Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for producing flavone of anoectochilus roxburghii by suspension cultivation of cells

A technology of suspension culture cells and lotus flavonoids, applied in the direction of fermentation, can solve the problems of long production cycle, low biomass growth rate, low target product yield, etc., to improve product yield, uniform supply of nutrients and dissolved oxygen, The effect of shortening the cell culture cycle

Inactive Publication Date: 2015-03-25
WEST ANHUI UNIV
View PDF4 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The technology of using cell engineering technology to produce clematis flavonoids has been studied at home and abroad for a long time, and many achievements have been made, but there are many difficult problems in the production process, such as low biomass growth rate, low yield of target products, and production cycle. long wait
There is no report on the production of clematis flavonoids by suspension culture of clematis cells in an air-lift stirred tank, so it is very meaningful to develop a method for efficiently producing natural products of clematis flavonoids

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Examples

Experimental program
Comparison scheme
Effect test

specific Embodiment 1

[0016] 1. Training stage

[0017] (1) Single cell preparation

[0018] Take clematis capsules, sterilize them according to the requirements of tissue culture technology, and carry out aseptic sowing. The sowing medium is an improved 1 / 2MS solid medium. Take them out after they germinate into protocorms. Choose protocorms with bright color, large individuals and vigorous vitality. For bulbs, separate and prepare single cells by enzymatic hydrolysis, filter with a microporous filter with a pore size of 80 microns, separate the filtrate through a low-speed centrifuge, and collect the precipitated cells;

[0019] (2) Single cell line culture

[0020] Inoculate the collected single cells into a baffle shake flask containing improved 1 / 2MS liquid medium, the inoculum amount of fresh cells is 25g / L, the light intensity is 3000lux, the color temperature is 5000K, shake culture, adjust the pH value to 5.6 ±2, the temperature is 22±2°C, when the cell density reaches 10,000 cells / mL, t...

specific Embodiment 2

[0026] 1. Training stage

[0027] (1) Single cell preparation

[0028] Take clematis capsules, sterilize them according to the requirements of tissue culture technology, and then carry out aseptic sowing. The sowing medium is a modified 1 / 2MS solid medium. Take them out after they germinate into protocorms. Choose protocorms with bright color, large individuals and vigorous vitality. For bulbs, single cells were separated by enzymatic hydrolysis, filtered through a microporous filter with a pore size of 60 microns, the filtrate was separated by a low-speed centrifuge, and the precipitated cells were collected;

[0029] (2) Single cell line culture

[0030] Inoculate the collected single cells into a baffle shake flask containing improved 1 / 2MS liquid medium, the inoculum amount of fresh cells is 40g / L, the light intensity is 2000lux, the color temperature is 6000K, shake culture, adjust the pH value to 5.6 ±2, the temperature is 22±2°C, when the cell density reaches 20,000 c...

experiment example 1

[0039] The mensuration of experimental example 1 total flavonoid content

[0040] 1. The total flavonoid content extracted by the embodiment is analyzed, and the specific analysis method is as follows. Drawing of standard curve. Accurately draw 0.4, 0.8, 1.2, 1.6, 2.0, 2.4 ml of 0.2 mg / ml rutin solution, place them in cuvettes respectively, replace the rutin solution with distilled water as a zero tube, add water to 2.4 ml each, and add 5 0.4 ml of % NaNO2 solution, shake well, let stand for 6 min; add 0.4 ml of 10% Al(NO3)3 solution, shake well, let stand for 6 min; add 4 ml of 4% NaOH solution, shake well, let stand for 15 min; The absorbance was measured at 510 nm, and a standard curve was drawn with the absorbance as the ordinate and the rutin content as the abscissa. Regression calculation is performed on the concentration Y (mg / ml) from the absorbance X to obtain the standard curve equation: Y=0.9324X+0.004 R2=0.9993;

[0041] Determination of the sample: Accurately w...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides a method for producing flavone of anoectochilus roxburghii by suspension cultivation of cells, and particularly relates a method for producing flavone of anoectochilus roxburghii by utilizing a cell culture reactor to culture anoectochilus roxburghii cells. The method comprises the following steps: aseptically sowing anoectochilus roxburghii seeds, germinating to form protocorms, and separating undifferentiated single cells in the high-activity protocorms by utilizing an enzymolysis method; further culturing a single cell line; finally culturing cell mass, wherein a plant growth regulator and a bio-enzyme preparation are added into a culture medium to activate cell division and growth and prevent cell from growing in an aggregated or agglomerated way, and proper illumination intensity and color temperature are selected to regulate and control expression of a target product. The method can be used for greatly shortening the cell culture cycle and increasing the product yield of flavone of anoectochilus roxburghii, is simple and stable in process, and is suitable for industrial production.

Description

technical field [0001] The invention relates to the production technology of flavonoids, in particular to the production technology of Aureus flavones, in particular to a method for producing Aureus flavones by a cell culture method. Background technique [0002] Clematis clematis flavonoids are one of the main components of clematis clematis, which can treat bronchitis, nephritis, cystitis, diabetes, hematuria, rheumatoid arthritis, acute and chronic liver disease, hypertension, arteriosclerosis, cerebral thrombosis, etc. Both have auxiliary effects. The pharmacological effects of clematis flavonoids are mainly manifested in nutrition, two-way regulation of blood sugar and blood pressure, lowering blood lipids, and regulating the body's immunity. With the development of society, people pay more and more attention to health, and their demand will also increase. However, the traditional extraction method of clematis flavonoids is to use the roots, stems, leaves and other pla...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/06
Inventor 邓辉陈乃富刘文中陈存武韩邦兴余茂耘
Owner WEST ANHUI UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com