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目的:揭示百药煎传统炮制过程中的微生物菌群,同时初步考察所分离菌株鞣质水解能力。方法:应用经典微生物分离纯化方法,结合形态学考察和16S r DNA或18S r DNA基因序列分析,对百药煎炮制过程中样品进行微生物的分离及菌种初步分类鉴定。采用含单宁酸的固体培养基初步筛选具有鞣质降解能力的菌株。结果:共分离获得细菌7株、酵母菌7株、丝状真菌3株。7种细菌分别为枯草芽孢杆菌、蜡样芽孢杆菌、地衣形芽孢杆菌、黄海芽孢杆菌、巨大芽孢杆菌菌株、同温层芽孢杆菌、其他芽孢杆菌;7种酵母菌分别为伯顿丝孢毕赤酵母、2株克鲁维酵母菌株、弗比恩酵母菌、奥默柯达菌、异常威克汉姆酵母菌、异常毕赤酵母菌;3株丝状真菌分别为青霉菌、卷枝毛霉菌、黑曲霉菌。除枯草芽孢杆菌、黄海芽孢杆菌、芽孢杆菌、克鲁维酵母菌HMY3、弗比恩酵母菌外,其余菌株均具有鞣质降解能力。结论:百药煎传统发酵炮制工艺涉及多种微生物共同参与混合发酵过程,初步筛选证明5株细菌、5株酵母菌和3株丝状真菌具有鞣质降解能力,这一发现为研究百药煎现代炮制工艺奠定了微生物菌种基础。
OBJECTIVE: To reveal the microbial flora during the traditional processing of Chinese medicinal herbs, and to investigate the hydrolytic capacity of tannin in the isolated strains. Methods: The classical microorganism separation and purification method, combined with morphological investigation and 16S r DNA or 18S r DNA sequence analysis were used to separate the microorganisms in the samples during the course of the hundred herbs decocting and preliminary identification of the strains. The tannic acid-containing solid medium was used to screen the strains with tannin degradation ability. Results: Seven bacteria, seven yeasts and three filamentous fungi were isolated. The seven kinds of bacteria were Bacillus subtilis, Bacillus cereus, Bacillus licheniformis, Bacillus megaterium, Bacillus megaterium, Bacillus thuringiensis and other Bacillus. The seven yeasts were Bacillus Yeast, two Kluyveromyces strains, Phoebe yeast, Omekodak, abnormal Wickham yeast, Pichia anomali; three filamentous fungi were Penicillium, Mucor, Aspergillus niger. Except for Bacillus subtilis, Bacillus burgdorferi, Bacillus, Kluyveromyces HMY3 and Phoebe yeast, all the other strains had the capacity of tannin degradation. CONCLUSION: The traditional fermentation process of Hundreds of herbs involves multiple microorganisms participating in the mixed fermentation process. Preliminary screening shows that five strains of bacteria, five strains of yeast and three strains of filamentous fungi have the capacity of tannin degradation. Modern processing technology laid the foundation for microbial species.