花脸香蘑E70菌株发酵条件优化及其胞外多糖生物活性

Fermentation Condition Optimization and Immunoactivity of Extracellular Polysaccharide from Lepista sordida E70

该研究以达州市农业科学研究院野外采集的花脸香蘑子实体经分离的E70菌株为研究对象,结合形态学鉴定和分子生物学鉴定方式对其进行分类鉴定,采用单因素实验研究非营养因子对其液体发酵产胞外多糖的影响,并以细胞增殖-毒性检测试剂盒(cell counting kit-8,CCK-8)探寻其发酵液胞外多糖对B细胞、T细胞和巨噬细胞RAW264.7增殖的影响。结果显示,该野生菌株为花脸香蘑Lepista sordida,适宜其胞外多糖合成的最佳非营养因子为接种量20个(0.1 cm^(2)/个)、温度25℃、转速180 r/min,此条件下胞外多糖含量高达(6.07±0.26)g/L。花脸香蘑E70菌株胞外多糖对小鼠B细胞、T细胞和巨噬细胞在药物组各浓度下皆具有明显增殖效果,对B细胞效果最显著,在药物浓度为40μg/mL时,最高增殖率可达65.89%(P<0.0001)。

The fruiting body of the wild strain E70 suspected as Lepista sordida collected and isolated by the Dazhou Academy of Agricultural Sciences was identified based on morphological and molecular biological evi⁃dence.Single factor tests were conducted to study the influences of non⁃nutritional factors on the production of extracellular polysaccharide by liquid fermentation.The effects of extracellular polysaccharide on the prolifera⁃tion of B cells,T cells,and RAW 264.7 macrophages were investigated by the cell counting kit⁃8 method.The wild strain E70 was identified as L.sordida.The optimal non⁃nutritional factors for the synthesis of extracellular polysaccharide were inoculation amount of 20 PC(0.1 cm^(2)/PC),temperature of 25℃,and rotation speed of 180 r/min.Under these conditions,the extracellular polysaccharide yield was as high as(6.07±0.26)g/L.The extracellular polysaccharide of L.sordida E70 at different concentrations had significant proliferative effects on the proliferation of B cells,T cells,and macrophages in mice.Particularly,it exerted the strongest promoting ef⁃fect on the proliferation of B cells,which showed the highest proliferation rate of 65.89%at the extracellular polysaccharide concentration of 40μg/mL(P<0.0001).