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Crypthecodinium cohnii mutant strain with high yield of DHA (docosahexaenoic acid), as well as fermentation method and application thereof

A technology of Crypthecodinium korii and mutant strains, which can be applied in the fields of biotechnology, bioengineering and oil and fat engineering, and can solve the problems of high cost and low yield, etc.

Active Publication Date: 2015-05-06
WUHAN POLYTECHNIC UNIVERSITY +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] In research practice, because Cryptidinium koii is often a mixture of macrocysts and microcysts, it is difficult to purify and separate them into a single body. The large and small cysts always grow in the same algal colony, but only the large cysts Only cysts have high fatty acid and DHA content, thus causing great technical difficulties in fermentation production, resulting in low yield and high cost

Method used

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  • Crypthecodinium cohnii mutant strain with high yield of DHA (docosahexaenoic acid), as well as fermentation method and application thereof
  • Crypthecodinium cohnii mutant strain with high yield of DHA (docosahexaenoic acid), as well as fermentation method and application thereof
  • Crypthecodinium cohnii mutant strain with high yield of DHA (docosahexaenoic acid), as well as fermentation method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Pre-test

[0077] From the Crypthecodinium cohnii (seligo) JavoruicKy preserved in our laboratory, the 6 0 The pre-experiment of Co-γ rays, through each tube containing 5ml cultured algae liquid, after 1.2KGy, 1.8KGy, 2.4KGy, 3.0KGy, 4.2KGy and 5.4KGy 60 Co-γ-ray radiation mutagenesis treatment. The treated algae liquid was diluted and separated and the number of live bacteria was counted to obtain the single algae strain and the lethal rate (Table 5)

[0078] Table 5 Cryptidnodinium kelpii No. 6 treated with different doses 60 Mortality rate of Co-γ-ray irradiation

[0079] Processing (KGY)

Embodiment 2

[0081] Strain activation

[0082] After ATCC30772 was activated by liquid medium and agar plate, the ATCC30772 strains with large algae colonies in pure culture, typical characteristics, and microscopic examination were selected. Put 5 individual plants into 500ml Erlenmeyer flasks with 100ml of culture medium, culture them at 16-200rpm, 24-28°C for 7 days, and select those with fast growth, large biomass and Sudan black B staining from 5 bottles that pass the microscopic examination. The one with the best depth was put into 12 sterile tubes, each containing 5ml, 3 tubes were used as the control (starting strain), and the other 9 tubes were divided into three groups, 3 tubes in each group, and sent to Hubei Academy of Agricultural Sciences for radiation. according to the center 60 Co-gamma radiation.

Embodiment 3

[0084] radiation mutagenesis

[0085] Three groups of prefabricated 9 algae liquid tubes were irradiated at doses of 0.8KGy, 1.6KGy, and 2.4KGy respectively, and then wrapped with black cloth for multiple layers to avoid exposure to light. That is, go to the laboratory for serial dilution. CK strain takes 10 -5 、10 -6 、10 -7 Dilution, 0.8KGy treatment group takes 10 -5 、10 -6 、10 -7 Dilution, take 10 for 1.6KGy treatment group -2 、10 -3 、10 -4 Dilution, 2.4KGy treatment group takes 10 -1 、10 -2 、10 -3 Dilution Each dilution is sprayed on four plates, and placed in a constant temperature and humidity box at 24-28°C and RH60-90% for 1-3 days.

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Abstract

The invention discloses a crypthecodinium cohnii mutant strain with high yield of DHA (docosahexaenoic acid), as well as a fermentation method and application thereof. The crypthecodinium cohnii mutant strain with the high yield of DHA, namely crypthecodinium cohnii 2.4K-2A2-52.4K-2A2-5 with CCTCCNO of M2012151 is screened out by 60Co-gamma ray radiation mutation of an ATCC30772 crypthecodinium cohnii strain. The grease quality and oil content of the strain are respectively improved by 51.66% and 66.37% in comparison with those of the original strain, the average DHA content achieves 39.04%, which is improved by 3.39% in comparison with that of the original strain, and the DHA yield which is calculated by g / L of a fermentation solution is improved by 56.92% in comparison with that of the original strain. The DHA produced by fermentation of the crypthecodinium cohnii ATCC30772 mutant strain has broad market prospects.

Description

technical field [0001] The present invention relates to biotechnology, bioengineering and oil engineering, in particular to a mutant strain of Crypthecodinium cohnii (seligo) JavoruicKy with high yield of DHA, a preparation method of the mutant strain, and the mutant strain Application in the preparation of DHA. Background technique [0002] Lipids are an essential part of the human body and have many important physiological functions. Fatty acids are the key components of lipids. According to the presence or absence of double bonds, they can be divided into saturated fatty acids and unsaturated fatty acids (polyunsaturated fatty acids, PUFAs); according to The position of the double bond away from the methyl end is further divided into ω-3, ω-6, ω-9 and other unsaturated fatty acids. In 1971, Danish scientist Dyeherg J. found in Greenland that people in Aisi had less cardiovascular disease, which may be due to the consumption of PUFAs containing omega-3 such as DHA. DHA i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/12C12P7/64C12R1/89
Inventor 何东平陈明锴胡传荣陈涛田华佘隽张四红姚理严玲华
Owner WUHAN POLYTECHNIC UNIVERSITY
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