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Heterologous expression of cellulases from Myceliophthora heterotalica F.2.1.4 in Pichia Pastoris with characterization and purification of the enzymes produced

Grant number: 12/00506-9
Support type:Scholarships in Brazil - Doctorate
Effective date (Start): May 01, 2012
Effective date (End): May 31, 2016
Field of knowledge:Biological Sciences - Microbiology - Applied Microbiology
Principal Investigator:Eleni Gomes
Grantee:Carolina Bezerra Bussoli
Home Institution: Instituto de Biociências, Letras e Ciências Exatas (IBILCE). Universidade Estadual Paulista (UNESP). Campus de São José do Rio Preto. São José do Rio Preto , SP, Brazil
Associated research grant:10/12624-0 - Application of physical-chemical and enzymatic methods in the saccharification of sugar cane bagasse: studies on microorganisms, fermentative processes and hydrolyses methods, AP.TEM
Associated scholarship(s):14/00752-5 - Understanding the influence of glycosylation sites on thermostability of endoglucanases from thermophilic Myceliophthora heterothallica, BE.EP.DR

Abstract

The lignocellulosic material, found mainly in agro-industrial waste, is one of the most abundant natural organic complexes in the plant biomass, which is mainly composed of three main constituents: cellulose, hemicellulose and lignin. Cellulose is the biopolymer most commonly found in the vegetable kingdom, featuring a linear homopolymer composed of D-glucose residues joined by connection type ²-1.4 and can be used for bioethanol production by enzymatic hydrolysis. The cellulase enzyme complex is able to hydrolyze cellulose into fermentable monomers and it is naturally secreted by several filamentous fungi. Although several times this enzymes are detected in the growth medium for filamentous fungi, these enzymes with interesting properties may be impracticable for industrial due to low expression. From the biotecnologiacal standview, heterologous expression in Pichia pastoris provides many advantages for the production of recombinant eukaryotic proteins, producing enzyme types unique, without the interference of other enzymes in addition to being able to obtain high expression of the same, especially on an industrial scale . Recent studies have shown that the thermophilic fungus M. heterotalica F.2.1.4 is capable of producing cellulases with interesting properties for application in saccharification of sugarcane bagasse, but without a significant expression. Thus, this project aims to identify the gene coding for an endoglucanase and its cloning and expression in yeast Pichia pastoris. We also intend to purify and characterize the native and heterologous enzymes.

Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
BUSSOLI, Carolina Bezerra. Expressão heteróloga de celulases de Myceliophthora heterothallica F.2.1.4 em Pichia pastoris e Escherichia coli com a caracterização e purificação das enzimas produzidas. 2016. 131 f. Doctoral Thesis - Universidade Estadual Paulista "Júlio de Mesquita Filho" Instituto de Biociências, Letras e Ciências Exatas..

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