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Yellow root plants that have many medicinal properties such as antidiabetic, anti-inflammatory, anticancer and antibiotic. Based on the phytochemical test results of yellow root extract containing flavonoids, alkaloids, saponins, tannins, and terpenoids which can inhibit the growth of bacteria and fungi. The purpose of this study was to determine the antibacterial and antifungal activity of yellow root extracts against Escherichia coli bacteria as Gram-negative and Staphylococcus aureus as Gram-positive and the fungus Candida albicans. This study used a disk diffusion method with concentrations of yellow root extracts of 10%, 15%, and 20%. Respectively, positive controls for bacteria and fungi were amoxicillin and ketoconazole and CMC as negative control. The results show the highest average inhibitory diameter of yellow root extracts with a concentration of 20% on the growth of Escherichia coli and Staphylococcus aureus bacteria are 26.5 mm and 21.5 mm, respectively. The highest average inhibitory diameter of the fungus Candida albicans in yellow root extracts with a concentration of 15% is 23.2 mm. The conclusion of the study is the yellow root extract has antibacterial and antifungal activity against the growth of Escherichia coli and Staphylococcus aureus bacteria and the fungus Candida albicans.