Method for promoting production of mortierella alpina and oil yield of arachidonic acid

A technology of Mortierella alpina and arachidonic acid, which is applied in the field of microbial fermentation, can solve the problems of genetic instability of strains, long production cycle, increased energy consumption, etc., to promote the yield of ARA oil, reduce production costs, and operate simple and easy effects

Active Publication Date: 2017-12-08
HUAZHONG UNIV OF SCI & TECH
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  • Description
  • Claims
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Problems solved by technology

[0003] Improving ARA production by optimizing fermentation conditions is the focus of current research. Chinese patent 201310475443.1 discloses a method for increasing ARA production through mutagenesis and medium optimization. Although ARA production has been greatly improved, the mutagenesis is random and The mutated strain is genetically unstable, easily leads to back mutation, and is not easy for industrial application
Chinese patent CN201410556081.3 discloses a method for producing arachidonic acid oil based on t

Method used

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  • Method for promoting production of mortierella alpina and oil yield of arachidonic acid
  • Method for promoting production of mortierella alpina and oil yield of arachidonic acid
  • Method for promoting production of mortierella alpina and oil yield of arachidonic acid

Examples

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Embodiment 1

[0035] Example 1 Preparation of Accelerator for Regulating Mortierella Alpina Growth and ARA Oil Yield

[0036] (1) Preparation of Mortierella alpina seed solution

[0037] Inoculate Mortierella alpina, a slant strain preserved in a 4°C refrigerator, into PDA medium for 168 hours of activation to prepare spores, add 50 mL of sterile saline with glass beads to prepare spore liquid, and inoculate the spore liquid according to the inoculation amount of 10%. Into the seed culture medium for 48 hours at 28°C to prepare the seed solution;

[0038] (2) The seed liquid is inserted into the fermentation medium according to the inoculum amount (v / v) of 10%, and cultured at 28°C, and the fermentation period is 192h;

[0039] (3) After the fermentation, use a suction filter bottle for suction filtration, and place the obtained wet bacteria in an oven at 60°C for drying. It takes about 6-8 hours until the bacteria have a constant weight and weighed.

[0040] (4) Break the dry cell wall, ...

Embodiment 2

[0043] Example 2 Preparation of accelerators for regulating the growth of Mortierella alpina and the yield of ARA oil

[0044] (1) Preparation of Mortierella alpina seed solution

[0045] The slant strain Mortierella alpina preserved in the refrigerator at 4°C was inoculated into the PDA medium and activated for 168h to prepare spores, and 50 mL of sterile saline with glass beads was added to prepare the spore liquid, and the spore liquid was inoculated according to 10% ( v / v) inserting into the seed medium and culturing for 48 hours at 28°C to prepare seed solution;

[0046] (2) Insert the seed liquid into a 7L fermenter equipped with 4L fermentation medium according to the inoculum amount (v / v) of 6%, at 150rpm, with an aeration rate of 1.5vvm, and cultivate at 28°C;

[0047] (3) Begin continuous sampling from Mortierella alpina fermentation 72h, sampling time point 72h, 96h, 120h, 144h, 168h, finish to 192h fermentation, according to the extraction mode of step (5) in the em...

Embodiment 3

[0048] Embodiment 3 Add accelerator to the regulation (shake flask level) of Mortierella alpina growth and ARA oil yield

[0049] (1) Preparation of Mortierella alpina seed solution

[0050] The slant strain Mortierella alpina preserved in the refrigerator at 4°C was inoculated into the PDA medium and activated for 168h to prepare spores, and 50 mL of sterile saline with glass beads was added to prepare the spore liquid, and the spore liquid was inoculated according to 10% ( v / v) inserting into the seed medium and culturing for 48 hours at 28°C to prepare seed solution;

[0051] (2) Insert the seed liquid into the fermentation medium according to the inoculum amount (v / v) of 10%, and add the accelerator prepared in Example 1 of 0.1% volume ratio respectively at the beginning of fermentation 0h, 24h, and 48h, at 28°C Cultivation, the fermentation period is 180h; each experiment has 5 parallel samples.

[0052] (3) After the fermentation, use a suction filter bottle to carry o...

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Abstract

The invention relates to a method for promoting production of mortierella alpina and oil yield of arachidonic acid. The method comprises the following steps that fermented clear liquid of the mortierella alpina is collected, and through extraction and concentration of hydrophilic organic solvent, an accelerating agent which can adjust and control the production of the mortierella alpine and the oil yield of the arachidonic aicd is obtained by screening; and in the 0-48 hours at the early stage of fermentation, the accelerating agent is added to a normal medium according to the ratio of 0.1-0.5%, and a conditioned medium is prepared to inoculate and cultured at the temperature of 28 DEG C. After the fermentation, in fermentation results of some embodiments, biomass is 43.2 g/L, oil content is 61.3%, ARA content is 60.7%, ARA yield is 16.1 g/L, and the fermentation period is shortened from 192 hours to 168 hours. The method for promoting the production of the mortierella alpina and the oil yield of the arachidonic acid is simple and effective, promotes the production of the mortierella alpine, improves the oil yield of the arachidonic acid, and is of great significance to industrial application.

Description

technical field [0001] The invention relates to the field of microbial fermentation, in particular to a method for promoting the growth of Mortierella alpina and the yield of arachidonic acid oil. Background technique [0002] Arachidonic acid (ARA) is a long-chain polyunsaturated fatty acid with very important physiological functions. It is an important nutrient that cannot be synthesized by the human body itself but is indispensable. ARA exists in large quantities in the nerves of the human retina and cerebral cortex. In the tissue, it is the basis of the brain development of fetuses and infants, and it has a promoting effect on their intelligence and cognitive ability. [0003] Improving ARA production by optimizing fermentation conditions is the focus of current research. Chinese patent 201310475443.1 discloses a method for increasing ARA production through mutagenesis and medium optimization. Although ARA production has been greatly improved, the mutagenesis is random a...

Claims

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Application Information

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IPC IPC(8): C12P7/64C12R1/645
CPCC12P7/6427
Inventor 余龙江朱圆敏金航
Owner HUAZHONG UNIV OF SCI & TECH
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