Surfactin, a lipopeptide biosurfactant, was used to stabilize the formation of silver nanoparticles. Synthesis of silver nanoparticles using a borohydrate reduction was performed at three pH levels (pH 5, 7 and 9) and two different temperatures in the presence of surfactin. The nanomaterials were characterized by UV–vis spectroscopy, X-ray diffraction and transmission electron microscopy. The nanoparticles were synthesized at different pH conditions and temperature and remained stable for 2 months. The UV–vis spectra showed a surface plasmon resonance vibration band at 428 nm for all samples. TEM micrographs revealed that the mean nanoparticle size decreased with increase of pH from 5 to 9 (i.e. 17.8 ± 9.8, 6.9 ± 1.5 and 4.3 ± 1.1 nm) at 4 °C. However, at room temperature, size increased with pH (4.9 ± 1.4, 6.5 ± 1.6, 9.7 ± 4.3 nm at pH 5, 7 and 9 respectively). This report describes the use of a renewable, environmentally compatible, biodegradable surfactant as a stabilizing agent for the synthesis of silver nanoparticles.