Method for separating and purifying rhamnolipid

A rhamnolipid, separation and purification technology, applied in the field of separation and purification, can solve the problems of multiple extractions, difficulty in separation, and unavoidable crossover of components, and achieve the effects of good repeatability, low purity and simple process

Active Publication Date: 2010-07-28
DAQING HUALI ENERGY BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main defect of the extraction method is that the number of extractions is too many, emulsification is easy to occur, and further separation is difficult. At the same time, the crossover of components in the extraction process is difficult to avoid, and the purity of the product is affected.
Arino S (Arino S, Marchal R, Vandecasteele J P. Identification and production of a rhamnolipidic biosurfactant by a Pseudomonas species. Appl Microbiol Biotechnol, 1996, 45: 162-168), Manso Pajarron (A. Manso Pajarron, C.G. De Koster, W. Heerma, M. Schmidt, J. Haverkamp, ​​Glycoconjugate Journal, Structure identification of natural rhamnolipid mixtures by fast atom bombardment tandem mass spectrometry 1993, 10:

Method used

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  • Method for separating and purifying rhamnolipid

Examples

Experimental program
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Embodiment 1

[0024] A method for separating and purifying rhamnolipids, the method comprising the following steps:

[0025] (1) Centrifuge the rhamnolipid-containing fermentation broth at 5000r / min for 30min, adjust the pH of the obtained supernatant to 2.0 with 6mol / L hydrochloric acid, leave it overnight at 4°C and centrifuge at 5000r / min for 30min, collect the precipitate Utilize diethyl ether to extract, after combining organic phase, blow-dry and freeze-dry to obtain rhamnolipid crude product 0.5g, adopt the mixed organic solvent obtained by mixing chloroform and methanol at a volume ratio of 25:1, take the mixed organic solvent 1.0 mL dissolves the rhamnolipid crude product to obtain a dissolved sample;

[0026] (2) Add the dissolved sample obtained in step (1) to a 200-mesh silica gel column 200mL of the first organic solvent obtained by mixing chloroform and methanol at a volume ratio of 25:1 was added to a silica gel column to elute the dissolved sample, and then chloroform and ...

Embodiment 2

[0030] A method for separating and purifying rhamnolipids, the method comprising the following steps:

[0031] (1) Centrifuge the rhamnolipid-containing fermentation broth at 5000r / min for 30min, adjust the pH of the obtained supernatant to 1.5 with 6mol / L hydrochloric acid, leave it overnight at 4°C and centrifuge at 5000r / min for 30min, collect the precipitate Utilize ethyl ether to extract, combine the organic phases and then blow dry and dry to obtain 6.0 g of rhamnolipid crude product, adopt the mixed organic solvent obtained by mixing chloroform and methanol at a volume ratio of 20:1, and take 12.0 mL of the mixed organic solvent Dissolving the rhamnolipid crude product to obtain a dissolved sample, the amount of the dissolved sample is 2% of the column volume of the silica gel column;

[0032] (2) Add the dissolved sample obtained in step (1) to a 300-mesh silica gel column, the diameter of the silica gel column is 3cm, and the length and diameter ratio of the silica ge...

Embodiment 3

[0036] A method for separating and purifying rhamnolipids, the method comprising the following steps:

[0037] (1) Centrifuge the rhamnolipid-containing fermentation broth at 5000r / min for 30min, adjust the pH of the obtained supernatant to 1.8 with 6mol / L hydrochloric acid, leave it overnight at 4°C and centrifuge at 5000r / min for 30min, collect the precipitate Utilize ethyl ether to extract, after merging the organic phases, blow-dry and freeze-dry to obtain 2.0Kg of rhamnolipid crude product, adopt the mixed organic solvent obtained by mixing chloroform and methanol in a volume ratio of 30:1, take the mixed organic solvent 4.0 L dissolves the rhamnolipid crude product to obtain a dissolved sample, and the amount of dissolved sample is 2% of the column volume of the silica gel column;

[0038] (2) Add the dissolved sample obtained in step (1) to a 300-mesh silica gel column. The diameter of the silica gel column is 30 cm, and the aspect ratio is 10:1. Chloroform and methanol...

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Abstract

The invention relates to a method for separating and purifying rhamnolipid, which comprises the following steps: dissolving the crude product of rhamnolipid in mixed organic solvent; adding the dissolved sample to a silica gel column; eluting the dissolved sample by using the mixed organic solvent; collecting the eluent and separating pigments from the eluent to obtain rhamnolipid solution; and finally, subjecting the rhamnolipid solution to reduced-pressure distillation and concentration to obtain the purified rhamnolipid. Compared with the prior art, the invention overcomes the defects in the prior art and has the characteristics of simple operation, high sample-loading capacity and high separation efficiency. Therefore, the method of the invention constitutes an extraction method advantageous in simple process and simple, economical and effective application and suitable for industrialized batch production.

Description

technical field [0001] The invention relates to a separation and purification method, in particular to a separation and purification method of rhamnolipid. Background technique [0002] Rhamnolipids are some amphiphilic compound molecules with certain surface activity and a combination of hydrophilic and hydrophobic structures produced by microorganisms under certain conditions. Compared with chemical surfactants, rhamnolipids have the characteristics of non-toxic, harmless, degradable, good biocompatibility, resistance to extreme harsh conditions, good selectivity and specificity, so they are used in oil exploration, environmental management, It has potential application value in food industry, paper industry, cosmetics, biomedicine and other fields. However, since the production cost of rhamnolipids is 3-6 times higher than that of chemical surfactants (the cost of separation, extraction and concentration in the middle and lower reaches accounts for 60%-70% of the product...

Claims

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Application Information

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IPC IPC(8): C07H15/04C07H1/06C07H1/08
Inventor 牟伯中杨世忠刘金峰王丹丹
Owner DAQING HUALI ENERGY BIOLOGICAL TECH
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