Banca de QUALIFICAÇÃO: GERCIANE SILVA DE OLIVEIRA

Uma banca de QUALIFICAÇÃO de MESTRADO foi cadastrada pelo programa.
STUDENT : GERCIANE SILVA DE OLIVEIRA
DATE: 29/07/2022
TIME: 14:00
LOCAL: Sessão pública realizada por videoconferência
TITLE:

Evaluation of the genotoxicity of the trypsin inhibitor isolated from tamarind seed (tamarindus indica L.) and the antibacterial potential of the hydrolyzed inhibitor

KEY WORDS:

Protease inhibitors. Antimicrobial peptides. Molecular Dynamics Simulation.

PAGES: 89
BIG AREA: Ciências da Saúde
AREA: Nutrição
SUMMARY:

Bacterial infections have become a global concern as pathogenic bacteria have acquired resistance to antibiotics. As a result, the search for natural therapeutic agents with antibacterial action is increasing, and protease inhibitors are strong candidates for naturally presenting mechanisms of protection against pathogens. In this study, the genotoxicity of the trypsin inhibitor isolated from Tamarindo seeds (ITT) was evaluated and the antibacterial effect of the inhibitor and its hydrolyzate were investigated in vitro and in silico, respectively. For this, initially, the ITT was obtained by trypsin-Sepharose 4B affinity chromatography and identified. The cytotoxicity of ITT was evaluated by the MTT method and genotoxicity by the CBMN in CHO-K1 cells, using concentrations of 0.3 and 0.6 mg/mL. The ITT was subjected to simulated digestion and in vitro proteolytic hydrolysis according to the INFOGEST protocol, however, digestion fluids were not used for hydrolysis. The digested/hydrolyzed ITT was subjected to molecular mass analysis and inhibitory activity on trypsin to monitor digestion and hydrolysis. For the in sílico hydrolysis of ITT, model number 56, conformation number 287 (ITTp 56/287) was used and subjected to cleavage with trypsin and chymotrypsin in combination using the PeptideCutter analysis tool on the ExPASy server. Subsequently, to select the peptide with antibacterial potential, these were aligned with the ITTp 56/287 to identify the positions of the amino acid residues using CLUSTAL W and after selecting the peptide, the molecular dynamics (DM) was carried out using GROMACS. The ITT (0.3 and 0.6mg/mL) did not cause cytotoxicity or genotoxicity in the cells (p<0.05), when digested/hydrolyzed in vitro, it remained intact in the oral and gastric phases, while the intestinal enzymes were effective in cleaving the ITT. And during in silico assays, Peptidotrypchymo59 (TVSQTPIDIPIGLPVR) showed amphipathic amino acid residues, hydrophobicity 0.636, α-helix structure and interaction potential energy (EPI) of -518.08 kcal.mol-1 with the membrane of Gram-positive bacteria and threonine and arginine residues showed the best EPI. Therefore, the results bring new perspectives of studies and applications for ITT and its peptides on the antibacterial aspect.

 

COMMITTEE MEMBERS:
Presidente - 2578619 - ANA HELONEIDA DE ARAUJO MORAIS
Externa à Instituição - RICHELE JANAINA ARAUJO MACHADO - UNICHRISTUS
Interna - 2275877 - THAIS SOUZA PASSOS