Box-Behnken响应面法优化六神曲两优势菌协同发酵工艺

Box-Behnken Response Surface Method Optimizes Co-fermentation Process of Massa Medicata Fermentata by Two Dominant Fungi

目的:利用Box-Behnken响应面法优化六神曲中2种优势菌的协同发酵工艺。方法:利用米根霉Rhizopus oryzae、扣囊复膜孢酵母Saccharomycopsis fibuligera 2种优势菌协同六神曲发酵,在《中华人民共和国卫生部药品标准·中药成方制剂》第19册中六神曲组方的基础上,以蛋白酶活力和淀粉酶活力为评价指标进行发酵工艺单因素考察,进而采用Box-Behnken响应面法设计四因素三水平试验,并结合CRITIC权重法计算2种酶活力的综合得分,优化筛选六神曲2种优势菌协同发酵工艺。结果:以米根霉和扣囊复膜孢酵母按1∶2接种比例和15%总接种量接种于六神曲基质,32.5℃温度下发酵6 d制得六神曲,与同一厂家的9批商品六神曲相比,自制六神曲的酶活力批次间更稳定。结论:优化后的两菌协同发酵六神曲批间质量更加稳定。

Objective: To optimize the co-fermentation process of Massa Medicata Fermentata(MMF) by two dominant fungi based on Box-Behnken response surface method. Methods: Rhizopus oryzae and Saccharomycopsis fibuligera, two dominant fungi in MMF, were used to jointly ferment MMF. Based on the prescription of MMF in Drug Standard of Ministry of Health of the People’s Republic of China-Traditional Chinese Medicine Prescription Preparation(Volume 19), this study used protease activity and amylase activity as the evaluation indicators to carry out the single-factor experiments in the fermentation process. The Box-Behnken response surface method was used to design the experiment with4 factors and 3 levels, and the comprehensive score for protease activity and amylase activity in MMF was calculated by criteria importance though intercrieria correlation(CRITIC) method. The co-fermentation process of MMF by two dominant fungi was finally optimized and selected. Results: R. oryzae and S. fibuligera were inoculated into the natural fermentation system of MMF with 1∶2 of inoculation ratio and 15% total inoculation amount. The self-made MMF was obtained after 6d of co-fermentation at 32.5 ℃. As compared with the 9 batches of commercial MMF produced by the same manufacturer,the enzyme activities of self-made MMF were more stable among batches. Conclusion: After optimization, the quality of batches of MMF fermented by two dominant fungi is stable.