Production of astaxanthin and use thereof

Abstract

To provide a method for culturing Haematococcus that allows Haematococcus to survive and produce astaxanthin even in environments with high salinity concentrations similar to seawater, to provide a method for producing astaxanthin, aquaculture feed, and a method for culturing marine animals.SOLUTION: Provided are: a method for culturing Haematococcus including a salinity acclimation step of gradually increasing the salinity concentration of a culture medium from 0% to 3.5% in a method for culturing Haematococcus in which Haematococcus is cultured in the culture medium to produce astaxanthin; a method for producing astaxanthin; feed for fish culture using astaxanthin obtained by these methods; and a method for culturing marine animals.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention provides an astaxanthin-producing method for efficiently producing astaxanthin using Haematococcus. The present invention relates to the production and use of lactic acid bacteria. and method for producing astaxanthin, and astaxanthin-accumulating Haematococcus - Patent application The present invention relates to aquaculture feed containing the Haematococcus, and a method for cultivating marine animals using the Haematococcus. [Background technology]

[0002] Astaxanthin, a carotenoid antioxidant, has a high Astaxanthin has antioxidant properties, and is therefore used in health foods, cosmetics, and farmed fish. Conventionally, astaxanthin has been produced by a method using freshwater The method used is to cultivate the algae Haematococcus. When exposed to external stresses such as nitrogen restriction and salinity (e.g., sodium chloride), the shape The enzyme changes to produce astaxanthin.

[0003] For example, Patent Document 1 describes how Haematococcus can be cultivated outdoors and absorb strong sunlight. stress to the Haematococcus, or by adding 0.3-0.4% sodium chloride, etc. It has been reported that Haematococcus encysts when the salt concentration increases due to heating. In addition, Patent Document 2 describes the cultivation of lactic acid bacteria by adding sodium chloride to the culture medium in an outdoor culture pond. , vegetative cells become dormant (i.e., encystment) and astaxanthin accumulates inside the cells. It has been reported to induce

[0004] However, none of the above documents mentions the use of Haematococcus in a medium containing the same salinity as seawater or in a medium containing chloride. There is no description of culturing in sodium concentrations (approximately 3.5%). Haematococcus, a type of phytoplankton, dies when exposed to high salinity levels similar to those of seawater. Therefore, when Haematococcus is stimulated by salt, it becomes encysted. Although it is known that astaxanthin can be produced, it is difficult to cultivate it in high salinity conditions similar to seawater. It was impossible to do so. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Japanese Patent Application Laid-Open No. 2000-60532 [Patent Document 2] International Publication No. 2005 / 116238 Summary of the Invention [Problem to be solved by the invention]

[0006] The present invention reduces the lethality of Haematococcus and kills it even in a high-salinity environment similar to seawater. Haematococcus can be cultivated without the use of fermentation, resulting in high yields of astaxanthin. Cultivation method, astaxanthin production method, and astaxanthin obtained by these methods The present invention aims to provide aquaculture feed containing the compound and a method for cultivating marine animals. [Means for solving the problem]

[0007] The present invention relates to a method for producing astaxanthin by culturing Haematococcus in a culture medium. In a method for culturing cocci, the salt concentration of the culture medium is gradually increased from 0% to 3.5%. A method for culturing Haematococcus is provided, which includes an increased salinity acclimation step.

[0008] The present invention also provides a method for culturing Haematococcus in a culture medium to induce astaxanthin production in Haematococcus. In a method for producing astaxanthin by cultivating astaxanthin, the salt concentration of the culture solution is adjusted to an initial concentration The salinity sequence consists of N stages (N = an integer between 3 and 10) that increase in stages from 0.05 to 3.5%. The present invention provides a method for producing astaxanthin, which comprises culturing Haematococcus by a method including a reaction step. This is what is done.

[0009] Furthermore, the present invention relates to the above-mentioned method for culturing Haematococcus or method for producing astaxanthin. The water containing the Haematococcus that has accumulated the astaxanthin obtained by the above method in its cells. It provides feed for aquaculture.

[0010] The present invention provides a method for culturing Haematococcus or a method for producing astaxanthin, as described above. The resulting Haematococcus bacteria that had accumulated astaxanthin in their cells were then used as zooplankton. The present invention provides a method for cultivating marine animals, the method comprising: [Effects of the Invention]

[0011] The present invention reduces the lethality of Haematococcus even under high salinity conditions similar to those of seawater, and Survival (survival) as intermediate cells or encysted cells (cysts) that produce staxanthin This makes it possible to obtain astaxanthin in high yield. This allows freshwater Haematococcus to be cultivated without dying even in high salinity concentrations equivalent to those of seawater. It is now possible to release Haematococcus bacteria, which accumulate astaxanthin in abundance, into seawater and use them for aquaculture. Food for zooplankton such as rotifers, copepods, and Artemia, which are food for aquaculture Furthermore, according to the present invention, it is possible to develop a hemolymph that accumulates astaxanthin in large amounts. By feeding Matcoccus to rotifers, copepods, Artemia and other zooplankton, This provides a method for cultivating marine animals (mainly fish and crustaceans) by Increasing fish survival rates can maximize adult aquaculture production. [Brief explanation of the drawings]

[0012] [Figure 1] 1 is a table summarizing the culture conditions for Examples 1 to 3, Comparative Example 1, and Reference Examples 1 to 4. The numbers in the figure represent salt concentrations (%). [Figure 2] 1 is a graph showing the cell density, encystment rate, astaxanthin content, and astaxanthin yield of Examples 1 to 3. In each Example, the bar graphs correspond, from left to right, to the cell density, encystment rate, astaxanthin content, and astaxanthin yield. [Figure 3] 1 is a graph showing the cell densities of motile cells (VC: vegetative cells), immotile cells (GCC: green coccoid cells), intermediate cells (IC: intermediate cells), and encysted cells (cysts) on the final day of culture in Reference Examples 1 to 4. [Figure 4] 1 is a table summarizing the culture conditions of Comparative Example 1 and Examples 1 to 3. DETAILED DESCRIPTION OF THE INVENTION

[0013] In this specification, "salt" refers to "sodium chloride" as the main component, but also includes "other salts" Examples of "other salts" include potassium chloride and magnesium chloride. In this specification, "salt concentration" refers to the total amount of salt. If the salt is sodium chloride, the concentration of sodium chloride is compared to the salt content of sodium chloride and "other salts" " indicates the total salt concentration. Unless otherwise specified, percentages are expressed by weight. Furthermore, in this specification, the terms "survival" and "viability" of cells are used interchangeably. Both indicate survived.

[0014] <Characteristics of Haematococcus> Haematococcus is an algae that lives in freshwater with almost no salt content. The morphology of the plant changes in response to external stresses such as strong light, nitrogen limitation, high temperature, dryness, and high salinity. The Haematococcus changes (encysts) and produces astaxanthin. This is because Haematococcus eliminates active oxygen produced by stress. This is because it produces astaxanthin, an antioxidant. There are several species, and the species that are easily adapted to the salt acclimation process of the present invention are the Heliotrope species used in the examples. In addition to Matococcus lacustris, Haematococcus pluvialis and Haematococcus -Nivalis is one example.

[0015] Encystment of Haematococcus can take several forms. In the absence of external stress, the vegetative cells have two equal-length flagella. At this stage, cells grow by cell division. When the cells are exposed to the sun, they stop cell division and become immobile cells called green coccoid cells. When Haematococcus is further subjected to external stress, it changes to an asta state. Intermediate cells (Interm) start to produce xanthine and contain small amounts of astaxanthin in their cells. When the cell is subjected to more external stress, it changes into a state called "diate cell." Encysted cells that produce large amounts of astaxanthin and contain large amounts of astaxanthin within the cells It changes into a state called (Cyst).

[0016] <Haematococcus cultivation method> One aspect of the present invention is to produce astaxanthin by culturing Haematococcus in a culture medium. In the method for culturing Haematococcus, the salt concentration of the culture medium is adjusted to 0% to 3.5%. A method for culturing Haematococcus, which includes a step of gradually increasing salinity acclimation.

[0017] The salt acclimation process involves culturing Haematococcus while gradually increasing the salt concentration of the culture solution. The salinity acclimation process involves increasing the salinity from the initial concentration (0%) to the next concentration. The first step is the step where the concentration is increased by one step, and the second step is the step where the concentration is increased by one step. The final concentration was increased to N steps (N = an integer between 3 and 10). The Nth stage refers to the final stage, and the salinity at this stage is set to about the same as that of seawater. The range of about 3.2 to 3.8%, or about 3.5%, .

[0018] Here, "N" is an integer and can be between 3 and 10. "N" is the salinity of each level. It can be set appropriately depending on the degree of increase in concentration. Note that the Nth stage refers to the final stage.

[0019] The salt concentration of the culture medium in the salt acclimation process is, for example, 0.058% to 0. 58%, or 0.29-0.44%, and in the second stage 0.29-2.3%, or 0.4 4 to 1.8%, in the third stage 1.8% to 3.5%, and in the fourth stage 2.8% to 3.5%. The salt concentration can be 3.5% in the Nth stage. Examples include: Example 1: 1st stage 0.29%, 2nd stage 0.44%, 3rd stage 1.8%, 4th stage 3.5% Example 2: 1st stage 0.29%, 2nd stage 1.8%, 3rd stage 3.5% Example 3: 1st stage 0.44%, 2nd stage 1.8%, 3rd stage 3.5%

[0020] The duration of each stage of the salinity acclimation process should be determined based on the degree of increase in salinity and the total number of stages. The duration of each stage can be, for example, 3 to 10 days, 3 to 7 days, Or it can be 3 to 5 days. If the total number of stages is 3 or more, the duration of each stage For example, the duration of the first stage may be 3 to 5 days, and the duration of the second stage may be 3 to 5 days. Phase 2 should last 3-5 days, Phase 3 should last 3-5 days, and Phase 4 should last 3-10 days. In particular, by setting the duration of the first stage to 3 days or more, it is possible to increase the number of surviving cells. The duration of each stage of the salinity acclimation process can be specified as follows: Examples include: Example 1: Phase 1: 3 days, Phase 2: 3 days, Phase 3: 5 days, Phase 4: 5 days Example 2: Phase 1: 3 days, Phase 2: 5 days, Phase 3: 8 days

[0021] The overall duration of the salt acclimation process varies depending on the degree of increase in salinity, the total number of stages, and the yield of astaxanthin. The total period can be set in accordance with the rate, etc. Specifically, the total period can be set to, for example, 5 days to 30 days. The period of time can be 10 days, 10 to 25 days, or 14 to 20 days. Suitable periods include 16 or 20 days.

[0022] In the method for culturing Haematococcus of the present invention, the conditions other than the salt adaptation step are the same as those conventionally known. The method can be used.

[0023] Haematococcus that survived the Haematococcus culture method of the present invention were able to grow in a high-salt-concentration stock. At least some of them have been transformed into intermediate somatic or encysted cells by stress. And half to most (70% or more than 80%) of the surviving Haematococcus They are characterized as intermediate somatic cells or encysted cells. In particular, encysted cells produce a large amount of astaxanthin, and therefore, the hematococcal cells of the present invention Astaxanthin can be efficiently obtained by the sludge culture method. Haematococcus is a phytoplankton-derived algae, but it survives through the Haematococcus culture method of the present invention. The resulting Haematococcus can survive in high-salinity environments such as seawater. .

[0024] <Astaxanthin manufacturing method> Another aspect of the present invention is to culture Haematococcus in a culture medium to induce asta In a method for producing astaxanthin by producing xanthin, the salt concentration of the culture solution is , a method for culturing Haematococcus including a step of adapting to a salt concentration that is gradually increased from 0% to 3.5% This is a method for producing astaxanthin by cultivating Haematococcus. The cultivation and salinity adaptation steps are as described above.

[0025] By the method for producing astaxanthin of the present invention, it is possible to produce viable intermediate cells of Haematococcus or Astaxanthin can be obtained in high yields from encysted cells. However, the method for producing astaxanthin of the present invention can produce It contains a large amount of astaxanthin in its cells, allowing it to survive in high-salinity environments such as water. Accumulated Haematococcus can be obtained.

[0026] <Aquaculture feed> Another aspect of the present invention is the above-mentioned method for culturing Haematococcus or the method for producing astaxanthin. The astaxanthin obtained by the method was accumulated in the cells of the Haematococcus. The feed for aquaculture is a feed for aquaculture that is produced by the method for culturing Haematococcus of the present invention. or astaxanthin-accumulating hematopoietic cells obtained by the astaxanthin production method. It may contain tococcus as a main ingredient or one of the ingredients, or as an additive. Good too.

[0027] Haematococcus that has accumulated astaxanthin in its cells can grow at high temperatures, similar to seawater. These Haematococcus species can survive in high salinity environments. It can be used as feed for aquaculture (marine animal farming). Haematococcus that has accumulated ATP is released into aquaculture farms to feed on zooplankton such as rotifers. Furthermore, the zooplankton that have ingested Haematococcus are eaten by larval fish, It is a useful aquaculture feed rich in astaxanthin.

[0028] <Marine animal farming method> Another aspect of the present invention is the above-mentioned method for culturing Haematococcus or the method for producing astaxanthin. The Haematococcus that has accumulated astaxanthin in its cells obtained by the method is then subjected to animal experiments. A method of marine animal farming that involves feeding plankton.

[0029] Haematococcus that has accumulated astaxanthin in its cells is then used to infect animals such as rotifers. By feeding plankton to marine animals (mainly fish and crustaceans), larvae and fry are cultivated. Increasing survival rates of adult aquaculture fish can maximize adult aquaculture production. [Example]

[0030] The present invention will be described below with reference to specific embodiments, but the present invention is not limited to these embodiments. It is understood that various changes and modifications therein will occur to those skilled in the art without departing from the spirit and scope of the appended claims. may be practiced without departing from the scope or spirit of the invention as defined in the appended claims. It is understood.

[0031] (Measurement of cell density and cell composition of microalgae) A predetermined amount of culture medium is taken and fixed with glutaraldehyde (final concentration 2%). A portion of the sample was transferred to a hemocytometer, and the number of cells and the state of the cells were counted using a biological microscope. The cell density was calculated using the obtained cell count according to the following formula. The cell composition was determined from the observed cell state. (Cell density cell / mL) = (Number of counted cells cell) ÷ (Volume of observation area mL )

[0032] (Measurement of encystment rate) In the Examples, Comparative Examples, and Reference Examples, "encystment" refers to intermediate somatic cells or encystment. This indicates that the cells have been transformed into cysts. The encystment rate was calculated as follows. (Encyst rate%) = (number of intermediate cells + number of encysted cells) / (number of counted cells) cell) x 100

[0033] (Measurement of dry weight of microalgae) A predetermined amount of culture medium was collected and placed in a GF / F glass tube that had been preheated in a muffle furnace at 550°C for 2 hours. The particles were collected on a fiber filter paper (Whatman), and the filter paper was transferred to a thermostatic oven at 60°C and dried for 24 hours. The filter paper was then dried for a period of time, and the weight (including ash) was measured using a precision balance. The cells were transferred to a furnace and heated at 550°C for 4 hours, after which the weight was measured and used as the ash weight. The dry weight was calculated using the following formula: (Dry weight mg) = (Weight with ash mg) - (Ash weight mg)

[0034] (Astaxanthin measurement) Five milliliters of culture medium was collected on a GF / F glass fiber filter (Whatman), and 5 1 mL of N,N-dimethylformamide (DMF) and leave it at 4°C in the dark for 48 hours. After extraction, the DMF was passed through a 0.2 μm PTFE silica gel filter. 0.5 mL or more was filtered through a filter (Millipore) and analyzed by LC-MS (Wate The analysis was performed using Acquity UPLC.

[0035] [Example 1] (Pre-culture: Proliferation of cells used for adaptation culture) Haematococcus lacus, which produces astaxanthin, a xanthophyll The Tris strain NIES-144 was used. A 4-gallon, sealed polycarbonate bottle (N C medium with the following composition was placed in a container (algene) and inoculated with the NEIS-144 strain. Using an artificial climate machine, the light intensity was 100 μmol / m2 / s, the light / dark cycle was 12 hours, and the culture temperature was The culture was carried out at 25°C for 10 days while aerating air at 0.1 L / min.

[0036] [Table 1]

[0037] (Cell encystment due to salt stress using an acclimation process) The culture medium in which the NIES-144 strain was grown by pre-cultivation was centrifuged, the supernatant was discarded, and Concentrated NIES was placed in a 500 mL Erlenmeyer flask with 300 mL of C medium. The NaC strain was inoculated at an initial concentration of 1.0 × 105 cells / mL. 1 was added to the medium to adjust the NaCl concentration to 0.29%, and the medium was cultured for 3 days (first stage). After that, fresh NaCl was added to the medium to raise the NaCl concentration to 0.44%, and the medium was incubated for another 3 days. Then, NaCl was added and the mixture was cultured at a NaCl concentration of 1.8% for 5 min. After that, NaCl was added to 3.5% and cultured for 5 days (stage 3). The culture was carried out for a total of 4 stages, 16 days in total. The incubation was carried out under the same light, temperature, and aeration conditions as the pre-culture. After cultivation, the NIES-144 strain changed from green at the initial stage to red, indicating encysted cells. It was confirmed that encysted NIES-144 strain produced 0. The astaxanthin content was 0.041% (w / w), and the astaxanthin content was 0.041% (w / w) at a 3.5% NaCl concentration. A cell density of 5.7 × 104 cells / mL was obtained, and 76% of the cells were translocated into the cytoplasm. The obtained astaxanthin yield was 0.87 mg / L per culture solution. Ta.

[0038] [Comparative Example 1] NIES-1 was cultured in the same manner as in Example 1, except that the NaCl concentration on the first day of culture was 3.5%. The 44 strains were cultured for 16 days. The results after the culture are shown below together with those of Example 1.

[0039] [Table 2]

[0040] From Table 2, when microalgae (Haematococcus) were cultured while being acclimatized, 3.5% Na They can survive in environments where survival is difficult (high salinity), such as Cl, and maintain a relatively high cell density. In addition, environmental stress (high salinity) causes the cells to encyst and the astaxanthin On the other hand, in Comparative Example 1, more than 90% of the cells died within a few days of cultivation. Destroyed.

[0041] [Example 2] (Encyst formation due to adaptation to a sudden increase in salinity) Compared to Example 1, a more rapid increase in salinity was investigated in the acclimation culture. The NIES-144 strain was grown under the conditions of medium, initial cell density, light, water temperature, and aeration, with initial NaCl After culturing for 3 days at a concentration of 0.29% (first stage), the NaCl concentration was increased to 1.8% and the incubation time was 5 days. The NaCl concentration was then increased to 3.5% and the culture was continued for 8 days (stage 3). The results are shown below.

[0042] [Table 3]

[0043] From Table 3, in the adaptation culture with a rapid increase in concentration from 0.29% to 1.8%, The cells showed a higher cell density, i.e., a higher survival rate, compared to the case where no reaction was performed (Comparative Example 1). The cell density was lower than that in Example 1, where the concentration was increased relatively slowly in four steps. The encystment rate and astaxanthin content were also lower than those in Example 1. These results indicate that the cells cannot adapt quickly to sudden changes in concentration, and It can be seen that there is a range (degree) of increase in concentration that is specific to each type of serotonin.

[0044] [Example 3] (Cell encystment by adaptive culture with high initial concentration) The initial concentration was increased compared to Example 2, and adaptive culture was carried out with a gradual increase in concentration. Under the same culture conditions as in Example 1, the NIES-144 strain was grown at an initial NaCl concentration of 0.44%. After culturing for 3 days (first stage), the NaCl concentration was increased to 1.8% and the culture was continued for 5 days (second stage). The NaCl concentration was then increased to 3.5% and cultured for 8 days (third stage) (a total of 3 The results are shown below.

[0045] [Table 4]

[0046] From Table 4, in the three-stage adaptation with an initial concentration of 0.44%, the three-stage adaptation with an initial concentration of 0.29% The cell density and encystment rate were higher than those of the acclimated strain (Example 2). Although the results were comparable, the astaxanthin content per dry weight was lower. Since the concentration was lower than in Example 1, the astaxanthin yield was lower than in Example 1. As shown in the figure, in Example 3, the astaxanthin yield was lower than in Example 1, but Since adaptation culture can be performed in a shorter period than in Example 1, it is suitable for industrial use in terms of shortening the process. It is useful in applications.

[0047] [Reference example 1] (Duration of the first stage (adaptation period to initial concentration) and encystment) Compared to Example 3, by shortening the duration of the first stage (the adaptation period due to the initial concentration), Under the same culture environment as in Example 1, NIES-144 The strain was cultured for 1 day at an initial NaCl concentration of 0.44% (first stage), and then The concentration was increased to 1.8% and cultured for 4 days (second stage) (final concentration 1.8%, 5 days). As a result, the cell density was 5.1×104 cells / mL and the encystment rate was 82%.

[0048] [Reference example 2] Under the same culture environment as in Reference Example 1, the initial NaCl concentration was 0.44%, and the culture was continued for 3 days (first stage). ), and then the NaCl concentration was increased to 1.8% and cultured for 2 days (second stage) (final concentration 1 0.8%, 5 days). The results are shown below together with Reference Example 1.

[0049] [Table 5]

[0050] From Table 5, when the duration of the first stage (the adaptation period to the initial concentration) is short, the encystment rate is Although the encystment rate was high, the number of surviving cells was low. If the acclimation period at the initial concentration is shorter than the cell density, acclimation will not be completed in time, resulting in high stress load. It can be seen that the bacteria are exposed to high temperatures (which increases the rate of encystment).

[0051] [Reference example 3] (First stage salinity (initial concentration), duration and encystment) Compared to Reference Example 1, the salt concentration (initial concentration) in the first stage is lowered, and the duration (initial The incubation period was shortened. After culturing the S-144 strain for 1 day (first stage) with an initial NaCl concentration of 0.29%, The NaCl concentration was increased to 0.44% and cultured for 1 day (second stage), and the NaCl concentration was further increased to The concentration was increased to 1.8% and cultured for 5 days (stage 3) (final concentration 1.8%, 7 days). The cell density was significantly reduced to 3.4 × 104 cells / mL.

[0052] [Reference example 4] Under the same culture environment as in Reference Example 3, the initial NaCl concentration was 0.29%, and the culture was continued for 3 days (first stage). ), then the NaCl concentration was increased to 0.44% and cultured for 1 day (second stage), and then Na The Cl concentration was increased to 1.8% and cultured for 3 days (stage 3) (final concentration 1.8%, 7 days). The results are shown below together with those of Reference Example 3.

[0053] [Table 6]

[0054] From Table 6, it can be seen that even at low initial concentrations, an adaptation period of about 3 days is required. An acclimatization period of at least one day proves insufficient. [Industrial Applicability]

[0055] According to the present invention, a hematococcus can be used to efficiently produce astaxanthin in high yields. It is also possible to provide a method for culturing dregs and a method for producing astaxanthin. To cultivate freshwater Haematococcus without killing it even at salinity levels comparable to those of seawater This allows for the development of high-value added aquaculture feeds containing a higher percentage of astaxanthin, and Furthermore, the present invention provides a method for cultivating marine animals such as larvae and juveniles. Increasing the survival rate of adult fish can maximize the production of adult fish in aquaculture.

Claims

1. Haematococcus is cultivated in a culture medium to produce astaxanthin. In the culture method, A salinity adaptation step is included in which the salt concentration of the culture solution is increased stepwise from 0% to 3.5%. , Haematococcus culture method.

2. The salinity acclimation process involves increasing the salinity from 0% to the next concentration in one step as the first step, and The next step is the next step, and the final step is the final concentration. The method according to claim 1, including up to N stages (N=an integer from 3 to 10). The method for culturing Haematococcus described above.

3. 2. The method according to claim 1, wherein the salt concentration in the first stage of the salt acclimation step is 0.058 to 0.58%.

3. The method for culturing Haematococcus according to 2.

4. The method according to any one of claims 1 to 3, wherein the duration of the first stage of the salinity adaptation step is 3 days or more. The Haematococcus culture method described above.

5. 5. The method according to claim 1, wherein the total duration of the salinity adaptation step is between 5 days and 30 days. The method for culturing Haematococcus described above.

6. The Haematococcus according to any one of claims 1 to 5, wherein the salt comprises sodium chloride. Culture method.

7. Haematococcus includes Haematococcus lacustris, Haematococcus pluvialis, and Haematococcus nivalis. A method for culturing Haematococcus according to claim 1.

8. Haematococcus is cultured in a culture medium to produce astaxanthin. In the method for producing astaxanthin, A salinity adaptation step is included in which the salt concentration of the culture solution is increased stepwise from 0% to 3.5%. A method for producing astaxanthin, which comprises culturing Haematococcus by a Haematococcus culturing method.

9. The method for culturing Haematococcus according to any one of claims 1 to 7, or the method according to claim 8. Astaxanthin obtained by the astaxanthin production method of the present invention is accumulated in the cells. An aquaculture feed containing Haematococcus.

10. The method for culturing Haematococcus according to any one of claims 1 to 7, or the method according to claim 8. Astaxanthin obtained by the astaxanthin production method of the present invention is accumulated in the cells. A method for cultivating marine animals, comprising feeding Haematococcus to zooplankton.

Images