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 Opportunities: Scholarships in Brazil - Master
• Start date: November 01, 2018
• End date: October 31, 2020
• Field of knowledge: Biological Sciences - Biophysics - Molecular Biophysics
• Agreement: Coordination of Improvement of Higher Education Personnel (CAPES)
• Principal Investigator: Wanius José Garcia da Silva
• Grantee: Dnane Vieira Almeida
• Host 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.

Scientific publications

(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)

SANTOS, VALDENICE F.; ARAUJO, ANA C. J.; FREITAS, PRISCILLA R.; SILVA, ANA L. P.; SANTOS, ANA L. E.; DA ROCHA, BRUNO ANDERSON MATIAS; SILVA, ROMERIO R. S.; ALMEIDA, DNANE V.; GARCIA, WANIUS; COUTINHO, HENRIQUE D. M.; et al. Enhanced antibacterial activity of the gentamicin against multidrug-resistant strains when complexed with Canavalia ensiformis lectin. Microbial Pathogenesis, v. 152, MAR 2021. (17/17275-3, 18/13500-5)