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Mortierella alpina mutant strain capable of producing high-yield arachidonic acid, and fermentation method and application thereof

A technology of Mortierella alpina and mutant strains, which is applied in the field of microbial technology, can solve problems such as genotoxicity and chronic toxicity, and achieve the effects of easy extraction, good growth characteristics, and increased oxygen-dissolving area

Active Publication Date: 2014-01-01
WUHAN POLYTECHNIC UNIVERSITY +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] 3.2.1 Chronic toxicity
[0014] 3.2.2 Subchronic toxicity
[0018] 3.2.4 Genotoxicity
However, some researchers have reported that the carbon / nitrogen ratio is above 20:1 or even 50:1. This is due to the fact that the carbon and nitrogen sources used by the researchers are not the same and are caused by regional differences.

Method used

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  • Mortierella alpina mutant strain capable of producing high-yield arachidonic acid, and fermentation method and application thereof
  • Mortierella alpina mutant strain capable of producing high-yield arachidonic acid, and fermentation method and application thereof
  • Mortierella alpina mutant strain capable of producing high-yield arachidonic acid, and fermentation method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] 60 Coγ-irradiation mutagenesis

[0087] In this experiment, the ARA-producing strain Mortierella alpina200012201-12 (M12 for short, the same below) isolated and screened from Mortierella alpina was used as the starting strain. 60 A high-yield mutant strain of Mortierella alpina was obtained by Co-γ-ray irradiation mutagenesis breeding method. The specific method is as follows:

[0088] First, because the strain seldom produced conidia, the cells that grew well on the plate were selected for mutagenesis. The starting strain M12 was activated, that is, the M12 strain stored on the slant of the test tube in the refrigerator was transferred to the plate medium and cultured at 26°C for 5 days to activate. Then, according to the previous preprocessing experiments, it was found that when 60 When the irradiation dose of Co-γ ray was less than 1kGy, the mutagenic effect was not obvious, the characters of the original strain did not change basically, and the probability of po...

Embodiment 2

[0095] Mutation breeding

[0096] 60 After the Co-γ-ray mutagenesis is over, wrap the plate with black cloth and put it back into the sterile room, separate it in the sterile operating table, pick the irradiated mycelium with an inoculation loop and insert it on the primary screening plate medium, Each mutagenized strain was connected to 5 primary screening plates. And because the plate after mutagenesis is still likely to continue to grow and survive, we numbered it and put it in a constant temperature incubator together with the primary screening plate for cultivation. Taking M12-1-1 as an example, put it in the incubator for cultivation The plate numbers are M12-1-1, M12-1-1-1, M12-1-1-2, M12-1-1-3, M12-1-1-4 and M12-1-1-5 , and so on for the numbers of other primary screening plates. Since there are 24 plates taken for irradiation, there are 144 plates cultured in the incubator. After cultivating for a period of time, first calculate the lethality of strains under diff...

Embodiment 3

[0106] Scanning electron microscope observation of mutant strains

[0107] The Mortierella alpina mutant strains were made into 6 samples respectively cultured for 2-7 days, and at least 5 spray-coated slices were made for each sample, and observed by S-3000N and JSM-6390 scanning electron microscopes (SEM) and take photos.

[0108] The mycelium of Mortierella alpina mutant strain is tubular, colorless and transparent, with well-developed mycelia and dense growth, thicker than that of the wild-type strain (the diameter of the wild-type mycelium is 10-20 μm). After 7 days of cultivation, the mycelium diameter up to 30-40μm, and the hyphae are in the shape of lotus root joints ( figure 1 ). The asexual reproduction of wild-type Mortierella alpina relies on asexual spores, and its asexual spores are divided into conidia, stalks, etc. The adult spores of the conidia, stalks, etc. are separated and matured, even under an optical microscope. Many spherical and elliptical conidia ...

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Abstract

The invention discloses a Mortierella alpina mutant strain capable of producing high-yield arachidonic acid, and a fermentation method and application thereof. The naturally screened Mortierella alpina is mutagenized to obtain a Mortierella alpina mutant strain 200012201-12-2-2, CCTCC NO:M2013392 capable of producing high-yield arachidonic acid. After the strain is subjected to long-term transformation, the hereditary characters are stable and are not mutated, the biological characteristics are typical, and the growth traits are good in a microscopy and fermentation tank, and the strain has the advantages of high C and N utilization ratio, high biomass, high oil production level, high ARA yield and good quality, and can easily extract grease and required fatty acids; and by using the fermentation method, the yield of arachidonic acid is 15.1g / L fermentation liquid.

Description

technical field [0001] The present invention relates to the fields of microbial technology, microbial engineering and oil engineering, in particular to a mutant strain of Mortierella alpina with high arachidonic acid production, a method for preparing the mutant strain, and a method for preparing arachidonic acid from the mutant strain Acid application. technical background [0002] 1. Physiological activity and market of ARA [0003] Today's world needs to solve the problem of insufficient market caused by population growth. In addition to increasing the output of grain, oil, food, meat, vegetables, etc., it can also provide people with food by producing substances that are necessary for human beings and have physiological activity, thereby reducing human dependence on agricultural products. This is the fourth-generation function that has set off worldwide. Sexual food is a functional food with clear active ingredients and physiological functions. The oils and fats and o...

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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