Yan-Lin Chen, Yun-Ting Wang, Kai-le Guan, Xu Zhou, Pei-Jie Han, Xue-Wei Wang, Fen-Yan Bai, Jie D U, Yu-Zhen Ren
China National Traditional Chinese Medicine Co., Ltd. Beijing 100195, China.
Beijing Institute for Drug Control Beijing 102206, China.
Zhongguo Zhong Yao Za Zhi. 2020 Nov;45(21):5219-5225. doi: 10.19540/j.cnki.cjcmm.20200819.301.
This study aimed to clarify the microbial diversity, dominant species and the change of community structures in the fermentation of Liushenqu(Massa Medicata Fermentata), and explore the material foundation of its pharmacodynamics effect. On the basis of standardizing the fermentation process, Massa Medicata Fermentata was prepared by screening and optimizing the recipes and the standard formula issued by the Ministry. The community structure and growth process of fungi and bacteria in the samples at five time points(0, 17, 41, 48, 65 h) in the fermentation process of Massa Medicata Fermentata were analyzed by using isolation and culture of eight different media and high-throughput DNA sequencing technology. The results indicated that the samples of the two recipes pre-sented high microbial diversity at the initial fermentation stage, with Aspergillus spp. as the dominant species. As the fermentation process goes forward, Saccharomycopsis fibuligera and Rhizopus oryzae soon became dominant species from 17 h after fermentation commencement point to the fermentation end, while the other species were inhibited at a lower level from 17 h. The species diversity of bacteria in the initial fermentation samples was also high, and Enterobacter was the dominant species. Enterobacter cloacae, Pediococcus pentosaceus and Cronobacter sakazakii became dominant bacterial species 17 h after fermentation commencement, while the species diversity was decreased. Our results will be a scientific basis for promoting the fermentation process of Massa Medicata Fermentata by using pure microbial cultures.
本研究旨在阐明六神曲发酵过程中的微生物多样性、优势菌种及群落结构变化，探讨其药效作用的物质基础。在规范发酵工艺的基础上，通过筛选优化配方及参照部颁标准配方制备六神曲。采用8种不同培养基分离培养结合高通量DNA测序技术，对六神曲发酵过程中5个时间点（0、17、41、48、65 h）样品中的真菌和细菌群落结构及生长过程进行分析。结果表明，两种配方样品在发酵初期微生物多样性较高，以曲霉属为优势菌种。随着发酵进程推进，扣囊复膜孢酵母和米根霉从发酵起始点后17 h至发酵结束很快成为优势菌种，而其他菌种从17 h起受到较低程度抑制。发酵初期样品中细菌的物种多样性也较高，以肠杆菌属为优势菌种。发酵起始后17 h，阴沟肠杆菌、戊糖片球菌和阪崎肠杆菌成为优势细菌菌种，而物种多样性降低。本研究结果将为利用纯微生物培养促进六神曲发酵过程提供科学依据。
