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Method for preparing arachidonic acid through fermentation with addition of product accelerating agent

A technology of arachidonic acid and adding products, which is applied in the field of microbial fermentation to achieve the effects of easy operation, increased fermentation cost, and improved synthesis and accumulation in large quantities

Inactive Publication Date: 2015-12-02
RUNKE BIOENG FUJIAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In the prior art, Zhou Pengpeng et al reported the effect of glutamic acid on the synthesis of arachidonic acid by Mortierella alpina in 2002 V.30, No.9, by adding 0.8g / L The output of glutamic acid ARA is 1.41g / L, which is 70% higher than that of the control group, and the influence of surfactant on ARA output was studied in 2003 V.31, No.5, It was found that by adding 0.3% Tween20, the yield of ARA increased by 30.9% to 1.61g / L. At present, there is no report on the combination of various product accelerators to further increase the fermentation yield.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] 1. Strain activation: inoculate Mortierella alpina (Mortierella alpinaATCC16266) on the PDA slant medium, activate and cultivate at 25-30°C for 7-10 days, select well-grown slant seeds and wash the spores with sterile saline to prepare spores Concentration about 10 6 / ml of spore suspension.

[0035] 2. Seed solution preparation: the spore suspension is inoculated into 100ml liquid seed culture medium with a capacity of 500ml baffle Erlenmeyer flask with 10% inoculum size according to the volume ratio, cultivated for 2 days at 25°C and 200r / m conditions, and prepared Get the seed solution. The seed medium includes the following raw materials: glucose 30g / l, yeast powder 5g / l, KH 2 PO 4 0.6g / l, MgSO4 7H 2 O1.3g / l, adjust pH=6.0.

[0036] 3. Fermentation culture: transfer the cultured seed liquid to a baffled Erlenmeyer flask with 100ml liquid fermentation medium and a capacity of 500ml according to the inoculum amount of 5% to carry out shaker culture at a temperatu...

Embodiment 2

[0038] Mortierella alpina (Mortierella alpina ATCC32223) was activated as a strain, and the activation of the strain and the preparation of the seed solution were the same as in Example 1.

[0039] Fermentation culture: transfer the cultured seed solution to a baffled Erlenmeyer flask with 100ml liquid fermentation medium and a capacity of 500ml at a temperature of 30°C and a rotation speed of 180r / m. conditions for 8 days. The fermentation medium includes the following raw materials: glucose 70g, yeast powder 10g, KH 2 PO 4 3.6g, MgSO 4 ·7H 2 O1g, NaHCO 3 0.7g, CaCl 2 2H 2 O1g; EDTA0.05g, ZnCl 2 0.5×10 -3 g, FeCl 3 ·6H 2 O5×10 -5 g, adjust pH=6.0. After 36 hours of fermentation, the concentration of each component of the added product accelerator in the fermentation medium is: biotin 1mg / l, glutamic acid 1.3g / l, citric acid 2g / l, hydrogen peroxide 3μmol / l, volume ratio 2% n-dodecane. Shake flask culture without product promoter added during the fermentation was ...

Embodiment 3

[0041] Mortierella alpina (Mortierella alpina ATCC32221) was activated as a strain, and the activation of the strain and the preparation of the seed solution were the same as in Example 1.

[0042] Fermentation culture: transfer the cultured seed liquid to a baffled Erlenmeyer flask with 100ml liquid fermentation medium and a capacity of 500ml at a temperature of 28°C and a speed of 220r / m. Under the conditions, the culture time was 10 days. The fermentation medium includes the following raw materials: glucose 80g / l, yeast powder 10g / l, KH 2 PO 4 3g, MgSO 4 ·7H 2 O1g, NaHCO 3 0.5g, CaCl 2 2H 2 O0.3g; EDTA0.02g, ZnCl 2 0.3×10 -3 g, FeCl 3 ·6H 2 O0.3×10 -4 g, pH of the culture medium = 6.0. After the fermentation culture reaches 40h, the concentration of each component of the product promoter added to the fermentation medium in the fermentation medium is: biotin 2mg / l, glutamic acid 0.5g / l, citric acid 1.5g / l, hydrogen peroxide 4μmol / l, 3% n-dodecane by volume. Sh...

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Abstract

The invention relates to microbe fermentation, in particular to a method for preparing arachidonic acid through fermentation with addition of a product accelerating agent. The method comprises the following steps: inoculating mortierella alpina in a sterile fermentation culture medium containing a silicon source, a nitrogen source and inorganic salts for aerobic culture, collecting thalluses after culture, and extracting arachidonic acid-containing grease from the thalluses, wherein each liter of the sterile fermentation culture medium contains the following raw materials of the product accelerating agent: 1 to 5 mg / l of biotin, 0.5 to 2 g / l of glutamic acid, 1 to 3 g / l of citric acid, 1 to 5 [mu]mol / l of hydrogen peroxide and 1-5% of n-dodecane according to concentration in the fermentation culture medium. The method has the advantages that the technical problems of low yield, relatively high fermentation culture medium cost and the like in the prior art are solved; the yield is high; the fermentation process is simple; the operation is easy; the industrialized prospect is good.

Description

technical field [0001] The invention relates to microbial fermentation, in particular to a method for preparing arachidonic acid by adding a product accelerator to ferment. Background technique [0002] Arachidonic Acid (ARA for short: ARA) in Chinese is called 5,8,11,14-eicosatetraenoic acid, which belongs to the omega-6 (or n-6) series of long-chain polyunsaturated fatty acids (PUFAs) , abbreviated as C20:4(n-6). [0003] Arachidonic acid is an important component of cell membranes of various tissues in organisms, and is a precursor for the synthesis of prostaglandins in the human body, and plays an important role in the physiological metabolism of the human body. ARA synthesizes a variety of physiologically active substances in the human body, such as prostaglandins, cycloprostaglandins, leukotrienes, and thromboxanes. important role in the nervous system. In addition, ARA is of great significance to the growth and development of the retina, nervous system and brain of...

Claims

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Application Information

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IPC IPC(8): C12P7/64
Inventor 姜悦陈峰陈璇郑志达
Owner RUNKE BIOENG FUJIAN
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