Method for accumulating grease mutant chlorella quickly by mutating pyrenoid freshwater chlorella by laser

A technology of pyrenoid and chlorella, which is applied in the biological field, can solve problems such as high cost, high equipment requirements, and complicated technical processes, and achieve the effects of light damage, strong safety, and low cost

Inactive Publication Date: 2012-11-21
SHANDONG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the above-mentioned patented technologies related to promoting the growth of chlorella and producing biodiesel are advanced, they do not involve the breeding of high-oil-yielding microalgae induced by laser irradiation, and require large-scale professional facilities such as photobioreactors and fermentation tanks. The requirements are relatively high, the technical process is relatively complicated, and the cost is high, which undoubtedly increases the production cost and the difficulty of technology promotion

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Step 1. Preparation of algae liquid: under the conditions of temperature 20°C, light intensity 1800lx, and light-dark ratio 12h / 12h, use fluorescent lamps as the light source to cultivate Chlorella pyrenoidosa cells to the logarithmic growth phase, so that the cell concentration reaches 10 8 per ml, to obtain the algae liquid that will be subjected to laser irradiation mutagenesis;

[0018] Step 2. Laser irradiation mutagenesis: take 20ml of logarithmic growth phase algae liquid and pour it into a small jar, and irradiate it with 808nm semiconductor laser for 0.9min. During the irradiation process, use magnetic stirring to keep the irradiation uniform, and obtain mutant algae strain;

[0019] Step 3. Cultivation of mutant algal strains: Rinse the laser irradiated cell liquid into a 50ml Erlenmeyer flask filled with BG11 nutrient salts, the final concentration of nutrient salts in the algae liquid is 1 / 4 times, at a temperature of 20°C and a light intensity of 1800lx 1....

Embodiment 2

[0022] Step 1. Preparation of algae liquid: under the conditions of temperature 23°C, light intensity 1900lx, and light-dark ratio 12h / 12h, use fluorescent lamps as the light source to cultivate Chlorella pyrenoidosa cells to the logarithmic growth phase, so that the cell concentration reaches 10 8 per ml, to obtain the algae liquid that will be subjected to laser irradiation mutagenesis;

[0023]Step 2. Laser irradiation mutagenesis: Take 20ml of logarithmic growth phase algae liquid and pour it into a small jar, and irradiate it with 808nm semiconductor laser for 1.0min. During the irradiation process, use magnetic stirring to keep the irradiation uniform, and obtain mutant algae strain;

[0024] Step 3. Cultivation of mutant algae strains: wash the laser irradiated cell fluid into a 50ml Erlenmeyer flask filled with BG11 nutrient salts, the final concentration of nutrient salts in the algae fluid is 1 / 4 times, at a temperature of 23°C and a light intensity of 1900lx 1. The...

Embodiment 3

[0027] Step 1. Preparation of algae liquid: under the conditions of temperature 25°C, light intensity 2000lx, and light-dark ratio 12h / 12h, use fluorescent lamps as the light source to cultivate Chlorella pyrenoidosa cells to the logarithmic growth phase, so that the cell concentration reaches 10 8 per ml, to obtain the algae liquid that will be subjected to laser irradiation mutagenesis;

[0028] Step 2, laser irradiation mutagenesis: take 20ml of logarithmic growth phase algae liquid and pour it into a small jar, and irradiate it with 808nm semiconductor laser for 1.1min to obtain mutant algal strains;

[0029] Step 3. Cultivation of mutant algae strains: irradiate the cell fluid with laser light into a 50ml Erlenmeyer flask filled with BG11 nutrient salts. The final concentration of nutrient salts in the algae fluid is 1 / 4 times. Cultivate for 20 days under the culture condition of 12h / 12h at a dark ratio; then transfer the above-mentioned algae liquid to a 300ml conical fl...

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Abstract

The invention provides a method for accumulating grease mutant chlorella quickly by mutating pyrenoid freshwater chlorella by laser, comprising the following steps: (1) culturing the pyrenoid freshwater chlorella cell to the logarithmic growth phase by using the fluorescent lamp as the light source; (2) taking and pouring 20ml of chlorella solution in the logarithmic growth phase into a small wide-mouth bottle, irradiating by using the semiconductor laser for 0.9-1.1 minutes, and stirring under magnetic force during irradiating to obtain a mutant chlorella; and (3) pouring the irradiated and mutated chlorella solution into a 50ml triangular flask filled with the BG11 nutrition salt with the final concentration of 1 / 4 times, culturing for 20 days at the temperature of 20-25 DEG C and the light intensity of 1,800-2,000lx, transferring the chlorella solution into a 300ml triangular flask, continuing to culture for 20 days, transferring the chlorella solution into a 1,000ml triangular flask, and continuing to culture for 26 days, wherein the BG11 nutrition salt with final concentration of 1 / 4 times is added every eight days, and the chlorella solution is shaken for 3 times every day in the culturing stage. The method is easy and practical, low in cost and high in efficiency.

Description

technical field [0001] The invention provides a method for laser mutagenesis of chlorella pyrenoidosa to rapidly accumulate lipid mutant strains, and belongs to the field of biotechnology. Background technique [0002] Chlorella (Chlorella) is a common aquatic single-celled algae, which is an important genus in the Chlorella family of green algae. It has a wide ecological distribution and is distributed in both seawater and freshwater. Currently, it includes about 10 species. Chlorella cells are rich in protein, essential amino acids, polysaccharides, lipids, chlorophyll, carotene and vitamins, etc. It is widely used in health food, aquaculture, cosmetics, medicine and other fields, and the domestic and foreign markets are in short supply. Compared with the United States, Japan, Israel and other countries, the level of industrialized large-scale cultivation of Chlorella in my country is relatively lagging behind. Chlorella pyrenoidosa is a spherical single-celled freshwater...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N13/00C12R1/89
Inventor 高政权孟春晓叶乃好付圣贵高宏正苏远丰王元元赵俞任
Owner SHANDONG UNIV OF TECH
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