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Production of human recombinant Peptidoglycan recognition protein (rPGLYRP3) and antimicrobial peptide (AMP-rPGLYRP3) in Pichia pastoris and analysis of antibacterial activity in opportunistic strains

Grant number: 21/00425-8
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): May 01, 2021
Effective date (End): April 30, 2022
Field of knowledge:Biological Sciences - Microbiology - Applied Microbiology
Principal researcher:Maria Cristina da Silva Pranchevicius
Grantee:Pedro Mendes Laprega
Home Institution: Centro de Ciências Biológicas e da Saúde (CCBS). Universidade Federal de São Carlos (UFSCAR). São Carlos , SP, Brazil

Abstract

Over the last few years, deaths caused by multidrug-resistant bacteria (MDRB) have been rising at an alarming rate, therefore, the search for new compounds capable of dealing with these microorganisms is highly important. Amongst the compounds with potential for new therapies, there are the peptidoglycan recognition proteins (PGLYRPs) and antimicrobial peptides (AMPs). PGLYRPs recognize conserved molecular patterns in infectious organisms and unleash innate immune responses, including phagocytosis, antimicrobial and pro-inflammatory genes expression and cytokines production. AMPs, including those deriving from host protein fragments, are multifunctional, being capable of mediating many anti-bacterial and immunomodulatory functions and stimulating angiogenesis. Considering the search for new solutions for the emerging MDRB problem, in this study we are proposing the heterologous expression of not only PGLYRP3, but also of an antimicrobial peptide (AMP-rPGLYRP3) (PVMPRKVCPNIIKRS) within this encoded protein, already identified by us, in a Pichia pastoris system. In addition, we intend to analyze the antimicrobial activity of both rPGLYRP3 and AMP-rPGLYRP3 on opportunistic bacteria. In silico analysis regarding the peptide AMP-PGLYRP3, also performed by us, shows that it is a possible non-toxic and non-allergenic anti-bacterial candidate, being, most likely, not active against human erythrocytes, while active against the bacteria species Klebsiella pneumoniae. Therefore, the results from this project may amplify our knowledge regarding the anti-bacterial activity of both PGLYRP3 and AMP-rPGLYRP3, assisting, in the future, the development of new medications and, consequently, control bacterial infections.

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