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Microplate-based screening method for Haematococcus pluvialis

A technology of Haematococcus pluvialis and a screening method, which is applied in the field of Haematococcus pluvialis screening based on microwell plates, can solve the problems of low screening efficiency, large workload of Haematococcus pluvialis, etc. Reliable culture results and reduced consumption

Active Publication Date: 2021-10-26
RUNKE BIOENG FUJIAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The object of the present invention is to provide a screening method for Haematococcus pluvialls based on micropore plates, which solves the problems of heavy workload and low screening efficiency in the breeding of Haematococcus pluvialls, and is simple, economical, efficient, and reliable in screening results. The advantages

Method used

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  • Microplate-based screening method for Haematococcus pluvialis
  • Microplate-based screening method for Haematococcus pluvialis
  • Microplate-based screening method for Haematococcus pluvialis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] 24 natural strains of Haematococcus pluvialis were screened.

[0051] The seed liquid culture medium comprises the raw material of following content:

[0052] Sodium nitrate 250mg / L; Potassium dihydrogen phosphate 200mg / L; Dipotassium hydrogen phosphate 125mg / L; Magnesium sulfate heptahydrate 125mg / L; Calcium chloride dihydrate 60mg / L; Disodium edetate 75mg / L; Ferric ammonium citrate 8mg / L and trace elements; trace elements include cobalt salts, copper salts, molybdenum salts, nickel salts, and iodine salts with a content of 0.05 μM; vanadium salts, tungsten salts, and chromium salts with a content of 0.01 μM; 0.1 μM zinc salt and 1.0 μM manganese salt; adjust pH=6.8. In addition, 0.5 g / L of sodium erythorbate, 0.6 g / L of cysteine ​​hydrochloride and 0.2 g / L of sodium ascorbate were added.

[0053] Fermentation medium comprises the raw material of following content:

[0054] Sodium acetate trihydrate 2.5g / L; Sodium nitrate 1.3g / L; Sodium glutamate 0.8g / L; Magnesium c...

Embodiment 2

[0066] Concentration Curve Fitting of Astaxanthin Determination by Microplate Method and GB / T 31520 Method

[0067] The algal strain used in this example is Haematococcus pluvialis UTEX 2505.

[0068] The seed liquid culture medium comprises the raw material of following content:

[0069] Sodium nitrate 200mg / L; Potassium dihydrogen phosphate 150mg / L; Dipotassium hydrogen phosphate 100mg / L; Magnesium sulfate heptahydrate 100mg / L; Calcium chloride dihydrate 40mg / L; Disodium edetate 50mg / L; Ferric ammonium citrate 6mg / L and trace element, trace element and pH value are identical with embodiment 1.

[0070] Fermentation medium comprises the raw material of following content:

[0071] Sodium acetate trihydrate 2.0g / L; Sodium nitrate 1.0g / L; Sodium glutamate 0.4g / L; Magnesium chloride hexahydrate 0.3g / L; Ferric ammonium citrate 6mg / L; Calcium chloride dihydrate 0.03g / L ; Sodium carbonate 0.1g / L and trace elements, trace elements and pH value are the same as the seed liquid cultu...

Embodiment 3

[0083] The algal strain used in this example is Haematococcus pluvialis UTEX 2505, and the fermentation medium, seed liquid medium and seed liquid culture process are the same as in Example 2. According to the 8% inoculum amount, the seed solution was inoculated in a 500ml Erlenmeyer flask equipped with 200ml of fermentation medium, and the algae were cultivated to the end of the logarithm under the condition that the temperature was 28°C and the light intensity was 1800lux, adding a final concentration of 4g / L of sodium acetate, 0.4g / L of ferrous sulfate heptahydrate and 7g / L of sodium chloride promote the accumulation of astaxanthin under the light conditions of 9000lux light intensity, and obtain the culture solution after 10 days. Take 2ml of the culture solution in a 12-well polypropylene microwell plate (the well volume is 6.9ml), centrifuge at 3000 rpm for 3 minutes and discard the supernatant. Add dimethyl sulfoxide (2ml) of 1 times the volume of the culture solution t...

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Abstract

The invention belongs to a screening method for microorganisms, in particular to a screening method for Haematococcus pluvialis based on a micropore plate. It includes process steps such as seed cultivation, fermentation cultivation, promotion of astaxanthin accumulation, astaxanthin extraction, and screening of Haematococcus pluvialis. The microporous plates in each step are made of light-transmitting materials. The present invention effectively solves the problems of difficult cultivation, cumbersome astaxanthin extraction and measurement steps, large result error and unreliable screening results in the high-throughput screening of Haematococcus pluvialis by the existing micro-orifice plate method. It has the advantages of improved culture efficiency, simple operation, effectively reducing or even eliminating the edge effect of orifice culture caused by poor gas exchange capacity when culturing Haematococcus pluvialis on micro-orifice plates.

Description

technical field [0001] The invention belongs to a screening method for microorganisms, in particular to a screening method for Haematococcus pluvialis based on a micropore plate. Background technique [0002] Astaxanthin is a red carotenoid with the chemical name 3,3'-dihydroxy-4,4'-diketo-β,β'-carotene, a biologically active ingredient with strong antioxidant function , studies have found that its antioxidant activity is 10 times higher than other carotenoids and 550 times higher than vitamin E. Astaxanthin can enhance the body's immunity, prevent cancer, prevent skin aging, maintain the health of the eyes and the central nervous system, and have the function of preventing and treating cardiovascular diseases. Astaxanthin also has a strong coloring function. At present, the main sources of astaxanthin are chemical synthesis and natural resources. Compared with chemically synthesized astaxanthin, natural astaxanthin is easier to be absorbed by the human body and has high sa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/12C12P23/00G01N21/31C12R1/89
CPCC12N1/12C12P23/00G01N21/31
Inventor 姜悦陈峰陈璇柳泽深
Owner RUNKE BIOENG FUJIAN
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