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Method for obtaining raphanin from broccoli hairy root culture system

A sulforaphane and culture system technology, applied in the directions of organic chemistry, fermentation, etc., can solve the problems of difficult control of intermediate products, complex chemical synthesis process, no research report on exocrine sulforaphane, etc. simple effect

Active Publication Date: 2016-06-29
甘肃泽华生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The chemical synthesis method is complicated and the intermediate product is difficult to control; the enzymatic method is currently the commonly used method for the preparation of sulforaphane
However, there are few reports on the extraction of sulforaphane from broccoli hairy root culture system. Most of the extraction of sulforaphane is concentrated in broccoli seedlings, flower buds, and seeds, and there is no sulforaphane in broccoli hairy root. exocrine research reports

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1 A method for obtaining sulforaphane from broccoli hairy root culture system, comprising the following steps:

[0025] (1) Inoculate the hairy roots of broccoli into 300mL liquid medium according to the inoculation amount of 0.1g fresh weight, and obtain the hairy roots of broccoli after 15 days; Take out the hairy roots of broccoli cultured in the liquid medium with the tweezers, put them on the filter paper, absorb the water on the root surface, grind them into powder after freeze-drying to constant weight, and obtain the freeze-dried hairy root powder; at the same time, the liquid culture The base seal was stored at 4°C.

[0026] Among them: the freeze-drying condition refers to pre-freezing the material first, vacuuming when the temperature reaches -35°C, and heating when the pressure reaches 25Pa; from -35 to 0°C, every 5°C is a stage, and the temperature is controlled and heated at each stage 0.5h, and finally heated at 5°C for 1h under temperature co...

Embodiment 2

[0037] Embodiment 2 A method of obtaining sulforaphane from broccoli hairy root culture system, comprising the following steps:

[0038] (1) Inoculate the hairy roots of broccoli into 300mL liquid medium according to the inoculation amount of 0.1g fresh weight, and obtain the hairy roots of broccoli after 15 days; Take out the hairy roots of broccoli cultured in the liquid medium with the tweezers, put them on the filter paper, absorb the water on the root surface, grind them into powder after freeze-drying to constant weight, and obtain the freeze-dried hairy root powder; at the same time, the liquid culture The base seal was stored at 4°C.

[0039] Among them: the freeze-drying condition refers to pre-freezing the material first, vacuuming when the temperature reaches -40°C, and heating when the pressure reaches 30Pa; from -35 to 0°C, every 5°C is a stage, and the temperature is controlled and heated at each stage 1h, and finally heated at 5°C for 2h under temperature contr...

Embodiment 3

[0050] Embodiment 3 A method for obtaining sulforaphane from broccoli hairy root culture system, comprising the following steps:

[0051] (1) Inoculate the hairy roots of broccoli into 300mL liquid medium according to the inoculation amount of 0.1g fresh weight, and obtain the hairy roots of broccoli after 15 days; Take out the hairy roots of broccoli cultured in the liquid medium with the tweezers, put them on the filter paper, absorb the water on the root surface, grind them into powder after freeze-drying to constant weight, and obtain the freeze-dried hairy root powder; at the same time, the liquid culture The base seal was stored at 4°C.

[0052] Among them: the freeze-drying condition refers to pre-freezing the material first, vacuuming when the temperature reaches -45°C, and heating when the pressure reaches 35Pa; from -35 to 0°C, every 5°C is a stage, and the temperature is controlled and heated at each stage 1.5h, and finally heated at 5°C for 3h under temperature co...

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PUM

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Abstract

The invention relates to a method for obtaining raphanin from a broccoli hairy root culture system. The method comprises the following steps: (1) inoculating broccoli hairy root to a liquid medium, carrying out proliferation culture and freeze drying, and grinding into powder, thus obtaining lyophilized hairy root powder; (2) grinding newly harvested mustard seeds, carrying out ultrasonic oscillation extraction, and carrying out suction filtration to remove impurities, thus obtaining a crude enzyme; (3) adding the crude enzyme into the lyophilized hairy root powder, carrying out enzymolysis and ultrasonic oscillation extraction, and carrying out vacuum filtration to remove impurities, thus obtaining raphanin crude extraction liquid; (4) concentrating the raphanin crude extraction liquid, and extracting to obtain an ethyl acetate phase A; (5) extracting the liquid medium, thus obtaining an ethyl acetate phase B; (6) mixing the ethyl acetate phase A and the ethyl acetate phase B, and carrying out vacuum concentration and dissolution on the mixture, thus obtaining raphanin methanol solution; (7) carrying out vacuum concentration on the raphanin methanol solution, thus obtaining a raphanin crude extract; and (8) drying the raphanin crude extract to obtain a raphanin crude product with the purity being 5-30%. The method is simple and efficient.

Description

technical field [0001] The invention relates to a method for obtaining sulforaphane, in particular to a method for obtaining sulforaphane from a broccoli hairy root culture system. Background technique [0002] Broccoli is rich in anti-cancer substance sulforaphane, and was rated as one of the top ten most nutritious vegetables by the US "Time Magazine". Broccoli can enhance the body's immunity, promote growth, enhance blood vessel toughness and other effects, and can prevent the risk of breast cancer, gastric cancer and other cancers. Broccoli seeds and seedlings are low in sulforaphane but high in hairy root cultures. [0003] At present, the preparation methods of sulforaphane mainly include chemical synthesis and enzymatic methods. The chemical synthesis method is complicated and the intermediate product is difficult to control; the enzymatic method is currently the commonly used method for the preparation of sulforaphane. However, there are few reports on the extract...

Claims

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Application Information

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IPC IPC(8): C12P13/00C07C331/22
CPCC07C331/22C12P13/00
Inventor 李胜马绍英俞颖张真李秉建
Owner 甘肃泽华生物科技有限公司
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