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Method for producing DHA through Crypthecodinium cohnii fermentation

A dinoflagellate and fermenter technology, which is applied in the field of mixed oils and fats, can solve the problems of low genome predictability, unsuitability for large-scale production, low long-term genetic stability, etc., so as to improve market competitiveness and reduce production. Cost, high oil yield effect

Active Publication Date: 2014-08-06
INNER MONGOLIA KINGDOMWAY PHARMA LTD +1
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Problems solved by technology

Although its DHA productivity has been greatly improved compared with the previous research, for the industrial production of docosahexaenoic acid by using microalgae, the production cost is greatly reduced, the unit yield is increased, and the method of microbial fermentation to produce DHA can be vigorously developed. Promotion and popularization are not enough
[0005] The patent with the publication number CN102391955A discloses a mutagenesis screening method for dinoflagellates, which uses ultraviolet light + LiCl and diethyl sulfate to carry out compound mutagenesis on dinoflagellates to obtain strains with better performance, but The long-term genetic stability of this mutagenesis is not high, and the output of bacterial strain producing DHA is unstable, which is not suitable for industrialized production. Purification puts forward higher technical requirements and is not suitable for large-scale production

Method used

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  • Method for producing DHA through Crypthecodinium cohnii fermentation
  • Method for producing DHA through Crypthecodinium cohnii fermentation
  • Method for producing DHA through Crypthecodinium cohnii fermentation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 130

[0021] Embodiment 1.30L fermentor dinoflagellates ferment and produce DHA

[0022] The fresh dinoflagellate slant strains were inserted into a 2L shaker flask with 350mL medium and cultured for 48h at a shaker speed of 180rpm to prepare a DHA seed solution with a cell density of 8g / L.

[0023] In a 30L fermenter, insert the DHA seed liquid into the sterilized medium according to the inoculum amount of 3%, the volume after connection is 20L, the tank pressure is 0.02MPa, the initial stirring speed is 120rpm, the ventilation rate is 1vvm, and the dissolved oxygen speed is passed. The coupling controls the DO at about 50%. The whole fermentation process is fed with 40% ammonia water to control the pH at about 6.8. During the process, the carbon source is fed to control the glucose concentration at 1-5g / L. After 68 hours of cultivation, the fermentation is stopped and the tank is placed. The dry cell weight in the fermentation broth is determined to be 84g / L, the oil content is 50...

Embodiment 2100

[0026] Embodiment 2.100L fermentation tank dinoflagellates ferment and produce DHA

[0027] The fresh dinoflagellate slant strains were inserted into a 2L shaker flask with 350mL medium and cultured for 48h at a shaker speed of 180rpm to prepare a DHA seed solution with a cell density of 7g / L.

[0028] In a 100L fermenter, insert the DHA seed liquid into the sterilized medium according to the inoculation amount of 3%, the volume after connection is 50L, the tank pressure is 0.02MPa, the initial stirring speed is 120rpm, the ventilation rate is 1vvm, and the dissolved oxygen speed is passed. The coupling controls the DO at about 50%. The whole fermentation process is fed with 40% ammonia water to control the pH at about 6.8. During the process, the carbon source is fed to control the glucose concentration at 1-5g / L. After 72 hours of fermentation, the fermentation is stopped and the tank is placed. The dry cell weight in the fermentation broth was determined to be 91g / L, the oi...

Embodiment 3

[0031] Example 3.100m 3 Fermentation of dinoflagellation algae to produce DHA in a fermenter

[0032] Carry out industrial expansion culture to dinoflagellates, the process is as follows:

[0033] Put the fresh dinoflagellate slant strains into a 2L shake flask with 350mL of medium, culture on a shaker at 28°C for 48h, and the shaker rotates at 180rpm to prepare DHA seed solution with a cell density of 9g / L.

[0034] In the first-level seed tank, the DHA seed solution in the shake flask was inserted into the sterilized first-level seed tank culture medium according to the inoculum amount of 0.4%, the ventilation rate was 1vvm, the pressure was 0.02MPa, the stirring speed was 20Hz, and cultivated for 30h , The measured dry cell weight in the fermentation broth was 5g / L.

[0035] In the secondary seed tank, insert the primary seeds into the sterilized medium of the secondary seed tank according to the inoculation amount of 3%, the pressure is 0.02MPa, the stirring speed is 25H...

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Abstract

The invention provides a method for producing DHA through Crypthecodinium cohnii fermentation. According to the method, Crypthecodinium cohnii is used as a strain and a culture medium including a carbon source, a nitrogen source, inorganic salt and trace elements is used, wherein 20% glycerol is added into the carbon source, DHA grease is prepared after three-stage fermentation, and in the fermentation process, a dissolved oxygen control strategy is adopted to control dissolved oxygen at 50%. In produced DHA fermentation liquid, the cell dry weight is 70-110 g / L, the grease content is 40-60%, the DHA content accounts for 45-55% of total grease, and the highest DHA producing rate is 8.7 g / (L.d) and is 1.89 times existing highest producing rate of DHA produced through the Crypthecodinium cohnii, which is reported so far. The technical indexes of the method provided by the invention are obviously better than the existing technical indexes; the produced DHA grease is high in yield, high in purity, low in cost and is beneficial to large-scale industrial production of the DHA.

Description

technical field [0001] The invention relates to a microbial fermentation production method, in particular to the fermentation production of mixed oil containing docosahexaenoic acid by dinoflagellation strains. Background technique [0002] Docosahexaenoic acid (cis-4, 7, 10, 13, 16, 19-docosahexaenoic acid, DHA) is an extremely important long-chain polyunsaturated fatty acid of omega-3 series, which has the function of promoting brain development in infants and young children. Important physiological functions such as growth, vision protection, prevention and treatment of cardiovascular diseases, and improvement of the body's immunity. [0003] The current commercial sources of DHA are mainly fish oil and microalgae. The annual output of domestic fish oil is about 40,000 tons. The price of refined and purified fish oil with a DHA content of 30% is 40,000-50,000 yuan / ton. 200,000 USD / ton. In recent years, due to the fact that the quality of domestic fish oil cannot meet t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/64C12R1/89
Inventor 文昌陈必钦詹光煌吴轶徐鲁明蒋四富王霄凤
Owner INNER MONGOLIA KINGDOMWAY PHARMA LTD
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