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Method for producing DHA by fermentation of schizochytrium limacinum

A technology of Schizochytrium and fermentation liquid, which is applied in the field of DHA production by fermentation of Schizochytrium, and can solve the problems of low production efficiency, contamination, complicated production line and the like

Inactive Publication Date: 2020-07-07
CABIO BIOTECH WUHAN CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The invention adopts the idea of ​​batch fermentation, but the consumption of manpower and material resources for batch fermentation is relatively large, and each batch of fermentation needs to carry out operations such as loading, sterilization, inoculation, discharging, cleaning, etc. The process is cumbersome and the fermentation cycle is long , lower production efficiency
[0006] CN104263770A discloses a semi-continuous stage fermentation of Schizochytrium, which adopts the traditional continuous fermentation idea, the main fermentation tank adopts continuous fermentation, and the post-fermentation adopts the mode of batch fermentation, but the problem of this method is that as long as a certain When bacteria enter a fermentation tank, a large area of ​​bacteria will appear, which will eventually lead to abnormal fermentation
At present, continuous and semi-continuous fermentation are produced by single-line or double-line, each line involves multiple fermentation tanks, and the production line is complex

Method used

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  • Method for producing DHA by fermentation of schizochytrium limacinum

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] The present embodiment provides a kind of method that Schizochytrium produces DHA, comprises steps as follows:

[0075] (1) Activation, expansion and fermentation of Schizochytrium: the preserved Schizochytrium CABIO-A-2-II (Schizochytrium sp.CABIO-A-2-II) (its preservation number is CCTCC NO: M2019990 ; The preservation address is China Center for Type Culture Collection (CCTCC), China, Wuhan, Wuhan University, postcode: 430070; the preservation date is December 2, 2019) After being cultured on activated medium, it was transferred to the seed tank for expansion , the expansion temperature is 30°C, and the culture is continued for 60 hours to obtain the seed liquid; then the seed liquid is inoculated into the fermenter, after the inoculation is completed, the fermenter contains 36L of fermentation liquid, and the fermentation is continued at 30°C for 96h;

[0076](2) Feed 18L of fermented liquid into the enzymolysis tank, inactivate it and cool it down to 55°C, add sodi...

Embodiment 2

[0081] The present embodiment provides a kind of method that Schizochytrium produces DHA, comprises steps as follows:

[0082] (1) Activation, expansion and fermentation of Schizochytrium: the preserved Schizochytrium was cultured in an activated medium and then transferred to a seed tank for expansion. The expansion temperature was 28°C for 72 hours to obtain seeds Then inoculate the seed liquid into the fermenter. After the inoculation, the fermenter contains 40L of fermented liquid, and continue to ferment at 30°C for 72h;

[0083] (2) Feed 28L of fermentation broth into the enzymolysis tank, inactivate and cool down to 55°C, adjust the pH to 7.0, and then add a final concentration of 0.7% protease (500,000 U / g) and 0.3% cellulase (10,000 U / g) enzymolysis for 10 hours to obtain enzymolysis solution;

[0084] Discharge the enzymatic solution into the three-phase drum centrifuge, collect the DHA crude oil into the collection tank, and discharge the bacteria residue into the ...

Embodiment 3

[0088] The present embodiment provides a kind of method that Schizochytrium produces DHA, comprises steps as follows:

[0089] (1) Activation, expansion and fermentation of Schizochytrium: the preserved Schizochytrium was cultured in an activated medium and then transferred to a seed tank for expansion. The expansion temperature was 25°C for 48 hours to obtain seeds After the inoculation, the fermenter contains 36L of fermentation broth, and the fermentation is continued at 25°C for 96h;

[0090] (2) After fermentation, 14.4L of fermented liquid is fed into the enzymolysis tank, cooled to 60°C after inactivation, and then added with a final concentration of 0.6% protease (500,000 U / g) and 0.2% cellulase (1 10,000 U / g) enzymolysis for 10 hours to obtain the enzymolysis solution; discharge the enzymolysis solution into a three-phase drum centrifuge, collect DHA crude oil to a collection tank, and discharge the bacterial meal to a waste tank for use;

[0091] (3) the remaining 6...

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Abstract

The invention provides a method for producing DHA by fermentation of schizochytrium limacinum. The method comprises the following steps: (1) activating the schizochytrium limacinum, and carrying out expanding culture and fermenting so as to obtain a fermentation liquor; (2) partially discharging the fermentation liquor into an enzymolysis tank for enzymolysis, transferring an obtained enzymolysisliquid to separation equipment, extracting crude DHA oil, and recovering schizochytrium limacinum meal for subsequent usage; and (3) supplementing a part of culture medium components and recovered schizochytrium limacinum meal for subsequent usage into the residual fermentation liquor, continuing fermentation, and repeating the step (2) until a fermentation period is finished. According to the fermentation method provided by the invention, the schizochytrium limacinum meal is used as a nutrient substance for cyclic utilization, so nutrition can be provided for microorganisms, and the production activity of the microorganisms is maintained; the production efficiency of traditional semi-continuous fermentation can be improved; meanwhile, the bacterial contamination risk can be reduced; the DHA production cost of the schizochytrium limacinum can be reduced; the concept of green production can be met; and environmental protection can be easily achieved.

Description

technical field [0001] The invention belongs to the technical field of microbial fermentation, and in particular relates to a method for producing DHA by microbial fermentation, in particular to a method for producing DHA by fermentation of Schizochytrium. Background technique [0002] Docosahexaenoic acid (DHA) is an Omega-3 polyunsaturated fatty acid, which has important physiological functions such as promoting brain cell development, protecting eyesight, improving intelligence and improving memory. It is indispensable but cannot be synthesized by the human body. Important nutrients, widely used in infant food and pharmaceutical industries. The DHA obtained through the extraction and purification of deep-sea fish oil is relatively small, and the supply is limited by the season, which is far from meeting the market demand. [0003] Because marine microorganisms such as Thraustochytrium (Thraustochytrium), Schizochytrium (Schizochytrium) and Crythecodinium cohnii (Crytheco...

Claims

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Application Information

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IPC IPC(8): C12P7/64C12N1/14C12R1/645
CPCC12P7/6427C12N1/14
Inventor 李翔宇陆姝欢周齐余超汪志明
Owner CABIO BIOTECH WUHAN CO LTD
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