Polysaccharides are ideal green synthetic raw materials to improve the biocompatibility of metal nanoparticles. However, polysaccharides generally have weak reducibilities, being challenging supports for the direct preparation of Ag nanoparticles. In this work, gold nanoclusters were prepared using a triple helix glucan (Lentinan) via microwave-assisted synthesis and subsequently employed as seeds for the synthesis of a series of Ag-Au alloy nanoparticles (Ag-AuNPs). The results showed that gold nanoclusters can remarkably speed up the preparation of Ag-AuNPs without the addition of any other chemicals. The particle size of Ag-AuNPs increased at increasing Ag contents in the alloy. Results of UV-vis, transmission electron microscope (TEM) and inductively coupled plasma mass spectrometry (ICP-MS) suggested that Ag was quickly reduced to irregular silver nanoparticles (with gold nanoparticles as seeds), then gradually form more regular nano-alloys. Additionally, X-ray diffraction (XRD) and zeta potential results suggested that Ag-AuNPs could entrap the hydrophobic cavity of triple helix polysaccharides during the renaturation process. The nanocomposites exhibited good antifungal activity and low cytotoxicity to RAW264.7, Hela and LO2 cell lines in vitro.