A method to avoid photoinhibition of microalgae to improve astaxanthin production

A technology of photoinhibition and astaxanthin, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of unfavorable astaxanthin production, achieve the effect of various methods and improve efficiency

Active Publication Date: 2022-07-01
云南保山泽元藻业健康科技有限公司
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Problems solved by technology

In order to overcome the defect that the astaxanthin production is unfavorable when the cell growth condition is switched from the cell growth condition to the astaxanthin accumulation condition in the two-stage culture process, the present invention designs a method for adjusting the cell state of Haematococcus pluvialis to increase the astaxanthin production. There are three ways: (1) directly induce the high-density cells obtained in culture, and then transfer to the normal second-stage induction; (2) dilute the high-density cells obtained in culture, perform low-light induction, and then transfer to to the normal second-stage induction; (3) transfer the cultured high-density cells to the normal second-stage induction, but by means of shading (including shading with transparent or opaque materials) and optimizing the time of inoculation (that is, in the Inoculate at a time point when the light intensity is weak within a day) to make the cells adapt to the stress environment, and then cancel the shading

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  • A method to avoid photoinhibition of microalgae to improve astaxanthin production
  • A method to avoid photoinhibition of microalgae to improve astaxanthin production
  • A method to avoid photoinhibition of microalgae to improve astaxanthin production

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Effect test

Embodiment 1

[0099] Example 1. Transition process changes the state of microalgal cells

[0100] like figure 1 As shown, in the first stage of culture, high-density Haematococcus pluvialis cells were obtained by the method of heterotrophic culture. After adding transition medium and water to 1L in a 1L bioreactor, steam sterilization, and then when the temperature drops to 25°C, add 12ml of high-density (26g / L) cells obtained by heterotrophic Haematococcus pluvialis, The transitional culture was started, the temperature was maintained at 28-38°C, the air flow was 1vvm, and the artificial light was illuminated for 24 hours, and the light intensity on each side was about 50klx.

[0101] During cultivation, 1% carbon dioxide was started intermittently, and the pH was kept constant at 8-9. Promoting the microalgae to adapt to the stressful environment. After 144h, the cells changed from thick-walled spores to green cells, and the transition process ended.

[0102] figure 2 The transition...

Embodiment 2

[0103] Example 2. Cultivation method via transition process increases dry cell weight and astaxanthin production

[0104] In this example, the microalgae cells are cultured in the first stage, and then undergo the transition process of the present invention, and then conduct cell induction, and compare with the direct cell induction without the transition process of the present invention.

[0105] image 3 Shown is a comparison of Haematococcus pluvialis cells cultured in a 1 L bioreactor induction culture for 10 days with and without the transition process. Depend on image 3 It can be seen that after 10 days of light-induced culture, the dry weight of the cells after the transition process reached 1.4 g / L, the astaxanthin increased to 4.6%, and the astaxanthin yield reached 64 mg / L. After 10 days of light-induced culture of Haematococcus pluvialis cells without the transition process, the dry weight of the cells after the transition process only reached 0.3g / L, the astaxan...

Embodiment 3

[0107] Example 3. Changes in the intracellular photosynthetic system of cells treated with the transition process

[0108] Figure 4 It shows that the difference between the cells treated by the transition process and the control in terms of the intracellular photosynthetic system is the difference in the electron transfer efficiency (ETR) of the intracellular photosynthetic system II (Figure A) and the non-photochemical quenching (NPQ) of each treated cell, respectively. Difference (Panel B), photosynthetic efficiency Y(II) (Panel C) and other non-photochemical energy (Y(NO), Panel D) of photosynthetic system II. The above results show that the efficiency of the intracellular photosynthetic system II is improved by the transition process, especially the photosynthetic system is remarkably restored by the increase of nitrogen element during the transition. The non-photochemically quenched fraction is relatively small, indicating that the cells are more tolerant of strong ligh...

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Abstract

The present invention relates to a new method for avoiding photoinhibition to improve astaxanthin production during microalgae cultivation. The cultivation of microalgae to produce astaxanthin usually adopts a two-stage method. The first stage realizes the mass reproduction of cells and the accumulation of biomass through heterotrophic, autotrophic or polytrophic methods, and the second stage adopts light stress supplemented by nutrient stress and other methods. Accumulation of intracellular astaxanthin is achieved. However, from the first stage to the second stage, the phenomenon of photoinhibition often occurs, which seriously affects the yield of astaxanthin. Therefore, the present invention regulates the cell state by controlling the culture conditions in the transformation process, such as: controlling the composition of the medium, shading, transition to low light, increasing the initial seeding density, gradually diluting, adjusting the outdoor seeding time, etc. Therefore, it fully overcomes the problem of photoinhibition of microalgae under strong light, which can greatly improve the efficiency of microalgae production of astaxanthin, and realize low-cost, high-efficiency and large-scale cultivation of microalgae to produce astaxanthin.

Description

technical field [0001] The invention belongs to the technical field of microalgae biology, and in particular relates to a method for adjusting the state of microalgae cells and improving the yield of astaxanthin. Background technique [0002] Astaxanthin, the chemical name is 3,3'-dihydroxy-4,4'-diketo-β,β'-carotene, the molecular formula is C40H52O4, the relative molecular mass is 596.86, also known as astaxanthin , astaxanthin, or lobster shell pigment, is a keto carotenoid. It is pink in color, fat-soluble, insoluble in water, and easily soluble in organic solvents such as chloroform, acetone, benzene and carbon disulfide. The chemical structure of astaxanthin is composed of 4 isoprene units linked by a conjugated double bond, and 2 isoprene units at both ends to form a six-membered ring structure. Since the chemical structure of astaxanthin contains a long conjugated unsaturated double bond system, it is easily damaged by light, heat, oxides, etc. The chemical structu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/12C12R1/89
CPCC12N1/12
Inventor 李元广万民熙章真黄建科樊飞王军
Owner 云南保山泽元藻业健康科技有限公司
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