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Mortierella alpina strain capable of realizing high-yield production of arachidonic acid oil and application thereof

A technology of Mortierella alpine and arachidonic acid, which is applied in microorganism-based methods, microorganisms, fungi, etc., can solve the problems of instability, high development cost, low content, etc., and achieves small occupied space and relatively large production scale. Easy to control, high ARA effect

Inactive Publication Date: 2018-11-16
JIANGSU YUANDA XIANLE PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] ARA widely exists in animals. In the past, commercial ARA was mainly derived from pig liver and fish oil, but the content was extremely low and unstable, and the development cost was expensive, which made it difficult to meet the needs of society, especially the large amount of ARA required by industries such as dairy products.

Method used

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  • Mortierella alpina strain capable of realizing high-yield production of arachidonic acid oil and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Analysis of Fatty Acid Composition of Mortierella alpina GM712

[0024] The fermentation strain is Mortierella alpina GM712, which is used for liquid fermentation, and the composition of the fermentation medium (w / v): glucose 50, yeast powder 20, sodium glutamate 4, potassium dihydrogen phosphate 1.5, pH 7.0±0.2 . During the fermentation process, glucose solution with a concentration of 600g / L was added to control the sugar concentration of the fermentation liquid to 10-15g / L. The temperature of the fermentation was 28°C, the fermentation time was 7 days, and the fermentation ended. The mycelium was collected by suction filtration, freeze-dried until the weight remained constant, and the mycelium was ground by a mortar.

[0025] Take 0.1g of dried mycelium and add it to a test tube with a lid. Add 5 mL of potassium hydroxide-methanol solution with a concentration of 0.4 mol / L to it, and keep it in a water bath at 50°C for 60 minutes, then add 5 mL of a 14% boron trifl...

Embodiment 2

[0032] Determination of optimal incubation time

[0033] Inoculate the Mortierella alpina GM712 strain preserved on the slope into solid PDA medium, activate at 30°C until the spores mature, rinse the slope with sterile water, and make 10 8 Inoculate the spore suspension with more than 1 / mL into the liquid seed medium with 2% inoculum amount, and culture at 28°C for 1 day. Inoculate into the fermentation medium with 10% inoculum amount, and cultivate at 28°C for 4-8 days.

[0034] Using a single factor to investigate the optimal culture time for arachidonic acid accumulation, samples were taken once on the 4th day, 5th day, 6th day, 7th day and 8th day, and the dry weight and arachidonic acid production were measured. Fermentation medium composition (w / v): glucose 80, ammonium sulfate 5, magnesium sulfate (MgSO 4 ·7H 2 O) 0.5, potassium dihydrogen phosphate 1.5, pH 6.5, sterilize at 115°C for 20 minutes. It can be seen from Table 5 that the optimal culture time for the fer...

Embodiment 3

[0038] Determination of the best carbon source

[0039] Inoculate the Mortierella alpina GM712 strain preserved on the slope into solid PDA medium, activate at 30°C until the spores mature, rinse the slope with sterile water, and make 10 8 Inoculate the spore suspension with more than 1 / mL into the liquid seed medium with 2% inoculum amount, and culture at 28°C for 1 day. Inoculate into the fermentation medium with 10% inoculum amount, and culture at 28°C for 7 days.

[0040] Using a single factor to investigate the best carbon source for arachidonic acid accumulation, use fructose, maltose, sucrose, lactose, D-galactose, glycerol, soluble starch or potato starch to replace glucose in the basic fermentation medium for liquid fermentation, and measure Dry weight and arachidonic acid production. Both the liquid seed medium and the basic fermentation medium consist of (w / v, the same below): glucose 80, ammonium sulfate 5, magnesium sulfate (MgSO 4 ·7H 2 O) 0.5, potassium dihy...

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Abstract

The invention discloses a Mortierella alpina strain capable of realizing high-yield production of arachidonic acid oil and application thereof. The Mortierella alpina strain is classified and named asMortierella alpine GM712 and has been preserved in China General Microbiological Culture Collection Center (CGMCC), with an accession number of CGMCC No. 15272. When the strain is used for fermentation production of arachidonic acid, the output of arachidonic acid can reach as high as 14.0 g / L, and the yield of arachidonic acid can reach 1.76 g / L / d. The strain of the invention can be used for industrial production of arachidonic acid.

Description

technical field [0001] The invention relates to a Mortierella alpina strain, in particular to a Mortierella alpina strain with high arachidonic acid oil production and application thereof. Background technique [0002] Arachidonic Acid (ARA, or 5,8,11,14-eicosatetraenoic acid) belongs to the omega-6 series of long-chain polyunsaturated fatty acids. ARA is an important essential fatty acid for the human body, and it is a component of many eicosanic acid derivatives such as prostaglandin E2 (PGE2), prostacyclin (PGI2), thromboxane A2 (TXA2), leukotriene LeukotireneB4 (LTB4) and C4 (LTC4 ), these biologically active substances have an important regulatory effect on lipoprotein metabolism, hemorheology, vascular elasticity, leukocyte function and platelet activation. ARA also has many important physiological functions such as regulating various ion channels in the body, maintaining cell membrane function, controlling cell membrane permeability, and maintaining body water retent...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P7/64C12R1/645
CPCC12P7/6427C12R2001/645C12N1/145
Inventor 杜艳张军桥赵百合刘辉王俊丰张卫红夏竹田闫闹闹刘娟玲刘年金
Owner JIANGSU YUANDA XIANLE PHARMA
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