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Method for promoting growth of haematococcus pluvialis and accumulation of astaxanthin

A technology of Haematococcus pluvialis and astaxanthin, which is applied in the agricultural field, can solve the limited effect of improving the carbon fixation rate of Haematococcus pluvialis, the influence of the physiological and metabolic activities of microalgae cells, and the low efficiency of light energy utilization and transformation. To improve the utilization and conversion efficiency of light energy, increase the content of astaxanthin, and improve the efficiency of light energy conversion

Active Publication Date: 2019-11-22
TIANJIN AGRICULTURE COLLEGE
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Problems solved by technology

[0005] Simply supplemented with CO 2 The improvement effect on the carbon fixation rate of Haematococcus pluvialis is limited, and the pH of the culture solution is gradually reduced, which is not conducive to the transformation of Haematococcus pluvialis from the green vegetative cell stage to the red sclerenchyma cell stage;
[0006] Ethanolamine promotes the accumulation of astaxanthin in Haematococcus pluvialis cells by increasing the supply of carbon sources and inhibiting the photosynthesis of microalgae. However, its biological safety is unknown, and it has certain risks as a regulator of microalgae product synthesis;
[0007] During the cultivation process, the light intensity is only set without distinguishing and managing the wavelength of light, and the utilization and conversion efficiency of light energy is not high;
[0008] Using nitrogen-deficient BBM medium to cultivate Haematococcus pluvialis at the stage of red sclerenchyma cells is beneficial to the accumulation of astaxanthin in microalgae cells, but the physiological and metabolic activities of microalgae cells will be affected to a certain extent

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  • Method for promoting growth of haematococcus pluvialis and accumulation of astaxanthin

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Experimental program
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Effect test

Embodiment 1

[0059] The cultivation process was carried out in a 3L large Erlenmeyer flask, and the volume of each bottle was 2L. Add NaHCO at a concentration of 0.02mol / L to the BBM medium 3 , a concentration of 0.6mmol / L ethyl 3-hydroxybutyrate. The culture temperature was set at 22±1°C. The light-to-dark ratio is set to 12hL:12hD, and the mixed light of "700nm red light + white light" is used, and the total light intensity is 50μmol / m 2 ·s, where red light intensity: white light intensity = 2:1. During the cultivation process, sterile mixed air (air volume fraction 98%, CO 2 volume fraction 2%), the ventilation rate is 0.5L / min. Haematococcus pluvialis was cultivated to the end of the logarithmic growth phase, and the cell density of microalgae reached 2.7×10 7 cells / mL.

[0060] Use NaNO 3 The low-nitrogen BBM medium with a concentration of 0.02mol / L diluted the algae liquid at the end of the logarithmic growth phase to 5×10 5 cells / mL. FeSO with a concentration of 0.2mol / L wa...

Embodiment 2

[0063] The cultivation process was carried out in a 3L large Erlenmeyer flask, and the volume of each bottle was 2L. Add NaHCO at a concentration of 0.03mol / L to the BBM medium 3 , a concentration of 0.8mmol / L ethyl 3-hydroxybutyrate. The culture temperature was set at 22±1°C. The light-to-dark ratio is set to 12hL:12hD, and the mixed light of "700nm red light + white light" is used, and the total light intensity is 55μmol / m 2 ·s, where red light intensity: white light intensity = 3:1. During the cultivation process, sterile mixed air (air volume fraction 98%, CO 2 volume fraction 2%), the ventilation rate is 0.5L / min. Haematococcus pluvialis was cultivated to the end of the logarithmic growth phase, and the cell density of microalgae reached 4.5×10 7 cells / mL.

[0064] Use NaNO 3 The low-nitrogen BBM medium with a concentration of 0.03mol / L diluted the algae liquid at the end of the logarithmic growth phase to 5×10 5 cells / mL. FeSO with a concentration of 0.3mol / L wa...

Embodiment 3

[0067] The cultivation process was carried out in a 3L large Erlenmeyer flask, and the volume of each bottle was 2L. Add NaHCO at a concentration of 0.04mol / L to the BBM medium 3 , a concentration of 1mmol / L ethyl 3-hydroxybutyrate. The culture temperature was set at 22±1°C. The light-to-dark ratio is set to 12hL:12hD, and the mixed light of "700nm red light + white light" is used, and the total light intensity is 60μmol / m 2 ·s, where red light intensity: white light intensity = 3:1. During the cultivation process, sterile mixed air (air volume fraction 98%, CO 2 volume fraction 2%), the ventilation rate is 0.5L / min. Haematococcus pluvialis was cultivated to the end of the logarithmic growth phase, and the cell density of microalgae reached 4.2×10 7 cells / mL.

[0068] Use NaNO 3 The low-nitrogen BBM medium with a concentration of 0.03mol / L diluted the algae liquid at the end of the logarithmic growth phase to 5×10 5 cells / mL. FeSO with a concentration of 0.3mol / L was ...

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Abstract

The invention relates to a method for promoting growth of haematococcus pluvialis and accumulation of astaxanthin. The method comprises the following steps: (1) haematococcus pluvialis is cultured ina culture medium at the culture temperature of 22+ / -1 DEG C, light illumination is carried out, in the cultivation process, sterile mixed air is continuously introduced, and haematococcus pluvialis iscultured to the end of logarithmic growth period; and (2) an algal solution at the end of logarithmic growth period is diluted with a low-nitrogen BBM culture medium with the NaNO3 concentration of 0.01-0.03 mol / L, and the culture temperature is 22+ / -1 DEG C; and light illumination is carried out, when the absorbance OD490 of the haematococcus pluvialis culture liquid at the wavelength of 490 nmdoes not change significantly, microalgae cells are collected and astaxanthin is extracted. The culture and transformation cycle of microalgae are shortened significantly, the efficiency of light energy utilization and transformation and the astaxanthin content of microalgae cells are increased significantly, the interference to cell adherency and physiological metabolism is reduced, and the addedmaterials are used optimally.

Description

technical field [0001] The invention belongs to the technical field of agriculture, in particular to a method for promoting the growth of Haematococcus pluvialis and accumulating astaxanthin. Background technique [0002] Astaxanthin is a fat-soluble carotenoid, which has the strongest antioxidant capacity found in natural organisms so far. Its ability to scavenge free radicals and singlet oxygen quenching is much higher than that of vitamin E. , and the antioxidant capacity is more than 10 times higher than common antioxidant substances such as zeaxanthin, lycopene and β-carotene. In addition, astaxanthin has a good coloring effect, can enter organisms and store in tissues, and make the muscles and skin of aquaculture animals bright in color. It was designated by the former Ministry of Agriculture as the only coloring agent for aquatic animals. Astaxanthin also has the effects of improving the body's immunity, delaying aging and eliminating inflammation, so it is widely us...

Claims

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Application Information

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IPC IPC(8): C12N1/12C12N1/38C12P23/00C12R1/89
CPCC12N1/12C12N1/38C12P23/00
Inventor 窦勇李嘉仪陈家宇吴琳任虹烨闫永芳周文礼邵蓬于士国高金伟贾旭颖
Owner TIANJIN AGRICULTURE COLLEGE
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