冠突散囊菌对三七和金银花成分的转化作用及机理研究

The essence of fahua-fermentation of fuzhuan brick tea is solid state fermentation process of tea by Eurotium cristatum, it secrete a variety of enzymes for biotransformation of natural products in tea, which highlight huge potentiality in fermentation of traditional chinese medicine. In this study, we seek for the main variant phytochemistry and novel bioactive compounds in biotransfomation products from Panax notoginseng (Burk.) F.H. Chen and Lonicera japonica Thunb fermented by E.cristatum. Chemical constituents difference between before and after fermentation have been identified using UPLC/Q-TOF MS metabolomics analysis. The biotransformation pathway of major different novel compounds were described using RNA-seq and real time PCR. These results would confirm the biotransfomation of P. notoginseng (Burk.) F.H. Chen and L. japonica Thunb fermented by E.cristatum, elucidate the mechanism of biotransfomation and enzyme catalysis, and promote the development of fermentation of traditional chinese medicine and drug discovery.

茯砖茶“发花”工艺的实质是冠突散囊菌（Eurotium cristatum）以茶叶为基质的固态发酵过程，其中冠突散囊菌分泌各种酶类对茶叶化学成分进行生物转化，凸显出冠突散囊菌在发酵炮制中药上的巨大潜力。本项目拟以三七、金银花为药性基质，冠突散囊菌为发酵菌种，采用固态发酵技术，HPLC指纹图谱叠加比对分析发酵前后化学成分差异；UPLC/Q-TOF MS代谢组学技术研究发酵前后物质的变化，结合RNA-seq与定量PCR解析固体发酵过程中主要差异化合物的生物转化途径。本项目实施，可明确冠突散囊菌对三七、金银花的生物转化作用，初步阐明生物转化途径及新颖的酶催化机制，也有助于促进发展中药发酵理论及中药新药研发。

散囊菌是广泛存在于发酵茶中的一大类分类含糊，容易混淆，很多疑虑亟待阐明的益生真菌，统称为“金花菌”，包括冠突散囊菌、阿姆斯特丹散囊菌、谢瓦散囊菌和蜡叶散囊菌等，分泌的各种酶类极为丰富，在发酵过程中对茶叶成分生物转化是发酵茶口感、滋味和营养价值提升、保健功效形成的基础。微生物的生物转化在中药材发酵炮制及药理活性提升上凸显出巨大潜力，已成为国际研究热点。本项目以三七、金银花、葛根、黄芪等为药性基质，冠突散囊菌、阿姆斯特丹散囊菌为发酵菌种，采用固态发酵，醇提法提取，进行体外抗氧化、降血糖检测，HPLC指纹图谱叠加比对分析发酵前后化学成分差异；重点选择三七，进一步以UPLC/Q-TOF MS代谢组学技术研究发酵前后物质的变化，RNA-seq与定量PCR解析三七发酵过程中主要差异化合物的生物转化途径与机制。在中药材发酵研究基础上形成以金银花、葛根和黄芪等为发酵基质，冠突散囊菌、阿姆斯特丹散囊菌和酵母菌等为发酵菌种的高活性微生态制剂，在山东省梁山县、青海、内蒙、贵州等地进行小范围推广。养殖户反馈结果表明，该产品促生长、提升免疫力，调节动物肠道菌群，抗病毒，抗应激，降低腹泻率及死淘率的效果显著，能部分代替抗生素。