Bioactivity of enzymes in the degradation of the biofilm matrix to control oral diseases

• Grant number: 20/15092-1
• Support Opportunities: Regular Research Grants
• Start date: October 01, 2021
• End date: September 30, 2023
• Field of knowledge: Health Sciences - Dentistry - Social and Preventive Dentistry
• Principal Investigator: Antônio Pedro Ricomini Filho
• Grantee: Antônio Pedro Ricomini Filho
• Host Institution: Faculdade de Odontologia de Piracicaba (FOP). Universidade Estadual de Campinas (UNICAMP). Piracicaba , SP, Brazil
• Associated researchers: Fernanda Cristina Petersen ; Igor Polikarpov
• Associated scholarship(s): 23/03199-4 - Enzyme bioactivity in biofilm matrix degradation - steps for obtaining enzymes (Year 2), BP.TT; 21/14123-3 - Enzyme bioactivity in biofilm matrix degradation: steps for obtaining enzymes (Year 1), BP.TT

Abstract

The accumulation of biofilm in the oral cavity promotes the development of caries and periodontal disease, negatively affecting people's quality of life, as well as serving as a reservoir of microorganisms capable of causing systemic diseases. The presence of glucans with alfa-(1-3) bonds in the extracellular matrix allows biofilms to be strongly structured and adhered to surfaces, not being disorganized due to the lack of enzymes that degrade these polysaccharides in the oral cavity. Therefore, the aims of this study are to synthesize the enzyme alfa-(1-3) endoglucanase of Paenibacillus curdlanolyticus, characterize the crystal structure and evaluate the enzymatic action in the degradation of biofilms, in isolation and in conjunction with other enzymes. The enzyme alfa-(1-3) endoglucanase will be synthesized by cloning the mutP gene (ADT91063.1) present in P. curdlanolyticus and high-throughput expression. Structural and biochemical characterization will be carried out to understand the molecular mechanisms of enzymatic degradation. In addition, it will be assessed whether the presence of other enzymes, alfa-(1-4) glucanase, alfa-(1-6) glucanase and DNAse, could contribute to optimizing the biofilm matrix degradation process. An in vitro study will be carried out to define the best treatment condition for degradation of the biofilm matrix using a cariogenic biofilm model, evaluating biofilm and dental demineralization. As result, five main achievements are expected: implementation of a research center for the synthesis of enzymes with biological interest having visibility in clinical practice and product development, quality scientific production on a relevant and internationally relevant topic, consolidation of methodologies of interest that enable constant technological innovation, consolidation of a network with international partnerships, and produce highly trained and qualified human resources. (AU)