14-33: Trichoderma harzianum P49P11 isolated form Amazon forest is a potential source of cellulases and xylanase

Monday, April 29, 2013
Exhibit Hall
Priscila da Silva Delabona1, Daniel Kolling2, Deise Juliana da Silva Lima1, Cristiane Sanchez Farina3 and José Geraldo da Cruz Pradella1, (1)Brazilian Laboratory of Science and Technology of Bioethanol - CTBE, Campinas, Brazil, (2)Brazilian Bioethanol Science and Technology Laboratory – CTBE, Campinas, Brazil, (3)Embrapa Instrumentation, São Carlos, Brazil
We have being working with the isolation, screening and selection of biomass-degrading fungi species from the Amazon forest and analyzed the enzymatic complex produced by selected strains. Those data demonstrated that a new strain of Trichoderma harzianum P49P11 (was a potential source of glycosyl hydrolases (GH) for lignocellulosic biomass deconstruction. Aiming the on-site GH production the influence of pretreated sugarcane bagasse (pSB), sucrose (SUC) and soybean flour (SF) was investigated in shaking flasks and controlled bioreactor experiments using experimental mixture design (EMD). It was found that pH 5.0 controlled with NH4OH aqueous solution and an optimized culture medium maximized Fpase biosynthesis rate up to 20 FPU/L h, 90.000 IU/L xylanase and 25.000 IU/L β-glucosidase activities using proper combination of pSCB, SF and SUC. The process was scale up to 20 L fermentor and influence of agitation rate on GH rate production was accessed. Complementary analysis of T. harzianum secretome was performed to identify enzymes that could be missing in the enzymatic complex to achieve high-yield pSB hydrolysis. These analyses enabled the selection of pectinase and α-L-arabinofuranosidase (AF) to be further investigated as supplements to the crude enzymatic extract produced by T. harzianum. The effect of these enzymes was then evaluated using experimental design methodology and supplementation with pectinase and AF increased the efficiency of enzymatic hydrolysis by up to 116% comparable with commercial enzymatic cellulase cocktail. Overall, it was demonstrated that T. harzianum P49P11 enzymes have a great potential to be used in the industrial deconstruction of biomass.