Method for producing EPA and DHA through schizochytrium limacinum fermentation

A technology for producing strains of Schizochytrium, which is applied in the field of bioengineering, can solve the problems of different PUFA content and the content needs to be further improved, and achieve the effect of increasing the output of DHA

Pending Publication Date: 2022-05-31
NANJING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Among the fatty acids produced by Schizochytrium HX-308, saturated fatty acids represented by hexadecanoic acid and tetradecanoic acid accounted for more than 30%, and PUFA accounted for about 60% of the total proportion, but the contents of various PUFAs were different. The content of EPA and DHA, which have higher application value in the development of pharmaceuticals and health products, needs to be further increased

Method used

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  • Method for producing EPA and DHA through schizochytrium limacinum fermentation
  • Method for producing EPA and DHA through schizochytrium limacinum fermentation
  • Method for producing EPA and DHA through schizochytrium limacinum fermentation

Examples

Experimental program
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Effect test

Embodiment 1

[0045] Embodiment 1 A kind of method of Schizochytrium HX-308 fermentation production EPA and DHA

[0046] This embodiment includes the following steps carried out in sequence:

[0047] S1. Preparation of strains for fermentation

[0048] Prepare the seed medium and weigh the raw materials according to the following proportions: glucose 50g / L, yeast extract powder 5g / L, sodium sulfate 5g / L, magnesium sulfate 2g / L, ammonium sulfate 6g / L, potassium chloride 1g / L, Calcium chloride 0.1g / L, potassium sulfate 0.6g / L, potassium dihydrogen phosphate 1g / L, sodium glutamate 10g / L, nickel sulfate 0.05mg / L, vitamin B 6 5mg / L and vitamin B 12 0.5mg / L, prepared as a seed medium, and adjusted the pH to 6.6.

[0049] Select a single colony of Schizochytrium HX-308 and inoculate it in 50 mL of seed medium, and culture it on a shaker at 28°C and 180 r / min for 24 hours to form the first-generation seed; take 1 mL of the first-generation seed culture solution and inoculate it into 50 mL of s...

Embodiment 2

[0054] The construction method of embodiment 2 Schizochytrium engineering bacteria ED and Schizochytrium engineering bacteria EDG

[0055] (1) Construction method of Schizochytrium engineering bacteria ED

[0056] The Schizochytrium engineering strain ED is based on the Schizochytrium HX-308 with the preservation number CCTCC No. M209059 as the wild type, and overexpresses the Schizochytrium C18 elongase gene and the Schizochytrium n3 desaturase gene Schizochytrium engineering bacterium; it is after the C18 elongase gene of cloned Schizochytrium HX-308 and the n3 desaturase gene of Schizochytrium HX-308 are connected into pBS-Zeo vector, and then transformed into Chytrium chytrium HX-308 obtained. The specific construction method is:

[0057] (I) Clone C18ELO and n3DES gene fragments

[0058] According to the sequence information of the Schizochytrium C18 elongase (C18ELO) gene and the n3 desaturase (n3DES) gene, design the primers P1 and P2, primers P3 and P4 shown in the ...

Embodiment 3～8

[0081] The method of embodiment 3~8 Schizochytrium fermentation production EPA and DHA

[0082] Examples 3 to 8 are respectively a method for producing EPA and DHA by fermentation of Schizochytrium fungus. The specific fermentation production method is basically the same as that of Example 1, the only difference being the specific production strain, inoculum size or fermentation production parameters in the process parameters. The settings are different, and the specific differences are shown in Table 1;

[0083] Table 1 embodiment 3~8 process parameter table

[0084]

[0085] Other parts are the same as in Example 1 in the fermentation production method of Examples 3-8.

[0086] The lipids produced by fermentation in Examples 3-8 were analyzed by gas chromatography, and the results showed that EPA and DHA were successfully produced.

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Abstract

The invention belongs to the technical field of bioengineering, and discloses a method for producing EPA (eicosapentaenoic acid) and DHA (docosahexaenoic acid) by fermentation of schizochytrium limacinum, which comprises the following steps: by taking schizochytrium limacinum as a production strain, adding ensidipine into a fermentation culture medium, and fermenting to produce polyunsaturated fatty acid. According to the method, the EPA content and the DHA content in a schizochytrium limacinum fermentation product are increased by activating a part of the ELO / DES pathway of the schizochytrium limacinum, and the EPA content in the fermentation product is increased by 8.73 times and the DHA content is increased by 1.15 times by adopting the schizochytrium limacinum engineering bacterium EDG as a production strain.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and relates to a fermentation method of Schizochytrium, in particular to a method for producing EPA and DHA by Schizochytrium fermentation. Background technique [0002] Schizochytrium sp. is a heterotrophic marine protist of the genus Thraustochytrium rich in oil. It is widely used in scientific research and commercial production because of its fast growth and easy cultivation. The fatty acid accumulation of Schizochytrium is mainly DHA (docosahexaenoic acid), DPA (Docosapentaenoic acid), hexadecanoic acid and tetradecanoic acid, and in different strains of Schizochytrium, Production of polyunsaturated fatty acids (PUFAs) such as EPA (eicosapentaenoic acid), which is more helpful for cardiovascular disease and recovery from COVID-19 infection, varied. [0003] Among the fatty acids produced by Schizochytrium HX-308, saturated fatty acids represented by hexadecanoic acid and tetradecanoic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/6432C12P7/6434C12N1/15C12N15/79C12N15/53C12N15/54C12N1/38C12R1/645
CPCC12P7/6427C12N9/1029C12N9/0071C12N15/79C12N1/38
Inventor 黄和马旺孙小曼贾雨雷黄鹏伟
Owner NANJING NORMAL UNIVERSITY
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