In the present work, the optimization of extraction, emulsifying properties, and biological activities of polysaccharides from Lentinula edodes Sing (LES) were studied. The results showed LES polysaccharides extracted by hot water or ultrasonication are a group of β-glucan. Among all the samples, the one extracted by hot water showed the best emulsifying capacity. In addition, the results demonstrated that LES polysaccharide had strong scavenging activities in vitro on DPPH and ABTS radicals, which reached the highest level for the one extracted by 90 min ultrasonication (p < 0.05). Overall, Lentinula edodes Sing polysaccharides (LESPs) may have potential applications as emulsifying agents in food industries.