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Mutant strain with high yield of Gamma-linolenic acid and preparation method and application of mutant strain

A technology of mutant strains and linolenic acid, which is applied in the field of bioengineering, oil engineering, and biotechnology, and can solve the problems of plant resource limitation and further research

Inactive Publication Date: 2014-01-01
WUHAN POLYTECHNIC UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] γ-linolenic acid can be obtained from some plants, but plant resources are limited, and wider plant sources need to be found and developed
[0013] The production of γ-linolenic acid by microbial fermentation technology has been industrialized, but the improvement of yield and purity still needs further research

Method used

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  • Mutant strain with high yield of Gamma-linolenic acid and preparation method and application of mutant strain
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  • Mutant strain with high yield of Gamma-linolenic acid and preparation method and application of mutant strain

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Screening for lipogenic strains

[0070] The strains used in the research of the present invention are shown in Table 1. The solid medium is PDA medium, the liquid medium is glucose 4-6%, starch 0.5-2.0%, yeast extract is 0.1-0.8% and inorganic salts, pH6.0-7.0, The fermentation medium is 5-8% glucose, 1.0-3.0% starch, 0.5-1.5% yeast extract, 1.0-3.0% soybean meal powder and inorganic salts, pH 6.0-7.0;

[0071] Described inorganic salt is sodium chloride 0.1%, magnesium sulfate 0.01%.

[0072] Table 1 The original strains and sources used in the research

[0073]

[0074]

[0075] The screening criteria are as follows:

[0076] Bacterial growth and development performance: on the PDA medium, including slant seeds, petri dish flat seeds and triangular flask flat seeds, they can germinate into mycelia 24 hours after inoculation of conidia, form spores in 5-7 days, and cultivate in shake flasks for 24 hours , you can see that there are more mycelium on the wall of ...

Embodiment 2

[0084] First round of mutagenesis

[0085] Mortierella chrysogenum AS3.3410 was cultured in a petri dish for 7 days, and prepared into a spore suspension, counted with a microscope hemocytometer, and the concentration was 10 8 spores / ml. According to literature Huang Jianzhong et al., Microbiology Bulletin, 1998, 25 (4): 187-191; Yang Ge, etc., Fungal System, 1998, 17 (1): 86-90; (3): 140-143 methods, screening procedures such as Figure 5 . Combined mutagenesis with ultraviolet light plus LiCL, dilution and separation for each mutagenesis, and excellent single colonies were selected for continuous mutagenesis and screening. At different stages of screening, different indicators are selected to facilitate the selection of strains for subsequent research in a simple, fast and ready manner. The filtered indicators are as follows:

[0086] Bacterial growth performance: on the PDA medium, including slant seeds, petri dish flat seeds and triangular flask flat seeds, they can ge...

Embodiment 3

[0096] Shake flask fermentation of 5-6L culture medium

[0097] The selected ten mutant strains were then cultured in shake flasks, and 5 mutant strains were screened out, bacterial numbers 3-45 1 -3-1-1-2-3-8-1 strain, 3-45 1 -3-1-1-2-3-1-1 strain, 5-45 1 -7-2-1-2-5-1-1 strain,

[0098] 5-45 1 -7-2-1-2-5-1-2 and 5-45 1 -7-2-1-2-5-1-6 strains are abbreviated as 3-8-1 strains, 3-1-1 strains, 5-1-1 strains, 5-1-2 strains and 5-1 strains -6 plants. Then the five mutant strains were shaken and fermented in 5-6L culture solution. The culture medium is 2-5% starch, 1-3% soybean meal powder, 1-2% corn steep liquor, and 2-4% glucose. Yeast extract 0.3-1.0%, cottonseed oil 0.5-1.0% and inorganic salts, adjust pH 6.5-7.0. Cultivate for 72-96 hours at a temperature of 25-27°C and 180 r / min, and collect mycelium with nylon cloth. Air-dried at about 50°C to obtain mycelia. The experimental results are shown in Table 2.

[0099] Table 2 Shake flask fermentation growth and lipid p...

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Abstract

The invention discloses a mutant strain with high yield of Gamma-linolenic acid and a preparation method and application of the mutant strain. Mortierellaisabellina AS3.3410, the original strain, is subject to chemical and physical mutation technology, so that the Mortierellaisabellina 2W-15-2W2-22 mutant strain, with the CCTCC No. as M2012152, is obtained; the Mortierellaisabellina 2W15-2W2-22 mutant strain is subject to fermentation in a fermentation tank of 5000 L, wherein, in each liter of fermentation liquid, the biomass dry weight is 27.5-34.1 g, and the oil content is 55.9-60.3 %; after refinement, the main active ingredients in the refined oil are ALA, with the content of 20-37.3%, GLA, with the content of 8.5-10.3%, and ARA, with the content of 3.5-4.6%.

Description

technical field [0001] The invention relates to the fields of biotechnology, bioengineering and oil engineering, in particular to a mutant strain of Mortierella chrysogenum with high yield of GLA, and to a preparation method of the mutant strain. The mutant strain can produce GLA by fermentation technology. Background technique [0002] Today's world, including China, needs to solve the three major problems of population, food and environment, reduce the demand for food in food, and increase the intake of meat, fat and protein, especially foods with human physiological functions, which are more popular. In recent years, the fourth generation of functional food has been launched worldwide, that is, functional food with clear active ingredients and physiological functions. The polyunsaturated fatty acids (PUFAs) oils and β-carotene we studied are the fourth-generation functional foods in the world. PUFAs mainly include gamma-linolenic acid (GLA), alpha-linolenic acid (ALA), ...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P7/64C12R1/645
Inventor 何东平陈明锴胡传荣陈涛田华佘隽张四红姚理严玲华肖勇
Owner WUHAN POLYTECHNIC UNIVERSITY
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