A multi-factor co-evolutionary method to increase the production of Cryptidinosa dha

A co-evolutionary, Cryptodinomorphic technology, applied in the directions of microorganism-based methods, methods using microorganisms, biochemical equipment and methods, etc., can solve problems such as inhibiting the growth of Cryptodininos cells

Active Publication Date: 2021-09-28
KUNMING ZAONENG BIOTECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, the addition of sesamol will also hinder the syn...

Method used

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  • A multi-factor co-evolutionary method to increase the production of Cryptidinosa dha
  • A multi-factor co-evolutionary method to increase the production of Cryptidinosa dha
  • A multi-factor co-evolutionary method to increase the production of Cryptidinosa dha

Examples

Experimental program
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Embodiment 1

[0044] The first round of directional domestication using sethoxydim to provide selective pressure

[0045] 1. Prepare the C9N2 solid plate culture medium (5μM-20μM) with gradient concentration of sethenhexidine.

[0046] 2. Select 50 μL of fresh cells with an OD490nm of 0.8 and spread evenly on the resistant plate, and culture them upside down for about 2 weeks to observe the growth and determine the initial concentration for acclimation.

[0047] 3. Using C9N2 medium as the medium for continuous passage, wherein the initial concentration of sethenoxydim is 10 μM, insert 1 mL of fresh wild-type cells with an OD490nm of 0.8, and culture to the logarithmic phase.

[0048] 4. Transfer the bacterial strain obtained in the previous step into fresh C9N2 containing the same concentration of sethenoxydim, culture it to the mid-log phase and continue to subculture until the Cryptodinoflagellate cells are no longer sensitive to 10 μM sethenoxydim.

[0049] 5. The strains obtained in t...

Embodiment 2

[0054] This example is basically the same as Example 1, except that the initial strain used is the mutant strain obtained in Example 1, and sesamol is used to provide selection pressure for the second round of directional domestication.

[0055] After the above two examples, the obtained dual-acclimated bacterial strain is desensitized to sesamol. Therefore, during the fermentation process of the double domesticated strain, by adding 1 mM sesamol, the yield of DHA increased by about 130% compared with the wild type.

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Abstract

The invention discloses a method for obtaining a high-yielding docosahexaenoic acid-yielding Cryptodinium ATCC30556 mutant strain through multi-factor co-evolution. It is characterized in that it comprises the following steps: (1) determining the sensitive concentration of cryptodinininosa cells to sethenoxydim and sesamol; (2) using the sethenoxydim as selection pressure to domesticate cryptodinininosa to obtain a mutant strain with high oil yield; (3) The strain obtained in the above step is the starting strain, and the second domestication is carried out with sesamol as the selection pressure to obtain a mutant strain insensitive to sesamol; (4) adding sesamol to the insensitive strain promotes the biomass of Cryptodinium accumulation. By utilizing the method of the invention to carry out directional acclimation to the Cryptodinium cells, a bacterial strain insensitive to sesamol under different culture conditions can be obtained, and the production rate of DHA can be further increased by adding sesamol in vitro. With the mutant strain obtained by the present invention, adding 1mM sesamol in the cultivation process can greatly increase the yield of DHA by 130%.

Description

technical field [0001] The invention belongs to the field of industrial microbes, and in particular relates to a method for obtaining a preferred algal strain with high yield of docosahexaenoic acid (DHA) and its application. Background technique [0002] Docosahexaenoic Acid (DHA for short), is a colorless to pale yellow oily liquid, and is a polyunsaturated fatty acid necessary for the human body. DHA is a main component of the growth and maintenance of nervous system cells, and an important component of the brain and retina. Its content in the human cerebral cortex is as high as 20%, and it accounts for the largest proportion in the retina of the eye, accounting for about 50%. Intellectual and vision development of the fetus is very important. In addition, studies have pointed out that DHA has very good preventive and therapeutic effects on various cardiovascular and cerebrovascular diseases. [0003] So far, the main source of commercially available DHA is the extracti...

Claims

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Application Information

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IPC IPC(8): C12N1/36C12P7/64C12R1/89
CPCC12N1/36C12P7/6427
Inventor 张卫文陈磊刘璐刁进进
Owner KUNMING ZAONENG BIOTECH CO LTD
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