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Method for preparing squalene by utilizing heterotrophic biological membrane adherent autotrophic culture of grape algae

A technology of grape algae and biofilm, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the slow growth of algae cells, the research on the production of grape algae squalene has not received much attention, and the limitation of grape algae Problems such as the medical application of squalene to achieve the effects of promoting synthesis, protecting the marine ecological environment, and increasing production

Pending Publication Date: 2021-02-23
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the slow growth of the algae cells, the research on the production of grape algae squalene has not received much attention, which also greatly limits the medical application of grape algae squalene

Method used

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  • Method for preparing squalene by utilizing heterotrophic biological membrane adherent autotrophic culture of grape algae
  • Method for preparing squalene by utilizing heterotrophic biological membrane adherent autotrophic culture of grape algae
  • Method for preparing squalene by utilizing heterotrophic biological membrane adherent autotrophic culture of grape algae

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1: the comparison of botrytis liquid culture and biofilm adherent culture squalene production

[0021] Put Botrytis (B.braunii) in BG 11 medium, 1% CO 2 Concentration (volume ratio), light intensity 3000lux, liquid culture and adherent culture were carried out under the condition of temperature 25 ℃ respectively ( figure 1 a and b). BG 11 (g.L -1 ) The composition ratio of the medium is as follows: potassium nitrate 1.5, magnesium sulfate heptahydrate 0.075, dipotassium hydrogen phosphate trihydrate 0.04, anhydrous calcium chloride 0.027, sodium carbonate 0.02, citric acid 0.006, ferric ammonium citrate 0.006, ethylene di Sodium ferric amine tetraacetate 0.001, A5 solution 5mL, pH7.5.

[0022] The photobioreactor (PBR) used for the B.braunii liquid suspension culture experiment consists of a plate reactor (length 50cm, width 20cm, thickness 1cm, volume 1L) with an illumination area of ​​0.1m 2 ; Adhesive culture is that botrytis is separated from the medi...

Embodiment 2

[0024] Example 2: Effects of Different Carbon Sources and Their Concentrations on the Growth of Botrytis under Heterotrophic Conditions

[0025] Under the same light intensity (3000lux) and the same conditions as in Example 1, botrytis was respectively cultivated at an added concentration of 2.0g.L -1 In the medium with different organic carbon, the organic carbon sources were glycerol, glucose and acetic acid. Culture results such as image 3 as shown in a. The results show that Botrytis grows best in the medium where the organic carbon source is glucose, and the biological yield is 1.01g.L -1 .d -1 .

[0026] Under the condition that the organic carbon source is determined to be glucose, set different carbon source concentrations of 0.5, 1.0, 2.5, 5.0g.L -1 To cultivate. The results showed that botrytis in the concentration of glucose 2.5g.L -1 Under the best growth conditions, the biological yield is 1.21g.L -1 .d -1 .

Embodiment 3

[0027] Embodiment 3: adjust the N concentration to improve the yield of squalene produced by botrytis under autotrophic conditions

[0028] Botrytis was adherently cultured in BG 11 medium with different initial N concentrations under the same conditions as in Example 1 for 7 days. The initial N concentration was set to normal BG 11 (1.5g.L -1 ), 1 / 2N (0.75g.L -1 ), 1 / 4N (0.375g.L -1 ), 1 / 8N (0.1875g.L -1 ), 0N.

[0029] The result is as Figure 4 As shown, Botrytis has little difference under the conditions of normal N concentration of BG 11, 1 / 2N, 1 / 4N, and the biological yield is 5.24, 5.08, 4.86g.m -2 .d -1 , while the growth of algae cells was inhibited under the conditions of N concentration of 1 / 8N and 0N, and the biological output was 3.32 and 2.83g.m -2 .d -1 .

[0030] As for the content of squalene, the content of squalene under low N conditions was significantly increased, and the content of squalene under different N concentration conditions (normal BG11,...

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Abstract

The invention provides a method for preparing squalene by culturing grape algae. The method comprises the following steps of: optimizing a culture mode of the grape algae for producing squalene, so asto improve the yield of squalene in the grape algae culture process; and further improving the medical application of grape algae squalene, reducing environmental destruction caused by squalene application and development, and making up the defects in the prior art. The invention firstly provides a method for culturing grape algae, and the method comprises the following steps of: adding glucose into a BG11 culture medium to culture the grape algae, so as to propagate the algae. The invention further provides a method for improving the yield of squalene by the grape algae, and the method comprises the following steps of: culturing the grape algae in the BG11 culture medium, and reducing the nitrogen concentration in the BG11 culture medium. According to the method disclosed by the invention, synthesis of squalene is promoted under the condition that growth of the grape algae is not influenced, the yield of squalene with medical value is further improved, the application prospects of grape algae squalene products are improved, and the catching and killing amount of sharks is expected to be reduced, so that the marine ecological environment is protected.

Description

technical field [0001] The invention belongs to the technical field of botrytis propagating cultivation and high-value product production thereof, and in particular relates to a method for preparing squalene by cultivating botrytis. Background technique [0002] As a biosynthetic precursor of plant and animal steroids, squalene is an unsaponified colorless oily triterpenoid hydrocarbon with significant antioxidant and antibacterial activities, and is widely used in biomedicine and food supplements , high-quality cosmetics and other industries. In 2020, according to British media reports, some COVID-19 vaccine candidates contain squalene. In 2014 alone, the global squalene market demand reached 260,000 tons, and the market value was as high as 240 million US dollars. [0003] However, the current commercial source of squalene is mainly the liver oil of deep-sea sharks, but due to the protection of marine wildlife and fishery resources, the continuous supply and future avail...

Claims

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Application Information

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IPC IPC(8): C12N1/12C12P5/00C12R1/89
CPCC12N1/12C12P5/026
Inventor 程鹏飞周成旭严小军褚蕊蕊刘天中王海霞
Owner NINGBO UNIV
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