Banca de QUALIFICAÇÃO: ELLYNES AMANCIO CORREIA NUNES

Uma banca de QUALIFICAÇÃO de DOUTORADO foi cadastrada pelo programa.
STUDENT : ELLYNES AMANCIO CORREIA NUNES
DATE: 02/10/2023
TIME: 14:00
LOCAL: Videoconferência
TITLE:

Purification and biochemical characterization of bioactive molecules from ophidian venions

KEY WORDS:

Bioactive molecules. Ophidian venoms. Bioprospecting.

PAGES: 69
BIG AREA: Ciências Biológicas
AREA: Bioquímica
SUMMARY:

Animal toxins have bioactive molecules, with several known biological activities that can be directed to the treatment of several diseases, such as toxins isolated from ophidian venoms, which present about 90% of their composition in peptides and proteins. These, in turn, present the most diverse biochemical mechanisms, as can be seen in envenomation’s caused by snakes of the genus Bothrops, belonging to the Viperidae family, which demonstrate proteolytic, coagulant, and hemorrhagic activities. These activities are related to the presence of proteins such as phospholipase A₂, metalloproteinases and serine proteases, as well as lectins, non-enzymatic proteins, with affinity for carbohydrate molecules and with innate ability to agglutinate erythrocytes. Many of these proteins have already described pharmacological properties. Thus, the objective was to isolate and characterize bioactive molecules from Bothrops moojeni and B. erythromelas venoms. To this end, in the search for bioactive molecules from snake venoms, Molecular Exclusion Chromatography and Reverse Phase High Performance Liquid Chromatography were used, followed by Hemagglutination Assays, enzymatic and hemolysis assays. Molecular exclusion applied to B. moojeni venom generated a graph with 6 fractions, which were rechromatographed in RP-HPLC. Fractions from molecular exclusion were subjected to hemagglutination assays and only fraction 5 caused hemagglutination in ABO blood. Subsequently, the samples were subjected to a phospholipase activity assay and only fraction 2 showed substrate consumption, indicating a possible phospholipase fraction. The hemolytic assay showed that all fractions were below 20% hemolysis. Reverse phase chromatography was performed for B. erythromelas venom, resulting in 3 fractions monitored by SDS-PAGE electrophoresis gel. After lyophilization, the samples were tested in the hemagglutination test and only fraction 3 was able to agglutinate the blood of the ABO system. The fraction was rechromatographed in reverse phase. In view of these results, new fronts of biochemical and biophysical characterization will be carried out for all fractions with activity, with sequencing being the next step for confirmation of protein fractions.

COMMITTEE MEMBERS:
Externo à Instituição - CAIO FERNANDO RAMALHO OLIVEIRA
Presidente - 1149356 - ELIZEU ANTUNES DOS SANTOS
Interno - 1544647 - MATHEUS DE FREITAS FERNANDES PEDROSA