CN107326058B - Method for producing astaxanthin using haematococcus pluvialis - Google Patents
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- CN107326058B CN107326058B CN201710719236.4A CN201710719236A CN107326058B CN 107326058 B CN107326058 B CN 107326058B CN 201710719236 A CN201710719236 A CN 201710719236A CN 107326058 B CN107326058 B CN 107326058B
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P23/00—Preparation of compounds containing a cyclohexene ring having an unsaturated side chain containing at least ten carbon atoms bound by conjugated double bonds, e.g. carotenes
-
- 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 relates to the technical field of astaxanthin production, in particular to a method for producing astaxanthin by using haematococcus pluvialis. The method for producing astaxanthin by using haematococcus pluvialis has the following advantages: compared with the existing method for producing astaxanthin in an immersion mode, the method can reduce the using amount of the culture solution, reduce the production operation cost, particularly reduce the dehydration and drying cost of the harvested biomass and the operation cost of a culture system, has lower requirements on the culture systems such as culture equipment and the like, and is easy for industrial production; in the culture process, the method for producing astaxanthin by using haematococcus pluvialis adopts high-intensity illumination to ensure that haematococcus pluvialis biomass and astaxanthin are quickly accumulated together.
Description
Technical Field
The invention relates to the technical field of astaxanthin production, and discloses a method for producing astaxanthin by using haematococcus pluvialis.
Background
Astaxanthin is a red carotenoid and has strong antioxidant ability. The antioxidant capacity of the vitamin E is more than 10 times stronger than that of other carotenoids such as zeaxanthin, lutein, canthaxanthin and the like, and the free radical scavenging capacity of the vitamin E is 38 times stronger than that of carotene and 500 times stronger than that of vitamin E. Therefore, the astaxanthin can obviously reduce the morbidity risk of age-related degenerative diseases and ischemic diseases, can enhance the immune system and reduce the incidence rate of cancers, and has obvious effects on aging resistance, ultraviolet resistance, immunity enhancement, oxidation resistance and inflammation resistance when passing through a blood brain barrier and the like. At present, astaxanthin is widely applied to the industries of nutritional health products, cosmetics, foods and feeds. Currently, astaxanthin is mainly prepared from four sources, namely 1) artificially synthesized astaxanthin; 2) extracting crustacean; 3) producing Phaffia yeast; 4) and (5) producing haematococcus pluvialis. Wherein chemically synthesized astaxanthin occupies a major market share, costing $ 2500 per kilogram, with a worldwide market share estimated to exceed $ 2 billion per year, but is less bioavailable and safer due to the presence of non-natural conformational components in chemically synthesized astaxanthin. Chemically synthesized astaxanthin is therefore banned by the U.S. Food and Drug Administration (FDA) from entering the health care market. Therefore, the production and extraction of natural astaxanthin is an important development direction of the current market. Under appropriate conditions, the astaxanthin content of Haematococcus pluvialis can reach 5% of its dry weight, which is the most widely distributed astaxanthin producer in nature with the highest yield. Currently, Haematococcus pluvialis astaxanthin has been approved by the United states Food and Drug Administration (FDA) for market entry and is recognized by the Dietary Supplement Health and Education Act (DSHEA) as a new dietary ingredient. Therefore, the most market-potential astaxanthin production method at present is the large-scale production of natural astaxanthin by using Haematococcus pluvialis.
Although Haematococcus pluvialis is the best industrial source of astaxanthin, it grows slowly and is easily contaminated by other organisms during the cultivation process, and in order to optimize the industrial production process and increase the yield of astaxanthin, it is necessary to provide an excellent cultivation method for the production of natural astaxanthin so as to increase the industrial yield. Currently, the industrialized haematococcus pluvialis culture system which is put into production adopts two-section liquid immersion culture. Particularly, the culture process is divided into two sections, and weak light (not higher than 200 mu mol) is used in the first section-2s-1) And after the biomass concentration reaches a certain value, the biomass is turned into high-intensity illumination to accumulate the astaxanthin. Carbon dioxide (CO) in view of the submerged growth of microalgae cells2) The microalgae cells can be reached only by the culture solution, so the carbon supplement efficiency of the immersion culture system is low. At the same time, the large volume of culture liquid leads to high operating costs of the submerged culture system: a great deal of energy consumption in the links of stirring circulation, temperature control, ventilation and the like of the culture solution; the culture broth itself, including the large consumption of inorganic nutrients and water. At the same time, in view of in-leachingIn a submerged culture system, microalgae cells are dispersed in a culture solution (the biomass content is less than 1 percent), and a large amount of energy is consumed to dehydrate a harvested suspension (the total operation cost can reach 30 percent) during harvesting. Meanwhile, the culture time is prolonged by the stepwise accumulation of the biomass and the astaxanthin, and the production efficiency is further reduced.
The Chinese literature discloses a method for producing astaxanthin by cultivating haematococcus pluvialis in a one-stage immersion system, namely a method for producing astaxanthin by cultivating haematococcus pluvialis (Chinese patent literature with application number of 02138827. X). While the problems of submerged culture are not solved. The Chinese literature discloses an indirect immersion type culture system, namely a conversion method for culturing haematococcus pluvialis to produce astaxanthin (Chinese patent document with application number of 201210561399.1), although the effect of inducing astaxanthin synthesis is obvious by using the method, the system is stable and reliable in operation. However, this method requires preparation of immobilized cells, is very cumbersome, and is not suitable for large-scale culture. The chinese literature discloses a non-submerged biofilm system, namely, "a semi-solid culture method for industrial production of microalgae" (chinese patent application No. 201010250866. X), in which microalgae are inoculated on a material having water storage capacity, so that microalgae cells can be kept moist and grow without being submerged, the above-described system reduces the amount of culture fluid required for system operation, and reduces the operation cost while increasing the biomass concentration. However, the yield of astaxanthin in the chinese patent document with the application number 201010250866.X is to be improved.
Disclosure of Invention
The invention provides a method for producing astaxanthin by using haematococcus pluvialis, overcomes the defects of the prior art, and can effectively solve the problem that the yield of astaxanthin is to be improved in the existing astaxanthin production method.
The technical scheme of the invention is realized by the following measures: method for producing freshwater shrimps by using haematococcus pluvialisThe method of the element is carried out as follows: inoculating haematococcus pluvialis cells on a solid culture medium, adding a liquid culture medium into haematococcus pluvialis biomass after inoculation, but not submerging the haematococcus pluvialis biomass, wherein the culture process of the haematococcus pluvialis cells is divided into two stages, and illumination intensity of 400 mu mol lm is provided on the surface of the haematococcus pluvialis biomass in the culture process of the two stages-2s-1To 5000 mu mol-2s-1The method comprises the following steps of (1) providing high-intensity illumination, providing a gas phase containing carbon dioxide to haematococcus pluvialis cells, culturing the haematococcus pluvialis by using a liquid culture medium which satisfies the growth of the haematococcus pluvialis in a first stage, simultaneously accumulating biomass and astaxanthin, entering culture in a second stage when the concentration of the astaxanthin stops increasing and/or nitrogen and phosphorus in the liquid culture medium are completely consumed, changing the components of the liquid culture medium and/or increasing ultraviolet light in the second stage, and harvesting the haematococcus pluvialis biomass when the concentration of the astaxanthin in the second stage is not lower than 2.5% of the dry weight of the biomass.
The following is further optimization or/and improvement of the technical scheme of the invention:
the high-intensity illumination refers to sunlight and/or intensity of 400 mu mol-2s-1To 5000 mu mol-2s-1The artificial illumination and/or the intensity of the artificial illumination are not lower than 400 mu mol-2s-1To 5000 mu mol-2s-1The mixed illumination of (1).
The ultraviolet light is light with wavelength shorter than 400nm, and the intensity is not less than 50 μmol-2s-1(ii) a And/or the light source for illumination is a natural and/or artificial light source.
The above-mentioned modification of the composition of the liquid medium means addition of an inducing factor and/or a stress factor to the liquid medium.
The induction factor is sodium acetate, and the concentration of the sodium acetate is 5mM to 500 mM; and/or the induction factor is sodium chloride, and the concentration of the sodium chloride is 0.2-5%.
The stress factor is obtained by removing inorganic nitrogen source and/or inorganic phosphorus source from liquid culture medium.
The gas phase is air or other gas containing carbon dioxide.
The solid culture medium is a nontoxic solid material with a rough surface, and is made of more than one of filter paper, filter membrane, fiber fabric, sponge and plastic; and/or the surface of the solid medium is a plane or a curved surface.
The inoculation refers to a method for placing haematococcus pluvialis cells on the surface of a solid culture medium, and the inoculation method adopts filtration or smearing or spraying or injection.
The harvesting refers to a method for collecting haematococcus pluvialis cells from the surface of a solid culture medium, and the harvesting method adopts a spatula and/or washing.
The method for producing astaxanthin by using haematococcus pluvialis has the following advantages: compared with the existing method for producing astaxanthin in an immersion mode, the method can reduce the using amount of the culture solution, reduce the production operation cost, particularly reduce the dehydration and drying cost of the harvested biomass and the operation cost of a culture system, has lower requirements on the culture systems such as culture equipment and the like, and is easy for industrial production; in the culture process, the method for producing astaxanthin by using haematococcus pluvialis adopts high-intensity illumination to ensure that haematococcus pluvialis biomass and astaxanthin are quickly accumulated together.
Detailed Description
The present invention is not limited by the following examples, and specific embodiments may be determined according to the technical solutions and practical situations of the present invention. The various chemical reagents and chemical articles mentioned in the invention are all the chemical reagents and chemical articles which are well known and commonly used in the prior art, unless otherwise specified; the percentages in the invention are mass percentages unless otherwise specified; the normal temperature and room temperature in the present invention generally mean a temperature of 15 ℃ to 25 ℃, and are generally defined as 25 ℃.
The invention is further described below with reference to the following examples:
examples1: the method for producing astaxanthin by using haematococcus pluvialis is carried out according to the following method: inoculating haematococcus pluvialis cells on a solid culture medium, adding a liquid culture medium into haematococcus pluvialis biomass after inoculation, but not submerging the haematococcus pluvialis biomass, wherein the culture process of the haematococcus pluvialis cells is divided into two stages, and illumination intensity of 400 mu mol lm is provided on the surface of the haematococcus pluvialis biomass in the culture process of the two stages-2s-1To 5000 mu mol-2s-1The method comprises the following steps of (1) providing high-intensity illumination, providing a gas phase containing carbon dioxide to haematococcus pluvialis cells, culturing the haematococcus pluvialis by using a liquid culture medium which satisfies the growth of the haematococcus pluvialis in a first stage, simultaneously accumulating biomass and astaxanthin, entering culture in a second stage when the concentration of the astaxanthin stops increasing and/or nitrogen and phosphorus in the liquid culture medium are completely consumed, changing the components of the liquid culture medium and/or increasing ultraviolet light in the second stage, and harvesting the haematococcus pluvialis biomass when the concentration of the astaxanthin in the second stage is not lower than 2.5% of the dry weight of the biomass.
The liquid culture medium is a conventional liquid culture medium for culturing haematococcus pluvialis. Such as BBM medium and BG-11 medium.
Compared with the existing immersed astaxanthin production method, the method for producing astaxanthin by using haematococcus pluvialis adopts a non-immersed culture method to culture the haematococcus pluvialis, can reduce the using amount of a culture solution, reduce the production operation cost, particularly reduce the cost of dehydration and drying of harvested biomass and the operation cost of a culture system, has lower requirements on culture systems such as culture equipment and the like, and ensures that the production method is easy for industrial production.
In the culture process, the method for producing astaxanthin by using haematococcus pluvialis described in this example adopts high-intensity illumination to enable haematococcus pluvialis biomass and astaxanthin to be rapidly accumulated together, and in the second stage, the synthesis of astaxanthin is further enhanced through liquid medium components and/or ultraviolet light addition. By using the method for producing astaxanthin by using haematococcus pluvialis, the biomass concentration during harvesting can be obviously improved, and the astaxanthin concentration and the astaxanthin yield during harvesting are also improved, namely when the method is used for harvesting, the dry weight of the biomass is about 20-22% of the total weight of the harvested biomass, and when the traditional immersion system is used for producing astaxanthin, the dry weight of the biomass is 1% of the total weight of the harvested biomass; compared with the existing non-submerged culture method (Chinese patent document with application number of 201010250866.X, a semi-solid culture method for industrial production of microalgae), the method for producing astaxanthin by using haematococcus pluvialis in the embodiment can improve the yield of astaxanthin.
Example 2: the method for producing astaxanthin by using haematococcus pluvialis is carried out according to the following method: inoculating haematococcus pluvialis cells on a solid culture medium, adding a liquid culture medium into haematococcus pluvialis biomass after inoculation, but not submerging the haematococcus pluvialis biomass, wherein the culture process of the haematococcus pluvialis cells is divided into two stages, and illumination intensity of 400 mu mol lm is provided on the surface of the haematococcus pluvialis biomass in the culture process of the two stages-2s-1Or 5000 mu mol-2s-1The method comprises the following steps of (1) providing high-intensity illumination, providing a gas phase containing carbon dioxide to haematococcus pluvialis cells, culturing the haematococcus pluvialis by using a liquid culture medium which satisfies the growth of the haematococcus pluvialis in a first stage, simultaneously accumulating biomass and astaxanthin, entering culture in a second stage when the concentration of the astaxanthin stops increasing and/or nitrogen and phosphorus in the liquid culture medium are completely consumed, changing the components of the liquid culture medium and/or increasing ultraviolet light in the second stage, and harvesting the haematococcus pluvialis biomass when the concentration of the astaxanthin in the second stage is not lower than 2.5% of the dry weight of the biomass.
Example 3: as optimization of the above embodiment, the high intensity illumination refers to sunlight and/or intensity of 400 μmolm-2s-1To 5000 mu mol-2s-1The artificial illumination and/or the intensity of the artificial illumination are not lower than 400 mu mol-2s-1To 5000 mu mol-2s-1The mixed illumination of (1).
The source of the high-intensity illumination is wide, and the source of the high-intensity illumination can be flexibly selected according to actual conditions in industrial production.
Example 4: as optimization of the above embodiment, the ultraviolet light refers to light with a wavelength shorter than 400nm, and the intensity of the light is not less than 50 μmol-2s-1(ii) a And/or the light source for illumination is a natural and/or artificial light source.
The setting of the wavelength and the intensity of ultraviolet light can meet the requirement of further strengthening the synthesis of astaxanthin.
Example 5: as an optimization of the above embodiment, changing the composition of the liquid medium means adding an inducing factor and/or a stress factor to the liquid medium.
By adding high-intensity illumination, the requirement for further enhancing the synthesis of astaxanthin can be met.
Example 6: as optimization of the above example 5, the inducing factor is sodium acetate, the concentration of sodium acetate is 5mM to 500 mM; and/or the induction factor is sodium chloride, and the concentration (mass percentage) of the sodium chloride is 0.2-5%.
Example 7: as an optimization of the above example 5, the stress factor is to remove the inorganic nitrogen source and/or inorganic phosphorus source in the liquid medium.
Example 8: as an optimization of the above embodiment, the gas phase is air or other gas containing carbon dioxide.
Example 9: as for optimization of the above embodiment, the solid medium is a non-toxic solid material with a rough surface, and the solid medium is made of one or more of filter paper, filter membrane, fiber fabric, sponge and plastic; and/or the surface of the solid medium is a plane or a curved surface.
Example 10: as an optimization of the above embodiment, inoculation refers to a method for depositing haematococcus pluvialis cells on the surface of a solid medium, and the inoculation method adopts filtration or smearing or spraying or injection.
Example 11: as an optimization of the above examples, harvesting refers to a method that can collect Haematococcus pluvialis cells from the surface of a solid medium, the harvesting method using a spatula and/or a wash.
Example 12: the method for producing astaxanthin by using haematococcus pluvialis is carried out according to the following method: make rain grow into red ballInoculating algae cells on a solid culture medium, adding a liquid culture medium into haematococcus pluvialis biomass after inoculation, but not submerging the haematococcus pluvialis biomass, dividing the culture process of the haematococcus pluvialis cells into two stages, and providing illumination intensity of 400 mu mol on the surface of the haematococcus pluvialis biomass in the culture process of the two stages-2s-1The method comprises the following steps of (1) providing high-intensity illumination, providing a gas phase containing carbon dioxide to haematococcus pluvialis cells, culturing the haematococcus pluvialis by using a liquid culture medium which satisfies the growth of the haematococcus pluvialis in a first stage, simultaneously accumulating biomass and astaxanthin, entering culture in a second stage when the concentration of the astaxanthin stops increasing and/or nitrogen and phosphorus in the liquid culture medium are completely consumed, changing the components of the liquid culture medium and/or increasing ultraviolet light in the second stage, and harvesting the haematococcus pluvialis biomass when the concentration of the astaxanthin in the second stage is not lower than 2.5% of the dry weight of the biomass.
When harvested using the method described in this example, the biomass was approximately 20% dry weight of the total weight harvested.
Example 13: the method for producing astaxanthin by using haematococcus pluvialis is carried out according to the following method: inoculating haematococcus pluvialis cells on a solid culture medium, adding a liquid culture medium into haematococcus pluvialis biomass after inoculation, but not submerging the haematococcus pluvialis biomass, wherein the culture process of the haematococcus pluvialis cells is divided into two stages, and illumination intensity of 5000 mu mol lm is provided on the surface of the haematococcus pluvialis biomass in the culture process of the two stages-2s-1The method comprises the following steps of (1) providing high-intensity illumination, providing a gas phase containing carbon dioxide to haematococcus pluvialis cells, culturing the haematococcus pluvialis by using a liquid culture medium which satisfies the growth of the haematococcus pluvialis in a first stage, simultaneously accumulating biomass and astaxanthin, entering culture in a second stage when the concentration of the astaxanthin stops increasing and/or nitrogen and phosphorus in the liquid culture medium are completely consumed, changing the components of the liquid culture medium and/or increasing ultraviolet light in the second stage, and harvesting the haematococcus pluvialis biomass when the concentration of the astaxanthin in the second stage is not lower than 2.5% of the dry weight of the biomass.
When harvested using the method described in this example, the biomass was approximately 22% dry weight of the total weight harvested.
Example 14: the method for producing astaxanthin by using haematococcus pluvialis is carried out according to the following method: two sides of a glass fiber felt with the height of 1.5m, the width of 1m and the thickness of 1mm are covered by nylon filter membranes with the height of 1.5m and 1m, the thickness of 0.2mm and the pore diameter of 0.2 mu m, and the surface of the nylon filter membrane of the assembled system is hung vertical to the horizontal plane. BG11 culture solution is added into the glass fiber mat from the upper end of the glass fiber mat by an agricultural drip irrigation system, the culture solution is uniformly supplied to the length of 1m by 10 drip irrigation heads, and the total flow rate of the culture solution is 0.5L.min-1,The culture solution had a total volume of 4L and was stored in a vessel having a volume of 4L. Collecting the flowing culture solution at the lower end of the glass fiber felt by a liquid collecting tank, and returning the collected culture solution to the culture solution storage container. 4 culture systems (system nylon filter membranes) are parallelly placed on the ground at an interval of 25cm, and the total occupied area of the system is 1m2. After the nylon filter membrane is completely soaked, the haematococcus pluvialis is inoculated on the surface of the nylon filter membrane in a smearing mode, and the inoculation biomass is about 5g m-2. After inoculation, a sodium light lamp is used for providing illumination for the surface of the inoculated nylon filter membrane, and the light intensity on the surface of the nylon filter membrane is 500 mu mol-2s-1And a 16:8 light-dark cycle (near natural light) was used, with ambient temperatures from 28 ℃ to 30 ℃ during culture. During the culture process, compressed air with carbon dioxide content (volume percentage) of 5% is supplied into the culture solution storage container, and the flow rate is 1L min-1. After ten days of culture, when nitrogen and phosphorus in the culture solution were depleted, sodium chloride was added to the culture solution to a final concentration of 0.4% and sodium acetate was added to a final concentration of 20mM, and 50 μmol m strength was additionally provided to the biomass, as measured-2s-1The culture was continued for 5 days under ultraviolet irradiation.
The results showed that the yield of astaxanthin on the surface of the nylon filter membrane reached 0.75gm from the first day to the tenth day of the culture-2d-1The average yield of the nylon filter membrane is 6gm after being converted into the floor area yield-2d-1Than using PBR (using natural light)The highest record at present (0.12 gm)-2d-1) 50 times higher; by the fifteenth day of culture, the astaxanthin content of the biomass reached 3.2% of the dry weight and the total yield reached 1.16gm-2d-1After the conversion of the nylon filter membrane into the floor area yield, the average total yield of the astaxanthin reaches 8.25gm-2d-1Higher by a factor of 68 than the current highest recording using PBR (using natural light). At the same time, the directly harvested biomass was 22% of the total weight, much greater than the PBR system (less than 1%). Compared with the astaxanthin yield of the existing non-submerged culture method (Chinese patent document with the application number of 201010250866.X, a semi-solid culture method for microalgae industrial production) with a similar scale, the total yield of astaxanthin is improved by 2.75 times by using the culture method provided by the invention.
In summary, the method for producing astaxanthin by using haematococcus pluvialis of the present invention has the following advantages: compared with the existing method for producing astaxanthin in an immersion mode, the method can reduce the using amount of the culture solution, reduce the production operation cost, particularly reduce the dehydration and drying cost of the harvested biomass and the operation cost of a culture system, has lower requirements on the culture systems such as culture equipment and the like, and is easy for industrial production; in the culture process, the method for producing astaxanthin by using haematococcus pluvialis adopts high-intensity illumination to ensure that haematococcus pluvialis biomass and astaxanthin are quickly accumulated together.
The technical characteristics form an embodiment of the invention, which has strong adaptability and implementation effect, and unnecessary technical characteristics can be increased or decreased according to actual needs to meet the requirements of different situations.
Claims (12)
1. A method for producing astaxanthin by using Haematococcus pluvialis is characterized by comprising the following steps: inoculating Haematococcus pluvialis cells on a solid culture medium, and adding a liquid culture medium to Haematococcus pluvialis cells after inoculationThe method is characterized in that the haematococcus pluvialis biomass is not immersed in the material, the culture process of haematococcus pluvialis cells is divided into two stages, and in the culture process of the two stages, illumination intensity of 400 mu mol m & m is provided on the surface of the haematococcus pluvialis biomass-2· s-1To 5000 mu mol. m-2· s-1The haematococcus pluvialis is cultured by using a liquid culture medium meeting the growth of the haematococcus pluvialis in the first stage, biomass and astaxanthin are accumulated at the same time, when the concentration of the astaxanthin stops increasing and/or nitrogen and phosphorus in the liquid culture medium are depleted, the haematococcus pluvialis biomass enters the second stage of culture, and when the concentration of the astaxanthin in the second stage is not lower than 2.5 percent of the dry weight of the biomass, the haematococcus pluvialis biomass is harvested by changing the components of the liquid culture medium and/or increasing ultraviolet light in the second stage; wherein the change of the components of the liquid culture medium refers to the addition of an induction factor and/or a stress factor to the liquid culture medium, the induction factor is sodium acetate and/or sodium chloride, the concentration of the sodium acetate is 5mM to 500mM, the concentration of the sodium chloride is 0.2% to 5%, and the high-intensity illumination refers to sunlight and/or the intensity is 400 [ mu ] mol · m%-2· s-1To 5000 mu mol. m-2· s-1The artificial illumination and/or the intensity of the artificial illumination is not lower than 400 mu mol-m-2· s-1To 5000 mu mol. m-2· s-1The ultraviolet light is light with wavelength shorter than 400nm, and the intensity of the ultraviolet light is not less than 50 mu mol.m-2· s-1The light source for illumination is natural and/or artificial light source.
2. The method for producing astaxanthin using Haematococcus pluvialis as claimed in claim 1, wherein the stress factor is removal of inorganic nitrogen source and/or inorganic phosphorus source from the liquid medium.
3. The method for producing astaxanthin using Haematococcus pluvialis as claimed in claim 1 or 2, wherein the gas phase is air or other gas containing carbon dioxide.
4. The method for producing astaxanthin using Haematococcus pluvialis according to claim 1 or 2, wherein the solid medium is a non-toxic solid material having a rough surface, and the solid medium is one or more of filter paper, filter membrane, fiber fabric, sponge, and plastic; and/or the surface of the solid medium is a plane or a curved surface.
5. The method of producing astaxanthin using Haematococcus pluvialis as claimed in claim 3, wherein the solid medium is a non-toxic solid material having a rough surface, and the solid medium is one or more of filter paper, filter membrane, fiber fabric, sponge, and plastic; and/or the surface of the solid medium is a plane or a curved surface.
6. The method for producing astaxanthin using Haematococcus pluvialis as claimed in claim 1, 2 or 5, wherein the inoculation is a method capable of depositing the Haematococcus pluvialis cells on the surface of the solid medium by filtration or painting or spraying or injection.
7. The method of producing astaxanthin using Haematococcus pluvialis as claimed in claim 3, wherein the inoculation is a method of depositing the Haematococcus pluvialis cells on the surface of the solid medium by filtration or painting or spraying or injection.
8. The method of producing astaxanthin using Haematococcus pluvialis as claimed in claim 4, wherein the inoculation is a method of depositing the Haematococcus pluvialis cells on the surface of the solid medium, and the inoculation is performed by filtration or spreading or spraying or injection.
9. The method for producing astaxanthin using Haematococcus pluvialis as claimed in claim 1, 2, 5, 7 or 8, wherein the harvesting means a method capable of collecting Haematococcus pluvialis cells from the surface of the solid medium, the harvesting method using a spatula and/or a washing.
10. The method of producing astaxanthin using Haematococcus pluvialis as claimed in claim 3, wherein the harvesting means a method capable of collecting Haematococcus pluvialis cells from the surface of the solid medium, the harvesting method using a spatula and/or a wash.
11. The method of producing astaxanthin using Haematococcus pluvialis as claimed in claim 4, wherein the harvesting means a method capable of collecting Haematococcus pluvialis cells from the surface of the solid medium, the harvesting method using a spatula and/or a wash.
12. The method of producing astaxanthin using Haematococcus pluvialis as claimed in claim 6, wherein the harvesting means a method capable of collecting Haematococcus pluvialis cells from the surface of the solid medium, the harvesting method using a spatula and/or a wash.
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| CN109825546B (en) * | 2019-04-08 | 2020-12-18 | 睿藻生物科技（苏州）有限公司 | Method for producing astaxanthin by haematococcus pluvialis |
| CN110283867A (en) * | 2019-05-24 | 2019-09-27 | 华南理工大学 | A method of astaxanthin is produced using Zuo Fuse green alga |
| CN110577898B (en) * | 2019-10-25 | 2023-01-31 | 四川轻化工大学 | Method for producing astaxanthin by culturing haematococcus pluvialis through discontinuous two-step method |
| CN111286462A (en) * | 2020-02-12 | 2020-06-16 | 佛山市鼎科科技发展有限公司 | Astaxanthin microcapsule preparation method based on haematococcus pluvialis culture |
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