Mortierella alpine strain for producing arachidonic acid

A technology of Mortierella alpina and arachidonic acid, applied in the field of bioengineering, can solve the problems of uneven arachidonic acid production and different fermentation cycles, and achieve the effect of easier control of the fermentation process and shortened fermentation time

Inactive Publication Date: 2012-12-19
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] However, in the existing reports, due to the influence of factors such as the quality of the strain, fermentation conditi

Method used

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  • Mortierella alpine strain for producing arachidonic acid

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Embodiment 1

[0029] Analysis of Fatty Acid Composition of Mortierella alpina

[0030] Mortierella alpina Mortierella alpina JN168 adopts liquid fermentation and adopts standard PDA medium. The fermentation temperature was 30°C, and the fermentation ended after 7 days. The mycelium was collected by suction filtration, freeze-dried to constant weight, and the mycelium was ground with a mortar. Take about 0.1g of dried mycelium and add it to a test tube with a cover. Add 5 mL of 0.4 mol / L potassium hydroxide-methanol solution to it, keep it in a water bath at 50°C for 1 hour, then add 5 mL of 14% boron trifluoride-methanol solution, keep it in a water bath at 50°C for 1 hour, add 5 mL of n-hexane Extract with alkane, and finally add 2 mL of saturated sodium chloride solution to wash with water, pipette the upper layer solution and use GC-MS for detection. The analysis results are shown in Table 1. The results showed that arachidonic acid was the main oil produced by the bacteria.

[0031...

Embodiment 2

[0036] Determination of the best carbon source

[0037] Mortierella alpina Mortierella alpina The JN168 strain was inoculated into solid PDA medium, activated at 30°C until the spores matured, and the slope was washed with sterile water to make 10 6 Each / mL spore suspension was inoculated into liquid seed medium at 2% inoculum size, and cultured at 30°C for 1 day. Inoculate into the fermentation medium with 10% inoculum amount, and incubate at 30°C for 80 hours. 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 liquid seed medium and 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 dihydrogen phosphate 1.5...

Embodiment 3

[0041] Determination of the best nitrogen source

[0042] Mortierella alpina Mortierella alpina The JN168 strain was inoculated into solid PDA medium, activated at 30°C until the spores matured, and the slope was washed with sterile water to make 10 6 Each / mL spore suspension was inoculated into liquid seed medium at 2% inoculum size, and cultured at 30°C for 1 day. Inoculate into the fermentation medium with 10% inoculum amount, and incubate at 30°C for 80 hours. Using a single factor to investigate the best nitrogen source for arachidonic acid accumulation, use ammonium nitrate, yeast powder, yeast extract, peptone, peanut cake powder or bean cake powder to replace ammonium sulfate in the basic fermentation medium for liquid fermentation, and measure the dry weight and arachidonic acid production. Basic 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 6.0, sterilize at ...

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Abstract

The invention relates to a mortierella alpine strain for producing arachidonic acid, belonging to the technical field of bioengineering. The strain is classified and named as mortierella alpine JN168, and is preserved in China center for type culture collection (CCTCC) with the CCTCC No. M2012073. The mortierella alpine strain for producing the arachidonic acid is obtained by separating and screening soil in forests of Hyesan in Wuxi of Jiangsu Province, is stable in characters and can produce the arachidonic acid with high yield by a fermentation method.

Description

technical field [0001] A Mortierella alpina strain producing arachidonic acid belongs to the technical field of bioengineering. The invention relates to a Mortierella alpina strain with high arachidonic acid production. The strain has fast growth rate and high yield, and is of great significance for the industrialized production of arachidonic acid. Background technique [0002] The arachidonic acid system is called all-cis △-5,8,11,14-eicosatetraenoic acid, which belongs to the n-6 series of polyunsaturated fatty acids and is the metabolic hub of this series of polyunsaturated fatty acids. As the most abundant and widely distributed polyunsaturated fatty acid in the human body, arachidonic acid has significant physiological and pharmacological effects. Studies have shown that arachidonic acid has good biological activities in promoting brain development, lowering blood fat, anti-inflammation and anti-lipid oxidation. Based on these important functions of arachidonic acid,...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12R1/645
Inventor 蔡宇杰丁彦蕊周立树金大勇朱建航廖祥儒张大兵李枝玲罗军侠
Owner JIANGNAN UNIV
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