基于黄酮类化合物生物合成的工业大麻内生真菌HMY07次生代谢产物研究

doi:10.11924/j.issn.1000-6850.casb2024-0271

中国农学通报 ›› 2025, Vol. 41 ›› Issue (5): 56-62.doi: 10.11924/j.issn.1000-6850.casb2024-0271

基于黄酮类化合物生物合成的工业大麻内生真菌HMY07次生代谢产物研究

丛添玥¹(), 邢翔宇¹, 尤梦瑶¹, 程丽²(), 郑春英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分子生物学重点实验室，哈尔滨 150080

收稿日期:2024-04-27 修回日期:2024-10-15 出版日期:2025-02-15 发布日期:2025-02-11
通讯作者:
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方面的研究。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；
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方面的研究。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。
作者简介:
丛添玥，女，2000年出生，黑龙江哈尔滨人，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：0451-86608586，E-mail：cty000817@163.com。
基金资助:
黑龙江省种业创新发展资金项目“寒地龙药重点中药材种子种苗DNA指纹图谱库、特征库及质量控制体系研发与推广”(20230202); 黑龙江大学横向项目“工业大麻内生真菌及其次生代谢物提取、分离、鉴定”(21192); 黑龙江大学横向项目“一株产大麻酚的大麻内生真菌及其应用”(23114); 黑龙江大学横向项目“一种利用甘草内生真菌发酵工业大麻的方法”(23113)

Studies on Secondary Metabolites of Cannabis sativa L. Endophytic Fungus HMY07 Based on Flavonoid Biosynthesis

CONG Tianyue¹(), XING Xiangyu¹, YOU Mengyao¹, CHENG Li²(), ZHENG Chunying¹()

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 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 Molecular Biology, College of Heilongjiang Province / School of Life Sciences, Heilongjiang University, Harbin 150080

Received:2024-04-27 Revised:2024-10-15 Published:2025-02-15 Online:2025-02-11

摘要/Abstract

摘要：

基于黄酮类化合物的生物合成途径，对工业大麻内生真菌HMY07(Alternaria alternate)发酵液中的黄酮类化合物进行了提取和分离。采用牛津杯法对工业大麻内生真菌HMY07发酵液抑菌活性进行分析；同时采用DPPH法对其发酵液抗氧化活性进行测定；采用HPLC法对工业大麻内生真菌HMY07次生代谢产物进行分析后，以D₁₀₁大孔吸附树脂柱、硅胶柱色谱、聚酰胺柱及重结晶等方法对内生真菌发酵液中次生代谢产物进行分离。工业大麻内生真菌HMY07发酵液具有较好的抑菌及抗氧化活性，从工业大麻内生真菌HMY07发酵液中分离得到7个次生代谢产物，分别为：槲皮素(A)、山奈酚(B)、木犀草素(C)、肉桂酸(D)、阿魏酸(E)、芹菜素(F)及金丝桃苷(G)。工业大麻内生真菌HMY07能够产生多种黄酮类次生代谢产物，是一株具有深入开发应用价值的内生真菌，且所有化合物均为首次从该菌株中分离。

关键词: 工业大麻, 内生真菌, Alternaria alternata HMY07, 抑菌活性, 抗氧化活性, 黄酮类化合物

Abstract:

Based on the flavonoids biosynthesis, the flavonoids in the fermentation of Cannabis sativa L. endophytic fungus HMY07 (Alternaria alternata) were extracted and separated. The antimicrobial activities of the fermentation of Cannabis sativa L. endophytic fungus HMY07 was analyzed using Oxford cup method. Additionally, the antioxidant activities of the fermentation were measured by DPPH method. After analyzing the secondary metabolites of endophytic fungus HMY07 using HPLC, D₁₀₁ macroporous adsorption resin column, silica gel chromatography, polyamide column, and recrystallization were applied to separate the secondary metabolites in the endophytic fungal fermentation. The fermentation of Cannabis sativa L. endophytic fungus HMY07 showed good antimicrobial and antioxidant activities, and seven secondary metabolites were isolated from the fermentation. These metabolites are quercetin (A), kaempferol (B), luteolin (C), cinnamic acid (D), ferulic acid (E), apigenin (F), and hyperoside (G). The Cannabis sativa L. endophytic fungus HMY07 can produce a variety of flavonoid secondary metabolites, making it a valuable endophytic fungus for further development and application, with all isolated compounds reported for the first time from this strain.

Key words: Cannabis sativa L., endophytic fungus, Alternaria alternata HMY07, antimicrobial activities, antioxidant activities, flavonoids

丛添玥, 邢翔宇, 尤梦瑶, 程丽, 郑春英. 基于黄酮类化合物生物合成的工业大麻内生真菌HMY07次生代谢产物研究[J]. 中国农学通报, 2025, 41(5): 56-62.

CONG Tianyue, XING Xiangyu, YOU Mengyao, CHENG Li, ZHENG Chunying. Studies on Secondary Metabolites of Cannabis sativa L. Endophytic Fungus HMY07 Based on Flavonoid Biosynthesis[J]. Chinese Agricultural Science Bulletin, 2025, 41(5): 56-62.

图/表 3

参考文献 33

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样品	A	B	C	D	E	F
HMY07发酵液	12.25±0.20	7.25±0.22	12.96±0.13	9.50±0.22	11.72±0.23	11.25±0.23
甲醇	-	-	-	-	-	-
Str (100 μg/mL)	22.00±0.30	20.50±0.32	18.50±0.25	21.00±0.21	22.55±0.15	24.00±0.55
Nys (100 μg/mL)	-	-	-	-	-	-
样品	G	H	I	J	K
HMY07发酵液	14.88±0.15	8.50±0.22	10.07±0.18	9.02±0.35	12.25±0.30
甲醇	-	-	-	-	-
Str (100 μg/mL)	19.25±0.51	22.00±0.20	20.50±0.22	17.25±0.41	-
Nys (100 μg/mL)	-	-	-	-	22.00±0.30

样品	A	B	C	D	E	F
HMY07发酵液	12.25±0.20	7.25±0.22	12.96±0.13	9.50±0.22	11.72±0.23	11.25±0.23
甲醇	-	-	-	-	-	-
Str (100 μg/mL)	22.00±0.30	20.50±0.32	18.50±0.25	21.00±0.21	22.55±0.15	24.00±0.55
Nys (100 μg/mL)	-	-	-	-	-	-
样品	G	H	I	J	K
HMY07发酵液	14.88±0.15	8.50±0.22	10.07±0.18	9.02±0.35	12.25±0.30
甲醇	-	-	-	-	-
Str (100 μg/mL)	19.25±0.51	22.00±0.20	20.50±0.22	17.25±0.41	-
Nys (100 μg/mL)	-	-	-	-	22.00±0.30

基于黄酮类化合物生物合成的工业大麻内生真菌HMY07次生代谢产物研究

Studies on Secondary Metabolites of Cannabis sativa L. Endophytic Fungus HMY07 Based on Flavonoid Biosynthesis

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