles showed several enzyme bonds such as CBHs, EGs and BGLs. The T.harzianumhad both enzyme bonds of Cel7A (CBHI) and Cel7B (EG). Finally, the results of the 2D PAGE analysis showed that the profile of protein in optimium conditions of enzyme production had several enzymes such as CBHs, EGs and the high values of cellulose activity due to synergism that occurred between the CBH and EG.
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