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Ocimum gratissimum (OG) is a species rich in essential oils (EO), which is known for its antimicrobial and antioxidant properties. This study aimed to encapsulate the essential oil of Ocimum gratissimum (OGE), determine its chemical composition, and evaluate its antioxidant and antimicrobial activities against six pathogenic bacteria, comparing it with the free essential oil (OGF). The EO was extracted by hydrodistillation using a Clevenger-type apparatus, and an oil-in-water emulsion was prepared using a combination of biopolymers: maltodextrin (MA), cashew gum (CG), and inulin (IN). The chemical profile was identified using gas chromatography–mass spectrometry (GC–MS). Antioxidant activity was assessed using the Oxygen Radical Absorbance Capacity with fluorescein (ORAC-FL) method, while the Minimum Inhibitory Concentrations (MIC) and Minimum Bactericidal Concentrations (MBC) were determined by the microdilution method. Microparticles were formed using the spray-drying method, achieving an encapsulation efficiency of 45.2%. The analysis identified eugenol as the main compound both before and after microencapsulation. The OGE microparticles demonstrated high inhibitory and bactericidal effects against S. aureus, S. choleraesuis, and E. coli, with MIC values of 500 µg·mL−1 and MBC values of 1000 µg·mL−1, as well as antioxidant activity of 1914.0 µmol-TE·g−1. Therefore, it can be inferred that the EO of OG maintained its antimicrobial and antioxidant effects even after microencapsulation by spray-drying, making it a promising natural ingredient. [ABSTRACT FROM AUTHOR]