In this study, we introduce a simple and green method for synthesis of gold nanoparticles (AuNPs) using microbial glycolipid mannosylerythritol lipid (MEL) produced from
Ustilago maydis
CGMCC 5.203 and to evaluate their biomedical activities. MEL was found 10.3 g/L using sunflower oil. The formation of MEL-AuNPs was verified using UV–visible spectrum, XRD, TEM, FTIR, SEM, and EDX. In the biomedical examinations, MEL-AuNPs demonstrated potential cytotoxicity against HepG2 cells, and IC
50
values were found to be 100 and 75 µg/mL for 24 h and 48 h of exposure, respectively, which indicates its good performance against cancer cells. The IC
50
value of MEL-AuNPs was found to be 115 and 124 µg/mL for DPPH and ABTS scavenging activities, respectively. The biosynthesized MEL-AuNPs significantly inhibited cell growth of pathogenic Gram-positive and Gram-negative bacteria. These findings indicated that MEL plays a crucial role in the rapid biofabrication method of metallic NPs possessed the potential of biomedical activities.