Pythium irregulare strain for producing eicosapentaenoic acid

A technology of eicosapentaenoic acid and Pythium, applied in the field of bioengineering, can solve the problems of unstable quantity and quality of polyunsaturated fatty acids, reduce consumer acceptance, etc., and achieve the effect of stable properties

Inactive Publication Date: 2013-09-25
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, fish oil contains cholesterol and other substances that are unhealthy for the human body. In addition, the amount and type of fish caught will be affected by changes in the season, location and food chain in the ocean, resulting in unstable quantity and quality of polyunsaturated fatty acids
And fatty acids refined from fish oil often have an unbearable fishy smell, reducing consumer acceptance

Method used

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  • Pythium irregulare strain for producing eicosapentaenoic acid
  • Pythium irregulare strain for producing eicosapentaenoic acid
  • Pythium irregulare strain for producing eicosapentaenoic acid

Examples

Experimental program
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Effect test

Embodiment 1

[0027] Analysis of Fatty Acid Composition of Pythium maloestrogens

[0028] Pythium malatus Python irregular JN168 adopts liquid fermentation and adopts standard PDA medium. The fermentation temperature was 25°C, and the fermentation ended after 4 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.

[0029] Chromatographic conditions: Finnigan Trace MS (USA) gas chromatography-mass spectrometer. A DB-MAX capillary column (30m×0.25m...

Embodiment 2

[0034] Determination of the best carbon source

[0035] Pythium slant Python irregular The JN168 strain was inoculated on the plate, and then cultured in an incubator at 25°C. The plate medium composition is: glucose 20g / L, yeast powder 5g / L, agar 20%. After 3 days, scrape the mycelia on the plate into sterile water, shake and break the mycelium, and inoculate it into the seed medium with a 10% inoculum amount, cultivate it on a shaker at 25°C for 48 hours, and then inoculate it with a 10% inoculum amount. In the fermentation medium, culture on a shaker at 25°C for 96 hours. Using a single factor to investigate the best carbon source for accumulating eicosapentaenoic acid, using fructose, maltose, sucrose, lactose, D-galactose, glycerol, corn flour, or potato starch instead of glucose in the basic fermentation medium for liquid Fermentation, dry weight and eicosapentaenoic acid production were measured. Basic fermentation medium composition (w / v): glucose 60g / L, yeast pow...

Embodiment 3

[0039] Determination of the best nitrogen source

[0040] Pythium slant Python irregular The JN168 strain was inoculated on the plate, and then cultured in an incubator at 25°C. The plate medium composition is: glucose 20g / L, yeast powder 5g / L, agar 20%. After 3 days, scrape the mycelia on the plate into sterile water, shake and break the mycelium, and inoculate it into the seed medium with a 10% inoculum amount, cultivate it on a shaker at 25°C for 48 hours, and then inoculate it with a 10% inoculum amount. In the fermentation medium, culture on a shaker at 25°C for 96 hours. Using a single factor to investigate the best nitrogen source for the accumulation of eicosapentaenoic acid, use ammonium nitrate, ammonium sulfate, yeast extract, peptone, peanut cake powder or bean cake powder to replace the yeast powder in the basic fermentation medium for liquid fermentation, and measure Dry weight and eicosapentaenoic acid production. Basic fermentation medium composition (w / v)...

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Abstract

The invention relates to a Pythium irregulare strain for producing eicosapentaenoic acid, belonging to the technical field of biological engineering. The strain is named Pythium irregulare JN168 and is preserved in China Center for Type Culture Collection (CCTCC), and the preserve number of the strain is CCTCC NO: M2012421. By carrying out isolation and screening on soil of forests in Huishan of Wuxi of Jiangsu province, the Pythium irregulare strain which is stable in characters and capable of producing the eicosapentaenoic acid is obtained and can be used for producing eicosapentaenoic acid (EPA) through liquid state fermentation; and the yield of EPA can be above 5.5 g/L, and meanwhile, EPA accounts for more than 30% of total grease.

Description

technical field [0001] A strain of Pythium terrestris producing eicosapentaenoic acid (EPA) belongs to the technical field of bioengineering. The invention relates to a high-yielding eicosapentaenoic acid Pythium strain. The strain has fast growth and high yield, and is of great significance for the industrial production of eicosapentaenoic acid. Background technique [0002] Eicosapentaenoic acid (5,8,11,14,17-Eicosapentaenoic acid EPA) is a 20-carbon straight-chain unsaturated fatty acid (PUFA) with 5 non-conjugated double bonds. The first double bond starts at the third carbon atom at the methyl end, which belongs to the omega-3 series of unsaturated fatty acids. It is named 20: 5n-3 by n, Ω, shorthand nomenclature, and 5, 8, 11, 14, 17-cis-eicosapentaenoic acid by IUPAC system nomenclature, and its molecular formula is C 20 h 30 o 2 , the relative molecular weight is 302.6. At room temperature, it is a light yellow oily liquid, easily soluble in non-polar solvents s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12P7/64C12R1/645
Inventor 蔡宇杰陈卫刘飞翔丁彦蕊金大勇朱建航廖祥儒李枝玲罗军侠张大兵温志友
Owner JIANGNAN UNIV
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