化风丹药母发酵过程中优势菌种分离鉴定及不同温度和pH值下菌株生长情况  

Isolation and identification of dominant strains in the fermentation process of HuaFengDan YaoMu and investigation of their growth under different temperature and pH value

作　　者：曹国琼 刘敏 刘耀 徐剑 陈有丽 吴静澜 张永萍 CAO Guo-qiong;LIU Min;LIU Yao;XU Jian;CHEN You-li;WU Jing-lan;ZHANG Yong-ping(School of Pharmacy,Guizhou University of Traditional Chinese Medicine,Guiyang 550025,China;National Engineering Research Center for Ethnic Miao Medicine,Guiyang 550025,China;Guizhou Engineering Research Center for Processing and Preparation of Traditional Chinese Medicines,Guiyang 550025,China;Zunyi Liaoyuanhetang Pharmaceutical Co.,Ltd.,Zunyi 563000,China)

机构地区：[1]贵州中医药大学药学院,贵州贵阳550025 [2]国家苗药工程技术研究中心,贵州贵阳550025 [3]贵州中药炮制与制剂工程技术研究中心,贵州贵阳550025 [4]遵义廖元和堂药业有限公司,贵州遵义563000

出　　处：《中成药》2023年第12期4011-4017,共7页Chinese Traditional Patent Medicine

基　　金：国家自然科学基金(81860704,82360774);贵州省科学技术厅科学技术基金(黔科合基础-ZK[2021]一般517);贵州省高等学校中药民族药(苗药)新剂型新制剂工程研究中心(黔教技[2022]022号);贵州省高层次创新型人才培养计划(“千”层次)(2020)。

摘　　要：目的对化风丹药母发酵过程中优势菌种进行分离鉴定,考察不同温度和pH值对优势菌株生长的影响。方法用选择性培养基对化风丹药母发酵过程中0周至6周样品中的细菌、霉菌、酵母菌分别进行培养、菌落计数、分离纯化优势菌种;采用16S rDNA,26S rDNA序列对优势细菌和真菌进行鉴定;分别考察阿氏芽孢杆菌、汉逊德巴利酵母菌、米曲霉的最适宜生长温度、pH值。结果化风丹药母发酵过程中细菌0周至4周数量增加,4周至6周数量趋于稳定;酵母菌在发酵0周至3周时数量增加,4周至6周数量不断减少;霉菌在0周时没有生长,1周至4周数量平稳增加,5周至6周霉菌数量呈现下降趋势。通过构建系统发育树,鉴定筛选出化风丹药母发酵过程中的优势菌种,以阿氏芽孢杆菌、汉逊德巴利酵母菌、米曲霉为研究对象,其生长的最适温度分别为35、25~30、35℃;最适pH值分别为8、6、7。结论化风丹药母发酵过程中优势微生物分别为阿氏芽孢杆菌、汉逊德巴利酵母菌、米曲霉。AIM To isolate and identify the dominant strains in the fermentation process of the HuaFengDan YaoMu,and to investigate the effects of different temperatures and pH values upon their growth.METHODS During the 0-6 weeks fermentation process of HuaFengDan YaoMu in the selective medium,the cultured bacteria,the molds and the yeasts had the colonies counted,and the dominant strains isolated and purified,particularly the dominant bacteria and fungi determined by 16S rDNA and 26S rDNA sequences.The optimal growth temperature and pH value of Bacillus aryabhattai,Debaryomyces hansenii,Aspergillus oryzae were investigated respectively.RESULTS HuaFengDan YaoMu found its bacterial counts increased from 0 to 4 weeks and stabilized from 4 to 6 weeks;its yeast counts increased during 0-3 weeks,and decreased continuously during 4-6 weeks;and its mold counts remained unchanged at 0 week,but increased steadily from 1 to 4 weeks,and decreased at 5 to 6 weeks of the fermentation.In the fermentation process of HuaFengDan YaoMu,the dominant strains of Bacillus aryabhattai,Debaryomyces hansenii and Aspergillus oryzae,identified and screened out by the phylogenetic tree,adapted best to the growth temperature of 35℃,25-30℃and 35℃,at pH value of 8,6 and 7,respectively.CONCLUSION In the fermentation process of HuaFengDan YaoMu,the dominant microorganisms of Bacillus aryabhattai,Debaryomyces hansenii,Aspergillus oryzae are identifiable and separable.

关 键 词：化风丹药母 分离与鉴定 阿氏芽孢杆菌 汉逊德巴利酵母菌 米曲霉 

分 类 号：R282[医药卫生—中药学]