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A method for regulating the cell cycle synchronization of Haematococcus pluvialis and its application

A Haematococcus pluvialis and cell cycle technology is applied in the field of regulation of Haematococcus pluvialis cell cycle synchronization, which can solve problems such as inability to meet research needs, inability to achieve synchronous culture, and the like, and achieves a simple, easy-to-implement and synchronized method. high degree of effect

Active Publication Date: 2019-05-03
云南中科雨虹生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, most of them only achieve partial synchronization in the germination stage of zoospores, especially cannot achieve complete synchronization culture, which cannot meet the needs of research.

Method used

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  • A method for regulating the cell cycle synchronization of Haematococcus pluvialis and its application
  • A method for regulating the cell cycle synchronization of Haematococcus pluvialis and its application
  • A method for regulating the cell cycle synchronization of Haematococcus pluvialis and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Haematococcus pluvialis NIES-144 algae strain from NIES algal species bank in Japan, in nitrogen-free BG11 medium and 200 μmol / m 2 / s under continuous light for 6-30 days; after that, inoculate the chlamydospores obtained at different time points into 3N-BBM containing 4.0mmol / L sodium nitrate, the initial cell density was 0.8g / L, and the 2 / s continuous light with 1.5% CO 2 After 48 hours, observe and measure the germination rate of Firmicutes spores. With the prolongation of culture time, the maturity of Firmicutes gradually increased, and the carotenoid / chlorophyll content ratio gradually increased, from 2.2 at the beginning to 12.5 on the 17th day. After 48 hours of germination, it first increased and then decreased, and the highest The germination rate reached 97% within 48 hours. When the culture medium was supplemented with ferrous sulfate with a final concentration of 450 μmol / L, the content ratio of carotenoid / chlorophyll in the obtained chrystrophic spores w...

Embodiment 2

[0056] Haematococcus pluvialis SAG 34-1b strain from the German SAG collection, in nitrogen-free BBM medium and 150 μmol / m 2 / s under continuous light for 8 days, the ratio of total carotenoids / total chlorophyll in the cells was 2.5. Centrifuge to discard the supernatant and harvest the spores, then resuspend them in BBM medium containing 2.0mmol / L ammonium carbonate and 2.0mmol / L urea, the initial cell density is 1.0g / L, and place them in the Erlenmeyer flask Cultivate, observe and measure the germination rate of Firmicutes after 48 hours, and the synchronous germination rates are 90% and 80% respectively. By 72 hours, the cells in the culture system rapidly cystized and formed chlamydospores.

Embodiment 3

[0058] Haematococcus pluvialis K-0084 strain from the Danish SCCAP strain bank, under nitrogen starvation and 200 μmol / m 2 The ratio of total carotenoids / total chlorophyll in the cells was 4.1 when cultured under the double stress of continuous light for 6 days. Centrifuge to discard the supernatant and harvest the chlamydospores, then resuspend in the BG11 medium containing 0-8.0mmol / L sodium nitrate at a cell density of 0.25g / L, 2 1.0% CO2 is fed under continuous light 2 After 48 hours, observe and measure the germination rate of Firmicutes spores. For the chrystrophic spores resuspended in the medium without sodium nitrate, the 48-hour germination rate was 0; for the chrystrophic spores resuspended in the medium containing 0.5-2.0mmol / L sodium nitrate, the 48-hour germination rate exceeded 90% %; resuspended in the culture medium containing 4.0, 8.0mmol / L sodium nitrate, the germination rate decreased instead in 48 hours, but due to the residual nitrogen in the medium, th...

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Abstract

The invention discloses a regulating method of cell cycle synchronization of haematococcus pluvialis. The method includes the following steps that mature haematococcus pluvialis chlamydospores synchronously germinate to produce red flagellated cells in a nitrogen limited culture medium and in the presence of strong light; the red flagellated cells undergo synchronous sporangium formation to form chlamydospores, and the process repeats periodically to realize that the haematococcus pluvialis is in the synchronous growth state in multiple cell cycles. The invention further provides an application. The application includes that the mature haematococcus pluvialis chlamydospores are put in the nitrogen limited culture medium, strong light and air rich in carbon dioxide are provided, and the chlamydospores form chlamydospores again via synchronous germination and synchronous sporangium formation; part of the formed chlamydospores are harvested directly while the other part is added into a new culture medium for continuous culture; periodically repeated culture is conduced according to the method. The method is simple, haematococcus pluvialis cell masses high in synchronization degree in multiple cell cycles can be obtained, and haematococcus pluvialis chlamydospores high in astaxanthin content can be obtained quickly.

Description

technical field [0001] The invention relates to the field of microalgae biotechnology, in particular to a method for regulating and controlling the synchronization of the cell cycle of Haematococcus pluvialis and its application. Background technique [0002] Astaxanthin is an oxidized carotene. Astaxanthin has strong free radical scavenging ability and coloring performance, so astaxanthin and its derivatives have been widely used in the fields of medicine, health care products, skin care products and high-end feed additives. [0003] Cultivating Haematococcus pluvialis to produce astaxanthin is the main way to produce natural astaxanthin, in which the content of astaxanthin is as high as 2% to 7% of the dry cell weight, and it is the most potential new resource for producing astaxanthin. It has attracted widespread attention from academic circles and business circles at home and abroad. Astaxanthin products derived from Haematococcus pluvialis have successively obtained G...

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 刘天中陈林王俊峰张维汪辉高莉丽周文俊张兰兰
Owner 云南中科雨虹生物科技有限公司
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