In order to realize the resource utilization of the Pinus koraiensis pine cone, the polysaccharides were obtained by a traditional hot water extraction process, and the optimal extraction process was confirmed. The effects of a traditional hot water, alkali, and ultrasound assisted extraction on the yield and properties of polysaccharides were studied. The structure, monosaccharide composition, and content of polysaccharides obtained by different extraction methods were analyzed by means of Fourier infrared chromatography and liquid chromatography. Effects of polysaccharides extracted by different methods on the antimicrobial activity of Staphylococcus aureus and Escherichia coli were studied. The scavenging effect of DPPH free radicals were also examined. The optimum condition of the hot water extraction was identified as an extraction temperature of 100 °C, a ratio of material to water at 1:16 (w/v), an extraction time of 4 h, and a yield of polysaccharide at 10.2%. In comparison, the yield of the pine cone polysaccharide was improved significantly to 29.1% by using NaOH solution extraction method. The FTIR spectra of polysaccharides showed that the samples possessed the typical chemical structure of a polysaccharide.