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Gingko cell culture system-based study for biosynthesizing ginkgo terpene lactones from levopimaradiene

A technology of ginkgolidene and pipine diene, which is applied to the determination/testing of microorganisms, microorganisms, plant cells, etc., and can solve the problems of complex chemical total synthesis steps, long plant growth cycle, and difficulty in regulation

Pending Publication Date: 2017-08-15
于荣敏 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the large market demand, the traditional large-scale planting has various shortcomings such as long plant growth cycle, low yield, easy to be affected by climate and environment conditions, and difficult to control, which restricts the development of this subject. At the same time, traditional extraction, separation and purification methods cannot To meet the requirements, the chemical total synthesis steps are complex and costly and have no industrial production value, so scientists turn their attention to modern biotechnology

Method used

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  • Gingko cell culture system-based study for biosynthesizing ginkgo terpene lactones from levopimaradiene
  • Gingko cell culture system-based study for biosynthesizing ginkgo terpene lactones from levopimaradiene
  • Gingko cell culture system-based study for biosynthesizing ginkgo terpene lactones from levopimaradiene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Effects of levopipine diene (LP) on the growth of ginkgo stem cells (CMCs) and dedifferentiated cells (DDCs) and the content of ginkgolidene

[0023] (1) Cultivation of Ginkgo CMCs and DDCs

[0024] The medium used for Ginkgo suspension cells is MS medium, supplemented with vitamin C (50.0mg / L), sucrose (30g / L), and DDCs with naphthaleneacetic acid (NAA, 2.0mg / L) and indolebutyric acid (IBA, 2.0 mg / L), 2.0mg / L NAA and 2,4-dichlorophenoxyacetic acid (2,4-D, 2.0mg / L) were added to CMCs, the pH value of the medium was 5.75-5.80, and the contents were divided into 250mL cones In a shaped bottle (containing 100mL culture medium), sterilize by high temperature steam at 121°C for 20 minutes, cool to room temperature after sterilization, subculture the amount of cells to 50g / L, place on a shaker after subculture, rotate at 110rpm, 25°C Cultivate in the dark.

[0025] (2) The setting of the experimental group

[0026] Ginkgo suspension cells were pre-cultured (100mL medium) f...

Embodiment 2

[0034] Detection of Expression Levels of Key Enzymes in Ginkgo Suspension Cells by PCR

[0035] (1) Cultivation of Ginkgo CMCs and DDCs

[0036] The medium used for Ginkgo suspension cells is MS medium, supplemented with vitamin C (50.0mg / L), sucrose (30g / L), and DDCs with naphthaleneacetic acid (NAA, 2.0mg / L) and indolebutyric acid (IBA, 2.0 mg / L), 2.0mg / L NAA and 2,4-dichlorophenoxyacetic acid (2,4-D, 2.0mg / L) were added to CMCs, the pH value of the medium was 5.75-5.80, and the contents were divided into 250mL cones In a shaped bottle (containing 100mL culture medium), sterilize by high temperature steam at 121°C for 20 minutes, cool to room temperature after sterilization, subculture the amount of cells to 50g / L, place on a shaker after subculture, rotate at 110rpm, 25°C Cultivate in the dark.

[0037] (2) The setting of the experimental group

[0038] Ginkgo suspension cells were pre-cultured (100mL culture medium) for 13 days, and levopipinediene dissolved in DMSO was...

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Abstract

The invention relates to an application of ginkgo suspension cell biotransformation of levopimaradiene. It is found that the levopimaradiene can significantly promote the synthesis of the ginkgo terpene lactones in gingko cells, and the ginkgo terpene lactones in stem cells have more kinds and higher content than ginkgo terpene lactones in dedifferentiated cells; and the levopimaradiene has stronger influences on the synthesis of the ginkgo terpene lactones in the stem cell system than the dedifferentiated cells. Additionally, the levopimaradiene can promote the synthesis of bilobalide in the stem cell system and the dedifferentiated cell systems, so the bilobalide can be metabolized by ginkgolides in an MEP pathway. A PCR result shows that the levopimaradiene affects the biosynthesis of the ginkgo terpene lactones through regulating the expression level of key enzymes in the MEP pathway.

Description

technical field [0001] The present invention relates to the effect of levopipine adiene on the biosynthesis of ginkgolidene based on ginkgo cell culture system. Background technique [0002] Ginkgo belongs to the gymnosperm door and is a plant of the genus Ginkgo in the family Ginkgoceae. Ginkgo biloba contains a variety of medicinal secondary metabolites. Ginkgolide is a natural and specific antagonist of platelet activating factor receptors, and it is also an excellent natural medicine for the treatment and prevention of cardiovascular and cerebrovascular diseases. Due to the large market demand, the traditional large-scale planting has various shortcomings such as long plant growth cycle, low yield, easy to be affected by climate and environment conditions, and difficult to control, which restricts the development of this subject. At the same time, traditional extraction, separation and purification methods cannot To meet the requirements, the chemical total synthesis st...

Claims

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Application Information

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IPC IPC(8): C12N5/04C12P17/18C12Q1/68
CPCC12N5/04C12P17/181C12Q1/6895C12N2500/38C12N2500/34C12N2500/30C12Q2600/158
Inventor 于荣敏朱建华王溥
Owner 于荣敏
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