A method based on reactive oxygen species to regulate the fermentation of Mortierella alpina to produce arachidonic acid oil

A technology of Mortierella alpine and arachidonic acid, which is applied in the field of microbial fermentation, can solve the problems that products and biomass cannot be accumulated at the same time, reduce the total energy consumption of stirring, and enhance the production intensity, so as to reduce the total energy consumption of stirring and shorten the Fermentation cycle, the effect of improving production intensity

Active Publication Date: 2021-07-23
NANJING TECH UNIV
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Problems solved by technology

[0004] Therefore, in view of the disadvantages of slow growth in the early stage of fermentation, inability to simultaneously accumulate products and biomass in the middle and late stages, and the accumulation of a large amount of active oxygen, the present invention aims at the intracellular active oxygen concentration of Mortierella alpina, and implements regulation in stages to increase the arachidonic The fermentation time is shortened while the acid production is increased, the production intensity is enhanced, and the total energy consumption of stirring is reduced, providing a simple and effective new fermentation process of Mortierella alpina arachidonic acid oil, which can be used for reference by similar oil-producing microorganisms

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  • A method based on reactive oxygen species to regulate the fermentation of Mortierella alpina to produce arachidonic acid oil

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Embodiment 1

[0029] Example 1. Based on active oxygen regulation and control of 7.5 L tank Mortierella alpina fermentation to produce arachidonic acid oil

[0030] 1. Activation of strains and preparation of seed solution: Mortierella alpina strain R807 (CCTCC M 2012118) was selected as the starting strain, and the preserved strains were inserted into PDA slant medium, and placed in an incubator at 25°C Cultivate for 10 days, transfer to a 500mL grooved bottle containing 100mL seed medium and culture in a constant temperature shaker, the inoculum size is 10% (v / v, 10mL), the culture conditions are 125rpm, the temperature is 25°C, and the culture time is 1 -2 days.

[0031]2. Inoculate the seed liquid obtained in step 1 into a 7.5L tank (New Brunswick Scientific, USA), the liquid volume is 5L, the inoculum volume is 10% (v / v, 500mL), and the culture temperature is controlled at 25°C. During the 0-60 hour period of fermentation, it is a relatively high active oxygen period. At the beginnin...

Embodiment 2

[0033] Example 2. Arachidonic acid oil produced by fermentation of Mortierella alpina in a 25 m³ tank based on active oxygen regulation

[0034] 1. Activation of strains and preparation of seed solution: Mortierella alpina strain R807 (CCTCC M 2012118) was selected as the starting strain, and the preserved strains were inserted into PDA slant medium, and placed in an incubator at 25°C Cultured for 10 days, transferred to a 500 mL grooved bottle containing 100 mL seed medium and cultured in a constant temperature shaker, the inoculum volume was 10% (v / v, 10 mL), the culture conditions were 125 rpm, the temperature was 25°C, The cultivation time is 2 days, and the primary seed liquid is prepared. Then, according to the inoculum volume of 10% (v / v), the primary seed solution was inserted into a 150 L seed tank, the cultivation conditions were 175 rpm, the temperature was 25°C, and the cultivation time was 1 day to prepare the secondary seed solution. Next, all the prepared secon...

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Abstract

The invention relates to a method for regulating Mortierella alpine fermentation to produce arachidonic acid oil based on active oxygen. The Mortierella alpina strain is amplified and cultivated step by step through a test tube slant medium and a shake flask medium to prepare a seed liquid, and then inoculated in a fermenter for fermentation. The entire fermentation process is divided into two stages: a relatively high active oxygen period and a relatively low active oxygen period; in the relatively high active oxygen period, the active oxygen source is added 8 to 12 hours after the fermentation starts, and the intracellular active oxygen is controlled at 100 to 125 RFLU to stimulate rapid cell proliferation; during the period of relatively low reactive oxygen species, the intracellular reactive oxygen species are controlled at about 200 RFLU by intermittent hypoxia or adding antioxidants to reduce the loss of intracellular products. A total of 168 hours of fermentation, at the end of the Mortierella alpina cell dry weight, oil content, the percentage of arachidonic acid in the total oil, and the unit yield of arachidonic acid can reach 59.8 g / L, 60.1%, respectively. 65.7%, 23.6 g / L, the production intensity of arachidonic acid can reach 3.37 g / (L*d).

Description

technical field [0001] The invention relates to a method for producing arachidonic acid oil by regulating the fermentation of Mortierella alpina based on active oxygen, and belongs to the technical field of microbial fermentation. Background technique [0002] In the past thirty years, people have paid more and more attention to dietary nutrition, and long-chain polyunsaturated fatty acids have gradually come into people's sight. Omega-6 long-chain polyunsaturated fatty acid arachidonic acid (ARA) not only plays an important role in the composition of membrane phospholipids, but also accumulates in large quantities in the brain and other nerve tissues, and can be used as a synthetic signal molecule eicosanoid It is the precursor of acid (including prostaglandin, thromboxane and leukotrienes, etc.), which is widely used in medicine and cosmetics. Especially in the field of infant food and nutritional products, since arachidonic acid oil cannot be synthesized in infants and y...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P7/64C12R1/645
CPCC12P7/6472
Inventor 纪晓俊石焜黄和任路静王维鉴
Owner NANJING TECH UNIV
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