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Preparation method for MEL (Mannosylerythritol lipids)

A technology of sugar erythritol and sugar alcohol lipid, which is applied in the field of preparation of mannose erythritol lipid, can solve the problems of low yield of mannose erythritol lipid and the like, and achieves the effect of increasing yield

Inactive Publication Date: 2015-07-22
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In view of the deficiencies in the prior art, the object of the present invention is to provide a method for high mannose erythritol lipid production, which solves the problem of low production of mannose erythritol lipid

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] (1) Strain activation: use the sterilized maltose agar plate as the activation medium, insert the refrigerated strain Pseudozyma aphidis DSMZ70725, activate and cultivate at 28°C for 3 days, and wait until the plate is covered with bacteria spare. The components of the maltose agar medium include 1% glucose, 0.5% soybean peptone, 0.3% malt extract powder, 0.3% yeast powder, 1.5% agar powder and distilled water.

[0038] (2) Seed culture: Pick a ring of bacteria from the plate culture medium and insert it into a 250mL Erlenmeyer flask containing 50mL seed culture solution, and cultivate it at 28°C and 220rpm for 24h; put the cultured seed liquid at 4°C Centrifuge at 3000rpm for 15min, remove the supernatant, separate the bacterial cells, take the bacterial cells, and wash 3 times with 0.9% sodium chloride solution.

[0039] Wherein, the seed culture fluid components include 4.0% of glucose, 0.3% of sodium nitrate, 0.03% of magnesium sulfate, 0.03% of potassium dihydroge...

Embodiment 2

[0044] Step (1) and step (2) are carried out according to the implementation described in Example 1.

[0045] (3) Fermentation culture: the OD 600 =2.0 bacterial cells were inserted into a 500mL Erlenmeyer flask equipped with 100mL fermentation medium at 20mL / L, and cultured at 28°C and 220rpm for 10 days;

[0046] Among them, the components of the fermentation medium include: vegetable oil 9.68%, yeast powder 0.15%, magnesium sulfate 0.06%, calcium chloride 0.003%, manganese sulfate 0.01%, pH natural; that is, the C / N ratio is 59:1.

[0047] (4) Separation and purification: after the fermentation and cultivation were completed, the fermented liquid was extracted and separated twice with an equal volume of ethyl acetate, the organic phases were combined, and concentrated under reduced pressure with a rotary evaporator to remove the extraction solvent to obtain the mannose erythritol lipid. Crude. Using methanol and cyclohexane to further extract, separate and purify the crud...

Embodiment 3

[0049] Step (1) and step (2) are carried out according to the implementation described in Example 1.

[0050] (3) Fermentation culture: the OD 600 =2.0 bacterial cells were inserted into a 500mL Erlenmeyer flask equipped with 100mL fermentation medium at 20mL / L, and cultured at 28°C and 220rpm for 10 days;

[0051] Wherein, the composition of fermentation medium comprises: 6.32% of vegetable oil, 0.15% of yeast powder, 0.06% of magnesium sulfate, 0.003% of calcium chloride, 0.01% of manganese sulfate, pH natural; That is to say, the C / N ratio in the culture system is 37:1 .

[0052] (4) Separation and purification: after the fermentation and cultivation were completed, the fermented liquid was extracted and separated twice with an equal volume of ethyl acetate, the organic phases were combined, and concentrated under reduced pressure with a rotary evaporator to remove the extraction solvent to obtain the mannose erythritol lipid. Crude. Using methanol and cyclohexane to fur...

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Abstract

The invention discloses a preparation method for high-yield MEL (Mannosylerythritol lipids). The preparation method comprises the following steps: inoculating a fermentation culture medium with Pseudozyma aphidis DSMZ70725, and performing fermentation cultivation, so as to obtain the MEL, wherein vegetable oil is adopted as a substrate and a carbon source in the fermentation cultivation, and the ratio of C to N in the fermentation culture medium is (30-60):1. According to the invention, the yield of the MEL is increased through adjusting the ratio of C to N in the cultivation system, and the difficulty of low yield of the MEL prepared by adopting the preparation method is solved.

Description

technical field [0001] The invention relates to the field of microbial fermentation, in particular to a preparation method of mannose erythritol lipid. Background technique [0002] Mannosylerythritol lipids (MELs) is a glycolipid biosurfactant. At present, the strains used to transform and produce MELs mainly include Ustligo maydis and Pseudazyma. [0003] Compared with chemical surfactants, MELs not only have the characteristics of biocompatibility, biodegradability, non-toxicity or low toxicity, but also have good surface activity, emulsification, biodegradability, and lower critical micelle concentration. It also has many special physiological activities, such as inhibiting microbial growth, inducing cell mutation, differentiating human myelogenous leukemia cell lines and melanoma cells, improving gene transfection efficiency, and having a strong coordination ability with glycoproteins, etc. These The characteristics make MELs have great application prospects in indust...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/44C12R1/645
Inventor 陈启和范琳琳董亚晨牛永武李宏吉
Owner ZHEJIANG UNIV
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