粗毛纤孔菌液体发酵工艺优化及胞外多糖的抗菌和抗肿瘤活性

唐少军; 雷平; 邵晨霞; 吴胜莲; 杨祎; 靳磊; 刘宇波; 贺月林; 许隽

doi:10.13386/j.issn1002-0306.2020050184

粗毛纤孔菌液体发酵工艺优化及胞外多糖的抗菌和抗肿瘤活性

Optimization of Liquid Fermentation Process of Inonotus hispidus and Antibacterial and Antitumor Activities of Exopolysaccharides

摘要

摘要: 目的：优化粗毛纤孔菌液体发酵工艺，提高胞外多糖产量，以利于其在抑菌和抗肿瘤活性方面进行开发利用。方法：利用单因素实验结合正交试验优化粗毛纤孔菌液体发酵工艺；采用牛津杯法对胞外多糖进行抑菌活性实验；采用体外细胞实验分析胞外多糖的抗肿瘤活性。结果：粗毛纤孔菌的最适发酵工艺为：葡萄糖35 g/L，酵母粉10 g/L，KH₂PO₄ 1.5 g/L，MgSO₄·7H₂O 1.0 g/L，VB₁ 0.05 g/L，培养温度26℃，转速140 r/min，培养时间12 d，pH6.5，该发酵工艺下的菌丝体和胞外多糖的产量分别为1.75 g/100 mL、167 mg/100 mL，较优化前菌丝体和胞外多糖分别提高了1.16、1.85倍。胞外多糖能明显的抑制金黄色葡萄球菌、白色念珠菌、酵母菌，抑菌圈直径分别为11、12、11 mm，且对宫颈癌细胞Hela有明显的促凋亡作用。结论：优化了粗毛纤孔菌液体发酵工艺，可为粗毛纤孔菌发酵液中胞外多糖的开发利用提供参考。

Abstract: Objective:To optimize the liquid fermentation process and increase the yield of exopolysaccharides,which could be used in the antibacterial and antitumor activities. Methods:The single factor experiment combined with orthogonal experiment was used to optimize the liquid fermentation process of I.hispidus.The oxford cup method was used to test the antibacterial activity of exopolysaccharides. The antitumor activity of exopolysaccharide was analyzed in vitro. Results:The results showed that the optimum fermentation process of trichoderma was glucose 35 g/L,yeast powder 10 g/L,KH₂PO₄ 1.5 g/L,MgSO₄·7H₂O 1.0 g/L,VB₁ 0.05 g/L,culture temperature 26 ℃,rotate speed 140 r/min,incubation time 12 d,pH6.5.The yield of mycelia and exopolysaccharides under this fermentation process was 1.75 g/100 mL and 167 mg/100 mL,respectively,which were 1.16 and 1.85 times higher than those before optimization. Exopolysaccharide could inhibit staphylococcus aureus,candida albicans and yeast,the diameter of bacteriostatic circle was 11,12 and 11 mm,respectively. It also had a significant pro-apoptotic effect on cervical cancer cells.Conclusions:The liquid fermentation process of I. hispidus was optimized,which can provide reference for the development and utilization of exopolysaccharides in the fermentation broth of I. hispidus.