Tujuan penelitian untuk mengetahui aktivitas antibakteri fraksi polar ekstrak etanol daun pepaya (Carica papaya L.) terhadap bakteri Staphylococcus aureus dan Escherichia coli. Proses ekstraksi dilakukan menggunakan metode maserasi dengan pelarut etanol 70%. Ekstrak etanol selanjutnya difraksinasi menggunakan pelarut etanol-air. Uji aktivitas antibakteri yang digunakan adalah metode difusi agar cara Kirby Bauer. Variasi konsentrasi fraksi etanol-air ekstrak etanol daun pepaya 10, 15, 20, 25, 30, dan 100%. Zona hambat pertumbuhan bakteri Staphylococcus aureus dan Escherichia coli diukur menggunakan jangka sorong. Hasil penelitian menunjukkan zona hambat fraksi etanol-air terhadap bakteri Staphylococcus aureus yang paling optimum yaitu pada konsentrasi 100% sebesar 14,75 mm; sedangkan pada bakteri Escherichia coli yang paling optimum yaitu pada konsentrasi 100% sebesar 11,53 mm. Namun tidak efektif jika dibandingkan dengan zona hambat kontrol positif kloramfenikol. Kesimpulan fraksi etanol-air ekstrak etanol daun pepaya (Carica papaya L.) dapat menghambat pertumbuhan bakteri Staphylococcus aureus dan Escherichia coli.Antibacterial activities of polar fraction of papaya leaf ethanolic extract (Carica papaya Linn.) against Escherichia coli and Staphylococcus aureusThis study aimed to determine the antibacterial activities polar fraction of papaya leaf ethanol extract against Escherichia coli and Staphylococcus aureus. Simplicia was extracted by ethanol 70% with the maceration method. Ethanolic extract was fractionated with ethanol water. Kirby Bauer diffusion method agar was used for antibacterial activities. Variation concentration of the ethanol-water fraction of papaya leaf ethanolic extract 10, 15, 20, 25, 30, and 100%. The growth inhibition zones of Escherichia coli and Staphylococcus aureus are measured using a caliper. The zone of inhibition of the ethanol-water fraction against Staphylococcus aureus was the most optimum at a concentration of 100% at 14.75 mm, while the most optimum for Escherichia coli was at a concentration of 100% at 11.53 mm, but did not effective than positive control Chloramphenicol. The conclusion of this study is that the ethanol-water fraction of papaya leaf ethanol extract could inhibit the growth of Staphylococcus aureus and Escherichia coli bacteria.