CN103834567B - A kind of microalgae culture method - Google Patents
A kind of microalgae culture method Download PDFInfo
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- CN103834567B CN103834567B CN201410067347.8A CN201410067347A CN103834567B CN 103834567 B CN103834567 B CN 103834567B CN 201410067347 A CN201410067347 A CN 201410067347A CN 103834567 B CN103834567 B CN 103834567B
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Abstract
The present invention relates to a kind of microalgae culture method, micro-algae Immobilized culture and liquid culture combine by described method, are specifically inoculated on solid material by micro-algae and carry out Immobilized culture, are then inoculated into by the micro-algae through Immobilized culture in liquid and continue to cultivate.Use method of the present invention to cultivate micro-algae and do not need harsh condition; energy consumption is lower; not easily pollute, and highdensity frustule can be produced, can cultivate by solid-liquid the industrial scale production that the method combined realizes the secondary metabolites such as pigment, lipid, hydro carbons and carbohydrate.
Description
Technical field
The present invention relates to microdisk electrode technical field, be specifically related to a kind of microalgae culture method.
Background technology
Micro-algae is the photosynthetic lower hydrobiont having chlorophyll, can carry out putting oxygen, and its structure is simple, and growth and breeding is rapid, high to solar energy utilization ratio, strong to environmental compatibility, is therefore subject to people and more and more payes attention to.Active substance contained by micro-algae and the secondary metabolite of production have important economic worth, at energy field, micro-algae is expected to become the third generation biomass energy raw material after food crop bio-ethanol, Mierocrystalline cellulose bio-ethanol and Lu Sheng crop biofuel; At environmental area, micro-algae has the ability of GHG (Greenhouse Gases) emissions mitigation carbonic acid gas, and has broad application prospects in process life and industrial sewage; At field of food, micro-algae rich in proteins, grease, carotenoid and unsaturated fatty acids, can be used for food or foodstuff additive; At field of medicaments, the research also common reporter of microbiotic new in seeking from micro-algae Biological resources, anticancer and antiviral.
Certain slightly algae can synthesize secondary metabolite in a large number as grease, polysaccharide or carotenoid etc. under specific growth conditions, and the accumulation of secondary metabolite is controlled process, and these features accelerate the commercial development that micro-algae realizes various secondary metabolite.We can utilize change and the breeding technique upgrading of envrionment conditions, realize the efficient accumulation of micro-algae secondary metabolite.
At present to the many employings of micro-algae production secondary metabolite cultivation is liquid culture, first strict control its pH value, temperature and nutritional condition, make it under optimum condition, produce as far as possible many vegetative cells, and then utilize stress conditions to induce vegetative cell to produce secondary metabolite.The cultivation of Haematocoocus Pluvialls abroad more adopts this method, and the company of Hawaii, America adopts the bioreactor closed to carry out liquid cultivation in conjunction with open cultivation pool; Dunaliella salina produces that carotenoid also more adopts this method, green frustule biomass accumulation to a certain extent after, under transferring to high salt height luminous environment, the accumulation of induction frustule β-carotene.
In traditional micro-algae liquid cultivation, a large amount of energy expenditures is in water body operation, stirring, ventilation, carbonic acid gas supply and results, and algae cell density not high (about 1-3g/L); Need condition harsh, necessary strictly controlled environment condition, centre has link to go wrong, and may cause the failure of cell proliferation; If once there is protozoon or assorted algae pollution in water body, administer and more difficultly maybe cannot to administer, can only discard, thus produce a large amount of material resources and human cost.In a word, the problem that prior art existence condition harshness, high, the easy pollution of energy consumption and cell density are low.
Summary of the invention
The present invention is directed to the problem that traditional micro-algae liquid cultivation conditional harshness, high, the easy pollution of energy consumption and cell density are low, a kind of microalgae culture method is provided, use method of the present invention to cultivate micro-algae and do not need harsh condition, energy consumption is lower, not easily pollute, and highdensity frustule can be produced.
Technical scheme of the present invention is as follows:
A kind of microalgae culture method is combined at micro-algae Immobilized culture and liquid culture.
Can be specifically micro-algae is inoculated on solid material and carries out Immobilized culture, then the micro-algae through Immobilized culture is inoculated in liquid and continue to cultivate.
In microalgae culture method of the present invention, solid material is the solid material with wetting ability and/or water-retentivity, such as, in non-woven fabrics, cellulose acetate film, coarse cloth, sponge, bolting silk and cotton strap one or more, or other any solid material that can adhere to micro-algae.The shape of described solid material can be plane, cubic, spherical or linear etc.
In microalgae culture method of the present invention, solid material can be only any one in above-mentioned materials, also can be used in combination, typical but the example combinations of indefiniteness such as cellulose acetate film and non-woven fabrics combine, cellulose acetate film and coarse cloth combination, sponge and bolting silk combination, non-woven fabrics and cotton strap combination, etc.
In microalgae culture method of the present invention, solid material has wetting ability and/or water-retentivity, can keep liquid nutrient medium well, for micro-algae provides the nutritive ingredient of growth needs; Meanwhile, solid material also provides the supporting dielectric of micro algae growth.Owing to adopting Immobilized culture, do not relate to stirring in liquid culture and water body operation etc., therefore energy consumption is lower, and reduces the risk of exogenous pollution.
In microalgae culture method of the present invention, substratum can be any substratum that can be used in microdisk electrode, and the substratum that those skilled in the art can be suitable for according to each micro-algae concrete is selected.Even so, in the embodiment of the present invention, used medium is BG11 substratum.
The concrete formula of BG11 substratum is as shown in table 1:
The formula of table 1BG11 substratum
| Composition | Content |
| NaNO 3 | 1.5g/L |
| K 2HPO 4·3H 2O | 0.04g/L |
| MgSO 4·7H 2O | 0.075g/L |
| CaCl 2·2H 2O | 0.036g/L |
| Citric acid | 0.006g/L |
| FeCl 3·6H 2O | 0.00315g/L |
| Na 2EDTA·2H 2O | 0.00436g/L |
| Na 2CO 3 | 0.02g/L |
| A 5Trace element * | 1ml/L |
Wherein, A
5the formula of trace element is as shown in table 2:
Table 2A
5the formula of trace element
| Composition | Content |
| H 3BO 3 | 2.86g |
| MnCl 2·H 2O | 1.81g |
| ZnSO 4·7H 2O | 0.222g |
| CuSO 4·5H 2O | 0.079g |
| Na 2MoO 4·2H 2O | 0.390g |
| Co(NO 3) 2·6H 2O | 0.0494g |
| Deionized water | 1L |
In microalgae culture method of the present invention, carry out gathering step after micro-algae Immobilized culture, micro-algae density when gathering is 20g-200g/m
2, such as 25g/m
2, 30g/m
2, 50g/m
2, 100g/m
2, 180g/m
2or 190g/m
2.
In microalgae culture method of the present invention, the Initial seeding density micro-algae being inoculated into described solid material can be 5-50g/m
2, such as 10g/m
2, 15g/m
2, 20g/m
2, 25g/m
2, 30g/m
2, 35g/m
2, 45g/m
2, 45g/m
2, 48g/m
2or 50g/m
2.
In microalgae culture method of the present invention, the intensity of illumination of described solid material surface can be 10-600 μm of ol/m
2/ s, such as 15 μm of ol/m
2/ s, 20 μm of ol/m
2/ s, 30 μm of ol/m
2/ s, 40 μm of ol/m
2/ s, 50 μm of ol/m
2/ s, 100 μm of ol/m
2/ s, 150 μm of ol/m
2/ s, 200 μm of ol/m
2/ s, 250 μm of ol/m
2/ s, 300 μm of ol/m
2/ s, 350 μm of ol/m
2/ s, 400 μm of ol/m
2/ s, 450 μm of ol/m
2/ s, 500 μm of ol/m
2/ s, 550 μm of ol/m
2/ s or 580 μm ol/m
2/ s, preferred 15-300 μm ol/m
2/ s, most preferably 50-200 μm of ol/m
2/ s.
In microalgae culture method of the present invention, during micro-algae Immobilized culture, pass into carbonated gas.Wherein, the volume percent of described carbon dioxide in gas can be 0.1-15%, such as 0.2%, 0.5%, 0.8%, 1%, 1.2%, 1.5%, 2%, 3%, 4%, 5%, 8%, 10%, 12% or 14%, and preferred 0.1-5%, most preferably 0.1-2%.
In microalgae culture method of the present invention, the temperature of microdisk electrode can be 15-50 DEG C, such as 15 DEG C, 17 DEG C, 19 DEG C, 21 DEG C, 23 DEG C, 25 DEG C, 27 DEG C, 29 DEG C, 31 DEG C, 33 DEG C, 34 DEG C, 35 DEG C, 38 DEG C, 40 DEG C, 42 DEG C, 45 DEG C, 48 DEG C or 49 DEG C, preferred 20-40 DEG C, most preferably 25-30 DEG C.
In microalgae culture method of the present invention, the time of micro-algae Immobilized culture can be 2-10 days, such as 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days or 10 days, preferred 3-7 days.
In microalgae culture method of the present invention, described liquid culture liquid used can be the micro-algae culture medium of all kinds, concentration or composition, containing the algae liquid of one or more micro-algaes, and water, containing the solution of promotive factor, containing one or more in the solution of the stress-inducing factor.Wherein, described promotive factor is as tethelin, the organic carbon source of the growth promoting effects added in substratum, and the stress-inducing factor is as the NaCl etc. of the high density of adding in substratum.
In microalgae culture method of the present invention, described micro-algae includes but not limited to micro-algae of Cyanophyta, Chlorophyta, Xanthophyta, different whip algae door and rhodophyta.Wherein, described Cyanophyta includes but not limited to unicellular algae and multicellular algae, and such as cellulous filamentous cyanobacteria (Filamentous Cyanobacteria), resistant to elevated temperatures Thermal Cyanophytes are as seat algae (Phormidium); Described Chlorophyta includes but not limited to chlorella (Chlorella), Dunaliella salina (Dunaliella), grid algae (Scenedesmus), Haematocoocus Pluvialls (Haematococcuspluvialis), Chlorococcum (Chlorococcum), discoctaster algae (Pediastrum) and Botryococcus braunii (Botryococcus braunii), preferred grid algae, Haematocoocus Pluvialls and Botryococcus braunii; Described different whip algae door includes but not limited to intend Nannochloropsis oceanica (Nannochloropsis), Phaeodactylum tricornutum (Phaeodactylumtricornutum), boat-shaped algae (Navicula) and Chaetoceros (Chaetoceros); Described rhodophyta includes but not limited to Porphyridium cruentum (Porphyridium) and resistant to elevated temperatures hot spring red algae (Galdieria sulphuraria).
In microalgae culture method of the present invention, the Initial seeding density micro-algae of Immobilized culture being inoculated into liquid can be 0.5-20g/L, such as 0.6g/L, 0.8g/L, 1.2g/L, 1.5g/L, 2g/L, 4g/L, 5g/L, 7g/L, 9g/L, 11g/L, 13g/L, 14g/L, 15g/L, 16g/L, 18g/L or 19g/L, preferred 1-5g/L, more preferably 1-2g/L.
In microalgae culture method of the present invention, during described liquid culture, the intensity of illumination on culture surface can be 100-2500 μm of ol/m
2/ s, such as 150 μm of ol/m
2/ s, 250 μm of ol/m
2/ s, 350 μm of ol/m
2/ s, 500 μm of ol/m
2/ s, 700 μm of ol/m
2/ s, 900 μm of ol/m
2/ s, 1100 μm of ol/m
2/ s, 1200 μm of ol/m
2/ s, 1400 μm of ol/m
2/ s, 1500 μm of ol/m
2/ s, 1700 μm of ol/m
2/ s, 1900 μm of ol/m
2/ s, 2100 μm of ol/m
2/ s, 2300 μm of ol/m
2/ s, 2400 μm of ol/m
2/ s or 2500 μm ol/m
2/ s.
In microalgae culture method of the present invention, the one or more inducing culture that carries out that can adjust when described liquid culture in the concentration of intensity of illumination, temperature, liquid component and concentration, carbonic acid gas and pH value parameters produces secondary metabolite.Wherein, described secondary metabolite include but not limited in pigment, lipid, hydro carbons and carbohydrate one or more, pigment includes but not limited to astaxanthin and/or carotenoid; The time of described inducing culture can be 2-10 days, such as 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days or 10 days, preferred 3-4 days.
In microalgae culture method of the present invention, described cultivation can be carried out at closed, not exclusively closed or open environment.Wherein, the reactor etc. of the transparent materials such as the environment such as glass reactor or polyvinyl chloride (Polyvinyl chloridepolymer, PVC) closed, the glass booth etc. that the environment not exclusively closed such as ventilates, open environment such as raceway pond etc.
Adopt microalgae culture method of the present invention, in the liquid culture stage, in conjunction with the advantage of traditional liquid cultivation, according to light intensity and temperature adjustment frustule concentration and in conjunction with stress conditions such as nitrogen stress and/or phosphorus, interpolation salt and/or organic carbon sources, the synthesis of efficient induction secondary metabolite, the processing ease of induction secondary metabolite realizes, induced efficiency significantly improves.
Beneficial effect of the present invention is: microalgae culture method of the present invention, micro-algae Immobilized culture and liquid culture are combined, Immobilized culture technology is adopted in the microalgae cell multiplicative stage, compare traditional liquid cultivation, energy consumption is low, frustule growth is polluted soon, not easily, light utilising efficiency is high, does not need harsh condition just can produce highdensity frustule; At micro-algae secondary metabolite induction period, adopt liquid culture efficiently can induce the synthesis of secondary metabolite.
Accompanying drawing explanation
Fig. 1 is the change curve of Haematocoocus Pluvialls with incubation time growth frustule biomass of the embodiment of the present invention 1 Immobilized culture.
Embodiment
Below in conjunction with embodiment, embodiment of the present invention are described in detail.It will be understood to those of skill in the art that following examples are only the preferred embodiments of the present invention, so that understand the present invention better, thus should not be considered as limiting scope of the present invention.
Experimental technique in following embodiment, if no special instructions, is ordinary method; Experiment material used, if no special instructions, is and is purchased available from routine biochemistry chemical reagent work.
Embodiment 1: the cultivation of Haematocoocus Pluvialls
Be placed on non-woven fabrics by Haematocoocus Pluvialls (Haematococcus pluvialis) frustule, utilize vacuum pump to remove moisture, frustule is trapped on non-woven fabrics; In bottom glass reactor non-woven fabrics being placed on the 0.3m*0.3m*0.04m that can close cultivation, whole reactor obliquity is that 5%(is horizontal by 4.5 ° of angles) place, reactor top has a plastics filling pipe, for nutrient solution entrance, nutrient solution can be made to continue wetting whole culture system, one nutrient solution outlet is arranged at reactor another section of bottom, passes into and a certain amount ofly contain 0.5% (V/V) CO in reactor
2pressurized gas.Adopt the luminescent lamp of artificial end face irradiation, intensity of illumination is 250 μm of ol/m
2/ s, 24h continuous light; Temperature is about 25 DEG C, and initial inoculation concentration is at 10g/m
2, substratum is BG11 substratum.As shown in Figure 1, under Immobilized culture, the cell proliferation stage biomass accumulation ability of frustule is strong, and cultivate 6 days, the biomass of frustule almost linearly increases, the 6th day time, reach 62g/m
2.By the frustule of above-mentioned Immobilized culture, under above-mentioned same culture condition, be seeded in the BG11 substratum of liquid, adopt the pillar reaction of internal diameter 3cm, high 60cm, initial inoculation concentration controls, for 2g/L, to cultivate 4 days biomasss and reach 6g/L.Utilize two stage culture Haematocoocus Pluvialls 10 days, biomass (calculating with dry weight) grows into 15g from 0.9g, and output reaches 17.6g/m
2/ d.
Embodiment 2: utilize haematococcus pluvialis to produce astaxanthin
Be placed on cellulose acetate film by Haematocoocus Pluvialls (Haematococcus pluvialis) frustule, utilize vacuum pump to remove moisture, frustule is trapped on film; Film is taken off, be positioned on the wetting non-woven fabrics of prior substratum, in bottom the glass reactor that non-woven fabrics is placed on the 0.3m*0.3m*0.04m that can close cultivation, whole reactor obliquity is that 5%(is horizontal by 4.5 ° of angles) place, reactor top has a plastics filling pipe, is nutrient solution entrance, and nutrient solution can be made to continue wetting whole culture system, one nutrient solution outlet is arranged at reactor another section of bottom, passes into and a certain amount ofly contain 0.2% (V/V) CO in reactor
2pressurized gas.Adopt the luminescent lamp of artificial end face irradiation, illumination is 200 μm of ol/m
2/ s, 24h continuous light; Temperature is about 25 DEG C, and initial inoculation concentration is 10g/m
2, within first 3 days, substratum is BG11 substratum, the rear BG11 substratum changing nitrogen stress for 3 days into.Result shows, and Immobilized culture 6 days, every day, average dry weight output was 9.1g/m
2/ d, phototranstormation efficiency (calculation formula: PE=E1*100%/E, wherein E1 and E is the energy of biomass and photon respectively, 1mol photon is containing 217kJ energy, 1g micro algae biomass is containing 21.7kJ energy) be 5.3%, the phototranstormation efficiency (phototranstormation efficiency is 3.46%) being the column reactor of 4cm than internal diameter under same light intensity improves 53%, and content astaxanthin is 1.5% of dry weight.After by immobilization algae kind with a small amount of de-ionised water in the column reactor of 3cm internal diameter, add nitrogen stress BG11 liquid nutrient medium continue induction, Initial seeding density controls at 1g/L, 350 μm of ol/m
2under the lasting 24h light intensity of/s, liquid induces 3 days, and its content astaxanthin is that the 4%(of dry weight adopts Boussiba, 1992 measuring methods), adopt the training method of two step couplings, Haematocoocus Pluvialls is cultivated 9 days content astaxanthins and is reached 4%.
Embodiment 3: the cultivation of grid algae
Be placed on cellulose acetate film by grid algae (Scenedesmus) frustule, utilize vacuum pump to remove moisture, frustule is trapped on film; Film is taken off, be positioned on the wetting coarse cloth of prior substratum, in bottom the glass reactor that this coarse cloth is placed on the 0.3m*0.3m*0.04m that can close cultivation, whole reactor obliquity is that 5%(is horizontal by 4.5 ° of angles) place, reactor top has a plastics filling pipe, is nutrient solution entrance, and nutrient solution can be made to continue wetting whole culture system, one nutrient solution outlet is arranged at reactor another section of bottom, passes into and a certain amount ofly contain 1% (V/V) CO in reactor
2pressurized gas.Adopt the luminescent lamp of artificial end face irradiation, illumination is 250 μm of ol/m
2/ s, 24h continuous light; Temperature is about 25 DEG C, and initial inoculation concentration is 10g/m
2, substratum is BG11 substratum.Result shows, and Immobilized culture 6 days, every day, average dry weight output was 78g/m
2, phototranstormation efficiency is 6.0%, and the phototranstormation efficiency (phototranstormation efficiency is 4%) being the column reactor of 4cm than internal diameter under same light intensity improves 50%, and fat content is 12% of dry weight.To cultivate a small amount of de-ionised water of algae kind after 6 days in the column reactor of 5cm internal diameter, high 60cm, add liquid B G11 substratum, Initial seeding density controls at 0.8g/L, 250 μm of ol/m
2under the lasting 24h light intensity of/s, cultivate 4 days, utilize two stage culture grid algae 10 days, output reaches 38g/m
2/ d, fat content is that the 20%(of dry weight adopts Inna, 2005 measuring methods).
Embodiment 4: the cultivation of Botryococcus braunii
Utilize the PVC reactor that length, width and height are 1.5m*0.5m*0.05m, tiling water-absorbing material (as sponge) in reactor, sponge upper berth one deck 350 object bolting silk, be evenly sprayed on bolting silk by Botryococcus braunii (Botryococcus braunii), inoculum density is 12g/m
2.Under outdoor conditions, by 7%(horizontal by 6.3 ° of angles) obliquity be positioned on the through metal that length, width and height are 1.5m*0.5m*2m, reactor top has a plastics liquid supplementation pipe, nutrient solution can be made to continue wetting whole culture system, one nutrient solution outlet is arranged at reactor another section of bottom, passes into and a certain amount ofly contain 0.8% (V/V) CO in the tank of fluid infusion
2pressurized gas, will the intensity control of algae kind be radiated at 300 μm of ol/m by sun visor
2about/s, temperature is 20-30 DEG C, and substratum is BG11 substratum.Continuous Immobilized culture 7 days, biomass reaches 49g/m
2.Results cultured cells, places it in the plate-type reactor of 50 × 50 × 5cm, and adopt BG11 substratum, initial algae kind inoculum density is 1.5g/L, and now sun average intensity every day is about 400 μm of ol/m
2/ s, proceeds to cultivate, cultivates 4 days.Utilize two stage culture Wild Vitis species 10 days, output reaches 11g/m
2/ d.The 20%(that total hydrocarbon content reaches dry weight adopts Inna, 2005 measuring methods).
Embodiment 5: the cultivation of filamentous cyanobacteria
Utilize the common glass sheet that length and width are 1m × 1m, ordinary filter paper is pasted on two sides, as mending the layer of nutrient solution, after on filter paper, paste one deck printer paper, be spread evenly across on printer paper by filamentous cyanobacteria (Filamentous Cyanobacteria), inoculum density is about 10g/m
2.Under outdoor conditions, the above-mentioned sheet glass scribbling frond is placed in transparent vessel, totally 10, every sheet interval 10cm, there is a plastics liquid supplementation pipe of evenly punching on every sheets top, and can make to continue wetting whole culture system, has the container collecting nutrient solution bottom sheet glass, be placed with liquid circulating pump in container, pass into and a certain amount ofly contain 0.5% (V/V) CO
2pressurized gas, the intensity control of every sheet is at 50-400 μm of ol/m
2between/s, temperature is 25-35 DEG C, and substratum is BG11 substratum.Continuous Immobilized culture 7 days, every day, biomass yield was about 80g/m
2/ d.Results cultured cells, places it in the outdoor raceway pond that level is 0.05m, and adopt BG11 substratum, Initial seeding density is 2.5g/L, and now sun average intensity every day is 450 μm of ol/m
2/ s, liquid culture 3 days, utilize two stage culture filamentous cyanobacteria 10 days, output reaches 90g/m
2/ d.
Embodiment 6: chlorella production lipid
In the good glass booth of ventilation, utilize 10m*1m raceway pond, inoculation chlorella (Chlorellasp.) algae kind, inoculation liquid level is 10cm, and starting point concentration is 0.5g/L, will adhere to the good cotton strap of chlorella (length and width are: 1m*0.8m), vertical hanging is in raceway pond, constantly supplement the algae liquid wetting cloth in raceway pond, one end of cloth has about 5cm to be immersed in raceway pond algae liquid, cloth interval 0.2m.Wherein, raceway pond circular arc sky stays, and places a stirring rake.Be provided with blowdown pipe bottom raceway pond, inside pass into containing 1.5% (V/V) CO
2pressurized gas, now sun average intensity every day is 550 μm of ol/m
2/ s, temperature is 15-25 DEG C, and substratum is BG11 substratum.Cultivate 7 days, on cloth, biomass yield is about 15g/m every day
2/ d, liquid bio volume production amount is 3.1g/m
2/ d, the ultimate production that raceway pond is cultivated 7 days is 18.1g/m
2the 15%(that/d fat content reaches dry weight adopts Inna, 2005 measuring methods).
Applicant states, the present invention illustrates detailed features of the present invention and method detailed by above-described embodiment, but the present invention is not limited to above-mentioned detailed features and method detailed, namely do not mean that the present invention must rely on above-mentioned detailed features and method detailed could be implemented.Person of ordinary skill in the field should understand, any improvement in the present invention, to equivalence replacement and the interpolation of ancillary component, concrete way choice etc. that the present invention selects component, all drops within protection scope of the present invention and open scope.
Claims (13)
1. a microalgae culture method, is characterized in that, is inoculated into by micro-algae on solid material and carries out Immobilized culture, is then inoculated into by the micro-algae through Immobilized culture in liquid and continues to cultivate.
2. method according to claim 1, is characterized in that, carries out gathering step after micro-algae Immobilized culture, and micro-algae density when gathering is 20g-200g/m
2.
3. method according to claim 1, is characterized in that, the Initial seeding density micro-algae being inoculated into described solid material is 5-50g/m
2.
4. method according to claim 1, is characterized in that, the intensity of illumination of described solid material surface is 10-600 μm of ol/m
2/ s.
5. method according to claim 1, is characterized in that, passes into carbonated gas during described Immobilized culture, and the volume percent that described carbonic acid gas accounts for described gas is 0.1-15%.
6. method according to claim 1, is characterized in that, the temperature of described Immobilized culture is 15-50 DEG C.
7. method according to claim 1, is characterized in that, described liquid is the micro-algae culture medium of all kinds, concentration or composition, and containing the algae liquid of one or more micro-algaes, water, containing the solution of promotive factor, containing one or more in the solution of the stress-inducing factor.
8. method according to claim 1, is characterized in that, described micro-algae comprises micro-algae of Cyanophyta, Chlorophyta, Xanthophyta, different whip algae door and rhodophyta.
9. method according to claim 1, is characterized in that, the Initial seeding density micro-algae of Immobilized culture being inoculated into liquid is 0.5-20g/L.
10. method according to claim 1, is characterized in that, during described liquid culture, the intensity of illumination on culture surface is 100-2500 μm of ol/m
2/ s.
11. methods according to claim 9 or 10, it is characterized in that, the one or more inducing culture that carries out adjusted when described liquid culture in intensity of illumination, temperature, liquid component and concentration, the concentration of carbonic acid gas and pH value parameters produces secondary metabolite.
12. methods according to claim 11, is characterized in that, the time of described inducing culture is 2-10 days.
13. methods according to claim 11, is characterized in that, the time of described inducing culture is 3-4 days.
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| CN105132351A (en) * | 2015-10-12 | 2015-12-09 | 山东大学 | Method for rapidly accumulating lipid by high-temperature stress on micro-algae |
| CN106635810B (en) * | 2016-12-06 | 2020-02-14 | 沈阳化工研究院有限公司 | Microalgae and culture method of microalgae |
| CN108728364B (en) * | 2017-04-17 | 2022-01-04 | 天津大学 | Microalgae culture method based on paper culture device |
| CN108277162A (en) * | 2018-01-24 | 2018-07-13 | 新奥科技发展有限公司 | A kind of cultural method of microalgae |
| CN109679853A (en) * | 2019-01-18 | 2019-04-26 | 昆明理工大学 | The method for improving haematococcus pluvialis biomass and astaxanthin yield using fulvic acid |
| CN112575049A (en) * | 2019-09-27 | 2021-03-30 | 中南民族大学 | Method for producing astaxanthin by utilizing immobilized haematococcus pluvialis two-step method |
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