Effects of Endophytic Fungus RP4 Fermentation on Contents of Three Major Cannabinoids in Cannabis sativa L.

doi:10.11924/j.issn.1000-6850.casb2023-0645

Abstract

Abstract:

In order to efficiently produce cannabidiol (CBD) raw materials, Cannabis sativa L. was fermented by using the endophytic fungus RP4 strain of licorice with good biological activity. Three major cannabinoids, CBD, CBDA and THC were selected as the research objects to study the dynamic changes of CBD, CBDA and THC contents before and after fermentation. The dynamic changes of CBD, CBDA and THC contents in Cannabis sativa L. during fermentation were analyzed by HPLC. Results showed that during the fermentation process, the contents of CBD and THC increased first and then decreased. When the fermentation time was 3 days, both the contents of CBD and THC reached the maximum value of 5.8643±0.0726 and 0.4878±0.0002 (mg/g) respectively, and the increase of THC was always kept within the scope of national control (THC<0.3%), the content of CBDA decreased continuously, especially in the first day of fermentation. After fermentation in different regions and different parts of Cannabis sativa L., the content of cannabinoid components increased differently. The above results indicated that the microbial fermentation technology could be used to produce CBD in Cannabis sativa L. directly and efficiently, which greatly improved the utilization rate of Cannabis sativa L. resources and laid a foundation for the extraction and separation of CBD and other secondary metabolites.

Key words: Cannabis sativa L., cannabidiol, tetrahydrocannabinol, cannabidiolic acid, fermentation, microbial fermentation, high-performance liquid chromatography (HPLC), secondary metabolites

GUAN Xin, ZHANG He, QI Kexiang, LI Wanru, JIANG Shuo, ZHENG Chunying. Effects of Endophytic Fungus RP4 Fermentation on Contents of Three Major Cannabinoids in Cannabis sativa L.[J]. Chinese Agricultural Science Bulletin, 2024, 40(18): 150-156.

Figures/Tables 9

References 34

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仪器	型号规格	生产厂家
电子精密天平	PL303	梅特勒-托利多仪器有限公司
超净工作台	SZX	南通科学仪器有限公司
空气浴振荡器	HZQ-C	哈尔滨市东联电子技术开发有限公司
旋转蒸发仪	R-201	上海申胜生物技术有限公司
超声波清洗器	KQ-100DE	昆山市超声仪器有限公司
紫外可见波长检测器	FL2000	浙江温岭福立分析仪器有限公司
高效液相色谱仪	FL2000	浙江温岭福立分析仪器有限公司
400MH固体超导核磁共振波谱仪	AVANCE Ⅲ	美国Thermo Fisher公司

仪器	型号规格	生产厂家
电子精密天平	PL303	梅特勒-托利多仪器有限公司
超净工作台	SZX	南通科学仪器有限公司
空气浴振荡器	HZQ-C	哈尔滨市东联电子技术开发有限公司
旋转蒸发仪	R-201	上海申胜生物技术有限公司
超声波清洗器	KQ-100DE	昆山市超声仪器有限公司
紫外可见波长检测器	FL2000	浙江温岭福立分析仪器有限公司
高效液相色谱仪	FL2000	浙江温岭福立分析仪器有限公司
400MH固体超导核磁共振波谱仪	AVANCE Ⅲ	美国Thermo Fisher公司

参数	设置条件
色谱柱	Venusil XBP-C₁₈柱(4.6 mm×250 mm，5 μm，USA)
流动相	①乙腈-水-0.1%甲酸溶液(75:25)，②甲醇-乙腈-水-0.1%磷酸溶液(40:30:30)
波长	①220 nm，②254 nm
流速	1 mL/min
柱温	25℃
进样量	10 μL

参数	设置条件
色谱柱	Venusil XBP-C₁₈柱(4.6 mm×250 mm，5 μm，USA)
流动相	①乙腈-水-0.1%甲酸溶液(75:25)，②甲醇-乙腈-水-0.1%磷酸溶液(40:30:30)
波长	①220 nm，②254 nm
流速	1 mL/min
柱温	25℃
进样量	10 μL

组分	回归方程	相关系数r	线性范围/(mg/mL)
CBD	Y=6×10⁶X+264482	0.9998	0.0625~1.0000
CBDA	Y=2×10⁷X-5841	0.9996	0.0625~1.0000
THC	Y=8.5206×10⁵X+47649	0.9998	0.0625~1.0000