CN107326058A - Use the method for Haematococcus pluvialis production astaxanthin - Google Patents
Use the method for Haematococcus pluvialis production astaxanthin Download PDFInfo
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- CN107326058A CN107326058A CN201710719236.4A CN201710719236A CN107326058A CN 107326058 A CN107326058 A CN 107326058A CN 201710719236 A CN201710719236 A CN 201710719236A CN 107326058 A CN107326058 A CN 107326058A
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- haematococcus pluvialis
- astaxanthin
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- JEBFVOLFMLUKLF-IFPLVEIFSA-N Astaxanthin Natural products CC(=C/C=C/C(=C/C=C/C1=C(C)C(=O)C(O)CC1(C)C)/C)C=CC=C(/C)C=CC=C(/C)C=CC2=C(C)C(=O)C(O)CC2(C)C JEBFVOLFMLUKLF-IFPLVEIFSA-N 0.000 title claims abstract description 103
- 239000001168 astaxanthin Substances 0.000 title claims abstract description 103
- 235000013793 astaxanthin Nutrition 0.000 title claims abstract description 103
- MQZIGYBFDRPAKN-ZWAPEEGVSA-N astaxanthin Chemical compound C([C@H](O)C(=O)C=1C)C(C)(C)C=1/C=C/C(/C)=C/C=C/C(/C)=C/C=C/C=C(C)C=CC=C(C)C=CC1=C(C)C(=O)[C@@H](O)CC1(C)C MQZIGYBFDRPAKN-ZWAPEEGVSA-N 0.000 title claims abstract description 103
- 229940022405 astaxanthin Drugs 0.000 title claims abstract description 103
- 241000168517 Haematococcus lacustris Species 0.000 title claims abstract description 101
- 238000000034 method Methods 0.000 title claims abstract description 68
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 66
- 239000002028 Biomass Substances 0.000 claims abstract description 51
- 235000015097 nutrients Nutrition 0.000 claims abstract description 42
- 238000005286 illumination Methods 0.000 claims abstract description 28
- 238000003306 harvesting Methods 0.000 claims abstract description 26
- 238000011534 incubation Methods 0.000 claims abstract description 20
- 238000009825 accumulation Methods 0.000 claims abstract description 13
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 claims description 25
- 239000012530 fluid Substances 0.000 claims description 22
- 239000007787 solid Substances 0.000 claims description 21
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical group [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 16
- 238000011081 inoculation Methods 0.000 claims description 15
- 229910002092 carbon dioxide Inorganic materials 0.000 claims description 13
- 239000001569 carbon dioxide Substances 0.000 claims description 12
- 239000007789 gas Substances 0.000 claims description 12
- 230000001939 inductive effect Effects 0.000 claims description 9
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical group [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 8
- 238000009630 liquid culture Methods 0.000 claims description 8
- 239000001632 sodium acetate Substances 0.000 claims description 8
- 235000017281 sodium acetate Nutrition 0.000 claims description 8
- 239000011780 sodium chloride Substances 0.000 claims description 8
- YUWBVKYVJWNVLE-UHFFFAOYSA-N [N].[P] Chemical compound [N].[P] YUWBVKYVJWNVLE-UHFFFAOYSA-N 0.000 claims description 7
- 239000012528 membrane Substances 0.000 claims description 6
- 230000008859 change Effects 0.000 claims description 5
- 239000004744 fabric Substances 0.000 claims description 3
- 238000011010 flushing procedure Methods 0.000 claims description 3
- BHEPBYXIRTUNPN-UHFFFAOYSA-N hydridophosphorus(.) (triplet) Chemical compound [PH] BHEPBYXIRTUNPN-UHFFFAOYSA-N 0.000 claims description 3
- 231100000252 nontoxic Toxicity 0.000 claims description 3
- 230000003000 nontoxic effect Effects 0.000 claims description 3
- 239000004033 plastic Substances 0.000 claims description 3
- 229920003023 plastic Polymers 0.000 claims description 3
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- 238000002347 injection Methods 0.000 claims description 2
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- 238000005507 spraying Methods 0.000 claims description 2
- 239000000835 fiber Substances 0.000 claims 1
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- 230000008901 benefit Effects 0.000 abstract description 4
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- 239000004677 Nylon Substances 0.000 description 10
- 238000005457 optimization Methods 0.000 description 10
- 238000002386 leaching Methods 0.000 description 8
- 229920001778 nylon Polymers 0.000 description 8
- 239000000306 component Substances 0.000 description 7
- 239000007788 liquid Substances 0.000 description 7
- 238000012549 training Methods 0.000 description 7
- 241000168525 Haematococcus Species 0.000 description 5
- 239000000463 material Substances 0.000 description 5
- 239000001963 growth medium Substances 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
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- 239000000126 substance Substances 0.000 description 3
- 241000238557 Decapoda Species 0.000 description 2
- FDSDTBUPSURDBL-LOFNIBRQSA-N canthaxanthin Chemical compound CC=1C(=O)CCC(C)(C)C=1/C=C/C(/C)=C/C=C/C(/C)=C/C=C/C=C(C)C=CC=C(C)C=CC1=C(C)C(=O)CCC1(C)C FDSDTBUPSURDBL-LOFNIBRQSA-N 0.000 description 2
- 235000021466 carotenoid Nutrition 0.000 description 2
- 150000001747 carotenoids Chemical class 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 238000005728 strengthening Methods 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- ZAKOWWREFLAJOT-UHFFFAOYSA-N DL-alpha-tocopherylacetate Chemical compound CC(=O)OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C ZAKOWWREFLAJOT-UHFFFAOYSA-N 0.000 description 1
- -1 Huang Zhi Chemical class 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 241000081271 Phaffia rhodozyma Species 0.000 description 1
- OOUTWVMJGMVRQF-DOYZGLONSA-N Phoenicoxanthin Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)C(=O)C(O)CC1(C)C)C=CC=C(/C)C=CC2=C(C)C(=O)CCC2(C)C OOUTWVMJGMVRQF-DOYZGLONSA-N 0.000 description 1
- 230000002292 Radical scavenging effect Effects 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 239000012190 activator Substances 0.000 description 1
- 238000013019 agitation Methods 0.000 description 1
- 230000003712 anti-aging effect Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 230000008499 blood brain barrier function Effects 0.000 description 1
- 210000001218 blood-brain barrier Anatomy 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 235000012682 canthaxanthin Nutrition 0.000 description 1
- 239000001659 canthaxanthin Substances 0.000 description 1
- 229940008033 canthaxanthin Drugs 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 230000035567 cellular accumulation Effects 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
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- 238000005265 energy consumption Methods 0.000 description 1
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- 230000002708 enhancing effect Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 235000012041 food component Nutrition 0.000 description 1
- 239000003365 glass fiber Substances 0.000 description 1
- 210000001822 immobilized cell Anatomy 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 230000000302 ischemic effect Effects 0.000 description 1
- 235000012680 lutein Nutrition 0.000 description 1
- 239000001656 lutein Substances 0.000 description 1
- KBPHJBAIARWVSC-RGZFRNHPSA-N lutein Chemical compound C([C@H](O)CC=1C)C(C)(C)C=1\C=C\C(\C)=C\C=C\C(\C)=C\C=C\C=C(/C)\C=C\C=C(/C)\C=C\[C@H]1C(C)=C[C@H](O)CC1(C)C KBPHJBAIARWVSC-RGZFRNHPSA-N 0.000 description 1
- 229960005375 lutein Drugs 0.000 description 1
- ORAKUVXRZWMARG-WZLJTJAWSA-N lutein Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CCCC1(C)C)C=CC=C(/C)C=CC2C(=CC(O)CC2(C)C)C ORAKUVXRZWMARG-WZLJTJAWSA-N 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000012533 medium component Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 208000015122 neurodegenerative disease Diseases 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 238000007790 scraping Methods 0.000 description 1
- 235000015170 shellfish Nutrition 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- KBPHJBAIARWVSC-XQIHNALSSA-N trans-lutein Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CC(O)CC1(C)C)C=CC=C(/C)C=CC2C(=CC(O)CC2(C)C)C KBPHJBAIARWVSC-XQIHNALSSA-N 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 238000009423 ventilation Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- FJHBOVDFOQMZRV-XQIHNALSSA-N xanthophyll Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CC(O)CC1(C)C)C=CC=C(/C)C=CC2C=C(C)C(O)CC2(C)C FJHBOVDFOQMZRV-XQIHNALSSA-N 0.000 description 1
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- 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
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/12—Unicellular algae; Culture media therefor
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Abstract
It is a kind of method of use Haematococcus pluvialis production astaxanthin the present invention relates to production of astaxanthin technical field, in incubation, illumination uses high-intensity illumination.The method of use Haematococcus pluvialis production astaxanthin of the present invention, has following advantages concurrently：For existing immersion production astaxanthin method, the usage amount of nutrient solution can be reduced, reduce production run cost, cost and the operating cost of culture systems that particularly biomass is dehydrated after reduction harvest, and the requirement to culture systems such as culture devices is relatively low, this production method is easy to industrialized production；In incubation, the method of use Haematococcus pluvialis production astaxanthin of the present invention uses high-intensity illumination, make haematococcus pluvialis biomass and the common Rapid Accumulation of astaxanthin, use the method for use Haematococcus pluvialis production astaxanthin of the present invention, biomass concentration during harvest can be obviously improved, astaxanthin concentration and astaxanthin yield when also just improving harvest.
Description
Technical field
It is a kind of method of use Haematococcus pluvialis production astaxanthin the present invention relates to production of astaxanthin technical field.
Background technology
Astaxanthin is a kind of red carotenoid, with strong oxidation resistance.Its oxidation resistance compares corn
Other carotenoid such as Huang Zhi, lutein, canthaxanthin are strong more than 10 times, and on radical scavenging activity, it is stronger than carrotene
It is 38 times, stronger than vitamin E 500 times.Therefore, astaxanthin can significantly reduce age-related degenerative disease and ischaemic
Property disease onset risk, while immune system can also be strengthened, cancer incidence is reduced, in anti-aging, uvioresistant, enhancing
Immunity, pass through in terms of the anti-oxidant anti-inflammatory of blood-brain barrier and also have remarkable effect.At present, astaxanthin is widely used to nutrition
Health products, cosmetics, food and feedstuff industry.Current astaxanthin mainly has four kinds of sources:1) it is artificial synthesized；2) shellfish
Extract；3) phaffia rhodozyma is produced；4) Haematococcus pluvialis production.The astaxanthin that wherein chemical synthesis is obtained occupies main market
Share, price reaches 2500 dollars of per kilogram, and the annual global market share is estimated to exceed 200,000,000 dollars, but due to chemistry conjunction
Into astaxanthin in there is non-native conformation composition, cause it that there is relatively low bioavailability and security.Therefore chemistry is closed
Into astaxanthin health-product market is prohibited from entering by food and medicine Surveillance Authority of the U.S. (FDA).So, to natural astaxanthin
Production and extraction are the important development directions of Vehicles Collected from Market.It is most extensive as being distributed in nature, yield highest astaxanthin
The producer, under suitable conditions, the content astaxanthin of haematococcus pluvialis is up to the 5% of its dry weight.At present, haematococcus pluvialis shrimp is blue or green
Element enters market by food and medicine Surveillance Authority of the U.S. (FDA) approval, and by dietary supplements health and Education Act
(DSHEA) it is recognized as a kind of new dietary ingredient.Therefore, the Production method of astaxanthin of most market potential is to utilize rain life at present
Haematococcus mass produces natural astaxanthin.
Although haematococcus pluvialis are astaxanthin industrial production sources best at present, it is slow-growing, in incubation
Easily polluted by other biological, in order to optimize industrial processes, improve the yield of astaxanthin, it is necessary to be the life of natural astaxanthin
Production provides excellent cultural method, to improve industrial output.The current industrialization haematococcus pluvialis training system that production has been put into
System uses two-part liquid submersion culture.Incubation is specifically divided into two sections, weak light is used in first paragraph（No
Higher than 200 μm olm-2s-1）To carry out the accumulation of biomass, switch to high-intensity illumination after biomass concentration reaches certain value, with
Carry out the accumulation of astaxanthin.In view of the characteristics of microalgae cell submergence growth, carbon dioxide（CO2）It need to be got to by nutrient solution
Microalgae cell, therefore the benefit carbon efficiencies of immersion culture systems are relatively low.Meanwhile, the nutrient solution of large volume result in immersion culture
The high operating cost of system：Mass energy consumption in the links such as agitation cycle, temperature control, the ventilation of nutrient solution；Nutrient solution in itself,
A large amount of consumption including inorganic nutrients and water.Meanwhile, in view of in immersion culture systems, microalgae cell is scattered in nutrient solution
（Biomass content is less than 1%）, need to expend substantial amounts of energy during harvest the suspension of harvest be dehydrated（Up to total operation
The 30% of expense）.The substep accumulation of biomass and astaxanthin extends incubation time simultaneously, reduce further production efficiency.
Chinese document discloses a kind of method of one-part form immersion system culture Haematococcus pluvialis production astaxanthin, i.e., " one
The method for planting culture Haematococcus pluvialis production astaxanthin "（Application No. 02138827.X Chinese patent literature）Though, the method
It is relatively simple using a step cultivation, but only induced by carbon dioxide, Cellular Accumulation astaxanthin is slow, and the method harvested is more
Trouble.The problem of immersion culture simultaneously is produced is not resolved.Chinese document discloses a kind of indirect immersion training
The system of supporting, i.e. " a kind of method for transformation for cultivating Haematococcus pluvialis production astaxanthin "（In Application No. 201210561399.1
State's patent document）Although inducing the effect of chemical activators obvious using this method, system operation is relatively stable reliable.But should
Method needs to prepare immobilized cell, very cumbersome, is unsuitable for large-scale culture.Chinese document discloses a kind of non-immersion
Biofilm system, i.e. " a kind of semisolid cultural method produced for microalgae industry metaplasia "（Application No. 201010250866.X
Chinese patent literature）, in this culture systems, microalgae is seeded on the material with water retention, in this way, microalgae cell is not
Needing to be submerged can also keep moistening and growing, and system as described above reduces the Culture liquid measure needed for system operation, carry
Operating cost is reduced while high biomass concentration.But, Application No. 201010250866.X Chinese patent literature
Astaxanthin yield has to be hoisted.
The content of the invention
The invention provides a kind of method of use Haematococcus pluvialis production astaxanthin, overcome above-mentioned prior art no
Foot, it can effectively solve existing Production method of astaxanthin and there is the problem of astaxanthin yield has to be hoisted.
The technical scheme is that realized by following measures：A kind of use Haematococcus pluvialis production astaxanthin
Method, is carried out as follows：Haematococcus pluvialis cell is seeded on solid medium, fluid nutrient medium is added after inoculation
Into haematococcus pluvialis biomass, but haematococcus pluvialis biomass is not submerged, the incubation of haematococcus pluvialis cell is divided into
In two stages, in the incubation in two stages, it is 400 μ to provide intensity of illumination in haematococcus pluvialis biological surface
molm-2s-1To 5000 μm of olm-2s-1High-intensity illumination, and to haematococcus pluvialis cell provide the gas containing carbon dioxide
Phase, the first stage is cultivated haematococcus pluvialis using the fluid nutrient medium for meeting haematococcus pluvialis growing, while being given birth to
The accumulation of material and astaxanthin, after the nitrogen phosphorus that the concentration of astaxanthin stops in increase and/or fluid nutrient medium is depleted, enters
Enter the culture of second stage, second stage is by changing Liquid Culture based component and/or increase ultraviolet light, when in second stage
After astaxanthin concentration is not less than the 2.5% of biomass dry weight, haematococcus pluvialis biomass is harvested.
Here is the further optimization and/or improvements to foregoing invention technical scheme：
Above-mentioned high-intensity illumination refers to sunlight and/or intensity is 400 μm of olm-2s-1To 5000 μm of olm-2s-1Artificial lighting and/
Or intensity is not less than 400 μm of olm-2s-1To 5000 μm of olm-2s-1Mixed light shine.
Above-mentioned ultraviolet light refers to that wavelength is shorter than 400nm light, not less than 50 μm olm of its intensity-2s-1；And/or, illumination
Light source be nature and/or artificial light source.
Above-mentioned change Liquid Culture based component is directed to fluid nutrient medium addition inducible factor and/or stress factors.
Above-mentioned inducible factor is sodium acetate, and the concentration of sodium acetate is 5mM to 500mM；And/or, inducible factor is sodium chloride,
The concentration of sodium chloride is 0.2% to 5%.
Above-mentioned stress factors are the inorganic nitrogen-sourced and/or inorganic phosphorous sources in removing fluid nutrient medium.
Above-mentioned gas are mutually air or other gases for containing carbon dioxide.
Above-mentioned solid medium refers to solid material nontoxic and with rough surface, and solid medium is using filter paper, filter
One or more of film, fabric, sponge, plastics；And/or, solid culture primary surface is plane or curved surface.
Above-mentioned inoculation refers to the method that haematococcus pluvialis cell can be placed to solid culture primary surface, and inoculation method is used
Filtering is smeared or spraying or injection.
Above-mentioned harvest refers to can be from the method for solid medium acquisition surface haematococcus pluvialis cell, and harvesting method is using scraping
Shovel and/or flushing.
The method of use Haematococcus pluvialis production astaxanthin of the present invention, has following advantages concurrently：Relative to existing leaching
Do not have for formula production astaxanthin method, the usage amount of nutrient solution can be reduced, production run cost is reduced, harvest is particularly reduced
Biomass is dehydrated afterwards cost and the operating cost of culture systems, and requirement to culture systems such as culture devices compared with
It is low, this production method is easy to industrialized production；In incubation, use Haematococcus pluvialis production shrimp of the present invention is blue or green
The method of element uses high-intensity illumination, makes haematococcus pluvialis biomass and the common Rapid Accumulation of astaxanthin, uses institute of the present invention
The method for the use Haematococcus pluvialis production astaxanthin stated, can be obviously improved biomass concentration during harvest, also just improve
Astaxanthin concentration and astaxanthin yield during harvest.
Embodiment
The present invention is not limited by following embodiments, can technique according to the invention scheme and actual conditions it is specific to determine
Embodiment.Various chemical reagent and chemical article are previously mentioned in the present invention unless otherwise specified, is public in the prior art
Know public chemical reagent and chemical article；Percentage in the present invention is as being mass percent without specified otherwise；This hair
Normal temperature, room temperature in bright refer generally to 15 DEG C to 25 DEG C of temperature, are commonly defined as 25 DEG C.
With reference to embodiment, the invention will be further described：
Embodiment 1：This uses the method for Haematococcus pluvialis production astaxanthin, carries out as follows：By haematococcus pluvialis cell
It is seeded on solid medium, fluid nutrient medium is added in haematococcus pluvialis biomass after inoculation, but not submergence rain life
Haematococcus biomass, the incubation of haematococcus pluvialis cell is divided into two stages, in the incubation in two stages, exists
It is 400 μm of olm that haematococcus pluvialis biological surface, which provides intensity of illumination,-2s-1To 5000 μm of olm-2s-1High-intensity illumination, and
The gas phase containing carbon dioxide is provided to haematococcus pluvialis cell, the first stage uses the liquid for meeting haematococcus pluvialis growing
Culture medium is cultivated haematococcus pluvialis, while the accumulation of biomass and astaxanthin is carried out, when the concentration of astaxanthin stops increasing
Plus and/or fluid nutrient medium in nitrogen phosphorus it is depleted after, into the culture of second stage, second stage by change liquid train
Based component and/or increase ultraviolet light are supported, after the astaxanthin concentration in second stage is not less than the 2.5% of biomass dry weight, to rain
Raw haematococcus biomass is harvested.
Fluid nutrient medium is the conventional liq culture medium for haematococcus pluvialis culture.Such as BBM culture mediums and BG-11 trainings
Support base.
The method of use Haematococcus pluvialis production astaxanthin described in the present embodiment is using non-immersion cultural method to rain
Raw haematococcus is cultivated, and it can reduce the usage amount of nutrient solution for existing immersion production astaxanthin method,
Biomass is dehydrated after reduction production run cost, particularly reduction harvest cost and the operating cost of culture systems, and
And the requirement to culture systems such as culture devices is relatively low, this production method is easy to industrialized production.
In incubation, the method for the use Haematococcus pluvialis production astaxanthin described in the present embodiment uses high intensity
Illumination, makes haematococcus pluvialis biomass and the common Rapid Accumulation of astaxanthin, second stage, by Liquid Culture based component and/or
Increase ultraviolet light, further strengthen the synthesis of astaxanthin.Use the use Haematococcus pluvialis production astaxanthin described in the present embodiment
Method, biomass concentration during harvest can be obviously improved, astaxanthin concentration and astaxanthin production when also just improving harvest
Amount, i.e., when using the present embodiment methods described harvest, biomass dry weight is about harvest gross weight 20% to 22%, uses tradition leaching
When not having formula system production astaxanthin, the 1% of gross weight when its biomass dry weight is harvest；Compared to existing non-immersion culture side
Method（Application No. 201010250866.X Chinese patent literature, a kind of semisolid culture side produced for microalgae industry metaplasia
Method）, the method for the use Haematococcus pluvialis production astaxanthin described in the present embodiment can improve the yield of astaxanthin.
Embodiment 2：This uses the method for Haematococcus pluvialis production astaxanthin, carries out as follows：By haematococcus pluvialis
Cell is seeded on solid medium, and fluid nutrient medium is added in haematococcus pluvialis biomass after inoculation, but does not submerge
Haematococcus pluvialis biomass, the incubation of haematococcus pluvialis cell is divided into two stages, in the incubation in two stages,
It is 400 μm of olm to provide intensity of illumination in haematococcus pluvialis biological surface-2s-1Or 5000 μm of olm-2s-1High-intensity illumination,
And the gas phase containing carbon dioxide is provided to haematococcus pluvialis cell, the first stage is using meeting haematococcus pluvialis growing
Fluid nutrient medium is cultivated haematococcus pluvialis, while the accumulation of biomass and astaxanthin is carried out, when the concentration of astaxanthin is stopped
Only increase and/or fluid nutrient medium in nitrogen phosphorus it is depleted after, into the culture of second stage, second stage is by changing liquid
Body medium component and/or increase ultraviolet light, after the astaxanthin concentration in second stage is not less than the 2.5% of biomass dry weight,
Haematococcus pluvialis biomass is harvested.
Embodiment 3：As the optimization of above-described embodiment, high-intensity illumination refers to sunlight and/or intensity is 400 μm of olm-2s-1To 5000 μm of olm-2s-1Artificial lighting and/or intensity be not less than 400 μm of olm-2s-1To 5000 μm of olm-2s-1Mixed light
According to.
High-intensity illumination source is wide, is easy in industrialized production, according to actual conditions, flexibly selects high-strength light
According to source.
Embodiment 4：As the optimization of above-described embodiment, ultraviolet light refers to that wavelength is shorter than 400nm light, and its intensity is not small
In 50 μm of olm-2s-1；And/or, the light source of illumination is nature and/or artificial light source.
The setting of ultraviolet wavelength and intensity, disclosure satisfy that the synthesis requirement for further strengthening astaxanthin.
Embodiment 5：As the optimization of above-described embodiment, change Liquid Culture based component is directed to fluid nutrient medium addition and lured
Inducement and/or stress factors.
By adding high-intensity illumination, the synthesis requirement for further strengthening astaxanthin disclosure satisfy that.
Embodiment 6：As the optimization of above-described embodiment 5, inducible factor is sodium acetate, the concentration of sodium acetate for 5mM extremely
500mM；And/or, inducible factor is sodium chloride, the concentration of sodium chloride（Mass percent）For 0.2% to 5%.
Embodiment 7：As the optimization of above-described embodiment 5, stress factors for remove inorganic nitrogen-sourced in fluid nutrient medium and/
Or inorganic phosphorous sources.
Embodiment 8：As the optimization of above-described embodiment, gas phase is air or other gases for containing carbon dioxide.
Embodiment 9：As the optimization of above-described embodiment, solid medium refers to solid material nontoxic and with rough surface
Material, solid medium is using one or more of filter paper, filter membrane, fabric, sponge, plastics；And/or, solid culture base table
Face is plane or curved surface.
Embodiment 10：As the optimization of above-described embodiment, inoculation refers to haematococcus pluvialis cell can be placed to solid training
The method for supporting primary surface, inoculation method is using filtering or smears or sprays or injects.
Embodiment 11：As the optimization of above-described embodiment, harvest refers to that red ball can be given birth to from solid medium acquisition surface rain
The method of frustule, harvesting method uses spatula and/or flushing.
Embodiment 12：This uses the method for Haematococcus pluvialis production astaxanthin, carries out as follows：By haematococcus pluvialis
Cell is seeded on solid medium, and fluid nutrient medium is added in haematococcus pluvialis biomass after inoculation, but does not submerge
Haematococcus pluvialis biomass, the incubation of haematococcus pluvialis cell is divided into two stages, in the incubation in two stages,
It is 400 μm of olm to provide intensity of illumination in haematococcus pluvialis biological surface-2s-1High-intensity illumination, and give birth to red ball to rain
Frustule provides the gas phase containing carbon dioxide, and the first stage is using meeting the fluid nutrient medium of haematococcus pluvialis growing to rain
Raw haematococcus is cultivated, while the accumulation of biomass and astaxanthin is carried out, when the concentration of astaxanthin stops increase and/or liquid
After nitrogen phosphorus in culture medium is depleted, into the culture of second stage, second stage by change Liquid Culture based component and/
Or increase ultraviolet light, after the astaxanthin concentration in second stage is not less than the 2.5% of biomass dry weight, haematococcus pluvialis are given birth to
Material is harvested.
When being harvested using the present embodiment methods described, biomass dry weight is about harvest gross weight 20%.
Embodiment 13：This uses the method for Haematococcus pluvialis production astaxanthin, carries out as follows：By haematococcus pluvialis
Cell is seeded on solid medium, and fluid nutrient medium is added in haematococcus pluvialis biomass after inoculation, but does not submerge
Haematococcus pluvialis biomass, the incubation of haematococcus pluvialis cell is divided into two stages, in the incubation in two stages,
It is 5000 μm of olm to provide intensity of illumination in haematococcus pluvialis biological surface-2s-1High-intensity illumination, and to rain give birth to it is red
Ball frustule provides the gas phase containing carbon dioxide, and the first stage uses the fluid nutrient medium pair for meeting haematococcus pluvialis growing
Haematococcus pluvialis are cultivated, while the accumulation of biomass and astaxanthin is carried out, when the concentration of astaxanthin stops increase and/or liquid
After nitrogen phosphorus in body culture medium is depleted, into the culture of second stage, second stage is by changing Liquid Culture based component
And/or increase ultraviolet light, after the astaxanthin concentration in second stage is not less than the 2.5% of biomass dry weight, to haematococcus pluvialis
Biomass is harvested.
When being harvested using the present embodiment methods described, biomass dry weight is about harvest gross weight 22%.
Embodiment 14：This uses the method for Haematococcus pluvialis production astaxanthin, carries out as follows：By high 1.5m, width
The two sides for the glass mat that 1m, thickness are 1mm is with high a width of 1.5m and 1m, the nylon that thickness is 0.2mm, aperture is 0.2 μm
Filter membrane is covered, and the system nylon filter membrane surface after assembling is hung perpendicular to horizontal plane.Using agricultural dropleting enemaing system from glass fibre
Felt upper end adds BG11 nutrient solutions into glass mat, and nutrient solution is uniformly supplied in 1m length by 10 drip heads,
Nutrient solution total flow is 0.5L.min-1,Nutrient solution cumulative volume is 4L, and is stored in the container that a volume is 4L.In glass
The lower end of fibrofelt collects the nutrient solution of outflow with collecting tank, and the nutrient solution of collection is flowed back in nutrient solution storage container.By 4
Above-mentioned culture systems（System nylon filter membrane）It is spaced 25cm laid parallels on the ground, the system footprint gross area is 1m2.Treat nylon
After filter membrane soaks completely, haematococcus pluvialis are seeded in the surface of nylon leaching film using the mode of smearing, inoculation biological quality is about
For 5g m-2.Illumination is provided to the nylon leaching film surface of inoculation using sodium lamp after inoculation, the light intensity on nylon leaching film surface is 500 μ
molm-2s-1, and use 16:8 illumination-dark cycle（Close to natural light）, environment temperature is 28 C to 30 C during culture.Training
During supporting, carbon dioxide content is provided into nutrient solution storage container（Percent by volume）For 5% compressed air, flow is
1L min-1.After culture ten days, through measurement, now the nitrogen phosphorus in nutrient solution is depleted, and sodium chloride is added in nutrient solution
It is 0.4% to sodium chloride ultimate density, and it is 20mM to add sodium acetate to sodium acetate ultimate density, and be additionally provided to biomass
Intensity is 50 μm of olm-2s-1Ultraviolet illumination, continue cultivate 5 days.
As a result show, the yield of nylon leaching film surface astaxanthin just reached from first day of culture to the tenth day
0.75gm-2d-1Nylon leaching film, is scaled after floor space yield, average product reaches 6gm-2d-1, than using PBR（Use nature
Light）Current tidemark（0.12gm-2d-1）It is high 50 times；To culture the 15th day, the content astaxanthin of biomass, which has reached, to be done
The 3.2% of weight, total output reaches 1.16gm-2d-1Nylon leaching film, is scaled after floor space yield, the average total output of astaxanthin
Reach 8.25gm-2d-1, than using PBR（Use natural light）Current tidemark it is high 68 times.Meanwhile, the life after directly harvesting
Material dry weight is the 22% of gross weight, is far longer than PBR systems（Less than 1%）.Compared to the existing non-immersion training of similar size
The method of supporting（Application No. 201010250866.X Chinese patent literature, a kind of semisolid training produced for microalgae industry metaplasia
The method of supporting）Astaxanthin yield, using cultural method of the present invention, the total output of astaxanthin improves 2.75 times.
In summary, the method for use Haematococcus pluvialis production astaxanthin of the present invention, has following advantages concurrently：Relatively
For existing immersion production astaxanthin method, the usage amount of nutrient solution can be reduced, production run cost is reduced, particularly
Biomass is dehydrated after reduction harvest cost and the operating cost of culture systems, and to culture systems such as culture devices
It is required that it is relatively low, this production method is easy to industrialized production；In incubation, use haematococcus pluvialis of the present invention are given birth to
The method for producing astaxanthin uses high-intensity illumination, makes haematococcus pluvialis biomass and the common Rapid Accumulation of astaxanthin, uses this
The method of the described use Haematococcus pluvialis production astaxanthin of invention, can be obviously improved biomass concentration during harvest, also
Astaxanthin concentration and astaxanthin yield when improving harvest.
Above technical characteristic constitutes embodiments of the invention, and it has stronger adaptability and implementation result, can basis
The non-essential technical characteristic of increase and decrease is actually needed, to meet the demand of different situations.
Claims (10)
1. a kind of method of use Haematococcus pluvialis production astaxanthin, it is characterised in that carry out as follows：Rain is given birth into red ball
Frustule is seeded on solid medium, and fluid nutrient medium is added in haematococcus pluvialis biomass after inoculation, but does not soak
Do not have haematococcus pluvialis biomass, the incubation of haematococcus pluvialis cell is divided into two stages, in the incubation in two stages
In, it is 400 μm of olm to provide intensity of illumination in haematococcus pluvialis biological surface-2s-1To 5000 μm of olm-2s-1High-strength light
According to, and the gas phase containing carbon dioxide is provided to haematococcus pluvialis cell, the first stage is given birth to using haematococcus pluvialis are met
Long fluid nutrient medium is cultivated haematococcus pluvialis, while carry out the accumulation of biomass and astaxanthin, dense when astaxanthin
Degree stop increase and/or fluid nutrient medium in nitrogen phosphorus it is depleted after, into the culture of second stage, second stage is by changing
Become Liquid Culture based component and/or increase ultraviolet light, when the astaxanthin concentration in second stage is not less than biomass dry weight
After 2.5%, haematococcus pluvialis biomass is harvested.
2. the method for use Haematococcus pluvialis production astaxanthin according to claim 1, it is characterised in that high-intensity illumination
It is 400 μm of olm to refer to sunlight and/or intensity-2s-1To 5000 μm of olm-2s-1Artificial lighting and/or intensity be not less than 400 μ
molm-2s-1To 5000 μm of olm-2s-1Mixed light shine.
3. the method for use Haematococcus pluvialis production astaxanthin according to claim 1 or 2, it is characterised in that ultraviolet light is
Refer to the light that wavelength is shorter than 400nm, not less than 50 μm olm of its intensity-2s-1；And/or, the light source of illumination is nature and/or artificial
Light source.
4. the method for the use Haematococcus pluvialis production astaxanthin according to claim 1 or 2 or 3, it is characterised in that change
Liquid Culture based component is directed to fluid nutrient medium addition inducible factor and/or stress factors.
5. the method for use Haematococcus pluvialis production astaxanthin according to claim 4, it is characterised in that inducible factor is
Sodium acetate, the concentration of sodium acetate is 5mM to 500mM；And/or, inducible factor is sodium chloride, the concentration of sodium chloride for 0.2% to
5%。
6. the method for the use Haematococcus pluvialis production astaxanthin according to claim 4 or 5, it is characterised in that stress factors
To remove the inorganic nitrogen-sourced and/or inorganic phosphorous sources in fluid nutrient medium.
7. the method for the use Haematococcus pluvialis production astaxanthin according to claim 1 or 2 or 3 or 4 or 5 or 6, its feature
It is air or other gases for containing carbon dioxide to be gas phase.
8. the method for the use Haematococcus pluvialis production astaxanthin according to claim 1 or 2 or 3 or 4 or 5 or 6 or 7, its
It is characterised by that solid medium refers to solid material nontoxic and with rough surface, solid medium uses filter paper, filter membrane, fibre
One or more of dimensional fabric, sponge, plastics；And/or, solid culture primary surface is plane or curved surface.
9. the method for the use Haematococcus pluvialis production astaxanthin according to claim 1 or 2 or 3 or 4 or 5 or 6 or 7 or 8,
It is characterized in that inoculation refers to the method that haematococcus pluvialis cell can be placed to solid culture primary surface, inoculation method was used
Filter is smeared or spraying or injection.
10. use Haematococcus pluvialis production astaxanthin according to claim 1 or 2 or 3 or 4 or 5 or 6 or 7 or 8 or 9
Method, it is characterised in that harvest refers to adopt from the method for solid medium acquisition surface haematococcus pluvialis cell, harvesting method
With spatula and/or flushing.
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| CN110577898A (en) * | 2019-10-25 | 2019-12-17 | 四川轻化工大学 | Method for producing astaxanthin by culturing haematococcus pluvialis through discontinuous two-step method |
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