内生真菌发酵对刺五加异嗪皮啶合成的影响

doi:10.11924/j.issn.1000-6850.casb2020-0378

中国农学通报 ›› 2021, Vol. 37 ›› Issue (13): 122-126.doi: 10.11924/j.issn.1000-6850.casb2020-0378

所属专题：生物技术

• 食品·营养·检测·安全 • 上一篇下一篇

内生真菌发酵对刺五加异嗪皮啶合成的影响

万璐¹^,²(), 闫佳佳¹^,², 郑春英¹^,²()

¹黑龙江大学/农业微生物技术教育部工程研究中心,哈尔滨 150500
²黑龙江大学生命科学学院/黑龙江省普通高校微生物重点实验室,哈尔滨 150080

收稿日期:2020-08-19 修回日期:2021-01-16 出版日期:2021-05-05 发布日期:2021-05-18
通讯作者: 郑春英
作者简介:万璐,女,1997年出生,黑龙江佳木斯人,硕士研究生,研究方向为食品和药物生物活性挖掘及研发。通信地址：150080 黑龙江省哈尔滨市南岗区学府路74号黑龙江大学生命科学学院,Tel：0451-86608586,E-mail：1157647243@qq.com。
基金资助:
黑龙江大学校企合作横向项目“一株产原儿茶酸的枯草芽孢杆菌”(20170223)

Effects on the Biosythesis of Isofraxidin in Fermenfed Acanthopanax senticosus by Endophytic Fungus

Wan Lu¹^,²(), Yan Jiajia¹^,², Zheng Chunying¹^,²()

¹Engineering Research Center of Agricultural Microbiology Technology, Ministry of Education/Heilongjiang University, Harbin 150500
²Key Laboratory of Microbiology, College of Heilongjiang Province/School of Life Sciences, Heilongjiang University, Harbin 150080

Received:2020-08-19 Revised:2021-01-16 Online:2021-05-05 Published:2021-05-18
Contact: Zheng Chunying

摘要/Abstract

摘要：

采用内生真菌发酵刺五加以提高其活性成分异嗪皮啶的含量,并对其进行分离纯化。分别选取4株内生真菌对刺五加进行发酵,以异嗪皮啶为对照,采用HPLC法分析发酵样品中异嗪皮啶的含量变化,确定能够定向提高发酵样品中异嗪皮啶含量的目的菌株;采用硅胶柱色谱法分离纯化发酵样品中的异嗪皮啶;采用形态学观察和ITS rDNA序列同源性分析对目的菌株进行种属鉴定。结果表明,内生真菌AJ14能显著提高刺五加发酵样品中异嗪皮啶成分含量。采用硅胶柱色谱法从刺五加发酵样品中分离到异嗪皮啶;经鉴定,目的菌株AJ14为赭曲霉(Aspergillus ochraceus)。采用内生真菌发酵刺五加可定向提高异嗪皮啶的含量,研究结果可为微生物发酵法生产异嗪皮啶提供参考。

关键词: 刺五加, 内生真菌, 发酵, 异嗪皮啶, 生物合成, HPLC, 分离纯化, 活性成分

Abstract:

Endophytic fungi were used to ferment Acanthopanax senticosus to increase the contents of isofraxidin, then, isofraxidin was separated and purified. With isofraxidin as reference substance, four endophytic fungi were chosen to ferment A. senticosus, and the target strain which had the ability of improving the contents of isofraxidin was selected by HPLC. Meanwhile, isofraxidin was separated and purified by silica gel column chromatography method from the fermented A. senticosus, and the target strain was identified by its morphology characters and the ITS rDNA sequence homology. Isofraxidin in fermented A. senticosus by AJ14 was increased significantly which was separated and purified by silica gel column chromatography, and endophytic fungus AJ14 was identified as Aspergillus ochraceus. Fermented A. senticosus using endophytic fungi could biotransform isofraxidin directly, and this lays the foundation for the production of isofraxidin by microbial fermentation.

Key words: Acanthopanax senticosus, endophytic fungus, fermentation, isofraxidin, biosynthesis, HPLC, isolation and purification, active constituent

中图分类号:

R931

万璐, 闫佳佳, 郑春英. 内生真菌发酵对刺五加异嗪皮啶合成的影响[J]. 中国农学通报, 2021, 37(13): 122-126.

Wan Lu, Yan Jiajia, Zheng Chunying. Effects on the Biosythesis of Isofraxidin in Fermenfed Acanthopanax senticosus by Endophytic Fungus[J]. Chinese Agricultural Science Bulletin, 2021, 37(13): 122-126.

图/表 9

图1. 异嗪皮啶化学结构

图2. 发酵刺五加中异嗪皮啶成分的分离工艺流程

图3. 内生真菌AJ14发酵刺五加后的HPLC色谱图

组别	刺五加茎发酵样品	刺五加根发酵样品
刺五加生药	0.0611±0.0012	0.0598±0.0010
空白对照组	0.0740±0.0005	0.0725±0.0009
AJ7发酵样品	0.1075±0.0013^＊＊△△	0.0941±0.0008^＊＊△△
AJ14发酵样品	0.2215±0.0008^＊＊△△	0.1507±0.0012^＊＊△△
AJ15发酵样品	0.0645±0.0007^＊＊△△	0.0627±0.0011^＊＊△△
AJ19发酵样品	0.0957±0.0011^＊＊△△	0.0734±0.0010^＊＊△△

表1. 刺五加发酵样品中异嗪皮啶含量测定结果 mg/g

图4. 刺五加茎发酵样品异嗪皮啶含量测定结果

图5. TLC检测结果

图6. 分离得到的异嗪皮啶结晶HPLC色谱图

图7. AJ14菌落形态及孢子显微结构

图8. 内生真菌AJ14菌株的系统发育树

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内生真菌发酵对刺五加异嗪皮啶合成的影响

Effects on the Biosythesis of Isofraxidin in Fermenfed Acanthopanax senticosus by Endophytic Fungus

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