青黛炮制浸泡发酵环节微生物群落结构与吲哚类成分转化研究  

Microbial community structure and transformation of indoles in soaking and fermentation of Indigo Naturalis

作　　者：贺亚男[1] 马乐乐 吴意 陈露梦 张定堃[1] 阳向波 许润春[1] 王芳 韩丽[1] 杨明 HE Ya-nan;MA Le-le;WU Yi;CHEN Lu-meng;ZHANG Ding-kun;YANG Xiang-bo;XU Run-chun;WANG Fang;HAN Li;YANG Ming(State Key Laboratory of Southwestern Chinese Medicine Resources,School of Pharmacy,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China;Key Laboratory of Modern Preparation of Traditional Chinese Medicine,Ministry of Education,Jiangxi University of Traditional Chinese Medicine,Narwhang 330004,China;Sichuan Liancheng Xunkang Pharmaceutical Co.,Ltd.,Yaan 625000,China)

机构地区：[1]成都中医药大学药学院西南特色中药资源国家重点实验室,四川成都611137 [2]江西中医药大学现代中药制剂教育部重点实验室,江西南昌330004 [3]四川联成迅康医药股份有限公司,四川雅安625000

出　　处：《中国中药杂志》2021年第13期3180-3187,共8页China Journal of Chinese Materia Medica

基　　金：国家重点研发计划项目(2018YFC1707205);国家自然科学基金面上项目(81773918);江西中医药大学创新药物与高效节能降耗制药设备国家重点实验室开放基金项目(GZSYS202003);成都中医药大学西南特色中药资源重点实验室开放研究基金项目(2020XSGG020)。

摘　　要：马蓝叶的浸泡发酵是青黛炮制的重要环节,主要目的在于促进靛蓝、靛玉红前体物质的合成、内源性酶以及其他有效成分的溶出,但忽略了微生物在发酵过程的作用。该研究基于16S扩增子测序和生物信息学分析技术对青黛浸泡发酵与石灰打靛过程微生物群落结构变化进行研究,并对发酵液中的靛蓝、靛玉红、靛红、色胺酮、吲哚苷等成分进行含量测定,研究微生物与主要成分变化的关联关系。结果表明,随着发酵的进行,微生物多样性逐渐下降,尤其是打靛后微生物多样性降至最低。变形菌门、拟杆菌门、厚壁菌门是发酵过程主要的优势群落,且随着发酵的进行,微生物结构不断变化,变形菌门随着发酵时间增加,相对丰度逐渐降低,打靛后降至最低;厚壁菌门相对丰度升高,拟杆菌门先下降后升高。青黛中有效物质的含量也表现出了不同的规律,随着发酵的进行吲哚苷含量逐渐下降,靛蓝呈现先上升后下降趋势,靛玉红、靛红呈现先下降后上升趋势,色胺酮含量逐渐升高,这可能与微生物在不同成分的合成过程所扮演的作用有关。该研究初步阐明了微生物在浸泡发酵过程中的重要作用,为青黛炮制工艺过程的控制与高质量青黛饮片的制备提供了科学依据。The soaking and fermentation of Baphicacanthus cusia(Nees),the important intermediate link of Indigo Naturalis processing,facilitates the synthesis of indigo and indirubin precursors and the dissolution of endogenous enzymes and other effective components,while the role of microorganisms in the fermentation is ignored.The present study investigated the changes of microbial community structure in Indigo Naturalis processing based on 16S amplicon sequencing and bioinformatics.Meanwhile,the contents of indigo,indirubin,isatin,tryptanthrin,indole glycoside,etc.were determined to explore the correlation between the microorganisms and the alterations of the main components.As demonstrated by the results,the microbial diversity decreased gradually with the fermentation,which bottomed out after the addition of lime.Proteobacteria,Bacteroidetes,and Firmicutes were the main dominant communities in the fermentation.The relative abundance of Proteobacteria declined gradually with the prolongation of fermentation time,and to the lowest level after the addition of lime.The relative abundance of Firmicutes increased,and that of Bacteroidetes decreased first and then increased.The contents of effective substances in Indigo Naturalis also showed different variation tendencies.As fermentation went on,indole glycoside decreased gradually;indigo first increased and then decreased;indirubin and isatin first decreased and then increased;tryptanthrin gradually increased.Those changes were presumedly related to the roles of microorganisms in the synthesis of different components.This study preliminarily clarified the important role of microorganisms in the soaking and fermentation and provided a scientific basis for the control of Indigo Naturalis processing and the preparation of high-quality Indigo Naturalis.

关 键 词：青黛 浸泡发酵 微生物群落结构 靛蓝 靛玉红 

分 类 号：R283[医药卫生—中药学]