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Method for increasing content of docosahexaenoic acid in schizochytrium limacinum grease

A technology of docosahexaenoic acid and Schizochytrium, which is applied in the field of microbial fermentation, can solve the problems of reducing fermentation costs and increasing production efficiency, and achieve the effects of improving quality, increasing DHA content, and promoting a large amount of synthesis

Active Publication Date: 2015-04-01
WUHAN HUASHITE IND BIOTECH DEV +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The above method has improved the process technology of Schizochytrium fermentation to produce DHA, but there is no technology to combine the unique DHA synthesis pathway in Schizochytrium to regulate its metabolic pathway, in order to realize the production of oil produced by Schizochytrium. The further promotion of the DHA content in the medium; moreover, the further promotion of the increase of the DHA content can effectively improve the quality of the oil produced by Schizochytrium, and indirectly reduce the fermentation cost and increase the production efficiency

Method used

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  • Method for increasing content of docosahexaenoic acid in schizochytrium limacinum grease
  • Method for increasing content of docosahexaenoic acid in schizochytrium limacinum grease
  • Method for increasing content of docosahexaenoic acid in schizochytrium limacinum grease

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] (1) Add 50 mL of seed medium to a 250 mL Erlenmeyer flask. Insert the cryopreserved Schizochytrium strains, culture them for 2 days at a culture temperature of 25°C and a shaker rotation speed of 200rpm, and activate the cryopreserved Schizochytrium strains into Schizochytrium seed liquid;

[0028] Seed medium components (g / L): glucose 15.0, yeast powder 6.0, beef extract 3.0, peptone 2.0, corn steep liquor 4.0, MgSO 4 ·7H 2 O 0.2, KH 2 PO 4 2.0, Sea Crystal 15.0.

[0029] (2) Add 50 mL of fermentation medium to a 250 mL Erlenmeyer flask, insert Schizochytrium into the fermentation medium with an inoculum size of 10%, and culture at a constant temperature of 26° C. for 3 days, with a rotation speed of 200 rpm;

[0030] Fermentation medium components (g / L): 1,2-propanediol 40.0, yeast extract 6.0, MgSO 4 ·7H 2 O 0.6, KH 2 PO 4 0.5, NaNO 3 1.0, Na 2 SO 4 5.0;

[0031] CK (g / L): glucose 40.0, yeast extract 6.0, MgSO 4 ·7H 2 O 0.6, KH 2 PO 4 0.5, NaNO ...

Embodiment 2

[0044] (1) Add 50 mL of seed medium to a 250 mL Erlenmeyer flask. Insert the cryopreserved Schizochytrium strains, culture them for 2 days at a culture temperature of 25°C and a shaker rotation speed of 200rpm, and activate the cryopreserved Schizochytrium strains into Schizochytrium seed liquid;

[0045] Seed medium components (g / L): glucose 25.0, yeast powder 8.0, peptone 5.0, corn steep liquor 2.0, MgSO 4 ·7H 2 O 0.6, KH 2 PO 4 1.5, sea crystal 10.0.

[0046] (2) Add 50 mL of fermentation medium to a 250 mL Erlenmeyer flask, insert Schizochytrium into the fermentation medium with an inoculum size of 8%, and culture at a constant temperature of 26° C. for 3 days at a speed of 200 rpm;

[0047] Fermentation medium components (g / L): 1,2-propanediol 50.0, yeast extract 10.0, sodium glutamate 5.0, MgSO 4 ·7H 2 O 0.2, KH 2 PO 4 2.0, (NH 4 ) 2 SO 4 3.0, NaNO 3 2.0, Na 2 SO 4 8.0;

[0048] CK (g / L): glucose 50.0, yeast extract 10.0, sodium glutamate 5.0, MgSO 4...

Embodiment 3

[0057] (1) Add 50 mL of seed medium to a 250 mL Erlenmeyer flask. Insert the cryopreserved Schizochytrium strains, culture them for 2 days at a culture temperature of 25°C and a shaker rotation speed of 200rpm, and activate the cryopreserved Schizochytrium strains into Schizochytrium seed liquid;

[0058] Seed medium components (g / L): glucose 30.0, yeast powder 4.0, beef extract 5.0, corn steep liquor 6.0, MgSO 4 ·7H 2 O 1.0, KH 2 PO 4 0.5, sea crystal 20.0.

[0059] (2) Add 50 mL of fermentation medium to a 250 mL Erlenmeyer flask, insert Schizochytrium into the fermentation medium with an inoculum size of 4%, and culture at a constant temperature of 26° C. for 3 days, with a rotation speed of 200 rpm;

[0060] Fermentation medium components (g / L): 1,2-propanediol 60.0, yeast extract 15.0, sodium glutamate 10.0, MgSO 4 ·7H 2 O 1.0, KH 2 PO 4 1.5, (NH 4 ) 2 SO 4 5.0, NaNO 3 3.0, Na 2 SO 4 12.0;

[0061] After the fermentation medium was sterilized, the exog...

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Abstract

The invention relates to a method for increasing content of docosahexaenoic acid (DHA) in schizochytrium limacinum grease. The method is characterized by comprising the following steps: lowering metabolic flux of a pentose phosphate pathway in thallus by selecting a fermentable carbon source, and lowering the metabolic flux of the pentose phosphate pathway in thallus by further adding an exogenous regulation factor; inhibiting the activity of malic enzyme in a tricarboxylic acid transfer system to greatly lower the content of nicotinamide adenine dinucleotide phosphate (NADPH) in the schizochytrium limacinum thaluus, promote the large-scale synthesis of DHA and enable the content of DHA in the grease of the obtained thallus to be as high as 55.3%, wherein the fermentation carbon source is 1,2-propylene glycol or any one or a mixture of 1,2-propylene glycol and acetate, the exogenous regulation factor is preferably sesamol which is added after filtering and degerming when a fermentation culture medium is sterilized, and the addition amount of the sesamol is 0.5-3.0mM / L. The method disclosed by the invention is simple and easy to perform, obvious in effect, capable of greatly increasing the content of DHA in the thallus grease and easy for industrial application.

Description

technical field [0001] The invention relates to the field of microbial fermentation, in particular to a method for increasing the content of docosahexaenoic acid (DHA) in Schizochytrium oil. Background technique [0002] Docosahexaenoic acid (DHA) is a polyunsaturated fatty acid with very important physiological functions. It is an important nutrient that the human body cannot synthesize in large quantities but is indispensable. DHA exists in a large amount in the nerve tissue of the human retina and cerebral cortex, which can promote the development of the brain and vision, improve intelligence, memory and vision; in addition, it can also prevent the deposition of cholesterol on the blood vessel wall, prevent or reduce atherosclerosis sclerosis and coronary heart disease. [0003] Schizochytrium is an excellent resource for the production of polyunsaturated fatty acid oils. At present, the production of polyunsaturated fatty acid oil by large-scale fermentation using Schi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/64C12R1/645
CPCC12P7/6427
Inventor 余龙江陈伟朱圆敏周蓬蓬金文闻余金龙
Owner WUHAN HUASHITE IND BIOTECH DEV
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