| Full text | |
| Author(s): |
Santos, Clelton A.
[1]
;
Zanphorlin, Let-Cia M.
[2]
;
Crucello, Aline
[1]
;
Tonoli, Celisa C. C.
[3]
;
Ruller, Roberto
[2]
;
Horta, Maria A. C.
[1]
;
Murakami, Mario T.
[3]
;
de Souza, Anete Pereira
[4, 1]
Total Authors: 8
|
| Affiliation: | [1] Univ Estadual Campinas, Ctr Biol Mol & Engn Genet, Campinas, SP - Brazil
[2] Ctr Nacl Pesquisa Energia & Mat, Lab Nacl Ciencia & Tecnol Bioetanol, Campinas, SP - Brazil
[3] Ctr Nacl Pesquisa Energia & Mat, Lab Nacl Biociencias, Campinas, SP - Brazil
[4] Univ Estadual Campinas, Inst Biol, Dept Biol Vegetal, Campinas, SP - Brazil
Total Affiliations: 4
|
| Document type: | Journal article |
| Source: | BIOTECHNOLOGY FOR BIOFUELS; v. 9, MAR 22 2016. |
| Web of Science Citations: | 9 |
| Abstract | |
Background: The conversion of biomass-derived sugars via enzymatic hydrolysis for biofuel production is a challenge. Therefore, the search for microorganisms and key enzymes that increase the efficiency of the saccharification of cellulosic substrates remains an important and high-priority area of study. Trichoderma harzianum is an important fungus known for producing high levels of cellulolytic enzymes that can be used for cellulosic ethanol production. In this context, beta-glucosidases, which act synergistically with cellobiohydrolases and endo-beta-1,4-glucanases in the saccharification process, are potential biocatalysts for the conversion of plant biomass to free glucose residues. Results: In the present study, we used RNA-Seq and genomic data to identify the major beta-glucosidase expressed by T. harzianum under biomass degradation conditions. We mapped and quantified the expression of all of the beta-glucosidases from glycoside hydrolase families 1 and 3, and we identified the enzyme with the highest expression under these conditions. The target gene was cloned and heterologously expressed in Escherichia coli, and the recombinant protein (rThBgl) was purified with high yields. rThBgl was characterized using a comprehensive set of biochemical, spectroscopic, and hydrodynamic techniques. Finally, we determined the crystallographic structure of the recombinant protein at a resolution of 2.6 angstrom. Conclusions: Using a rational approach, we investigated the biochemical characteristics and determined the three-dimensional protein structure of a beta-glucosidase that is highly expressed by T. harzianum under biomass degradation conditions. The methodology described in this manuscript will be useful for the bio-prospection of key enzymes, including cellulases and other accessory enzymes, for the development and/or improvement of enzymatic cocktails designed to produce ethanol from plant biomass. (AU) | |
| FAPESP's process: | 14/09720-9 - Studies of the role of phosphorylation and protein binding partners in regulating the function of human class V myosins |
| Grantee: | Mário Tyago Murakami |
| Support Opportunities: | Regular Research Grants |
| FAPESP's process: | 08/52197-4 - Genomic-assisted breeding of sugarcane: using molecular markers for understanding the genetic architecture of quantitative traits and to implement marker assisted selection |
| Grantee: | Anete Pereira de Souza |
| Support Opportunities: | Program for Research on Bioenergy (BIOEN) - Thematic Grants |
| FAPESP's process: | 14/18856-1 - Study of global gene expression and genomic regions associated with control expression of enzymes involved in the degradation of lignocellulosic compounds by Trichoderma harzianum |
| Grantee: | Maria Augusta Crivelente Horta |
| Support Opportunities: | Scholarships in Brazil - Post-Doctoral |
| FAPESP's process: | 13/13309-0 - Studies of the structural and functional behavior of enzymes evolutionarily specialized in the degradation of plant biomass with potential biotechnological applications |
| Grantee: | Mário Tyago Murakami |
| Support Opportunities: | Regular Research Grants |