CN114164128A - Algae symbiotic bacteria composition, algae bacteria co-culture system and microalgae culture method - Google Patents
Algae symbiotic bacteria composition, algae bacteria co-culture system and microalgae culture method Download PDFInfo
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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
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
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- 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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- 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
- C12N1/20—Bacteria; Culture media therefor
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Abstract
The invention provides an algae symbiotic bacteria composition, which comprises rhodotorula benthica, photosynthetic bacteria, marine nitrobacteria and vibrio phagostii; the proportion of the rhodotorula benthica, the photosynthetic bacteria strain, the marine nitrobacteria and the vibrio phagostii is (20-30) g: (100-200) ml: (10-50) g: (1-3) g; the density of the photosynthetic bacteria strain is 200-500 hundred million/ml. Compared with the prior art, the method has the advantages that different bacterium combinations and algae symbiotic culture are adopted, so that the problems of easy aging, rot, harmful bacterium breeding and the like in the microalgae culture process can be solved, microalgae and beneficial bacteria are symbiotically cultured, the growth speed of algae is faster and more active, and the yield of microalgae is increased.
Description
Technical Field
The invention belongs to the technical field of microalgae culture, and particularly relates to an algae symbiotic bacteria composition, an algae co-culture system and a microalgae culture method.
Background
The chlorella vulgaris is rich in proteins, lipids, polysaccharides, vitamins, antioxidants and other functional nutritional ingredients, and therefore has wide application in the fields of medical raw materials, health-care food and biological energy. At present, the seawater chlorella culture technology is mostly concentrated on a photoautotrophic system, and the culture system has the defects of low production efficiency, large floor area, high harvest cost and the like, thereby seriously restricting the development of the seawater chlorella industry. The key point of development and utilization of the seawater chlorella resources is to improve the biomass of the seawater chlorella by an artificial large-scale culture technology.
In recent years, a heterotrophic high-cell-density culture technology for marine chlorella 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 marine chlorella culture research in recent years.
Most aquatic product seedling raising enterprises in China have self-provided marine chlorella culture facilities in seedling raising production, and can produce various marine chlorella for baits. However, general seedling farms generally lack corresponding professional technical strength, can only utilize respective algae ponds and natural water bodies for extensive culture, and are respectively positive in the germplasm, production technology and application method of the bait chlorella vulgaris, so that the chlorella vulgaris is disordered in germplasm, unstable in supply, unbalanced in nutrient content, low in bait titer and lack of a multi-variety intensive production application technology; meanwhile, the method is limited by the limitations of high-density cultivation, harvesting technology and concentrated solution preservation technology of the marine chlorella, and domestic uniform and specialized bait marine chlorella quality standards and centralized supply points are almost not available. Therefore, the algae seeds need to be supplemented in time for industrial seedling raising, and the initial feeding is very important.
Like other organisms, algae are also infested with external pests during their growth. The main pathogenic organisms of the marine chlorella are viruses, bacteria, fungi and protists. These diseases may seriously affect the growth of algae, especially impact the economic algae production, and increase the production cost.
In the long-term evolution process of the nature, different bacteria live in the growth environment of algae, and the two groups have complex and unique interaction, so that the method is very critical to the circulating process of substances and energy in the water environment. Algae is a photoautotrophic organism living in fresh water and seawater, supplies oxygen to a water body through photosynthesis, increases dissolved oxygen of the water body, provides oxygen for the water body, continuously degrades organic matters by bacteria, and can degrade CO generated by the organic matters by the bacteria2The photosynthesis is carried out, and the system formed by the circulation is called algal bacteria symbiosis.
Algae and bacteria have a complex relationship, and each algae forms its own unique flora and there is a specific interspecific relationship between the two. The algae mainly promote the growth of bacteria through nutrient exchange, gas, pH value adjustment and the like, for example, chlorella can promote the growth of aerobic heterotrophic bacillus through directly absorbing and utilizing NH4+ -N in water and improving dissolved oxygen. The bacteria, as an important decomposer in a water ecosystem, participate in the decomposition and transformation processes of various substances in a water environment through complex metabolic activities, and thus serve as providers and processors of microalgae nutrient substances; meanwhile, the bacteria can also play a role in promoting microalgae by producing or secreting metabolic substances or extracellular products beneficial to the microalgae. For example, ammoniating bacteria and nitrifying bacteria in the water body can decompose nitrogen-containing organic substances and further convert the nitrogen-containing organic substances into NH3+ -N through nitrification, so that inorganic nitrogen is provided for the growth of algae; the aerobic bacteria provide a living environment with stronger reducibility for the algae by utilizing high-concentration dissolved oxygen in the water around the microalgae.
However, algae have a fundamental mechanism for the production of active secondary metabolites, including fatty acid, phenolic, polysaccharide species, which have the effect of inhibiting or destroying certain bacteria, such as bacillus licheniformis, by their extracellular products rather than by nutrient competition. Meanwhile, the algae directly compete with the bacteria for nutrients to play an indirect inhibition role, for example, the growth of the high population density microcystis has an obvious inhibition effect on the growth of nitrobacteria, and the competitive utilization of NH4+ -N and the like exists between the microcystis and the nitrobacteria. The inhibition or antagonism of algae by bacteria is also manifested in various aspects, mainly by means of nutrition competition, chemical toxin production, release of algicidal enzyme and the like. In certain aquatic environments, bacteria inhibit the growth of algae by competing for nutrients when not supplemented by exogenous nutrients. Pseudomonas, bdellovibrio, bacillus, and Pythium species all can produce chemical toxins to be released in water, thereby inhibiting the growth of some microalgae such as dinoflagellate and diatom, and even killing algae cells.
And pollutants in the environment can be naturally degraded within a certain time range through the mutual promotion and mutual inhibition of algae and bacteria in the water body, so that the natural ecological chain of the water body is maintained, namely the self-purification effect of the water body. When excessive nutrient substances are dissolved in the water body, the ecological chain is impacted, the symbiotic relationship of bacteria and algae is still continued in the range borne by the ecological chain, but due to the physiological characteristics of blue-green algae, the inhibition effect of the bacteria on the algae is in a disadvantage, the water body develops towards the direction of eutrophication, at the moment, microorganisms are added into the water body to compete with the algae for N, P and other nutrients, the microbial flora structure is effectively repaired, the inhibition capability of the bacteria is enhanced, the water body can be effectively prevented from developing towards the direction of eutrophication, and the basic ecological balance of the water body is maintained.
When the amount of nutrients exceeds the tolerance of the ecological chain, the ecological chain is destroyed. Combining a plurality of external factors including temperature, illumination, nutritive salt, hydrology and the like, the algae do not depend on the metabolic products of bacteria any more, the nutrient substances in the water are directly utilized for rapid propagation, the inhibiting effect of the bacteria on the algae is rapidly collapsed, and even the algae is damaged by algal toxins; prey speed of predators (aquatic animals) in water is far lower than the growth speed of algae, the growth of algae is not limited, and water becomes the day under the algae. In addition, when the algae is propagated in a large quantity, part of cells are aged and died, the died cells are broken and degraded, oxygen in a water body is consumed, dissolved oxygen is rapidly reduced, and broken blue algae can release toxic and harmful substances such as blue algae toxin, ammonia nitrogen, nitrite and the like, so that a series of serious consequences are brought to a water ecological system. At this time, the growth and reproduction of bacteria in the water body are inhibited, and the water body is in a dormant state or partially died, and although the algicidal bacteria and the algophagous body can survive, the effect of inhibiting the algae is very little. At this time, the effect of microorganism throwing in water is very little, so an ecological and comprehensive treatment method is required to restore the ecological balance of water.
Disclosure of Invention
In view of the above, the technical problem to be solved by the present invention is to provide an algal symbiotic bacteria composition, an algal bacteria co-culture system and a microalgae cultivation method, wherein the algal bacteria co-culture system can avoid the problems of easy aging, decay, harmful bacteria breeding and the like of microalgae in a microalgae cultivation process, so that the microalgae and beneficial bacteria are symbiotically cultured, and the growth speed of the algae is faster and more active.
The invention provides an algae symbiotic bacteria composition, which comprises rhodotorula benthica, photosynthetic bacteria, marine nitrobacteria and vibrio phagostii; the proportion of the rhodotorula benthica, the photosynthetic bacteria strain, the marine nitrobacteria and the vibrio phagostii is (20-30) g: (100-200) ml: (10-50) g: (1-3) g; the density of the photosynthetic bacteria strain is 200-500 hundred million/ml.
Preferably, the proportion of the rhodotorula benthica, the photosynthetic bacteria strain, the marine nitrobacteria and the vibrio phagostii is (20-25) g: 150 ml: (17-30) g: (1-2) g.
The invention also provides an algae-bacteria co-culture system, which comprises an algae symbiotic bacteria composition, a microalgae culture medium and microalgae; the algae symbiotic bacteria composition comprises rhodotorula benthica, photosynthetic bacteria strains, marine nitrobacteria and vibrio phagostii; the proportion of the rhodotorula benthica, the photosynthetic bacteria strain, the marine nitrobacteria and the vibrio phagostii is (20-30) g: (100-200) ml: (10-50) g: (1-3) g; the density of the photosynthetic bacteria strain is 200-500 hundred million/ml.
Preferably, the formula of the microalgae culture medium comprises:
preferably, the tween is tween 80; the vitamins include vitamin B12, vitamin B1 and biotin; the trace elements include Na2EDTA、FeCl3、CuSO4、ZnSO4、CoCl2、MnC12With Na2MoO4。
Preferably, the concentration of the rhodotorula benthamii in the phycomycete co-culture system is 20-30 g/L; the concentration of the photosynthetic bacteria strain in the algae bacteria co-culture system is 100-200 ml/L; the concentration of marine nitrifying bacteria in the algae bacterium co-culture system is 10-50 g/L; the concentration of vibrio phagostii in the phycomycete co-culture system is 1-3 g/L.
Preferably, the inoculation density of the microalgae in the phycomycete co-culture system is 50-100 ten thousand per ml.
The invention also provides a culture method of microalgae, which comprises the following steps:
sterilizing and cooling a microalgae culture medium, inoculating an algae symbiotic bacteria composition and microalgae, and culturing;
the algae symbiotic bacteria composition comprises rhodotorula benthica, photosynthetic bacteria strains, marine nitrobacteria and vibrio phagostii; the proportion of the rhodotorula benthica, the photosynthetic bacteria strain, the marine nitrobacteria and the vibrio phagostii is (20-30) g: (100-200) ml: (10-50) g: (1-3) g; the density of the photosynthetic bacteria strain is 200-500 hundred million/ml.
Preferably, the temperature of the culture is 30-35 ℃; the culture time is 48-96 h; the illumination intensity of the culture is 3000-12000 lux;
the microalgae is one or more selected from Chlorella salina, diatom, Nannochloropsis oculata and oocyst algae.
Preferably, a mixed gas of carbon dioxide and oxygen is introduced in the culture process; the ventilation volume of the mixed gas is 1-5 vvm; the volume ratio of carbon dioxide to oxygen in the mixed gas is 2: 1.
the invention provides an algae symbiotic bacteria composition, which comprises rhodotorula benthica, photosynthetic bacteria, marine nitrobacteria and vibrio phagostii; the proportion of the rhodotorula benthica, the photosynthetic bacteria strain, the marine nitrobacteria and the vibrio phagostii is (20-30) g: (100-200) ml: (10-50) g: (1-3) g; the density of the photosynthetic bacteria strain is 200-500 hundred million/ml. Compared with the prior art, the method has the advantages that different bacterium combinations and algae symbiotic culture are adopted, so that the problems of easy aging, rot, harmful bacterium breeding and the like in the microalgae culture process can be solved, microalgae and beneficial bacteria are symbiotically cultured, the growth speed of algae is faster and more active, and the yield of microalgae is increased.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious 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.
The invention provides an algae symbiotic bacteria composition, which comprises rhodotorula benthica, photosynthetic bacteria, marine nitrobacteria and vibrio phagostii; the proportion of the rhodotorula benthica, the photosynthetic bacteria strain, the marine nitrobacteria and the vibrio phagostii is (20-30) g: (100-200) ml: (10-50) g: (1-3) g; the density of the photosynthetic bacteria strain is 200-500 hundred million/ml.
Wherein the content of effective viable bacteria of the rhodotorula benthica is preferably more than or equal to 100 hundred million/g; the content of effective viable bacteria of the marine nitrifying bacteria is preferably more than or equal to 200 hundred million/g, and more preferably 300-400 hundred million/g; the content of the effective viable bacteria of the vibrio phagostimuli is preferably more than or equal to 50 multiplied by 108 CFU/g.
The invention is not limited to the types of the marine rhodotorula, photosynthetic bacteria, marine nitrobacteria and vibrio phagostii, and the examples provided by the invention are dry marine rhodotorula of the Chinese family Huijia biotechnology, photosynthetic bacteria of the Jiangxi Youyou biotechnology Limited, dry marine nitrobacteria of the Shanghai Qianyu biotechnology Limited and vibrio phagostii of the Nanjing Selt to illustrate the effects of the algal symbiotic bacteria composition provided by the invention.
In the invention, the proportion of the rhodotorula benthica, the photosynthetic bacteria strain, the marine nitrobacteria and the vibrio phagostii is preferably (20-28) g: (120-180) ml: (15-40) g: (1-3) g, more preferably (20-25) g: (140-160) ml: (15-30) g: (1-3) g, preferably (20-25): 150 ml: (17-30) g: (1-2) g.
The invention also provides a phycomycete co-culture system, which comprises the following components: algae symbiotic bacteria composition, microalgae culture medium and microalgae; the algae symbiotic bacteria composition comprises rhodotorula benthica, photosynthetic bacteria strains, marine nitrobacteria and vibrio phagostii; the proportion of the rhodotorula benthica, the photosynthetic bacteria strain, the marine nitrobacteria and the vibrio phagostii is (20-30) g: (100-200) ml: (10-50) g: (1-3) g; the density of the photosynthetic bacteria strain is 200-500 hundred million/ml.
The proportion and the effective bacteria content of each bacterium in the algae symbiotic bacteria composition are the same as those described above, and are not described again.
In the invention, the concentration of the rhodotorula benthamii in the phycomycete co-culture system is preferably 20-30 g/L, more preferably 20-28 g/L, and still more preferably 20-25 g/L.
In the invention, the concentration of the photosynthetic bacteria strain in the phycomycete co-culture system is preferably 100-200 ml/L, more preferably 120-180 ml/L, more preferably 140-160 ml/L, and even more preferably 150 ml/L.
In the invention, the concentration of the marine nitrifying bacteria in the phycomycete co-culture system is preferably 10-50 g/L, more preferably 15-40 g/L, more preferably 15-30 g/L, and even more preferably 17-30 g/L.
In the invention, the concentration of vibrio phagostii in the phycomycete co-culture system is preferably 1-3 g/L, and more preferably 1-2 g/L.
The formula of the microalgae culture medium in the phycomycete co-culture system provided by the invention preferably comprises the following components:
further preferably comprises:
further preferably comprises:
the vitamins preferably include vitamin B12, vitamin B1 and biotin; the mass ratio of the vitamin B12, the vitamin B1 and the biotin is preferably (0.5-2): (0.5-2): (0.5-2), more preferably (0.5-1.5): (0.5-1.5): (0.5 to 1.5), and preferably 1: 1: 1; the trace elements include Na2EDTA、FeCl3、CuSO4、ZnSO4、CoCl2、MnC12With Na2MoO4(ii) a The Na is2EDTA、FeCl3、CuSO4、ZnSO4、CoCl2、MnC12With Na2MoO4The mass ratio of (1-3): (0.1-0.3): (0.05-0.15): (0.05-0.15): (0.01-0.1): (0.01-0.1): 0.01, more preferably (1.5 to 2.5): (0.15-0.25): (0.08-0.12): (0.08-0.12): (0.03-0.08): (0.03-0.0): 0.01, more preferably 2: 0.2: 0.1: 0.1: 0.05: 0.05: 0.01.
the balance of the culture medium is water, preferably filtered seawater.
The inoculation density of the microalgae in the phycomycete co-culture system is preferably 50-100 ten thousand per ml, more preferably 60-90 ten thousand per ml, and further preferably 70-80 ten thousand per ml; the microalgae is preferably one or more of Chlorella salina, diatom, Nannochloropsis oculata and oocystis algae.
The invention also provides a culture method of microalgae, which comprises the following steps: sterilizing and cooling a microalgae culture medium, inoculating an algae symbiotic bacteria composition and microalgae, and culturing; the algae symbiotic bacteria composition comprises rhodotorula benthica, photosynthetic bacteria strains, marine nitrobacteria and vibrio phagostii; the proportion of the rhodotorula benthica, the photosynthetic bacteria strain, the marine nitrobacteria and the vibrio phagostii is (20-30) g: (100-200) ml: (10-50) g: (1-3) g; the density of the photosynthetic bacteria strain is 200-500 hundred million/ml.
The formula of the microalgae culture medium and the algae symbiotic bacteria composition are the same as those described above, and are not described again.
Firstly, sterilizing a microalgae culture medium; the temperature for sterilization is preferably 110-120 ℃, and more preferably 115 ℃; the time for sterilization is preferably 20-40 min, and more preferably 30 min.
Sterilizing, cooling, inoculating the algae symbiotic bacteria composition and microalgae to form an algae bacteria co-culture system; the concentrations of the algae symbiotic bacteria composition and the microalgae in the algae co-culture system are the same as those described above, and are not described again; the microalgae is preferably one or more of Chlorella salina, diatom, Nannochloropsis oculata and oocystis algae.
Inoculating the algae symbiotic bacteria composition and microalgae, and culturing; the culture temperature is preferably 30-35 ℃; the culture time is preferably 48-96 h; the illumination intensity during culture is preferably 3000-12000 lux, more preferably 4000-10000 lux, further preferably 5000-8000 lux, and most preferably 6000-7000 lux; preferably, a mixed gas of carbon dioxide and oxygen is also introduced in the culture process; the ventilation volume of the mixed gas is preferably 1-5 vvm, and more preferably 2-5 vvm; the volume ratio of carbon dioxide to oxygen in the mixed gas is preferably 2: 1.
according to the method, different bacterium combinations and algae symbiotic culture are adopted, and a special microalgae culture medium is further selected, so that the problems of easy aging, rot, harmful bacterium breeding and the like in the microalgae culture process can be solved, microalgae and beneficial bacteria are symbiotically cultured, the growth speed of algae is faster and more active, and the yield of microalgae are improved.
In order to further illustrate the present invention, the following will describe an algal symbiotic bacteria composition, algal co-culture system and microalgae culture method in detail with reference to the examples.
The reagents used in the following examples are all commercially available; the sources of the raw materials used in the examples are shown in Table 1.
TABLE 1 sources of raw materials
| Species of | Source |
| Dry powder of ocean rhodotorula | Chinese Huiyou biological technology, total bacteria number more than 100 hundred million/g |
| Photosynthetic bacteria strain | Youye Biotech limited Jiangxi fish with a total bacteria count of 100 hundred million/g or more |
| Marine nitrifying bacteria dry powder | Shanghai Qianzyu Biotechnology Limited company, the total number of bacteria is more than or equal to 200 hundred million/g |
| Vibrio phagi dry powder | Bdellovibrio bacteriovorus of Nanjing Selt, 99 (%) |
| Yeast extract powder | Angel yeast, BR |
| Peptone | Kwangtong ring, BR |
Example 1
1.1 Medium composition:
80g/L NaNO3、15g/L NH4HCO3、5g/L NaH2PO4·2H2O、5g/L K2HPO4·3H2O、2g/L KH2PO40.1 v/v% Tween 80, 30g/L NaCl, 15g/L yeast extract powder, 30g/L peptone, 10g/L sodium carboxymethylcellulose, 10g/L sodium diacetate, 100 mu g/L vitamin and 80mg/L trace elements.
The microorganisms comprise 1: 1: 1, vitamin B12, vitamin B1 and biotin.
Wherein the trace elements are added in the form of trace element solution, and the preparation method of the trace element solution is to dissolve the trace element solution in 1L of water according to the formula shown in the following table.
| Name of material | Rank of | Dosage of |
| Na2EDTA | AR | 2g |
| FeCl3 | AR | 0.2g |
| CuSO4 | AR | 0.1g |
| ZnSO4 | AR | 0.1g |
| CoCl2 | AR | 0.05g |
| MnC12 | AR | 0.05g |
| Na2MoO4 | AR | 0.01g |
The composition of the co-culture bacteria is as follows:
1.2 in a 2L triangular flask, preparing 1L of the culture medium by using filtered seawater, sterilizing at 115 ℃ for 30min, cooling, inoculating co-cultured bacteria and marine chlorella algae seeds (the inoculation density of the marine chlorella is 80 ten thousand per ml) in a super clean bench, culturing at 32 ℃ for 96h, wherein the illumination intensity is 6000lux in the culturing process, and introducing mixed gas of carbon dioxide and oxygen, the ventilation volume is 2vvm (volume per minute), and the mixing ratio of the carbon dioxide and the oxygen is 2: the density and activity of algae are measured daily by placing algae liquid into a blood counting chamber, and calculating the content under a microscope, and the results are shown in tables 2 and 3.
Example 2
2.1 Medium composition:
100g/L NaNO3、14g/L NH4HCO3、10g/L NaH2PO4·2H2O、5g/L K2HPO4·3H2O、1g/L KH2PO40.1 v/v% Tween 80, 30g/L NaCl, 20g/L yeast extract powder, 30g/L peptone, 15g/L sodium hydroxymethyl cellulose, 10g/L sodium diacetate, 150 mu g/L vitamin and 80mg/L trace elements. The composition of vitamins and trace elements was the same as in example 1.
The composition of the co-culture bacteria is as follows:
2.2 in a 400L vertical photobioreactor, preparing 350L of the culture medium by using filtered seawater, inoculating co-cultured bacteria and marine chlorella algae seeds (the inoculation density of the marine chlorella is 80 ten thousand per ml), carrying out aeration culture at 32 ℃ for 96h, introducing mixed gas of carbon dioxide and oxygen, wherein the ventilation volume is 5vvm (volume per minute), and the mixing ratio of the carbon dioxide to the oxygen is 2: the density and activity of algae are measured daily by placing algae liquid into a blood counting chamber, and calculating the content under a microscope, and the results are shown in tables 2 and 3.
Comparative example 1
1.1 Medium composition:
conventional microalgae f/2 medium: 75g/L NaNO3、5g/L NaH2PO4·2H2O、30g/L Na2SiO3·9H2O。
The composition of the co-culture bacteria is as follows:
1.2 in a 2L triangular flask, preparing 1L of the culture medium by using filtered seawater, sterilizing for 30min at 115 ℃, cooling, inoculating co-cultured bacteria and seawater chlorella seeds (the inoculation density of the seawater chlorella is 80 ten thousand per ml) in a super clean bench, culturing for 96h at 32 ℃, introducing mixed gas of carbon dioxide and oxygen, introducing the ventilation volume of 2vvm (volume per minute), and mixing the carbon dioxide and the oxygen according to the proportion of 2: the density and activity of algae are measured daily by placing algae liquid into a blood counting chamber, and calculating the content under a microscope, and the results are shown in tables 2 and 3.
Comparative example 2
2.1 Medium composition
The medium formulation was the same as in example 1.
2.2 in a 400L vertical photobioreactor, preparing 350L of the culture medium by using filtered seawater, carrying out aeration culture on marine chlorella species (the inoculation density of the marine chlorella is 80 ten thousand per ml) at 32 ℃ for 96h under the illumination intensity of 6000lux, introducing mixed gas of carbon dioxide and oxygen, wherein the ventilation volume is 5vvm (volume per minute), and the mixing ratio of the carbon dioxide to the oxygen is 2: the density and activity of algae are measured daily by placing algae liquid into a blood counting chamber, and calculating the content under a microscope, and the results are shown in tables 2 and 3.
TABLE 2 microalgae cell number variation (ten thousand/ml)
| Incubation time | Example 1 | Example 2 | Comparative example 1 | Comparative example 2 |
| 0h | 26 | 42 | 19 | 17.5 |
| 12h | 58 | 65 | 48 | 25.5 |
| 24h | 109.5 | 138 | 55 | 42.5 |
| 36h | 155 | 242 | 66 | 55 |
| 48h | 238 | 517 | 52.5 | 46 |
| 60h | 275.5 | 835 | 92 | 43.5 |
| 72h | 360.5 | 1073 | 96 | 37.3 |
Table 3 microalgae cell viability change (%)
| Incubation time | Example 1 | Example 2 | Comparative example 1 | Comparative example 2 |
| 0h | 100 | 100 | 100 | 100 |
| 12h | 99 | 100 | 85 | 89 |
| 24h | 99 | 99 | 78 | 81 |
| 36h | 98 | 98 | 66 | 73 |
| 48h | 98 | 98 | 52.5 | 67 |
| 60h | 95 | 97 | 49 | 61 |
| 72h | 95 | 97 | 45 | 54 |
Therefore, the culture medium and the culture mode provided by the invention can obviously improve the growth rate of microalgae, stabilize the activity of the microalgae and improve the yield of the microalgae.
Claims (10)
1. An algae symbiotic bacteria composition is characterized by comprising rhodotorula benthamii, photosynthetic bacteria strains, marine nitrobacteria and vibrio phagostii; the proportion of the rhodotorula benthica, the photosynthetic bacteria strain, the marine nitrobacteria and the vibrio phagostii is (20-30) g: (100-200) ml: (10-50) g: (1-3) g; the density of the photosynthetic bacteria strain is 200-500 hundred million/ml.
2. The algal symbiotic bacteria composition of claim 1, wherein the ratio of the marine rhodotorula benthica, the photosynthetic bacteria species, the marine nitrobacteria and the vibrio phagostii is (20-25) g: 150 ml: (17-30) g: (1-2) g.
3. An algae-bacteria co-culture system is characterized by comprising an algae symbiotic bacteria composition, a microalgae culture medium and microalgae; the algae symbiotic bacteria composition comprises rhodotorula benthica, photosynthetic bacteria strains, marine nitrobacteria and vibrio phagostii; the proportion of the rhodotorula benthica, the photosynthetic bacteria strain, the marine nitrobacteria and the vibrio phagostii is (20-30) g: (100-200) ml: (10-50) g: (1-3) g; the density of the photosynthetic bacteria strain is 200-500 hundred million/ml.
4. The phycomycete co-cultivation system according to claim 3, wherein the formulation of the microalgal culture medium comprises:
5. the phycomycete co-cultivation system according to claim 4, wherein the tween is tween 80; the vitamins include vitamin B12, vitamin B1 and biotin; the trace elements include Na2EDTA、FeCl3、CuSO4、ZnSO4、CoCl2、MnC12With Na2MoO4。
6. The phycomycete co-cultivation system according to claim 3, wherein the concentration of rhodotorula hai in the phycomycete co-cultivation system is 20 to 30 g/L; the concentration of the photosynthetic bacteria strain in the algae bacteria co-culture system is 100-200 ml/L; the concentration of marine nitrifying bacteria in the algae bacterium co-culture system is 10-50 g/L; the concentration of vibrio phagostii in the phycomycete co-culture system is 1-3 g/L.
7. The phycomycete co-cultivation system according to claim 3, wherein the inoculation density of the microalgae in the phycomycete co-cultivation system is 50 to 100 ten thousand/ml.
8. A method for culturing microalgae, comprising:
sterilizing and cooling a microalgae culture medium, inoculating an algae symbiotic bacteria composition and microalgae, and culturing;
the algae symbiotic bacteria composition comprises rhodotorula benthica, photosynthetic bacteria strains, marine nitrobacteria and vibrio phagostii; the proportion of the rhodotorula benthica, the photosynthetic bacteria strain, the marine nitrobacteria and the vibrio phagostii is (20-30) g: (100-200) ml: (10-50) g: (1-3) g; the density of the photosynthetic bacteria strain is 200-500 hundred million/ml.
9. The method according to claim 8, wherein the temperature of the culture is 30 to 35 ℃; the culture time is 48-96 h; the illumination intensity of the culture is 3000-12000 lux;
the microalgae is one or more selected from Chlorella salina, diatom, Nannochloropsis oculata and oocyst algae.
10. The culture method according to claim 8, wherein a mixed gas of carbon dioxide and oxygen is introduced during the culture; the ventilation volume of the mixed gas is 1-5 vvm; the volume ratio of carbon dioxide to oxygen in the mixed gas is 2: 1.
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