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Preparation method of arachidonic acid

A technology of arachidonic acid and strains, applied in the field of arachidonic acid preparation, can solve the problems of low biomass, low oil content, late start of research, etc., and achieve simple fermentation process, easy purchase of raw materials, and convenient preparation Effect

Active Publication Date: 2013-12-18
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, Japan, the United States and other countries have successively produced ARA fermentation products. Domestic research in this field started late. Although some research has been done on the production of ARA by microbial fermentation, there are still low biomass and low oil content. Low-level problems, there is still a certain distance from large-scale industrial production

Method used

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  • Preparation method of arachidonic acid
  • Preparation method of arachidonic acid
  • Preparation method of arachidonic acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1. utilize Mortierella alpina to produce ARA

[0039] Mortierella alpina was cultured in a 250mL shake flask with a carrier volume of 50mL activation medium at 150rpm at 28°C for 3 days. Then insert the activated strain into the basal medium, put 50mL of the basal medium into a shaker flask with a liquid volume of 250mL, and inoculate 10% (v / v), and cultivate it in a reciprocating shaker at 28°C for 5-6 days, with a rotation speed of 150rpm , 28°C, harvested after 5 days of cultivation, harvested biomass by suction filtration, dried it at constant temperature to constant weight, ground and extracted oil with n-hexane, and evaporated to remove the organic solvent to obtain a total oil of 5g / L.

[0040] Activation medium composition (g / L):

[0041]

[0042]

[0043] Basal medium composition (g / L):

[0044]

[0045] The rest is water, pH 6.0.

Embodiment 2

[0046] Embodiment 2. Utilize basal medium to cultivate Mortierella alpina to produce ARA in fermenter

[0047] Culture in a 2L fermenter with a liquid volume of 1.5L of basic medium (as in Example 1), set the initial glucose concentration at 60-65g / L, sterilize separately, maintain the fermentation temperature at 28°C, and initially stir at 150-200rpm. Mortierella alpina was fermented in batches under the condition of ventilation volume 1vvm, the pH was adjusted to 6.5-7.0 before sterilization, and the initial pH after sterilization was 6.0-6.5. After 12 hours of cultivation, the dissolved oxygen began to drop rapidly, and the growth entered In the logarithmic phase, after 48 hours of cultivation, the growth begins to enter the stable phase. During the entire growth cycle, the initial pH value is 6.0-6.5. During the process, the pH value first increases, then decreases and then increases again. Stable between 7.0 and 7.9; dissolved oxygen decreased rapidly with the increase of...

Embodiment 3

[0049] Embodiment 3. Amino acids, vitamins and trace elements are on the impact of Mortierella alpina biomass and ARA oil content

[0050] Growth factors (vitamins, amino acids) and trace elements were added to the basal medium in Example 2 to increase the yield.

[0051] Added growth factors include:

[0052]

[0053] The vitamins are made into a 500× concentrate, sterilized by filtration with a 0.2 μm sterile filter membrane, and stored in a refrigerator at 4°C, and added directly to the basic medium before inoculation, with an addition amount of 2mL / L. Glutamic acid is added when preparing the basal medium before sterilization, and is sterilized together with the medium.

[0054] The added trace elements include (g / L):

[0055]

[0056] Trace elements were made into 1000×concentrate (that is, 1mL trace element concentrate was added to 1L medium), and adjusted to pH7. Use a 0.2 μm sterile filter membrane to filter and sterilize, store in a refrigerator at 4°C, and a...

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Abstract

A preparation method of arachidonic acid relates to the arachidonic acid. The method is as follows: optimizing culturing conditions of mortierella alpina arachidonic (ARA), fermenting in batches on basal media, then fermenting in batches on the optimized media by means of using yeast powder and corn steep liquor as nitrogen source, performing a fermenter enlarging experiment by using the yeast powder as the nitrogen source, culturing the mortierella alpina on the optimized media by using the corn steep liquor and the yeast powder as the mixed nitrogen source, and obtaining the mixed nitrogen source optimized media by replacing the nitrogen source on the optimized media into the corn steep liquor and the yeast powder; fermenting the mortierella alpina in batches on the mixed nitrogen source optimized media; performing high-glucose acclimatization to the mortierella alpina and fermenting the acclimated strains in batches on the mixed nitrogen source media; when the concentration of the glucose is reduced to 10g / L, performing fed-batch operation by 600 g / L of glucose so as to control the residual glucose within about 10 g / L, and starting to harvest dry strains, grease and the ARA after culturing for 168 h.

Description

technical field [0001] The invention relates to arachidonic acid, in particular to a preparation method of arachidonic acid. Background technique [0002] Arachidonic Acid (ARA for short, namely 5,8,11,14-eicosatetraenoic acid) belongs to the omega-6 series of long-chain polyunsaturated fatty acids, because the human body does not have the corresponding enzyme system to synthesize Specific unsaturated double bonds, therefore considered essential fatty acids, are many eicosan derivatives such as prostaglandin E 2 (PGE 2 ), prostacyclin (PGI 2 ), Thromboxane A 2 (TXA 2 ), Leukotirene B 4 (LTB 4 ) and C 4 (LTC 4 ) immediate precursor. ARA also has many important physiological functions such as regulating various ion channels in the body, controlling the permeability of the cell membrane, and maintaining the body's water retention. At the same time, studies have shown that arachidonic acid is of great significance to the development of the baby's brain and the improvem...

Claims

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Application Information

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IPC IPC(8): C12P7/64C12R1/645
Inventor 卢英华曾思钰凌雪萍敬科举吴意珣陈翠雪
Owner XIAMEN UNIV
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