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Method for producing secondary metabolites by mixed culture of metabolically complementary plant cell lines

A technology for secondary metabolites, plant cell lines, applied in microorganism-based methods, plant cells, biochemical equipment and methods, etc., to achieve the effect of improving yield

Active Publication Date: 2018-11-27
安赛搏(重庆)生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The control of culture conditions mainly includes the control of physical parameters such as pH, temperature, air, light, shear force and ion solubility, so as to provide suitable culture conditions for the cultivation of plant cells; the screening of high-yield target product plant cell strains adopts induction , mutagenesis and transgenic methods, and the use of metabolic coupling culture between plant cell lines has not been reported and applied

Method used

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  • Method for producing secondary metabolites by mixed culture of metabolically complementary plant cell lines
  • Method for producing secondary metabolites by mixed culture of metabolically complementary plant cell lines
  • Method for producing secondary metabolites by mixed culture of metabolically complementary plant cell lines

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] (1) Taxus cell line 1 ( Taxus wallichiana var. mairei CGMCC no10001) can synthesize 10DAB, 10DAB is an intermediate product of paclitaxel synthesis, and is a key metabolite of paclitaxel synthesis in the metabolic pathway. The culture conditions are: medium is B5 medium, in which auxin is 2,4-D, content 10mg / L, the mitogen is 6-BA, the content is 1mg / L, the sucrose concentration is 15g / L, the fresh weight of the inoculated cells is 100g / L, the inoculated is 100% fresh membrane filter B5 medium, and the subculture growth cycle For 14 days, the culture temperature is 25°C, and the shaker speed is 100rpm;

[0032] (2) Taxus cell line 2 ( Taxus wallichiana var. mairei CGMCC no10002) can synthesize paclitaxel, and the culture conditions are the same as Taxus cell line 1;

[0033] (3) The yew cell line 1 and the yew cell line 2 were mixed according to the volume ratio of 1:1 and cultured. The culture conditions were as follows: the culture temperature was 25°C, the shak...

Embodiment 2

[0039] (1) Taxus cell line 1 ( Taxus wallichiana var. mairei CGMCC no.10001) can synthesize 10DAB, 10DAB is an intermediate product of paclitaxel synthesis, and is a key metabolite of paclitaxel synthesis in the metabolic pathway. The culture conditions are: the medium is B5 medium, and the auxin is 2,4-D , the content is 10mg / L, the mitogen is 6-BA, the content is 1mg / L, the sucrose concentration is 15g / L, the fresh weight of the inoculated cells is 100g / L, the inoculated is 100% fresh membrane-filtered B5 medium, subcultured The growth period is 14 days, the culture temperature is 25°C, and the shaker speed is 100rpm;

[0040] (2) Taxus cell line 2 ( Taxus wallichiana var. mairei CGMCC no.10002) can synthesize paclitaxel, and the culture conditions are the same as Taxus cell line 1;

[0041] (3) Taxus cell line 3 ( Taxus wallichiana var. mairei P-13-1) can synthesize BATIII, BAT III is an intermediate product of paclitaxel synthesis, and is a key metabolite of pacli...

Embodiment 3

[0046] (1) Grape cell line 1 ( Vitis vinifera L VV1000), capable of synthesizing resveratrol, the culture conditions are: the medium is B5 medium, in which the auxin is KT, the content is 2mg / L, the cleavage is NAA, the content is 1mg / L, and the sucrose concentration is 30g / L , the fresh weight of the inoculated cells is 100g / L, the inoculated is 100% fresh membrane filter B5 medium, the subculture growth period is 7 days, the culture temperature is 25°C, and the shaker speed is 100rpm;

[0047] (2) Taxus cell line 2 ( Taxus wallichiana var. mairei CGMCC no10002) can synthesize paclitaxel, and the culture conditions are as follows: the medium is B5 medium, in which the auxin is 2,4-D, the content is 10mg / L, the cleavage is 6-BA, the content is 1mg / L, and the sucrose concentration is 15g / L, the fresh weight of the inoculated cells is 100g / L, the inoculated is 100% fresh membrane filter B5 medium, the subculture growth cycle is 14 days, the culture temperature is 25°C, and...

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Abstract

The invention discloses a method for producing a secondary metabolite through hybrid culture of metabolism-complementary plant cell lines. On the basis that a target product is produced by independently adopting a cell line, two or more cell lines are adopted for hybrid culture, metabolism coupling is carried out in the target product production process, and the target product yield is improved. Experimental results show that after two or more cell lines are adopted for hybrid culture, the target product yield is improved by 13-90% compared with an original single cell line.

Description

technical field [0001] The invention belongs to the technical field of plant cell culture production, and in particular relates to a mixed culture method of two or more metabolically complementary plant cell lines. Background technique [0002] In the process system of producing secondary metabolites by plant cell culture, explants are mainly used to induce and screen cell lines producing secondary metabolites, and then the purebred cell lines are used to produce secondary metabolites through suspension culture. Under the system, it is difficult to increase the yield of target metabolites, and it is difficult to increase the yield of target metabolites in the screening of high-yielding cell lines and the limitation of key intermediates, and the cost of adding substrates and intermediates is high. The mixed culture technology of cell lines with complementary metabolic capabilities is used to help improve the production of plant secondary metabolites. [0003] The methods for...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P17/02C12P7/22C12N5/04C12R1/91
Inventor 张桂才付宝龙何俊雄何乐雨丁靖志张卫
Owner 安赛搏(重庆)生物技术有限公司