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

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. ...

AI Technical Summary

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 fragile, and the flagella are easily damaged and poisoned by ice crystals and refrigerants during cryopreservation. At the same time, the subsequent induction of astaxanthin requires stimulation by conditions such as high light, high salt, and nutrient deficiency. , the maintenance of cryopreserved cell activity and subsequent induction methods are key issues that need to be addressed

However, there are no relevant reports on the large-scale preservation of Haematococcus pluvialis algae mud and the subsequent astaxanthin induction method

However, the existing microalgae preservation methods have disadvantages such as high cost, low survival, limited to some algae species, and not suitable for large-scale production, so they cannot be applied to the production process of Haematococcus pluvialis