CN114032176A - Culture system and culture method for promoting growth of microalgae - Google Patents
Culture system and culture method for promoting growth of microalgae Download PDFInfo
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- 230000012010 growth Effects 0.000 title claims abstract description 23
- 238000012136 culture method Methods 0.000 title claims abstract description 13
- 230000001737 promoting effect Effects 0.000 title claims abstract description 8
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 claims description 22
- 238000000034 method Methods 0.000 claims description 13
- 239000013589 supplement Substances 0.000 claims description 13
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 13
- 239000004317 sodium nitrate Substances 0.000 claims description 11
- 235000010344 sodium nitrate Nutrition 0.000 claims description 11
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims description 10
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 claims description 10
- ZGTMUACCHSMWAC-UHFFFAOYSA-L EDTA disodium salt (anhydrous) Chemical compound [Na+].[Na+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O ZGTMUACCHSMWAC-UHFFFAOYSA-L 0.000 claims description 9
- 229940044631 ferric chloride hexahydrate Drugs 0.000 claims description 9
- NQXWGWZJXJUMQB-UHFFFAOYSA-K iron trichloride hexahydrate Chemical compound O.O.O.O.O.O.[Cl-].Cl[Fe+]Cl NQXWGWZJXJUMQB-UHFFFAOYSA-K 0.000 claims description 9
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 9
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 9
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 8
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 8
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 claims description 6
- 238000012258 culturing Methods 0.000 claims description 6
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 5
- 229910021529 ammonia Inorganic materials 0.000 claims description 5
- 239000001569 carbon dioxide Substances 0.000 claims description 5
- 229910002092 carbon dioxide Inorganic materials 0.000 claims description 5
- 239000008103 glucose Substances 0.000 claims description 5
- 230000002572 peristaltic effect Effects 0.000 claims description 4
- 241000195649 Chlorella <Chlorellales> Species 0.000 claims description 3
- 239000012531 culture fluid Substances 0.000 claims description 3
- 238000005286 illumination Methods 0.000 claims description 3
- 241000224474 Nannochloropsis Species 0.000 claims description 2
- 241000514008 Oocystis Species 0.000 claims description 2
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims description 2
- 230000001502 supplementing effect Effects 0.000 claims description 2
- 238000012364 cultivation method Methods 0.000 claims 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 abstract description 8
- 229910052799 carbon Inorganic materials 0.000 abstract description 8
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 abstract description 6
- 235000015097 nutrients Nutrition 0.000 abstract description 4
- 229910052757 nitrogen Inorganic materials 0.000 abstract description 3
- 238000011031 large-scale manufacturing process Methods 0.000 abstract description 2
- 239000000463 material Substances 0.000 abstract description 2
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 16
- 239000007788 liquid Substances 0.000 description 9
- 229960000583 acetic acid Drugs 0.000 description 7
- 239000001963 growth medium Substances 0.000 description 7
- 230000008569 process Effects 0.000 description 7
- 239000007789 gas Substances 0.000 description 5
- 239000012362 glacial acetic acid Substances 0.000 description 5
- 230000029553 photosynthesis Effects 0.000 description 5
- 238000010672 photosynthesis Methods 0.000 description 5
- 230000001105 regulatory effect Effects 0.000 description 5
- 241000195493 Cryptophyta Species 0.000 description 4
- -1 genetic engineering Substances 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 230000029219 regulation of pH Effects 0.000 description 4
- 238000003756 stirring Methods 0.000 description 4
- 238000009360 aquaculture Methods 0.000 description 3
- 244000144974 aquaculture Species 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 230000000052 comparative effect Effects 0.000 description 3
- 230000001651 autotrophic effect Effects 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 125000005587 carbonate group Chemical group 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 230000035764 nutrition Effects 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 230000029058 respiratory gaseous exchange Effects 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000002028 Biomass Substances 0.000 description 1
- 241001491691 Thalassiosira Species 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000019522 cellular metabolic process Effects 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000006052 feed supplement Substances 0.000 description 1
- 239000000446 fuel Substances 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 238000010899 nucleation Methods 0.000 description 1
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
- 230000000243 photosynthetic effect Effects 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
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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
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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
- C12N1/38—Chemical stimulation of growth or activity by addition of chemical compounds which are not essential growth factors; Stimulation of growth by removal of a chemical compound
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- Bioinformatics & Cheminformatics (AREA)
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- General Engineering & Computer Science (AREA)
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Abstract
The invention relates to the technical field of microalgae culture, in particular to a culture system and a culture method for promoting the growth of microalgae. In the culture system, the nutrient components in the culture solution and the supplementary materials are reasonably matched, and a proper carbon source and a proper nitrogen source are provided for the growth of the microalgae while the pH environment is stabilized, so that the growth of the microalgae is effectively improved, and the large-scale production of the microalgae is realized.
Description
Technical Field
The invention relates to the technical field of microalgae cultivation, in particular to a culture system and a culture method for promoting microalgae growth.
Background
Microalgae are autotrophic plants which are widely distributed on land and sea, rich in nutrition and high in photosynthetic efficiency, and polysaccharides, proteins, pigments and the like generated by cell metabolism, so that the microalgae have good development prospects in the fields of food, medicine, genetic engineering, liquid fuel and the like. In the process of microalgae cultivation, the concentration of ions in microalgae culture solution changes along with the strength of photosynthesis and respiration of microalgae, the concentration of ions required by growth is reduced, the nutrition required by microalgae production is insufficient, and the growth rate is obviously reduced along with the prolonging of time. At the same time, the aging and death of the microalgae can be caused, and the production rate and the product quality are reduced.
The culture medium adopted by the traditional microalgae culture method has higher cost, and mainly depends on manual feeding in the culture process, so that not only is the time consumption long and the working procedure reproduction, but also errors are easy to occur or the feeding is not timely, and the growth of the algae is influenced. Therefore, the culture solution and the culture method which can obviously improve the biomass of the microalgae have important practical significance.
Disclosure of Invention
In view of the above, the present invention provides a culture system and a culture method for promoting the growth of microalgae. The culture system can effectively promote the growth of microalgae and realize the large-scale production of the microalgae.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a culture system for promoting microalgae growth, which comprises a culture solution and a supplement;
the culture solution consists of sodium nitrate, monopotassium phosphate, EDTA-2Na, ferric chloride hexahydrate and water;
the feed consists of ammonia, carbon dioxide, glucose and water.
In the process of microalgae cultivation, due to different substances added in a culture medium, carbon sources or nitrogen sources in microalgae can be absorbed when the microalgae is subjected to autotrophic or heterotrophic cultivation, so that the substances are reduced, and the pH value of a culture solution is changed. The invention adopts specific culture solution to culture microalgae, and supplements the specific pH regulation solution (gas) through a pH regulation system. After the supplemented pH regulation liquid (gas) is utilized by the microalgae, the pH rises (falls), the pH regulation system supplements the relevant regulation liquid (gas), and the steps are repeated, so that the pH environment for microalgae cultivation is kept in a stable state, meanwhile, the supplement with specific components can be continuously added into the microalgae cultivation liquid, the collocation of all nutrient components is reasonable, proper carbon and nitrogen sources are provided for the growth of the microalgae while the pH is controlled, and stable bait concentration is provided for the growth environment of the microalgae, thereby effectively improving the growth amount of the microalgae.
In some embodiments, the concentration of ammonia in the feed is 2-5%; the glucose concentration is 1-3%.
In some embodiments the culture fluid consists of water and the following components:
800g/L of sodium nitrate 500-.
Preferably, the culture fluid consists of water and the following components: 500-700g/L sodium nitrate, 130-L, EDTA-2Na 100-130g/L potassium dihydrogen phosphate and 50g/L ferric chloride hexahydrate.
In a specific embodiment of the invention, the culture solution consists of water and the following components:
500g/L of sodium nitrate, 100g/L, EDTA-2Na 100g/L of monopotassium phosphate and 50g/L of ferric chloride hexahydrate;
or 700g/L of sodium nitrate, 130g/L, EDTA-2Na 120g/L of monopotassium phosphate and 50g/L of ferric chloride hexahydrate.
The invention also provides application of the culture system in microalgae culture.
Preferably, the microalgae is chlorella, nannochloropsis, oocystis or thalassiosira.
The invention provides a culture method of microalgae, which comprises the following steps:
inoculating microalgae into a culture solution in the culture system, introducing air, culturing for 48-96h at 30-35 ℃, culturing at the illumination intensity of 3000-: 1;
supplementing the supplementary material into the culture system to ensure that the pH of the culture system is 8.0-8.2 and the concentration of carbonate ions is more than or equal to 2 mg/L.
In the invention, the feeding time is not fixed, the feeding time is influenced by the photosynthesis of the microalgae, the pH of the microalgae liquid can be changed, the photosynthesis is strong, the microalgae absorbs carbon dioxide to enable the pH to rise, the respiration is strong, the algae releases carbon dioxide, and the pH drops. The system automatically feeds according to the pH change fed back by the sensor, if the pH rises (exceeds 8.2), carbon dioxide is added to reduce the pH, and if the pH continuously drops (is lower than 8.0), ammonia is added to increase the pH.
In some embodiments, the microalgae has a seeding density of 50 to 150 ten thousand per milliliter.
In some embodiments, the culturing conditions are 48-96h, 30-35 ℃ with illumination intensity of 3000-: 1.
in the invention, the pH is regulated and controlled by a pH sensor, a peristaltic pump and a control system.
In the invention, the culture device is a pipeline type or column type photobioreactor.
Compared with the prior art, the invention has the following beneficial effects:
(1) the nutrient components in the culture medium and the supplement are reasonably matched, and the requirement of the microalgae on the nutrient components in the growth process can be met, so that the growth of the microalgae is effectively promoted, and the growth amount of the microalgae is increased.
(2) The invention can effectively solve the problems of long time consumption, procedure propagation, easy error and the like of manual feeding in the existing breeding feeding technology.
(3) The invention can realize automatic control of feed supplement, ensures that feed liquid supplement is not limited by time and personnel, accurately, precisely and quantitatively feeds, and greatly improves the culture efficiency.
(4) The feed liquid replenishing system has low cost investment and simple and convenient operation, and is more suitable for common breeding families.
Detailed Description
The invention provides a culture system and a culture method for promoting microalgae growth. Those skilled in the art can modify the process parameters appropriately to achieve the desired results with reference to the disclosure herein. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the methods and applications of this invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications in the methods and applications described herein, as well as other suitable variations and combinations, may be made to implement and use the techniques of this invention without departing from the spirit and scope of the invention.
The test materials adopted by the invention are all common commercial products and can be purchased in the market.
The invention is further illustrated by the following examples:
example 1
Firstly, inoculating microalgae seeds into a pipeline type or column type photobioreactor, wherein a culture medium used by 1 ton of aquaculture water is as follows: 500g of sodium nitrate, 100g of monopotassium phosphate, 100g of EDTA-2Na and 50g of ferric chloride hexahydrate, and then continuously introducing air to continuously stir the culture solution. During the process of microalgae photoautotrophy, carbonate in the culture solution is continuously utilized for photosynthesis. The decrease in carbonate groups greatly increases the pH of the culture solution. The pH change is fed back to the pH controller through the pH sensor, the pH controller automatically supplements pH regulating liquid + gas (namely supplement) acetic acid (acetic acid, glacial acetic acid and glacial acetic acid) into the reactor through a peristaltic pump, the pH is regulated to be maintained within the range of 8.0-8.2, and simultaneously, a carbon source required by microalgae production is supplemented, so that the concentration of carbonate is kept above 2 mg/L.
Example 2
Firstly, inoculating marine chlorella algae seeds into a microalgae runway pool, wherein a culture medium used by 1 ton of culture water is as follows: 700g of sodium nitrate, 130g of monopotassium phosphate, 120g of EDTA-2Na and 50g of ferric chloride hexahydrate, and then continuously stirring the culture solution in the runway by a water pump to ensure that the culture solution is continuously and uniformly mixed. During the process of microalgae photoautotrophy, carbonate in the culture solution is continuously utilized for photosynthesis. The decrease in carbonate groups greatly increases the pH of the culture solution. The pH change is fed back to the pH controller through the pH sensor, the pH controller automatically supplements pH regulating liquid + gas (namely supplement) acetic acid (acetic acid, glacial acetic acid and glacial acetic acid) into the raceway pond through the peristaltic pump, the pH is regulated to be maintained within the range of 8.0-8.2, and simultaneously, carbon source required by microalgae production is supplemented, so that the concentration of carbonate is kept above 2 mg/L.
Comparative example 1
Firstly, inoculating microalgae seeds into a pipeline type or column type photobioreactor, wherein a culture medium used by 1 ton of aquaculture water is as follows: 500g of sodium nitrate, 100g of monopotassium phosphate, 100g of EDTA-2Na and 50g of ferric chloride hexahydrate, and then continuously introducing air to continuously stir the culture solution. At the same time, 500 ml/ton of 50% strength glucose solution was fed daily to supplement the carbon source.
Comparative example 2
Firstly, inoculating microalgae seeds into a pipeline type or column type photobioreactor, wherein a culture medium used by 1 ton of aquaculture water is as follows: 500g of sodium nitrate, 100g of monopotassium phosphate, 100g of EDTA-2Na and 50g of ferric chloride hexahydrate, and then continuously introducing air to continuously stir the culture solution. At the same time, 500 ml/ton of 20% glacial acetic acid solution is added daily to supplement the carbon source.
TABLE 1
The results in Table 1 show that the culture method can stably control the pH to 8.0-8.2, provide the optimal growth pH for the microalgae, stably control the concentration of the carbon source to 2-3mg/L, and finally obviously promote the growth of the microalgae, and the algae density reaches 2670-3570 ten thousand per milliliter after 4 days of growth, which is obviously superior to that of comparative examples 1-2.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that it is obvious to those skilled in the art that various modifications and improvements can be made without departing from the principle of the present invention, and these modifications and improvements should also be considered as the protection scope of the present invention.
Claims (10)
1. A culture system for promoting the growth of microalgae is characterized by comprising a culture solution and a supplement;
the culture solution consists of sodium nitrate, monopotassium phosphate, EDTA-2Na, ferric chloride hexahydrate and water;
the feed consists of ammonia, carbon dioxide, glucose and water.
2. The culture system of claim 1, wherein the concentration of ammonia in the feed is 2-5%; the glucose concentration is 1-3%.
3. The culture system of claim 1, wherein the culture fluid consists of water and the following components:
800g/L of sodium nitrate 500-.
4. Use of the culture system of any one of claims 1 to 3 in microalgae culture.
5. The use according to claim, wherein the microalgae is chlorella, nannochloropsis, oocystis or thalassiosia.
6. A method for culturing microalgae, comprising:
inoculating microalgae into the culture solution in the culture system of any one of claims 1-3, introducing air, culturing for 48-96h at 30-35 ℃, culturing with illumination intensity of 3000-: 1;
supplementing the supplementary material into the culture system to ensure that the pH of the culture system is 8.0-8.2 and the concentration of carbonate ions is more than or equal to 2 mg/L.
7. The cultivation method according to claim 6, wherein the microalgae are inoculated at a density of 50-150 ten thousand/ml.
8. The culture method according to claim 6, wherein the culture conditions are 30-35 ℃ culture, light intensity of 3000-15000LUX, light-to-dark ratio of 1: 1.
9. the culture method according to claim 6, wherein the pH is controlled by a pH sensor, a peristaltic pump and a control system.
10. The culture method according to claim 6, wherein the culture device is a pipe type or column type photobioreactor.
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| CN113278557A (en) * | 2021-05-25 | 2021-08-20 | 海南绿藻世界生物科技有限公司 | Symbiotic bacteria composition, preparation method thereof and microalgae culture method |
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