Improved Cellulase Production by Trichoderma reesei RUT C30 under SSF Through Process Optimization

Abstract

This work investigated the preparation of chitosan nanoparticles used as carriers for immobilized enzyme. The morphologic characterization of chitosan nanoparticles was evaluated by scanning electron microscope. The various preparation methods of chitosan nanoparticles were discussed and chosen. The effect of factors such as molecular weight of chitosan, chitosan concentration, TPP concentration, and solution pH on the size of chitosan nanoparticles was studied. Based on these results, response surface methodology was employed. The results showed that solution pH, TPP concentration, and chitosan concentration significantly affected the size of chitosan nanoparticles. The adequacy of the predictive model equation for predicting the magnitude orders of the size of chitosan nanoparticles was verified effectively by the validation data. Immobilization conditions were investigated as well. The minimum particles size was about 42 +/- 5 nm under the optimized conditions. The optimal conditions of immobilization were as follows: one milligram of neutral proteinase was immobilized on chitosan nanoparticles for about 15 min at 40 degrees C. Under the optimized conditions, the enzyme activity yield was 84.3%.

Trichoderma reesei RUT-C30 – thirty years of strain improvement

“…RUT-C30 outperformed a high cellulase-producing mutant MCG77 (Gallo et al, 1978) and its parent strain QM9414. From that time to the present day, RUT-C30 has been used as a paradigm for developing processes for the production of cellulolytic enzymes (Olsson et al, 2003;Bailey & Tähtiharju, 2003;Juhász et al, 2004;Singhania et al, 2007), engineering of hypercellulolytic strains (e.g. Collén et al, 2005;Limó n et al, 2011) and production of cellulases on agricultural and forest residues with a special emphasis of converting lignocellulosic biomass to ethanol (GyalaiKorpos et al, 2011;Choy et al, 2011).…”

Optimization of Medium Components for β-glucosidase Production in Schizophyllum commune KUC9397 and Enzymatic Hydrolysis of Lignocellulosic Biomass

“…However, enzymatic hydrolysis of cellulosic biomass requires high enzyme loadings, and cellulase production is considered to be the most expensive step of enzymatic hydrolysis. Significantly reduced cellulase cost is therefore critical to the development of an economical process for bioethanol production from lignocellulosic biomass (Singhania et al 2007). Several researchers have made an effort to reduce enzyme production costs or to enhance enzyme activity.…”

An Overview on Fungal Cellulases with an Industrial Perspective

“…The statistical tools such as Plackett-Burman, Box-Behnken, central composite designs, ANN alone or in their combination are the most commonly used methods for the screening of independent variables that significantly affect the enzyme production. For instance, Singhania et al [65] employed a combination of Plackett-Burman and central composite designs to maximize the production of cellulase produced by T. reesei RUT C30, which resulted in 6.2 folds increase in cellulase production in the presence of 37.5% moisture content and 30°C. Mixed cultures of T. reesei and A. phoenicis showed β-glucosidase activity and filter paper activity of 0.64 IU/mL and 1.54 FPU/mL, respectively on dairy manure at 27°C and pH 5, which was optimized using BoxBehnken design [66].…”