金针菇、赤芝混菌发酵工艺优化及其活性成分检测

Mixed fermentation technology and active substance detection of Flammulina velutipes and Ganoderma lucidum

采用金针菇和赤芝混菌发酵,对培养基组成、培养温度、培养时间和转速等因素进行了优化,摇瓶和发酵罐逐级扩大培养后,分析发酵液和菌丝体中三萜、多糖和锌元素等含量,并与单菌发酵进行对比分析,探讨混菌发酵对代谢产物合成生成转化的影响。通过单因素和正交试验确定了最佳工艺参数:山芋、马铃薯等天然培养基中添加10 g/L黄豆粉和30 g/L玉米浆对三萜和多糖等代谢产物的生成最有利,温度24℃、转速110 r/min条件下混菌发酵培养10 d,胞内三萜、胞外三萜和胞外多糖含量分别为3.45 mg/g、3.05 mg/mL、38.67 mg/mL。扩大培养有利于菌体生长和代谢产物合成,10 L发酵罐混菌培养时胞内和胞外三萜产量分别达到3.94 mg/g和3.49 mg/mL,胞外多糖52.71 mg/mL,胞内锌含量186.42μg/g,均高于单菌发酵培养。结果表明混菌发酵金针菇和赤芝菌体生长良好,且存在一定的协同增效作用,促进了三萜、多糖等活性成分的代谢合成和微量元素的富集转化,拓展了单一菌株发酵的应用范畴,具有良好的应用前景。

The fermentation technology of mixed fermentation was adopted to select the fermented Flammulina velutipes and Ganoderma lucidum as the fermentation object. The culture medium composition, culture temperature, culture time and shaker speed were optimized. The biomass, triterpenoids, polysaccharide and zinc content were detected after the gradual expansion of the shake flask and fermenting tank, and the comparative analysis with single bacteria fermentation was conducted to explore the effect of the mixed fermentation on the synthesis and transformation of metabolites. The optimum conditionsby single factor and orthogonal test were identified： addition of 10 g/L soybean powder and 30 g/L corn syrup in natural medium such as sweet potato and potato was beneficial to the generation of triterpenes and polysaccharides. The optimal mixed fermentation conditions were identified： culture temperature 24 ℃, shaker speed 110 r/min and culture time 10 days, the contents of intracellular triterpene, extracellular triterpene and extracellular polysaccharide were up to 3.45 mg/g, 3.05 mg/mL, 38.67 mg/mL respectively. The experimental results showed that mixed fermentation was beneficial to the growth of microorganism and the synthesis of metabolites. The experimental results of 10 L fermenting tank showed that some contents of active ingredients were higher than that of single bacteria fermentation, such as the yield of intracellular and extracellular triterpenoids was up to 3.94 mg/g and 3.49 mg/mL respectively, the content of extracellular polysaccharide was 52.71 mg/mL, and the content of intracellular trace element zinc was 186.42 μg/g. These results showed that mixed fermentation was beneficial to microbial growth, and there was a certain synergistic effect between Flammulina velutipes and Ganoderma lucidum. It could improved the deficiency of single strain fermentation and expand the application scope of the single strain fermentation, which was a good application prospect.