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Method for separating and extracting beta-sitosterol by using Bacillus megaterium

A technology of Bacillus megaterium and sitosterol, which is applied in the biological field, can solve the problems of unreachable separation results, cumbersome chemical reaction steps, and difficult operation, etc., and achieve the effects of easy cultivation and preservation, reliable source, and wide application range

Active Publication Date: 2019-08-06
GUIZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The chemical method has defects such as cumbersome reaction steps, difficult operation, high cost and low recovery rate, and cannot achieve ideal separation results, so it is difficult to achieve large-scale industrial production; the main applied physical methods include molecular distillation and high-efficiency liquid phase. , adsorption method, solvent crystallization method, etc. Among them, the solvent crystallization method has the advantages of simple operation, short and simple process flow, high product purity, easy solvent recovery, and suitable for industrial production, but it is only suitable for substances whose solubility does not change much with temperature

Method used

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  • Method for separating and extracting beta-sitosterol by using Bacillus megaterium
  • Method for separating and extracting beta-sitosterol by using Bacillus megaterium

Examples

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Effect test

Embodiment 1

[0023] A method utilizing Bacillus megaterium to isolate and extract β-sitosterol, comprising the following steps:

[0024] (1) Preparation of crude extract

[0025] The bacterial cells of Bacillus megaterium were spray-dried to make bacterial powder for later use. The bacterial powder and ethanol with a mass ratio of 1:3 were refluxed and extracted three times at a pressure of 0.04 MPa and a temperature of 40°C for 4 hours each time to obtain Bacillus megaterium extract, concentrated under reduced pressure to 1 / 10 of the total volume, then added an equal volume of water to dissolve, extracted the aqueous solution with an equal volume of ethyl acetate to obtain the upper layer ethyl acetate extract, and then concentrated under reduced pressure to ethyl acetate Evaporate to dryness to obtain crude extract;

[0026] (2) Atmospheric pressure silica gel column chromatography

[0027] Dissolve the crude extract with an equal volume of ethyl acetate, mix the sample with 40 mesh si...

Embodiment 2

[0037] A method utilizing Bacillus megaterium to isolate and extract β-sitosterol, comprising the following steps:

[0038] (1) Preparation of crude extract

[0039] The bacterial cells of Bacillus megaterium were spray-dried to make bacterial powder for later use. The bacterial powder and ethanol with a mass ratio of 1:3 were refluxed and extracted three times at a pressure of 0.05 MPa and a temperature of 50°C for 4 hours each time to obtain Bacillus megaterium extract, concentrated under reduced pressure to 1 / 8 of the total volume, then added an equal volume of water to dissolve, extracted the aqueous solution with an equal volume of ethyl acetate to obtain the upper layer ethyl acetate extract, and then concentrated under reduced pressure to ethyl acetate Evaporate to dryness to obtain crude extract;

[0040] (2) Atmospheric pressure silica gel column chromatography

[0041] Dissolve the crude extract with an equal volume of ethyl acetate, mix the sample with 60 mesh sil...

Embodiment 3

[0051] A method utilizing Bacillus megaterium to isolate and extract β-sitosterol, comprising the following steps:

[0052] (1) Preparation of crude extract

[0053] The bacterial cells of Bacillus megaterium were spray-dried to make bacterial powder for later use. The bacterial powder and ethanol with a mass ratio of 1:3 were refluxed and extracted three times at a pressure of 0.06 MPa and a temperature of 60°C for 4 hours each time to obtain Bacillus megaterium extract, concentrated under reduced pressure to 1 / 5 of the total volume, then added an equal volume of water to dissolve, extracted the aqueous solution with an equal volume of ethyl acetate to obtain the upper layer ethyl acetate extract, then concentrated under reduced pressure to ethyl acetate Evaporate to dryness to obtain crude extract;

[0054] (2) Atmospheric pressure silica gel column chromatography

[0055] Dissolve the crude extract with an equal volume of ethyl acetate, mix the sample with 80 mesh silica ...

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Abstract

The invention discloses a method for separating and extracting beta-sitosterol by using Bacillus megaterium. The method comprises: preparing a crude extract: carrying out spray drying on Bacillus megaterium cells to prepare bacterial powder so as to be spare, carrying out reflux extraction on the bacterial powder and ethanol for 4 h three times according to a mass ratio of 1:3 at a temperature of40-60 DEG C under a pressure of 0.04-0.06 MPa to obtain a Bacillus megaterium extraction liquid, carrying out pressure reducing concentration to achieve 1 / 10-1 / 5 of the total volume, adding an equal volume of water, dissolving, extracting the aqueous solution with ethyl acetate according to a volume ratio of 1:1 to obtain an upper layer ethyl acetate extraction liquid, and carrying out pressure reducing concentration until evaporating the ethyl acetate to obtain a crude extract; and carrying out normal pressure silica gel column chromatography three times, carrying out gel column chromatography, and carrying out re-crystallization. According to the present invention, the substrate is cheap and easy to obtain, the process is simple and easy to perform, and the production process is environmentally friendly.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for separating and extracting β-sitosterol by using Bacillus megaterium. Background technique [0002] β-sitosterol (β-sitosterol) is one of the main components of phytosterols. It belongs to tetracyclic and tricyclic compounds. It is a white crystal powder at room temperature, odorless and tasteless, and its melting point can reach 130-140 ° C. It is insoluble in water. It can be dissolved in certain organic solvents and has high nutritional value and biological activity. It is widely used in the fields of medicine, food, health care products and cosmetics. Studies in recent years have shown that β-sitosterol has obvious pharmacological effects such as lowering cholesterol, relieving cough, anti-cancer, anti-inflammation, anti-oxidation, and preventing and controlling hypertension. The separation methods mainly include chemical and physical methods. The chemical method h...

Claims

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Application Information

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IPC IPC(8): C07J9/00
CPCC07J9/00
Inventor 李祝谢毓丹肖洋吉玉玉赵妗颐
Owner GUIZHOU UNIV