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中国农学通报

中国农学通报 ›› 2023, Vol. 39 ›› Issue (27): 118-125.doi: 10.11924/j.issn.1000-6850.casb2022-0971

• 生物科学 • 上一篇    下一篇

基于黄酮类化合物生物合成的内生细菌HS8次生代谢产物的分离

祁可香1(), 关昕1, 尤梦瑶1, 张赫1, 王宇晴1, 程丽2(), 郑春英1()   

  1. 1 黑龙江大学生命科学学院，农业微生物技术教育部工程研究中心，黑龙江省寒区植物基因与生物发酵重点实验室，黑龙江省普通高校微生物重点实验室，哈尔滨 150080
    2 黑龙江省普通高校分子生物学重点实验室，哈尔滨 150080
  • 收稿日期:2022-11-23 修回日期:2023-03-15 出版日期:2023-09-25 发布日期:2023-09-22
  • 通讯作者: 郑春英，女，1968年出生，黑龙江哈尔滨人，教授，博士，主要从事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：zhengchunying68@163.com。程丽，女，1978年出生，黑龙江哈尔滨人，副教授，博士，主要从事食品营养与安全。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：chengli@hlju.edu.cn。
  • 作者简介:

    祁可香，女，1999年出生，甘肃白银市人，硕士研究生，研究方向：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：0451-886608586，E-mail：。

  • 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)

The Endophytic Bacterium HS8: Isolation of Secondary Metabolites Based on Flavonoids Biosynthesis

QI Kexiang1(), GUAN Xin1, YOU Mengyao1, ZHANG He1, WANG Yuqing1, CHENG Li2(), ZHENG Chunying1()   

  1. 1 Engineering Research Center of Agricultural Microbiology Technology, Ministry of Education/Heilongjiang Provincial Key Laboratory of Plant Genetic Engineering and Biological Fermentation Engineering for Cold Region/ Key Laboratory of Microbiology, College of Heilongjiang Province/ School of Life Sciences, Heilongjiang University, Harbin 150080
    2 Heilongjiang Provincial Key Laboratory of Molecular Biology, Harbin 150080
  • Received:2022-11-23 Revised:2023-03-15 Published-:2023-09-25 Online:2023-09-22

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摘要：

基于黄酮类化合物的生物合成，研究药用植物黄芪内生细菌HS8次生代谢产物，旨在综合利用内生细菌HS8。以黄芪内生细菌HS8为试材，对其发酵液抗氧化活性和次生代谢产物进行分析，并采用柱色谱分离法，对内生细菌HS8发酵液中黄酮类成分进行了分离；采用形态学观察、生理生化指标测定及16S rDNA序列分析，对内生细菌HS8进行了种属鉴定。实验结果表明，内生细菌HS8具有良好的抗氧化活性；经HPLC分析发现，在内生细菌HS8发酵液中存在芒柄花素和毛蕊异黄酮，因此，采用柱色谱分离法分离得到了芒柄花素和毛蕊异黄酮，经鉴定，黄芪内生细菌HS8为枯草芽孢杆菌(Bacillus subtilis)。上述结果表明，黄芪内生枯草芽孢杆菌HS8为黄酮类成分芒柄花素和毛蕊异黄酮产生菌。

关键词: 内生细菌HS8, 抗氧化活性, 黄酮类化合物, HPLC, 次生代谢产物, 提取分离

Abstract:

Based on the flavonoids biosynthesis, the secondary metabolites of endophytic bacterium HS8 from Astragalus membranaceus were studied to utilize endophytic fungi HS8 comprehensively. Endophytic bacterium HS8 from Astragalus membranaceus was chosen as the research object, the antioxidant activities and secondary metabolites of fermentation of HS8 were analyzed, and the flavonoids were separated from the endophytic bacteria HS8 fermentation broth by column chromatography. Endophytic bacterium HS8 was identified by the 16S rDNA sequence homology and their physiological and biochemical characteristics. The results showed that endophytic bacterium HS8 had good antioxidant activities. Under the guidance of HPLC analysis, formononetin and calycosin in the fermentation of endophytic bacterium HS8 were isolated by column chromatography methods, and the endophytic bacterium HS8 was identified as Bacillus subtilis. These results indicated that the endophytic bacterium HS8 from Astragalus astragalus could produce formononetin and calycosin, which was a flavonoids-producing strain.

Key words: endophytic bacterium HS8, antioxidant activities, flavonoids, HPLC, secondary metabolites, extraction and separation