茯茶冠突散囊菌发酵生产吲哚生物碱的研究

茶叶科学 ›› 2017, Vol. 37 ›› Issue (5): 503-512.

茯茶冠突散囊菌发酵生产吲哚生物碱的研究

刘丽萍^1,2, 唐雨薇^1,2, 王若娴¹, 李志冰¹, 刘仲华^1,2,3, 刘硕谦^1,2,3,*

1. 湖南农业大学园艺园林学院,湖南长沙 410128;
2. 国家植物功能成分利用工程技术研究中心,湖南长沙 410128;
3. 教育部茶学重点实验室,湖南长沙410128

收稿日期:2017-06-02 修回日期:2017-06-19 出版日期:2017-10-25 发布日期:2019-08-23
通讯作者: *shqianliu@sina.com
作者简介:刘丽萍,女,硕士研究生,主要从事茶叶化学生物学研究。
基金资助:
国家自然科学基金（31670689）

Production of Bioactive Indole Alkaloids through Fermention of Eurotium cristatum from Fuzhuan Tea

LIU Liping^1,2, TANG Yuwei^1,2, WANG Ruoxian¹, LI Zhibing¹, LIU Zhonghua^1,2,3, LIU Shuoqian^1,2,3,*

1. College of Horticulture and Hardening, Hunan Agricultural University, Changsha 410128, China,
2. National Research Center of Engineering Technology for Utilization of Functional Ingredients from Botanicals, Changsha 410128, China,
3. Key Lab of Tea Science, Ministry of Education, Changsha 410128, China

Received:2017-06-02 Revised:2017-06-19 Online:2017-10-25 Published:2019-08-23

摘要/Abstract

摘要： 冠突散囊菌是茯砖茶“发花”过程中的优势菌,其体内含有丰富的吲哚类生物碱类代谢产物。吲哚生物碱类化合物具有抗菌、抗过敏、抗辐射、抗紫外和抗肿瘤等重要的生物活性,开发前景广阔。为提高吲哚类生物碱类代谢产物的产量,本研究考察了冠突散囊菌繁殖、生长与发酵的影响因素,并利用薄层色谱（TLC）、高效液相色谱（HPLC）、液相色谱-质谱联用技术（LC-MS）对其发酵产物内含成分进行了分析。结果表明冠突散囊菌在察氏固体培养基上能快速繁殖,8βd内能产生大量的成熟孢子,液体培养的最佳条件与时长分别为pH 6.0、温度30℃、转速200βr·min^-1和培养时长6βd;鲜茶干粉、鲜茶提取物、茯茶粉、茯茶提取物和色氨酸等能显著诱导吲哚生物碱的合成,其中色氨酸诱导效应最强;内含成分分析表明,TLC可分离出至少10种冠突散囊菌代谢物质,HPLC与LC-MS检测分析其代谢产物的主要成分为吲哚类生物碱,产量达到2.45βg·L^-1。本研究为茯茶冠突散囊菌的高值化开发提供了技术支撑。

关键词: 冠突散囊菌, 发酵条件, 吲哚类生物碱, 内含成分分析

Abstract: Eurotium cristatum is the dominant fungi in Funzhuan tea, which produces indole alkaloids that have many important bioactivities, such as antibacterial, antiallergic, antiradiation, antiultraviolet, antitumor effects. In order to improve the production of indole alkaloids by E. cristatum, we investigated the factors affecting reproduction, growth and the fermentation of E. cristatum. The fermentation products of E. cristatum were analyzed using thin-layer chromatography, high-performance liquid chromatography and liquid chromatography-mass spectrum. The results showed that E. cristatum reproduced fast on Czapek-Dox solid medium with a large number of mature spores in 8 days, and the optimal growth conditions were obtained as following: pH 6.0, culture temperature 30℃, rotating speed 200 r·min^-1 and culture time 6 days. Moreover, fresh tea leaf powder, fresh tea leaf extract, Funzhuan tea powder, Funzhuan tea extract and tryptophan obviously improved the biosynthesis of indole alkaloids, with the best enhancing effect of tryptophan. Furthermore, at least 10 compounds were observed in thin-layer chromatography, most of which were identified to be indole alkaloids by high-performance liquid chromatography and liquid chromatography-mass spectrum, with a yield up to 2.45 g·L^-1. The present work provided technical support for utilization of E. cristatum in Fuzhuan tea.

Key words: Eurotium cristatum, fermentation condition, indole alkaloids, component analysis

中图分类号:

刘丽萍, 唐雨薇, 王若娴, 李志冰, 刘仲华, 刘硕谦. 茯茶冠突散囊菌发酵生产吲哚生物碱的研究[J]. 茶叶科学, 2017, 37(5): 503-512.

LIU Liping, TANG Yuwei, WANG Ruoxian, LI Zhibing, LIU Zhonghua, LIU Shuoqian. Production of Bioactive Indole Alkaloids through Fermention of Eurotium cristatum from Fuzhuan Tea[J]. Journal of Tea Science, 2017, 37(5): 503-512.

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