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Biophysical and biochemical studies of exoglucanases from Trichoderma harzianum involved in the biodegradation of cellulose


In nature, cellulose is degraded by a variety of micro-organisms. However, its high degree of crystallinity makes cellulose degradation a problem of important interest especially in biotechnological applications. Lignocellulosic biomass, such as sugar cane bagasse, holds a promise of environmentally friendly bioenergy production in Brazil. However, enzymatic hydrolysis, currently considered a method of choice in biomass saccharification, is hampered by considerable cell-wall recalcitrance. To make this technology sustainable and cost effective, our comprehension of cellulose enzymatic hydrolysis should be significantly improved. The filamentous fungus T. reesei secretes a mixture of cellulases, which have been shown to act synergistically, which has been widely used as model in studies of cellulose breakdown. However, cellulases from filamentous fungus T. harzianum are part of a less-studied class of the proteins which yet has not been structurally and biochemically characterized. Here, we propose to conduct a structural and biochemical studies of exoglucanases, CBHI and CBHII, from T. harzianum. CBHI and CBHII are very potent enzymes, which can achieve complete, although slow, hydrolysis of crystalline cellulose even without the help of endoglucanases. The secondary and tertiary structures will be analyzed using circular dichroism (CD) spectroscopy and intrinsic fluorescence spectroscopy, respectively, as a function of pH and temperature variation. The enzymes will be assayed for activity in the presence of soluble and insoluble substrates. (AU)

Scientific publications (7)
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
ROSSETO, FLAVIO RODOLFO; MANZINE, LIVIA REGINA; NETO, MARIO DE OLIVEIRA; POLIKARPOV, IGOR. Biophysical and biochemical studies of a major endoglucanase secreted by Xanthomonas campestris pv. campestris.. Enzyme and Microbial Technology, v. 91, p. 1-7, SEP 2016. Web of Science Citations: 2.
BERNARDINELLI, OIGRES DANIEL; LIMA, MARISA APARECIDA; REZENDE, CAMILA ALVES; POLIKARPOV, IGOR; DEAZEVEDO, EDUARDO RIBEIRO. Quantitative C-13 MultiCP solid-state NMR as a tool for evaluation of cellulose crystallinity index measured directly inside sugarcane biomass. BIOTECHNOLOGY FOR BIOFUELS, v. 8, AUG 5 2015. Web of Science Citations: 37.
TSUCHIDA, JEFFERSON ESQUINA; REZENDE, CAMILA ALVES; DE OLIVEIRA-SILVA, RODRIGO; LIMA, MARISA APARECIDA; D'EURYDICE, MARCEL NOGUEIRA; POLIKARPOV, IGOR; BONAGAMBA, TITO JOSE. Nuclear magnetic resonance investigation of water accessibility in cellulose of pretreated sugarcane bagasse. BIOTECHNOLOGY FOR BIOFUELS, v. 7, SEP 10 2014. Web of Science Citations: 23.
LIMA, MARISA A.; GOMEZ, LEONARDO D.; STEELE-KING, CLARE G.; SIMISTER, RACHAEL; BERNARDINELLI, OIGRES D.; CARVALHO, MARCELO A.; REZENDE, CAMILA A.; LABATE, CARLOS A.; DEAZEVEDO, EDUARDO R.; MCQUEEN-MASON, SIMON J.; POLIKARPOV, IGOR. Evaluating the composition and processing potential of novel sources of Brazilian biomass for sustainable biorenewables production. BIOTECHNOLOGY FOR BIOFUELS, v. 7, JAN 18 2014. Web of Science Citations: 30.
LIMA, MARISA A.; LAVORENTE, GABRIELA B.; DA SILVA, HANA K. P.; BRAGATTO, JULIANO; REZENDE, CAMILA A.; BERNARDINELLI, OIGRES D.; DEAZEVEDO, EDUARDO R.; GOMEZ, LEONARDO D.; MCQUEEN-MASON, SIMON J.; LABATE, CARLOS A.; POLIKARPOV, IGOR. Effects of pretreatment on morphology, chemical composition and enzymatic digestibility of eucalyptus bark: a potentially valuable source of fermentable sugars for biofuel production - part 1. BIOTECHNOLOGY FOR BIOFUELS, v. 6, MAY 9 2013. Web of Science Citations: 58.
COLUSSI, FRANCIELI; GARCIA, WANIUS; ROSSETO, FLAVIO RODOLFO; SOARES DE MELLO, BRUNO LUAN; DE OLIVEIRA NETO, MARIO; POLIKARPOV, IGOR. Effect of pH and temperature on the global compactness, structure, and activity of cellobiohydrolase Cel7A from Trichoderma harzianum. EUROPEAN BIOPHYSICS JOURNAL WITH BIOPHYSICS LETTERS, v. 41, n. 1, p. 89-98, JAN 2012. Web of Science Citations: 13.
REZENDE, CAMILA ALVES; DE LIMA, MARISA APARECIDA; MAZIERO, PRISCILA; DEAZEVEDO, EDUARDO RIBEIRO; GARCIA, WANIUS; POLIKARPOV, IGOR. Chemical and morphological characterization of sugarcane bagasse submitted to a delignification process for enhanced enzymatic digestibility. BIOTECHNOLOGY FOR BIOFUELS, v. 4, p. 1-18, NOV 28 2011. Web of Science Citations: 177.

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