Combined antibiotic application method for haematococcus pluvialis pure culture

A technology of Haematococcus pluvialis and an application method is applied in the field of combined antibiotic application of pure breeding Haematococcus pluvialis to achieve the effects of remarkable effect, high growth rate and biomass accumulation, and simple operation

Pending Publication Date: 2021-11-30
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The Haematococcus pluvialis species obtained by traditional methods are not pure species in the true sense, and the culture medium contains other phycobacteria. Or reduce the productivity of Haematococcus pluvialis has a potential role

Method used

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  • Combined antibiotic application method for haematococcus pluvialis pure culture

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] A kind of MCM culture medium preparation that is suitable for using combined antibiotics and cultivating pure species of Haematococcus pluvialis comprises the following steps:

[0027] (1) Prepare the mother liquor of solution A: weigh 50g of potassium nitrate and dissolve it in a small amount of solvent to completely dissolve it; weigh 2g of potassium dihydrogen phosphate and dissolve it in a small amount of solvent to completely dissolve it; weigh 10g of magnesium sulfate heptahydrate Dissolve it completely in the solvent; mix the dissolved potassium nitrate solution, potassium dihydrogen phosphate solution and magnesium sulfate heptahydrate solution together, and use the solvent to set the volume to 1 L to make A liquid mother liquor, so that the concentration of potassium nitrate is 50g / L, the concentration of potassium dihydrogen phosphate is 2g / L, and the concentration of magnesium sulfate heptahydrate is 10g / L;

[0028] (2) Preparation of solution B mother liquo...

Embodiment 2

[0036] Embodiment 2: the preparation and application method of the mother liquor of joint antibiotics of the present invention:

[0037] (1) Prepare 60g / L antibiotic mother solution: weigh 6g streptomycin and dissolve it in 100mL ultrapure water to dissolve it completely; weigh 6g kanamycin and dissolve it in 100mL ultrapure water to dissolve it completely; weigh 6g Dissolve gentamicin in 100mL ultrapure water to dissolve it completely; weigh 6g of ampicillin and dissolve it in 100mL ultrapure water to dissolve it completely; make streptomycin concentration 60g / L and kanamycin concentration 60g / L, the concentration of gentamicin is 60g / L, and the concentration of ampicillin is 60g / L; adopt suction filtration to sterilize;

[0038] (2) When in use, take 1mL streptomycin antibiotic stock solution, 1mL kanamycin antibiotic stock solution, 1mL gentamycin antibiotic stock solution, and 1mL ampicillin antibiotic stock solution, and dissolve them in 96mL MCM medium in order to make ...

Embodiment 3

[0039] Embodiment 3: the obtaining method of Haematococcus pluvialis pure species of the present invention:

[0040] (1) Preparation: six 100ml autoclaved clear glass conical flasks, add 95ml of the above-mentioned MCM medium into the transparent glass conical flasks, and add the above-mentioned 1mL streptomycin antibiotic mother solution, 1mL kanamycin antibiotic mother solution, 1mL gentamicin antibiotic stock solution, 1mL ampicillin antibiotic stock solution, mix them evenly;

[0041] (2) Inoculation: On the first day, each Erlenmeyer flask was inoculated with 1ml of Haematococcus pluvialis algae liquid, which contained Haematococcus pluvialis green vegetative growth stage algae;

[0042] (3) Cultivation: put 6 Erlenmeyer flasks on a shaker, the shaker speed is 180r / min; the shaker is placed in a light incubator for cultivation, using daylight as the light source, the light-dark cycle is 14:10, and the temperature is 20°C. Light intensity 60μmol·m -2 ·s -1 ;

[0043] (...

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Abstract

The invention relates to a combined antibiotic application method for haematococcus pluvialis pure culture. A haematococcus pluvialis culture medium MCM culture medium used in an experiment is as follows: a formula of a solution A comprises 500mg/L of potassium nitrate, 20mg/L of dipotassium phosphate and 100mg/L of magnesium sulfate heptahydrate; a formula of a solution B comprises 80 mg/L of calcium chloride hexahydrate; a formula of a solution C comprises 372 [mu]g/L of disodium ethylene diamine tetraacetate and 417 [mu]g/L of ferric sulfate heptahydrate; a formula of a solution D comprises 7.189 [mu]g/L of zinc sulfate, 1.237 [mu]g/L of phosphoric acid, 0.476 [mu]g/L of cobalt chloride dehydrate, 6.242 [mu]g/L of copper sulfate pentahydrate, 8.451 [mu]g/L of manganese sulfate monohydrate and 0.726 [mu]g/L of sodium molybdate dehydrate; formula solvents are tap water, and after high-pressure sterilization, the solution A, the solution B, the solution C and the solution D dissolve in the solvents in sequence; and four antibiotics, namely streptomycin, kanamycin, gentamicin and ampicillin are combined for use, the pH value is adjusted to 6.99-7.03 and the concentration of the antibiotics are 600mu g/ml, culturing is performed on a shaker in a light box for 3 days, and purifying and inspecting are performed by combining with a solid culture medium to obtain the pure haematococcus pluvialis.

Description

technical field [0001] The invention relates to a method for obtaining pure microalgae, in particular to a combined antibiotic application method suitable for the pure culture of Haematococcus pluvialis, and in particular to a method suitable for the vegetative growth of green cells of Haematococcus pluvialis The method of obtaining and cultivating purebreds at different stages. Background technique [0002] Haematococcus pluvialis ( Haematococcus pluvialis ) is a single-celled algae suitable for growth in fresh water, belonging to Chlorophyta, Chlorophyceae, Volvox, Haematococcus, and Haematococcus. When the environment of Haematococcus pluvialis is unfavorable, it produces thick-walled spores and accumulates a large amount of astaxanthin. Haematococcus pluvialis contains a large amount of astaxanthin (up to 7% of dry cell weight), which is an important molecular biology test object and raw material for the production of natural astaxanthin, and is recognized as the safes...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/38C12N1/12C12R1/89
CPCC12N1/38C12N1/12
Inventor 王巧晗凌滨峰李芸张静陈希凤刘岩李景玉宫庆礼
Owner OCEAN UNIV OF CHINA
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