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Method for producing rhamnolipid by virtue of fermentation and separation of pseudomonas aeruginosa

A technology for Pseudomonas aeruginosa and rhamnolipid, which is applied in the field of separation and production of rhamnolipid and fermentation of Pseudomonas aeruginosa, and can solve the problem that there is no public sale of pure rhamnolipid glycolipid and restrictions on rhamnolipid Commercial production process, high production costs and other problems, to achieve the effect of easy control of equipment, lower production costs, and reduce foaming

Inactive Publication Date: 2012-11-28
CHINA NAT OFFSHORE OIL CORP +3
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  • Abstract
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  • Application Information

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Problems solved by technology

[0005] The application range of rhamnolipids is very wide, but its production cost remains high, which limits the large-scale commercial production process of rhamnolipids. At present, there is no disclosure of pure rhamnolipid glycolipids in the international and domestic scope. Sales
Factors restricting the industrialized production of rhamnolipids mainly include the production capacity of strains, culture conditions, production methods, and separation and extraction methods. Among them, research on strain mutagenesis and optimization of culture conditions has been very common, and relatively good strains have also been found. It can achieve high and stable yield under suitable culture conditions, but overall, there is still a big gap between the existing technology and the low cost required for industrialized production

Method used

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  • Method for producing rhamnolipid by virtue of fermentation and separation of pseudomonas aeruginosa
  • Method for producing rhamnolipid by virtue of fermentation and separation of pseudomonas aeruginosa
  • Method for producing rhamnolipid by virtue of fermentation and separation of pseudomonas aeruginosa

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Embodiment 1

[0031] see figure 1 , a method for Pseudomonas aeruginosa fermentation, separation and production of rhamnolipids, comprising the following steps:

[0032] 1) The Pseudomonas aeruginosa strains were transferred into shake flasks for expansion culture by the slant inoculation method to obtain the Pseudomonas aeruginosa seed liquid.

[0033] 2) First add the sterilized fermentation medium into the fermenter, then put the Pseudomonas aeruginosa seed liquid into the fermenter according to the inoculum amount of 8%, and add 1‰ of foam into the medium before inoculating the seed liquid The concentration of each component in the above fermentation medium is (g / L): soybean oil 40; KH2PO4 3; K2HPO4 6; NaNO3 2; NaCl 0.5; KCl0.5; MgSO4·7H2O 0.1; Element solution 2mL / L; pH of trace element solution: 7.0, concentration of each solute in trace element solution (g / L): FeSO4·7H2O 12; ZnSO4·7H2O 3.0; CoSO4·7H2O 1.0; MnSO4·2H2O 3.0.

[0034] In this embodiment, the above fermenter is a defoam...

Embodiment 2

[0047] The difference with embodiment 1 is:

[0048] 1. In step 2), the inoculation amount of Pseudomonas aeruginosa seed solution is 12%; 5‰ of foam enemy is added in the medium; the concentration of each component in the fermentation medium is (g / L): large Soybean oil 120; KH2PO4 7; K2HPO4 10; NaNO3 4; NaCl 2; KCl 2; MgSO4 7H2O 1; anhydrous CaCl2 0.5; trace element solution 10mL / L; The concentration is (g / L): FeSO4·7H2O 12; ZnSO4·7H2O 3.0; CoSO4·7H2O 1.0; MnSO4·2H2O 3.0.

[0049] Two, in step 3), the pH of the pasty fermented liquid is adjusted to 3; the concentration of adding ethyl acetate for the first extraction is 60%, and the concentration of adding ethyl acetate for extraction again is 60%; obtained at the bottom of distillation tank 25 The paste rhamnolipid purity is 90%.

[0050] 3. In step 4), after drying the paste rhamnolipid, the purity of the obtained rhamnolipid is 99%.

[0051] Except that the above parameters are different from those in Example 1, the pro...

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Abstract

The invention discloses a method for producing rhamnolipid by virtue of fermentation and separation of pseudomonas aeruginosa. The method comprises the following steps: 1), transferring pseudomonas aeruginosa strains into a shake flask by using an inclined plane inoculation method to obtain a pseudomonas aeruginosa seed liquid; 2), adding a fermentation medium into a fermentation tank for sterilization, inoculating the pseudomonas aeruginosa seed liquid in the fermentation tank for fermentation according to 8%-12% of inoculation concentration, and fermenting the pseudomonas aeruginosa seed liquid for 120 hours to obtain pasty fermentation broth, wherein 0.1-0.5% glycerophosphorylethanolamine is added before the seed liquid is inoculated; and 3) carrying out twice extraction and distillation on the pasty fermentation broth to obtain paste rhamnolipid with the purity of 80%-90%. The method can meet the low-cast large-scale industrialized production requirements of rhamnolipid with the purity of more than 80%.

Description

technical field [0001] The invention relates to a method for producing rhamnolipid, in particular to a method for fermenting and separating producing rhamnolipid with Pseudomonas aeruginosa. Background technique [0002] In the process of oil exploration, development, refining, transportation and storage, due to accidents or operational errors, crude oil or oil products leak from the operation site or storage, and the spilled oil flows to the ground, water, beach or sea. Depending on the composition of the oil, an oil film of varying thickness is formed. This phenomenon is called oil spill. With the acceleration of the process of global economic integration, offshore oil transportation and offshore oil extraction have increased rapidly, and marine oil spill accidents have also continued to increase, which seriously threatens the ecological environment of the ocean and coastal areas. According to conservative estimates, 3.2 million tons of crude oil enters the ocean every ye...

Claims

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Application Information

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IPC IPC(8): C12P19/02C07H15/04C07H1/06C12R1/385
Inventor 曲良李广茹李明陈宇卢文玉朱零青杨雪
Owner CHINA NAT OFFSHORE OIL CORP
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