The goal of this study is to isolate and assess red betel leaf endophytic bacterial isolates for their ability to suppress pathogenic bacterial clinical isolates such as Staphylococcus aureus, Escherichia coli, and Klebsiella pneumoniae from red betel leaves. Molecular investigation of endophytic isolates based on 16S rRNA gene sequence comparisons with the GenBank genome database (NCBI). Tripto Soya agar was employed in isolation (TSA). Simultaneously, the antibacterial activity test was performed on Muller Hinton agar using the welling method (MHA). Two of the three endophytic bacteria isolates were positive for inhibitory activity against Staphylococcus aureus and Escherichia coli. Endophytic bacterial isolates were characterized using biochemical testing stages, DNA extraction stages, 16S rRNA gene amplification, electrophoresis, and bioinformatics techniques MEGA 7 software that uses 16S rRNA genes to produce phylogenetic trees, and screening using bioinformatics techniques MEGA 7 software that uses 16S rRNA genes to produce phylogenetic trees. Endophytic bacteria isolates with codes DSM 1 and DSM 2 were found to be related to Bacillus pumilus strain AUEC29 and Bacillus safensis strain NBRC 100820, respectively. The function of endophytic bacteria in environmental preservation initiatives provides an alternative to the use of plants as therapeutic extracts.

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