Abstract：Vanderbylia robiniophila is an important medicinal macrofungi. However, due to the lack of wild resources, the long period and the high cost of artificial cultivation, the fruit body of V. robiniophila is limited, so the extraction of polysaccharides from fermentation broth is an important way. In order to enrich the strain resources of V. robiniophila and improve the yield of polysaccharide in fermentation broth, a pure culture of V. robiniophila WH5 was obtained through the tissue isolation, it was identified by morphology and internal transcribed spacer (ITS) sequence analysis. The liquid fermentation technology of WH5 was optimized by single factor experiment and orthogonal experiment, and the anti-tumor activities of the exopolysaccharide in the fermentation broth were analyzed by cell experiment in vitro. The results showed that the length of ITS sequence from WH5 strain was 637 bp, and it was 99% similar to V. robiniophila (GenBank No. KX081122.1), clustered on one branch in the phylogenetic tree. Combining fruit body and mycelium morphology, strain WH5 was identified as V. robiniophila. The optimum liquid fermentation conditions of strain WH5 were as follows: Soluble starch 30 g/L, peptone 15 g/L, MgSO4•7H2O 1.0 g/L, KH2PO4 2.0 g/L, pH7.0, rotating speed 160 r/min, fermentation temperature 30 ℃, fermentation time 9 d, the yields of mycelium and exopolysaccharide were 27.52 and 1.91 g/L, respectively. It was found that exopolysaccharides from the fermentation broth could inhibit B16 and Hep-3B tumor cells with the half-inhibitory concentration (IC50) 24.12 and 13.25 mg/mL, respectively. This study is of great significance to enrich the fungus resources of V. robiniophila and promote the utilization of polysaccharides in the fermentation broth of V. robiniophila.
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