Glycoproteins; seeds; Hemagglutination; purification; anti-bacterial

 

 Lectins have the ability to bind carbohydrates and play crucial roles in cellular processes, making their potential in biotechnology vast. Gliricidia sepium is highly versatile, used in folk medicine, animal feed and much more. However, little has been explored about its lectin potential. This research aims to investigate G. sepium seeds as a possible source rich in lectins and explore their characteristics and applications in biotechnology. Aiming to achieve the objectives, the following methods were used: optimization of lectin extraction, biochemical characterization, purification using reverse micelles and ion exchange chromatography, electrophoretic analysis and evaluation of its biological activities, covering antibacterial, cytoprotective and anticoagulant actions. The most efficient method for lectin extraction was from seed coat flour in a buffer solution for 120 minutes. This extract revealed a remarkable ability to agglutinate different types of blood, especially rabbit erythrocytes, reaching a hemagglutinating activity (HA) of 4096 U.H./mL. Lectin HA varied significantly over a wide pH range, with the point of maximum activity at pH 8. Furthermore, HA remained stable over a temperature range of 10 to 80 °C. The lectin showed remarkable stability in the presence of metal ions, maintaining a consistent activity of 32 U.H./mL. Lectin activity was not affected by the presence of different carbohydrates. The three-step purification process concentrated the lectin up to 16-fold. The purified lectin had a molecular weight of 10 kDa and demonstrated remarkable efficacy in inhibiting the growth of bacteria associated with mastitis, with an inhibition rate of up to 89%. The lectin did not exhibit cytoprotective properties in human blood cells, suggesting its potential for treating conditions associated with cellular damage, such as cancer. It demonstrated anticoagulant potential, increasing the intrinsic pathway clotting time by up to 1.38 times after 2 hours of exposure. These results indicate a broad spectrum of biological activities and potential biotechnological applications of the G. sepium lectin, including its antibacterial, cytoprotective and anticoagulant properties. This paves the way for further investigation into the impact of this lectin on various medical conditions.