目的: 基于前期胆南星菌种分离及炮制原理研究,对优势菌种进行筛选并进一步优化混合菌种发酵胆南星的炮制工艺,为胆南星混合菌种发酵新工艺提供实验依据。方法: 将前期分离的菌种分别进行单菌种发酵制备胆南星,采用HPLC-ELSD法测定不同菌种制得的胆南星中猪胆酸、猪去氧胆酸、鹅去氧胆酸含量,筛选出胆南星发酵优势菌种。以游离型胆酸总量、牛磺酸、甘氨酸和胆红素为指标,以混合菌种的加入量、发酵温度、发酵时间为考察因素,采用正交试验联合多指标加权评分法对混合菌种发酵工艺进行考察,确定胆南星混合菌种发酵新工艺。结果: 筛选出铅黄肠球菌和肠球菌（厌氧）为优势菌种;混合菌种发酵炮制的新工艺为：在天南星细粉中加入含有10%~15%混合菌液[铅黄肠球菌液和肠球菌（厌氧）液1 ∶ 1混合]的猪胆汁1 ∶ 1混匀,置于32 ℃、80%湿度的恒温恒湿箱中发酵培养15 d,取出,蒸1 h至透,切块,晒干。结论: 优选出的胆南星混合菌种发酵工艺稳定可行,可提高胆南星产品质量及其稳定性,为后续发酵制胆南星的规范化生产提供科学依据。
Objective: To provide an experimental basis of the new fermentation process of multi-strain, based on the previous studies on the strain separation and processing principle of Arisaema Cum Bile, and the dominant strain is screened out and the process of multi-strain fermentation process was further optimized. Methods: Every strain isolated in the previous stage was single fermented to prepare a series of Arisaema Cum Bile. The content of hyocholic acid, hyodeoxycholic acid, and chenodeoxycholic acid was determined using HPLC-ELSD and screened for the advantageous strain. Taking the total amount of free bile acid and the content of taurine, glycine, and bilirubin as the indexes, making the addition amount, fermentation temperature, and fermentation time of the multi-strains as the factors, and combining the orthogonal test and multi-index weighted scoring method, the fermentation process was investigated and the new fermentation process of the multi-strains was determined. Results: The advantageous strains were identified as Lactobacillus plantarum and Enterococcus (anaerobic). A new process of multi-strain fermentation has been screened out: mix with a ratio (1 ∶ 1) of Arisaematis Rhizoma powder and bile containing 10%-15% multi-strains liquid [1 ∶ 1 mixture of Lactobacillus plantarum and Enterococcus (anaerobic)], ferment at 32 ℃ with 80% humidity for 15 days, take out, steam for 1 h to pass thoroughly, cut into diced, and dry in the sun. Conclusion: The optimized fermentation process for Arisaema Cum Bile is stable and feasible, which can improve the quality of the products and provide scientific basis for the standardized production of Arisaema Cum Bile.
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