The present study is focused on the production of a biosurfactant using Yarrowia lipolytica MTCC 9520 by employing inexpensive lipid waste, chicken tallow from slaughterhouses. Plackett–Burman and Box–Behnken Design analyses were adopted for preliminary screening of medium variables and further optimization. The maximal yield of 4.4 g L−1 of the biosurfactant was obtained from the optimized medium. The highest emulsification activity was found to be 55%, and the surface tension decreased to 37 mN m−1 at the end of 96 h. The critical micelle concentration of the biosurfactant was calculated as 1.2%. The produced biosurfactant was characterized as cationic lipoprotein in type, and the proteins present in the biosurfactant were observed to have molecular weights between 75 and 100 kDa. The fatty acids composition of the biosurfactant was detected by gas chromatography–mass spectrometry (GC–MS) analysis. Fourier transform infra red (FTIR) and nuclear magnetic resonance (NMR) analysis confirmed the lipoprotein nature of the extracted biosurfactant. Thermogravimetric (TG) and differential scanning calorimetry (DSC) analysis revealed the thermostable nature of the extracted biosurfactant. Surface plasmon resonance vibration peak at 421 nm was observed for the surfactant‐stabilized silver nanoparticles (AgNP) through UV–Vis spectrophotometry. The average particle size of the synthesized AgNP was calculated as 7.2 ± 0.4 nm from transmission electron microscopy (TEM) analysis. Energy dispersive x‐ray (EDX) spectroscopy exhibited the presence of silver in the synthesized nanoparticles. The zeta potential value of the synthesized AgNP was measured as −22.2 mV, and the polydispersity index was found as 2.3 through dynamic light scattering (DLS) analysis.

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