Enzymatic degradation of microbial biofilm exopolisacharides: structural biophysics, molecular biotechnology and synthetic biochemistry of CAZymes in search for new enzymatic tools for antimicrobial treatments

• Grant number: 21/08780-1
• Support Opportunities: Research Projects - Thematic Grants
• Duration: March 01, 2023 - February 29, 2028
• Field of knowledge: Biological Sciences - Biophysics - Molecular Biophysics
• Principal Investigator: Igor Polikarpov
• Grantee: Igor Polikarpov
• Host Institution: Instituto de Física de São Carlos (IFSC). Universidade de São Paulo (USP). São Carlos , SP, Brazil
• Associated researchers: Ana Carolina Migliorini Figueira ; André Luis Souza dos Santos ; Camila Alves de Rezende ; Cristina Kurachi ; Eduardo Ribeiro de Azevedo ; Fernando Segato ; Francisco Eduardo Gontijo Guimaraes ; Marcos Pileggi ; Mario de Oliveira Neto ; Natalia Mayumi Inada

Abstract

The consequences of microbial resistance to antibiotics may represent a catastrophe for public health in the near future with an estimated 700,000 deaths per year, in addition to prolonged hospitalizations and potentially intractable diseases. Considering only antimicrobial resistance, the estimated number of deaths will increase to more than 10 million deaths/year by 2050, if this situation is not controlled. Hospital infections are already an extremely serious public health problem in Brazil and worldwide, causing thousands of deaths every day in addition to the steady increase in medical treatment costs valued at $ 33 billion per year in the United States and E 13 to E 24 billion per year in the European Union.It is estimated that 99% of the microorganisms on Earth live in biofilms. The formation of bacterial biofilms can increase antibiotic resistance by 10 to 1,000 times. Biofilms have a complex structure that aggregates microbial cells within an extracellular matrix, composed mainly of exopolysaccharides, extracellular DNA and proteins, where exopolysaccharides represent the main fraction. Within the scope of this project, we are proposing to conduct systematic studies of enzymes active in complex carbohydrates (CAZymes) with potential in the degradation and prevention of the bacterial biofilm formation, using Structural Biophysics, Molecular Biotechnology and Synthetic Biochemistry techniques. Biochemical and structural characterizations of enzymes from recent and still poorly explored CAZymes families will be conducted and the enzymes with high specific activities for exopolysaccharides degradation will be identified. CAZymes with activities against different extracellular matrix biopolymers will be combined to achieve greater levels of degradation and prevention of biofilm formation. The enzymatic action will also be enhanced by the combined use with the existing antibiotics and/or photodynamic therapy. In addition, following Synthetic Biochemistry approaches, the enzymatic circuits will be developed and tested in order to interfere with the formation of biofilms. The enzymatic degradation of biofilms can strengthen our arsenal of antimicrobial therapy and provide important information for the development of revolutionary strategies for the treatment of persistent infections. (AU)