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Periplasmic expression, purification and characterization of the inorganic biochemistry of an oxidative enzyme of the insect Bombyx mori (silkworm)

Grant number: 18/13500-5
Support type:Scholarships in Brazil - Master
Effective date (Start): November 01, 2018
Effective date (End): October 31, 2020
Field of knowledge:Biological Sciences - Biophysics - Molecular Biophysics
Cooperation agreement: Coordination of Improvement of Higher Education Personnel (CAPES)
Principal Investigator:Wanius José Garcia da Silva
Grantee:Dnane Vieira Almeida
Home Institution: Centro de Ciências Naturais e Humanas (CCNH). Universidade Federal do ABC (UFABC). Ministério da Educação (Brasil). Santo André , SP, Brazil

Abstract

The Lytic Polysaccharide Monooxygenases (LPMOs) are a new discovered class of enzymes that assist glycoside hydrolases in the degradation of recalcitrant polysaccharides through oxidant activity. This class of metalloenzymes presents great potential for biotechnological applications, mainly in the area of degradation of structural polysaccharides. LPMOs can be used as complements to increase the enzymatic efficiency of lignocellulolytic cocktails. The objectives of this research project are to carry out the heterologous expression, purification and characterization of the inorganic biochemistry of an oxidative enzyme of the insect Bombyx mori (silkworm). The metalloenzyme from Bombyx mori (BmLPMOs) will be expressed in the periplasmic region and purified to homogeneity by chromatographic methods. Enzymatic activity studies will be performed to determine the specificity of the BmLPMOs in the oxidation of cellulose, chitin and starch. The importance of the metal ion (Cu2+) on the structural stability and in the oxidative process, as well as its interaction with the enzyme, will be studied through spectroscopic techniques. The results from this research project will enable the identification of polysaccharides and characterize the oxidative action of an insect metalozyme with potential for biotechnological applications.