蛹虫草-浙贝母-黑豆菌质固体双向发酵工艺及其抗肿瘤活性研究

Optimization of Solid Bidirectional Fermentation Process of Cordyceps militaris-Fritillaria thunbergii-Black Bean and Its Antitumor Activity

为实现浙贝母、黑豆高值化利用,开发一种具有抗肿瘤功效的蛹虫草菌质,将浙贝母-黑豆(浙贝母黑豆=21,质量比)作为蛹虫草(Cordyceps militaris)菌株发酵基质,以多酚含量为评价指标,采用单因素及响应面试验方法优化蛹虫草-浙贝母-黑豆菌质固体双向发酵工艺,采用MTT法测定发酵菌质体外抗肿瘤活性。结果表明,最佳发酵条件为料液比11.2(g/mL),接种量20 mL/100 g,培养温度21℃,培养时间29 d。优化条件下,菌质中多酚含量(6.24 mg/g)与模型预测值(6.35 mg/g)偏差为1.78%,且显著高于基质中多酚含量(1.19 mg/g);发酵菌质具有一定的体外抗肿瘤活性,其水提物对S180肿瘤细胞体外增殖抑制率表现出良好的量效及时效关系,以质量浓度相当于生药量200 mg/mL的水提液作用48 h条件下的细胞增殖抑制率最强,达到了73.08%,且在相同质量浓度及作用条件下,发酵菌质对S180肿瘤细胞的体外增殖抑制率是基质的1.10~1.88倍。

In order to achieve high-value utilization of Zhejiang beimu(Fritillaria thunbergii)and black beans,and to develop a kind of substance with anti-tumor effects about Cordyceps militaris,using F.thunbergii,black bean(21,mass ratio)as the fermentation substrate for C.militaris strains,the solid bi-directional fermentation technology of C.militaris-F.thunbergii-black bean was optimized by single factor test and response surface method.The antitumor activity of fermented mycoplasma in vitro was determined by MTT assay.The results showed that the optimal fermentation conditions were as follows:material-liquid ratio of 11.2 g/mL,inoculum volume of 20 mL/100 g,cultivation at 21℃,for 29 d.The deviation of the polyphenol content between the mycoplasma(6.24 mg/g)and the predicted value(6.35 mg/g)was 1.78%under optimized conditions,and the polyphenol content of the mycoplasma was significantly higher than the matrix(1.19 mg/g).The fermented mycoplasma could promote in vitro antitumor activities at certain extent,its water extract had a good dose-response and time-dependent relationship in the inhibition rate of S180 cells in vitro,and it showed the strongest activities under the condition of a mass concentration of 200 mg/mL and 48 hours of action,reaching 73.08%.The inhibition rate of S180 cells in vitro by fermenting bacterial substances was 1.10-1.88 times that of the substrate under the same mass concentration and action conditions.