On-scale production method for haematococcus pluvialis green cell preservation and astaxanthin induction

A technology of Haematococcus pluvialis and Haematococcus pluvialis algae liquid, applied in the biological field, can solve the problems of unsuitable for large-scale production, easy to cause pollution, low survival rate, etc., to improve production efficiency and increase the production of astaxanthin Yield, reduced equipment requirements and operating costs, effects of avoiding bleach deaths

Active Publication Date: 2017-05-31
BIOALGO CY CO LTD +1
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Problems solved by technology

The article "Research on Thalassiosira flocculation and post-coagulation preservation technology" reported that 10% glycerol was used to preserve chitosan flocculated Thalassiosira at -20°C. After preservation, the specific growth rate decreased slightly, but due to the viscosity of glycerol , not easy to remove, easy to cause pollution in subsequent cultivation
Patent 201010222005.0 discloses a method for preserving Chlorella, Botrytis, Chlorella, Nannochloropsis, and diatom algae species, using a process of freezing and vacuum drying after pre-freezing at a low temperature of -60°C, but the production cost of this method is high , only suitable for a small amount of strain preservation, not suitable for large-scale application
[0005] The green expanded cells of Haematococcus pluvialis are

Method used

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Embodiment 1

[0020] (1) Preparation of algae mud: the algae liquid of Haematococcus pluvialis in the logarithmic growth phase was collected in a 10 m 3 Stainless steel mixing tank, liquid volume is 7 m 3 . The stirred tank was left to settle at 4°C for 6-12 hours, and part of the supernatant medium was removed to obtain a concentrated algae liquid with a cell density of 10-20 g / L. Add DMSO with a concentration of 1-20% (v / v) pre-cooled at 4°C to the concentrated algae liquid at a volume ratio of 0.5-10:1, stir and mix evenly, the speed is 10-50 rpm, and the stirring time is 5 ~30 min. Then add skimmed milk powder with a mass ratio of 0.5%-20% (w / v), stir and mix well, then centrifuge at 1000-5000 rpm to remove the supernatant, and obtain algae mud for frozen preservation with a moisture content of 70-90% (w / w).

[0021] (2) Freezing preservation: quickly place the prepared algae mud at 4°C for 4-12 hours, and then transfer to -20°C for 90 days.

[0022] (3) Cell activation: Melt the f...

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Abstract

The invention relates to a method applicable to on-scale production for haematococcus pluvialis green cell preservation and later astaxanthin induction, and belongs to the field of biotechnology. The method provided by the invention comprises the following steps: firstly, performing concentration, mixing and centrifugation to prepare haematococcus pluvialis green cell alga slurry with a freeze-preservation agent and skimmed milk, performing pre-cooling at 4 DEG C, and performing freeze-preservation at -20 DEG C; performing water-bath unfreezing on preserved alga cells, performing freeze-preservation agent elution, performing low-nitrogen weak-light culture to activate alga cells, and performing nutrition deficiency and mixed carbon source addition, thereby achieving rapid induction of green cells and accumulation of astaxanthin. The method is simple to operate and applicable to large-scale production application, the dilemma that production of a production enterprise is halted in winter can be effectively solved, astaxanthin can be induced in spring from green alga cells produced in winter, the production efficiency of equipment and operators can be improved, and the yield of the astaxanthin can be increased.

Description

technical field [0001] The invention relates to the field of biotechnology, and in particular provides a method for preserving Haematococcus pluvialis green cells suitable for large-scale production and subsequent astaxanthin induction. Background technique [0002] In the large-scale cultivation of Haematococcus pluvialis, a two-step method is usually used for cultivation: the first step is green expansion cultivation, and Haematococcus pluvialis exists in the form of green vegetative cells under the condition of suitable environment and rich nutrition. During this process, algae cells divide and multiply, producing a large number of flagellar swimming cells. At this time, the cells are fragile and sensitive to environmental factors such as light and temperature. Therefore, green expansion is usually carried out in a room with well-controlled temperature and light. The second step is astaxanthin induction. The green cells are placed in unfavorable environmental conditions. ...

Claims

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Application Information

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IPC IPC(8): C12P23/00C12N1/04C12R1/89
CPCC12N1/04C12P23/00
Inventor 郑玉彬殷淑超郑玉瑞代云法王华陈树林
Owner BIOALGO CY CO LTD
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