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A kind of preservation method of Haematococcus pluvialis vegetative cells

A technology for the preservation of Haematococcus pluvialis, which is applied in the field of preservation of Haematococcus pluvialis vegetative cells, can solve the problems of easy aging, slow growth, and low conversion efficiency, and reduce labor costs and facility investment , the effect of not easy aging

Active Publication Date: 2015-11-04
LIJIANG CHENGHAI BAOER BIOLOGICAL DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, whether it is a one-step method or a two-step method, it requires the accumulation and preservation of a large number of Haematococcus pluvialis vegetative cells
The preservation and growth of vegetative cells are a pair of contradictions: rapid growth, easy to enter the plateau stage early, and the storage time is short; slow growth, the storage time is relatively long, which will affect the accumulation of vegetative cells
The preservation and aging of vegetative cells are also a pair of contradictory conditions: long storage time, prone to aging, affecting continued cultivation and transformation
Insufficient accumulation of vegetative cells in Haematococcus pluvialis will lead to low transformation efficiency

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Preparation of culture medium: 120mg / L NaNO 3 , 15mg / L of KH 2 PO 4 , 2.5mg / L of FeSO 4 ·7H 2 O, 0.25mg / L MnSO 4 , 2.5mg / L of EDTA-Na 2 .

[0016] The inoculated Haematococcus pluvialis has a vegetative cell density of 30,000-50,000 / ml. At 20° C., irradiated with natural light, the light intensity for cultivation is 2000 lx. After 5 days of cultivation, the density is 90,000-150,000 / ml, and it enters the preservation state.

[0017] The preparation of the preservation solution, the content of the main component in the preservation solution is 1000mg / L NaNO 3 , and 50mg / L of KH 2 PO 4 , 2.5mg / L FeSO 4 ·7H 2 O, 0.25mg / L MnSO 4 , 5.0mg / L of EDTA-Na 2 . Preservation solution can directly add NaNO in the culture solution 3 and KH 2 PO 4 made.

[0018] Control of storage conditions: add a red film with a transmitted light wavelength of 660-740nm between the irradiated light source and the culture medium; and the surface illumination condition of the storage s...

Embodiment 2

[0022] Preparation of culture medium: 130mg / L NaNO 3 , KH of 18mg / L 2 PO 4 , 2.5mg / L of FeSO 4 ·7H 2 O, 0.25mg / L MnSO 4 , 2.5mg / L of EDTA-Na 2 .

[0023] Haematococcus pluvialls vegetative cells inoculated in the prepared culture solution, so that the density of Haematococcus pluvialls vegetative cells in the culture solution after inoculation is 32,000 / ml, at 23 ° C, natural light irradiation, and the cultivation light intensity is 2000lx After 6 days of culture, the density was 135,000 cells / ml. At this time, NaNO was added to the culture medium 3 and KH 2 PO 4 As a preservation solution, make NaNO in the preservation solution 3 The content is 1200mg / L, KH 2 PO 4 It was 40 mg / L, and ampicillin was added to make the final concentration of ampicillin in the preservation solution 0.18 million u / L. Cover the culture medium with a red film so that the wavelength of the passing light is 660-740nm; and the surface illumination condition of the preservation medium is 13...

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Abstract

The invention provides a preservation method of haematococcus pluvialis nutritive cells. The preservation method comprises the following steps: placing haematococcus pluvialis in a culture solution to perform multiplication culture, wherein the concentration ratio of nitrogen to phosphorus in the culture solution is (20-30):1, and the culture temperature is 12-20 DEG C; allowing light emitted by a light source to pass through a red film with the thickness of 0.05-0.12 mm, wherein the illumination intensity is 500-1500 lx. Compared with the prior art, the preservation method has the advantages as follows: the preservation time is long and the possibility of ageing is low; the nutritive cells in the culture solution can keep nutritive growth; the time for culturing the nutritive cells to the maximum density in the plateau is 20-35 days; after the plateau, the nutritive cells can further be preserved for 20-35 days. The next step of nutritive cultivation or transformation is conducted, and the nutritive cells are continuously kept in a fast-growth state. The method is simple, convenient and feasible, can reduce facility investment for breed conservation, and lowers the labor cost.

Description

technical field [0001] The invention belongs to the technical field of algal cell culture and preservation, and in particular relates to a method for preserving vegetative cells of Haematococcus pluvialis. Background technique [0002] Natural astaxanthin is a kind of carotenoid, which has the strongest antioxidant capacity found in nature so far; it has the functions of protecting skin, improving immunity, inhibiting cancer, relieving fatigue, delaying aging, prevention and treatment The function of cardiovascular and cerebrovascular diseases is mainly used as a natural colorant for fish, and it is also widely used in aquatic products, cosmetics, medicine and other industries. Haematococcus pluvialis is a single-cell green algae that produces natural astaxanthin. High stability and safety; therefore, cultivating Haematococcus pluvialis has become the best way to obtain natural astaxanthin in the future, which has been vigorously promoted by various countries. [0003] Cur...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/04
Inventor 骆其君
Owner LIJIANG CHENGHAI BAOER BIOLOGICAL DEV
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