CN111607523A - High-yield industrial microalgae culture method - Google Patents
High-yield industrial microalgae culture method Download PDFInfo
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- CN111607523A CN111607523A CN202010629338.9A CN202010629338A CN111607523A CN 111607523 A CN111607523 A CN 111607523A CN 202010629338 A CN202010629338 A CN 202010629338A CN 111607523 A CN111607523 A CN 111607523A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/12—Unicellular algae; Culture media therefor
Abstract
The invention discloses a high-yield industrial microalgae culture method, which comprises the following steps: preparing facilities and equipment required by the industrial microalgae expansion culture; culturing the algae in a 250ml triangular flask until the algae grows to a logarithmic phase to obtain a seed solution; the first-stage culture was carried out in a 250 ml-4 l vessel, and air supplemented with carbon dioxide was supplied; taking one part of the culture medium for continuous primary culture, using the rest part of the culture medium for inoculation, and starting secondary culture, wherein the volume of a container is 20 liters to 120 liters; the third stage culture adopts a pipeline type photoreactor, a columnar photoreactor or a microalgae runway pool reactor, and the minimum volume of the container is 200 liters. The invention has the beneficial effects that: according to the invention, the three-stage culture method is adopted, and the nutrient solution is prepared according to the nutrient balance theory to culture the microalgae, so that the growth speed and the quantity of the microalgae can be greatly improved, the effect is obvious, the cost for cultivating the algae in a factory is greatly reduced, and the method is economical, practical and suitable for popularization and use.
Description
Technical Field
The invention relates to the technical field of microalgae culture, in particular to a high-yield industrialized microalgae culture method.
Background
Microalgae are rich in proteins, lipids, polysaccharides, vitamins, antioxidants and other functional nutrients, and therefore have wide applications in the fields of medical raw materials, health foods and bioenergy. At present, microalgae culture technology is mostly concentrated in photoautotrophic systems, and the culture systems have the defects of low production efficiency, large floor area, high harvesting cost and the like, thereby seriously restricting the development of microalgae industry. The improvement of microalgae biomass by an artificial large-scale culture technology is the key of microalgae resource development and utilization. In recent years, a heterotrophic high-cell-density culture technology for microalgae has been gradually developed, which can overcome the dependence of a light culture system on light, has the advantages of higher growth speed, capability of realizing pure culture, high cell yield, capability of greatly reducing downstream post-treatment cost, convenience for automatic control and the like, and has become a hotspot of microalgae culture research in recent years. There is a need in the art for a high-yielding industrial microalgae cultivation method.
Disclosure of Invention
The technical problem to be solved by the invention is to overcome the technical defects and provide a high-yield industrial microalgae culture method.
In order to achieve the purpose of the invention, the invention adopts the technical scheme that: a high-yield industrial microalgae culture method comprises the following steps:
(1) the facility and equipment required by the industrialized microalgae propagation are prepared, and the facility and the equipment comprise a greenhouse glass room, wherein the greenhouse glass room is internally provided with: automatic or semi-automatic temperature increasing and controlling equipment, air humidity controlling equipment and CO2The device comprises a generating and discharging device, a light supplementing device, a water distribution treatment system and a high-level water pool;
(2) culturing algae in a 250ml triangular flask with illumination of 5000lux at 22-26 deg.C, shaking the triangular flask 2-3 times per day, and obtaining seed solution when the algae grows to logarithmic phase;
(3) the first-stage culture was carried out in a 250 ml-4 l vessel, inoculated at a volume ratio of 10%, and supplied with air supplemented with carbon dioxide;
(4) taking one part of the culture medium to be continuously used for primary culture, using the other part of the culture medium to be inoculated, and starting secondary culture, wherein the secondary culture is carried out in a high and narrow upright cylindrical container, the volume of the container is 20 liters to 120 liters, and the algae liquid of the secondary culture is used as larva bait or is used for inoculation and starting tertiary culture;
(5) the third stage culture adopts a pipeline type photoreactor, a columnar photoreactor or a microalgae runway pool reactor, and the minimum volume of the container is 200 liters.
Furthermore, the greenhouse glass room is a 300-square-meter workshop, the floor space is 20 meters by 15 meters, and a double-roof connection type greenhouse is adopted.
Further, the primary culture is carried out at 20. + -. 10 ℃ under 2000. + -. 500lux for 7-14 days.
Further, the second-stage culture is carried out for 3-4 days at 30 +/-2 ℃ and 2000 +/-500 lux.
Further, the third-stage culture is carried out at 30 + -2 deg.C under 2000 + -500 lux for 3-4 days.
Further, the culture solution for algae seed, primary culture, secondary culture and tertiary culture comprises the following substances in parts by mass: 10-50 parts of water; 10-20 parts of agricultural compound amino acid powder; 10-50 parts of humate; 5-15 parts of monopotassium phosphate; 5-10 parts of urea; 1-5 parts of ferrous sulfate; 1-5 parts of calcium superphosphate; 1-5 parts of borax; 0.1-0.5 part of trace elements; 0.1-0.5 part of gibberellin.
The invention has the beneficial effects that: according to the invention, the three-stage culture method is adopted, and the nutrient solution is prepared according to the nutrient balance theory to culture the microalgae, so that the growth speed and the quantity of the microalgae can be greatly improved, the effect is obvious, the cost for cultivating the algae in a factory is greatly reduced, and the method is economical, practical and suitable for popularization and use.
Detailed Description
The technical solutions of the present invention will be described clearly and completely with reference to the following embodiments of the present invention, and it should be understood that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example one
A high-yield industrial microalgae culture method comprises the following steps:
(1) preparing facilities and equipment required by the propagation of industrial microalgae, wherein the facilities and the equipment comprise a greenhouse glass room, the greenhouse glass room is a 300-square-meter workshop, the occupied area is 20 meters by 15 meters, a double-roof connection type greenhouse is adopted, and equipment is arranged in the greenhouse glass room: automatic or semi-automatic temperature increasing and controlling equipment, air humidity controlling equipment and CO2The device comprises a generating and discharging device, a light supplementing device, a water distribution treatment system and a high-level water pool;
(2) culturing algae in a 250ml triangular flask with illumination of 5000lux at 22 deg.C, shaking the triangular flask 2 times per day, and growing to logarithmic phase to obtain seed solution;
(3) the primary culture was carried out in a 250ml vessel, inoculated at a volume ratio of 10%, supplied with air supplemented with carbon dioxide, and cultured at 20 ℃ for 7 days at 1500 lux;
(4) taking one part of the culture medium to be continuously used for primary culture, using the other part of the culture medium to inoculate, and starting secondary culture, wherein the secondary culture is carried out in a high and narrow upright cylindrical container with the volume of 20 liters, the secondary culture is carried out for 3 days under the conditions of 28 ℃ and 1500lux, and the algae liquid of the secondary culture is used as larval bait or is used for inoculating and starting tertiary culture;
(5) the third stage culture adopts a pipeline type photoreactor, a columnar photoreactor or a microalgae runway pool reactor, the minimum container is 200 liters, and the third stage culture is carried out for 3 days at the temperature of 28 ℃ and the temperature of 1500 lux.
The culture solution for algae strain, primary culture, secondary culture and tertiary culture comprises the following substances in parts by weight: 10 parts of water; 10 parts of agricultural compound amino acid powder; 10 parts of humate; 5 parts of monopotassium phosphate; 5 parts of urea; 1 part of ferrous sulfate; 1 part of calcium superphosphate; 1 part of borax; 0.1 part of trace elements; 0.1 part of gibberellin.
Example two
A high-yield industrial microalgae culture method comprises the following steps:
(1) preparing facilities and equipment required by the propagation of industrial microalgae, wherein the facilities and the equipment comprise a greenhouse glass room, the greenhouse glass room is a 300-square-meter workshop, the occupied area is 20 meters by 15 meters, a double-roof connection type greenhouse is adopted, and equipment is arranged in the greenhouse glass room: automatic or semi-automatic temperature increasing and controlling equipment, air humidity controlling equipment and CO2The device comprises a generating and discharging device, a light supplementing device, a water distribution treatment system and a high-level water pool;
(2) culturing algae in a 250ml triangular flask with the illuminance of 5000lux at 25 ℃, shaking the triangular flask 3 times every day, and obtaining seed liquid when the algae grow to the logarithmic phase;
(3) the primary culture was carried out in a 2-liter vessel, inoculated at a volume ratio of 10%, supplied with air supplemented with carbon dioxide, and cultured at 25 ℃ for 10 days at 2000 lux;
(4) taking one part of the culture medium to be continuously used for primary culture, using the other part of the culture medium to inoculate, and starting secondary culture, wherein the secondary culture is carried out in a high and narrow upright cylindrical container with the volume of 100 liters, the secondary culture is carried out for 3 days under the conditions of 30 ℃ and 2000lux, and the algae liquid of the secondary culture is used as larval bait or is used for inoculating and starting tertiary culture;
(5) the third stage culture adopts a pipeline type photoreactor, a columnar photoreactor or a microalgae runway pool reactor, the minimum container is 200 liters, and the third stage culture is carried out for 3 days at the temperature of 30 ℃ and the temperature of 2000 lux.
The culture solution for algae strain, primary culture, secondary culture and tertiary culture comprises the following substances in parts by weight: 30 parts of water; 15 parts of agricultural compound amino acid powder; 30 parts of humate; 10 parts of monopotassium phosphate; 8 parts of urea; 3 parts of ferrous sulfate; 3 parts of calcium superphosphate; 3 parts of borax; 0.3 part of trace elements; 0.3 part of gibberellin.
EXAMPLE III
A high-yield industrial microalgae culture method comprises the following steps:
(1) preparing facilities and equipment required by the propagation of industrial microalgae, wherein the facilities and the equipment comprise a greenhouse glass room, the greenhouse glass room is a 300-square-meter workshop, the occupied area is 20 meters by 15 meters, a double-roof connection type greenhouse is adopted, and equipment is arranged in the greenhouse glass room: automatic or semi-automatic temperature increasing and controlling equipment, air humidity controlling equipment and CO2The device comprises a generating and discharging device, a light supplementing device, a water distribution treatment system and a high-level water pool;
(2) culturing algae in a 250ml triangular flask with the illuminance of 5000lux at 26 ℃, shaking the triangular flask 3 times every day, and obtaining seed liquid when the algae grow to the logarithmic phase;
(3) the primary culture was carried out in a 4-liter vessel, inoculated at a volume ratio of 10%, supplied with air supplemented with carbon dioxide, and cultured at 30 ℃ for 14 days at 2500 lux;
(4) taking one part of the culture medium to be continuously used for primary culture, using the other part of the culture medium to inoculate, and starting secondary culture, wherein the secondary culture is carried out in a high and narrow upright cylindrical container with the volume of 120 liters, the secondary culture is carried out for 4 days under the conditions of 32 ℃ and 2500lux, and the algae liquid of the secondary culture is used as larval bait or is used for inoculating and starting tertiary culture;
(5) the third stage culture adopts a pipeline type photoreactor, a columnar photoreactor or a microalgae runway pool reactor, the minimum container is 200 liters, and the third stage culture is carried out for 4 days at the temperature of 32 ℃ and the temperature of 2500 lux.
The culture solution for algae strain, primary culture, secondary culture and tertiary culture comprises the following substances in parts by weight: 50 parts of water; 20 parts of agricultural compound amino acid powder; 50 parts of humate; 15 parts of monopotassium phosphate; 10 parts of urea; 5 parts of ferrous sulfate; 5 parts of calcium superphosphate; 5 parts of borax; 0.5 part of trace elements; 0.5 part of gibberellin.
The present invention and the embodiments thereof have been described above, but the description is not limited to the embodiments, but only one of the embodiments of the present invention, and the actual embodiments are not limited thereto. In conclusion, those skilled in the art should appreciate that they can readily use the disclosed conception and specific embodiments as a basis for designing or modifying other structures for carrying out the same purposes of the present invention without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (6)
1. A high-yield industrial microalgae culture method is characterized by comprising the following steps: the method comprises the following steps:
(1) the facility and equipment required by the industrialized microalgae propagation are prepared, and the facility and the equipment comprise a greenhouse glass room, wherein the greenhouse glass room is internally provided with: automatic or semi-automatic temperature increasing and controlling equipment, air humidity controlling equipment and CO2The device comprises a generating and discharging device, a light supplementing device, a water distribution treatment system and a high-level water pool;
(2) culturing algae in a 250ml triangular flask with illumination of 5000lux at 22-26 deg.C, shaking the triangular flask 2-3 times per day, and obtaining seed solution when the algae grows to logarithmic phase;
(3) the first-stage culture was carried out in a 250 ml-4 l vessel, inoculated at a volume ratio of 10%, and supplied with air supplemented with carbon dioxide;
(4) taking one part of the culture medium to be continuously used for primary culture, using the other part of the culture medium to be inoculated, and starting secondary culture, wherein the secondary culture is carried out in a high and narrow upright cylindrical container, the volume of the container is 20 liters to 120 liters, and the algae liquid of the secondary culture is used as larva bait or is used for inoculation and starting tertiary culture;
(5) the third stage culture adopts a pipeline type photoreactor, a columnar photoreactor or a microalgae runway pool reactor, and the minimum volume of the container is 200 liters.
2. The method for cultivating high-yield industrial microalgae according to claim 1, wherein the method comprises the following steps: the greenhouse glass room is a 300-square-meter workshop, the floor space is 20 meters by 15 meters, and a double-roof connection type greenhouse is adopted.
3. The method for cultivating high-yield industrial microalgae according to claim 1, wherein the method comprises the following steps: the primary culture is carried out for 7-14 days at 20 + -10 deg.C and 2000 + -500 lux.
4. The method for cultivating high-yield industrial microalgae according to claim 1, wherein the method comprises the following steps: the second-stage culture is carried out at 30 + -2 deg.C and 2000 + -500 lux for 3-4 days.
5. The method for cultivating high-yield industrial microalgae according to claim 1, wherein the method comprises the following steps: the third-stage culture is carried out at 30 +/-2 ℃ and 2000 +/-500 lux for 3-4 days.
6. The method for cultivating high-yield industrial microalgae according to claim 1, wherein the method comprises the following steps: the culture solution for algae strain, primary culture, secondary culture and tertiary culture comprises the following substances in parts by weight: 10-50 parts of water; 10-20 parts of agricultural compound amino acid powder; 10-50 parts of humate; 5-15 parts of monopotassium phosphate; 5-10 parts of urea; 1-5 parts of ferrous sulfate; 1-5 parts of calcium superphosphate; 1-5 parts of borax; 0.1-0.5 part of trace elements; 0.1-0.5 part of gibberellin.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101696389A (en) * | 2009-10-29 | 2010-04-21 | 新奥科技发展有限公司 | Microalgae culture method and photo-bioreactor system thereof |
| CN102206570A (en) * | 2010-03-31 | 2011-10-05 | 中国科学院青岛生物能源与过程研究所 | Apparatus and cultivating method for scaled cultivation of microalgae |
| CN105002094A (en) * | 2015-07-31 | 2015-10-28 | 临沂大学 | Method for semi-continuously culturing marine microalgae through solar greenhouse |
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- 2020-07-01 CN CN202010629338.9A patent/CN111607523A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101696389A (en) * | 2009-10-29 | 2010-04-21 | 新奥科技发展有限公司 | Microalgae culture method and photo-bioreactor system thereof |
| CN102206570A (en) * | 2010-03-31 | 2011-10-05 | 中国科学院青岛生物能源与过程研究所 | Apparatus and cultivating method for scaled cultivation of microalgae |
| CN105002094A (en) * | 2015-07-31 | 2015-10-28 | 临沂大学 | Method for semi-continuously culturing marine microalgae through solar greenhouse |
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