Continuous gradient supplementation method and device applied to efficient expanding propagation of haematococcus pluvialis

Abstract

The invention discloses a continuous gradient supplementation method and device applied to efficient expanding propagation of haematococcus pluvialis. The continuous gradient supplementation method comprises the steps of A, compounding different gradient supplementation culture mediums; B, performing activation and species expanding on the haematococcus pluvialis: transferring sterilized culture mediums into a conical flask on a superclean working table, and performing inoculation with haematococcus pluvialis algae species; C, performing inoculation of the haematococcus pluvialis into a fermenter: detecting the quantity of cells of algae liquid after species expanding, performing thorough cleaning with fresh sterilized culture mediums, performing resuspending, and performing inoculation into the fermenter; D, monitoring culturing parameters / performing pH control: monitoring the algae liquid through a main control unit; and E, performing directional adjustment of C / N ratio of the culture mediums. The device consists of the fermenter, an air inlet / outlet, a sample inlet / out, a supplementation opening, a pH / temperature / dissolved oxygen control unit and a continuous gradient supplementation unit. Through the adoption of the method, infection with bacteria in the transforming and culture process can be avoided, and the utilization efficiency of substrates is improved; produced cellsare used for subsequent astaxanthin induction, and the survival rate is high; and the yield of biomass in the stage is as high as 0.30g L<-1>d<-1>, and the yield of the astaxanthin in the stage is ashigh as 15.45mg L<-1>d<-1>.

Description

technical field [0001] The invention relates to the field of biological technology, more specifically to a continuous gradient feeding method applied to the high-efficiency propagation of Haematococcus pluvialis, and also relates to a continuous gradient feeding device, which is especially suitable for Haematococcus pluvialls. The stage of efficient multiplication of provenance in large-scale cultivation. Background technique [0002] Haematococcus pluvialis is a freshwater unicellular green alga with double flagella, belonging to the Chlorophyta, Volvox, and Haematococceae. Under stress conditions, such as strong light, nitrogen deficiency and other conditions, cells can accumulate carotenoids, of which more than 85% are astaxanthin and astaxanthin esters. [0003] Astaxanthin is a kind of keto-type secondary carotenoid which is a source of vitamin A. Its chemical name is 3,3'-dihydroxy-4,4'-diketo-β,β'-carotene. It is worth noting that the astaxanthin molecule contains l...

AI Technical Summary

Problems solved by technology

In the first stage of cell culture, there are three culture models in the existing literature reports. One is the photoautotrophic model, which uses light energy for photosynthesis. The model that has successfully achieved large-scale production in the world is mainly based on photosynthesis. The two-stage model of autotrophy, the traditional two-stage photoautotrophy has problems such as difficult expansion, low yield, large land occupation, great influence from seasonal changes, and low efficiency.

In addition, there are few reports on heterotrophic fermentation of Haematococcus pluvialis, and the biomass of heterotrophic fermentation can reach 27g / L

However, as can be seen from Table 1, due to the cells in the heterotrophic fermentation process under long-term dark conditions, the photosynthetic activity of the obtained cells is very low, and its maximum photosynthetic efficiency F v / F m If it is lower than 0.6, the cells obtained by heterotrophic growth will be under stress conditions such as high light and nitrogen deficiency in the second stage. Low yield (Table 1-BP s and AP s )

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