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Heterologous production, characterization and engineering of xylose isomerases for application in industrial fermentation of pentoses

Grant number: 12/50056-0
Support type:Research Grants - Research Partnership for Technological Innovation - PITE
Duration: March 01, 2013 - March 31, 2015
Field of knowledge:Biological Sciences - Biophysics
Cooperation agreement: Odebrecht Agroindustrial
Principal Investigator:Igor Polikarpov
Grantee:Igor Polikarpov
Home Institution: Instituto de Física de São Carlos (IFSC). Universidade de São Paulo (USP). São Carlos , SP, Brazil
Company: ETH Energia S/A
City: São Carlos

Abstract

To make the production of second generation ethanol economically sustainable, it is essential to use the hemicellulose fraction of a biomass. The pentoses (P5) are not fermentable by common yeasts (Saccharomyces cerevisiae) used by the Brazilian industry. However, the xylose, a predominant fraction of P5, can be robustly converted by the action of xylose isomerases (Xls) on xylulose, which, in turn, is easily fermented by S. cerevisiae. In this project we are proposing to use modern tools of bioinformatics, genetics, large scale heterologous expression and Structural Molecular Biology/Biochemistry to identify, clone and express novel and potentially patentable Xls. We also aim to conduct their enzymatic studies and to change the Xls using protein engineering, to produce enzymes with high enzymatic activity and pH and temperature profiles compatible to the Brazilian industrial process conditions. In addition, immobilization of the optimized Xls to increase their lifetime and enable recycling is contemplated. We will pre-treat sugarcane bagasse to produce hemicellulosic hydrolyzate and conduct tests of simultaneous isomerization and fermentation of PS using commercial and industrial yeasts in the fermentation conditions close to ones established by Brazilian industry. It is important to note that this process does not use genetically modified organisms and is therefore easily scalable and usable under industrial conditions in Brazil. (AU)

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