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Method for quickly separating purified haematococcus pluvialis species

A technology for the separation and purification of Haematococcus pluvialis, which is applied in the field of purification of microalgae species. It can solve the problems of difficulty in cultivating a single algae cell, affecting algae growth, and complicated operation, so as to reduce the maintenance cost of algae species and enhance the resistance , the effect of cell density and volume increase

Active Publication Date: 2018-09-28
NINGBO FUTIAN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The currently reported methods for purifying algae species mainly include picking algae cells under a microscope and adding antibiotics to the algae liquid to eliminate bacteria. The former is complicated to operate and has high technical requirements for operators, requiring certain experience in algae classification. Large and difficult to survive; the latter directly adds antibiotics to the algae liquid, although it can effectively inhibit the bacteria in the algae liquid, but the inability to completely eliminate bacteria will also affect the growth of algae and even induce mutations

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Natural water samples. Sampling in the field, Haematococcus pluvialis cells are the dominant population in a certain water area, and 500mL is sampled from this water area.

[0036] (1) Return to the laboratory, transfer the water sample into sterilized sterile water, and add the following nutrients: magnesium sulfate 0.02g / L, calcium chloride 0.01g / L, sodium citrate 0.1g / L , VB1 10μg / L, VB 12 0.1μg / L, ferric chloride 0.588mg / L, manganese chloride 0.108mg / L, zinc sulfate 0.078mg / L, cobalt chloride 0.012mg / L, sodium molybdate 0.0075mg / L; with a light intensity of 5000lx, 24h / d light, with CO 2 1% mixed gas (0.2vvm) for cultivation, the cultivation temperature is 23±2°C, and the cultivation time is 24h; after the cultivation is completed, let it stand for 1h, carefully remove the supernatant, and collect about 10mL of the algae liquid at the bottom of the cultivation container.

[0037] (2) Transfer the collected algae solution into a sterilized 15‰ sodium chloride so...

Embodiment 2

[0045] Natural water samples. Sampling in the field, there are Haematococcus pluvialis cells in a certain water area, and Haematococcus pluvialis cells, some green algae and diatoms are the dominant algal strains, and 500mL of samples are taken from this water area.

[0046] (1) Return to the laboratory, transfer the water sample into sterilized sterile water, and add the following nutritional ingredients: magnesium sulfate 0.02g / L, calcium chloride 0.01g / L, sodium citrate 0.1g / L , VB 1 10μg / L, VB 12 0.1μg / L, ferric chloride 0.588mg / L, manganese chloride 0.108mg / L, zinc sulfate 0.078mg / L, cobalt chloride 0.012mg / L, sodium molybdate 0.0075mg / L, using a light intensity of 5000lx, 24h / d light, with CO 2 1% mixed gas (0.2vvm) was used for cultivation, the cultivation temperature was 23±2°C, and the cultivation time was 48h; after the cultivation was completed, let stand for 30min, carefully remove the supernatant, and collect about 10mL of the algae liquid at the bottom of th...

Embodiment 3

[0055] Natural water samples. When sampling in the field, it was found that there were Haematococcus pluvialis cells in a certain water area, but the dominant algae species were other green algae and diatoms. A 500 mL sample was taken from the water.

[0056] (1) Return to the laboratory, transfer the water sample into sterilized sterile water, and add the following nutritional ingredients: magnesium sulfate 0.02g / L, calcium chloride 0.01g / L, sodium citrate 0.1g / L , VB 1 10μg / L, VB 12 0.1μg / L, ferric chloride 0.588mg / L, manganese chloride 0.108mg / L, zinc sulfate 0.078mg / L, cobalt chloride 0.012mg / L, sodium molybdate 0.0075mg / L; with a light intensity of 5000lx, 24h / d light, with CO 2 1% mixed gas (0.2vvm) for cultivation, the cultivation temperature is 23±2°C, and the cultivation time is 72h; after the cultivation is completed, let it stand for 15min, carefully remove the supernatant, and collect about 10mL of the algae liquid at the bottom of the cultivation container. ...

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Abstract

The invention discloses a method for quickly separating purified haematococcus pluvialis species. The method comprises the following steps that a water sample with haematococcus pluvialis cells or single-species haematococcus pluvialis liquid with infectious microbes is selected and added into an inorganic medium suitable for haematococcus pluvialis growing, and after induced culture treatment andstanding, cells at the bottom of a culture container are collected; the collected cells are re-dissolved in a prepared sterile sodium chloride solution, after standing, supernate is removed, and cells at the bottom are collected; the cells at the bottom are re-dissolved in a medium containing antibiotics for treatment of a certain period, and standing is carried out for sedimentation; after the supernate is removed, the cells at the bottom are re-dissolved in the sterile medium and are diluted to a certain concentration; the diluted liquid is positioned in a cultivation hole plate for cultivation; and after microscopy examination and bacteria examination, purified haematococcus pluvialis species are selected. According to the method for the quickly separating the purified haematococcus pluvialis species, the haematococcus pluvialis species with high purity can be obtained within a short time, the growth cycle is shortened, and succession probability of mass growth is improved.

Description

technical field [0001] The invention relates to the technical field of purification of microalgae species, in particular to a method for separating and purifying Haematococcus pluvialis species. Background technique [0002] Haematococcus pluvialis belongs to Chlorophyta, Chlorophyceae, Volvox, and Haematococcus, and is a freshwater diflagellated unicellular green algae. Four typical cell forms appear throughout the life cycle: small worm bodies, large worm bodies with flagella, gelatinous sheath bodies without motility, and red large cells with hard cell walls——red cysts (Haematocysts). In a clean environment with sufficient nutrients, the large worm body dominates; once the environment deteriorates, it will transform into a gelatinous body, and then into a red cyst. After the formation of red cysts, the cell density increases sharply (our study found that the density can be as high as 1.04-1.08g / cm 3 , while the rest of the cell density is usually not higher than 1.02g / c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/12C12N1/02C12R1/89
CPCC12N1/02C12N1/12
Inventor 王兆伟宋佳巍王闪
Owner NINGBO FUTIAN BIOTECH
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