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An efficient compound enzyme extraction process was developed and optimized to extract the polysaccharide from  Erythronium sibiricum  bulb via response surface methodology. The polysaccharide E2P was obtained. Then, the preliminary characteristics of E2P were determined via colorimetry and chromatography. Additionally, the immunoregulatory activities of E2P and another polysaccharide (ESBP, extracted using the hot water method) were compared. The optimized extraction results were as follows: temperature (54.56 °C), time (2.52 h), pH (6.53), and enzyme concentration ratio (0.5% cellulase:1.5% amylase). The yield (64.18% ± 2.91%) obtained under the aforementioned conditions was considerably higher than the yield of ESBP (37.25% ± 0.17%). The total sugar, uronic acid, starch, and protein contents of E2P were 81.77% ± 2.84%, 3.31% ± 0.45%, 3.29% ± 0.01%, and 0.24% ± 0.02%, respectively. The HPLC result suggested that the predominant monosaccharides of E2P included glucose, galactose, and arabinose, with a molar ratio of 543.2:1:1.8. The in vitro tests in RAW264.7 cells indicated that ESBP exhibited better immunomodulatory activities than E2P. In particular, ESBP can promote the proliferation, phagocytosis, and cytokine secretion abilities of cytokines, such as nitric oxide, tumor necrosis factor-α, and interleukin (IL)-1β of RAW264.6 cells. By contrast, E2P can only promote phagocytosis ability and the secretion of IL-1β.

Optimization of the composite enzyme extraction of polysaccharide from Erythronium sibiricum bulb and its immunoregulatory activities