CN102094061B - Method for producing lutein from microalgae - Google Patents
Method for producing lutein from microalgae Download PDFInfo
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Abstract
The invention relates to a method for quickly accumulating intracellular lutein of microalgae, comprising the following steps of: culturing by heterotrophism, diluting, culturing by photoinduction, collecting microalgae, extracting lutein and the like. With the method, the advantages of the quick accumulation of lutein of the microalga cell obtained by culturing by heterotrophism in the photoinduction stage can be fully exerted, and an important technical means is provided for the industrialization of the lutein derived from the microalgae.
Description
Technical field
The invention belongs to technical field of microalga biology, relate to a kind of method that microdisk electrode produces xenthophylls.
Background technology
Xenthophylls (Lutein) has another name called " Lutein ", is extensively be present in a kind of natural active matter in the plants such as vegetables, flowers, fruit and algae.Because of its to human body, there is vision protection, delay arteriosclerosis, multiple important physiological function such as anticancer, anti-oxidant, protection skin; so xenthophylls is widely used as food and feed additive at present, simultaneously vivid orange-yellow of xenthophylls makes it as the toner of family's poultry, aquatic animal and animal tissues.
Xenthophylls is application prospect widely in medicine and healthcare products, food and feed, makeup and aquaculture industry etc., and many domestic and international research institutions and company biosynthesizing, separation and Extraction, biochemical functions etc. to xenthophylls are studied.At present, xenthophylls mainly extracts and obtains from the plant such as Flower of Aztec Marigold, mary bush, but utilizes these plant production xenthophylls to there is some shortcoming.First, xenthophylls is mainly esterified form existence in these plants, need to carry out saponification process when therefore extracting xenthophylls from the plant such as Flower of Aztec Marigold, mary bush, this step not only reduces efficiency and yield, simultaneously remaining saponifying agent easily pollutes xenthophylls product, adds difficulty to purge process.Secondly, plant such as plantation Flower of Aztec Marigold, mary bush etc. needs a large amount of soils, and the growth cycle of these plants is generally 3 ~ 4 months (for micro-algae that can grow fast, the cycle is oversize).
Comparatively speaking, micro-algae is utilized to have some advantage to produce xenthophylls.First xenthophylls mainly exists in a free form in micro-algae, does not therefore need saponification step in process of production; Secondly, the growth cycle of micro-algae is short, production unit floor space is little, utilizes micro-algae to produce xenthophylls simultaneously and does not limit by season, weather and regional condition, quality product and output relatively stable; Finally, the product of micro-algae (as chlorella etc.) inherently a kind of high value, containing a large amount of protein, grease, polysaccharide isoreactivity composition, can these materials of separation and Extraction, realize the comprehensive utilization of microalgae cell.
So far, the training mode utilizing micro-algae to produce xenthophylls mainly contains light autotrophy and heterotrophism two kinds.The shortcoming that micro-algae light autotrophy is cultivated is that the microalgae cell speed of growth is slow, cell density and xenthophylls productive rate low.The current light autotrophy the highest volume productivity of xenthophylls of cultivating micro-algae cultivates 4.8mg/L/d (the Sanchez J F that Scenedesmus almeriensis obtains for Sanchez J F etc. in 2L column type bioreactor, Fernandez-Sevilla J M, Acien F G., et al.Biomass and lutein productivity ofScenedesmus almeriensis:influence of irradiance, dilution rate and temperature.Appl Microbiol Biotechnol, 2008, 79:719 ~ 729), the highest area yield of xenthophylls cultivates the 290mg/m that Scenedesmus almeriensis reaches for Fernandez etc. in 4000L pipeline reactor
2/ d (Del Campo J A, Mercedes Garcia Gonz á lez, Guerrero M G.Outdoorcultivation of microalgae for carotenoid production:current state and perspectives.Appl Microbiol Biotechnol, 2007,74:1163 ~ 1174).
The advantage of micro-algae Heterotrophic culture is that micro-algae can carry out high-density culture in bio-reactor, and cell growth rate is fast, but there is the shortcomings such as the pigment such as xenthophylls, chlorophyll and protein content in cell are low.Heterotrophic culture can obtain high-cell density and high vitro growth rates, thus the productive rate of xenthophylls is improved a lot relative to the cultivation of light autotrophy.
Micro-algae, except Heterotrophic culture and light autotrophy training mode, also has a kind of training mode of being of little use and Combined hardening model.So far, the most high-content (7.6mg/g) that microdisk electrode produces xenthophylls in the highest volume productivity (145mg/L/d) of xenthophylls and born of the same parents is all (the Martin obtained under the Combined hardening model condition of continuous illumination, Lucia.Process for obtaining lutein from algae.EP180843A1,2007).But this training mode can only be able to carry out in the closed photo bioreactor of steam sterilizing, and culturing process must ensure absolute aseptic, and need the reasonable disposition of light source, this cannot realize in actual production simultaneously.Therefore, utilize mixotrophism pattern to cultivate micro-algae production xenthophylls and not there is industrialization value.
From above-mentioned, no matter be adopt light autotrophy training mode or Heterotrophic culture pattern, lutein content and xenthophylls productive rate in lower born of the same parents, add the cost that micro-algae large scale culturing is higher simultaneously, constrains application microdisk electrode to produce the industrialization process of xenthophylls.Therefore, be necessary that the new microdisk electrode technique of exploration or method make xenthophylls productive rate and content increase substantially, make again the cost of micro-algae large scale culturing significantly decline simultaneously, the industrialized requirement utilizing microdisk electrode to produce xenthophylls can be met like this.
The relative merits of comprehensive above-mentioned several product xenthophylls microdisk electrode pattern, the present invention devises a kind of " heterotrophism-dilution-photoinduction series connection is cultivated " pattern for producing xenthophylls microdisk electrode, its flow process is as follows: first utilize bio-reactor Heterotrophic culture to produce the micro-algae of xenthophylls to obtain high-density cells, after in nutrient solution, organic carbon source is exhausted, with the substratum dilution algae liquid not containing organic carbon source, the light induction of (8 ~ 24h) within the short period of time, makes the xenthophylls rapid, high volume in frustule accumulate.The heterotrophism stage in this pattern can carry out in the bio-reactor of Heterotrophic culture at shaking flask, mechanical agitation type, air lift type, bubbling style etc., and object is the frustule in order to obtain higher density at short notice; The photoinduction stage can carry out in the system of micro-algae light autotrophy cultivation in any can be used for, and object improves lutein content in born of the same parents by photoinduction effect; Heterotrophism stage and photoinduction stage independently carry out, the heterotrophism stage release algae liquid first dilute with light inducing culture after proceed to photoinduction cultivation stage again.To need to guarantee to enter in the algae liquid in photoinduction stage not containing organic carbon source in process, the photoinduction stage can be avoided like this to grow too much miscellaneous bacteria; And can guarantee that the frustule in photoinduction stage can obtain sufficient illumination, realizes the quick raising of lutein content in born of the same parents by diluting effect.
Microdisk electrode method production xenthophylls is divided into frond growth in the present invention and product (xenthophylls) accumulates two stages, and namely Heterotrophic culture and photoinduction are cultivated.Can be obtained the microalgae cell of a large amount of produced xenthophylls at short notice by Heterotrophic culture, proceed to photoinduction after the dilution of algae liquid and cultivate, xanthophyll in algal cell content is promoted to rapidly initial twice even more than twice.The present invention has following advantage: (1) photoinduced algae cell density very high (2 ~ 10g/L), is about 10 times that conventional light autotrophy cultivates algae cell density (about 0.2 ~ 1g/L); (2) photoinduction time very short (about 1d ~ 2d), and micro-algae light autotrophy incubation time very long (about 7d ~ 14d), therefore, the production efficiency of xenthophylls greatly improves; (3) cultivate relative to micro-algae light autotrophy, algae cell density higher during photoinduction makes the floor space needed for photoinduction very little, and the high-cell density cost that makes to gather significantly reduces simultaneously; (4) impact of Heterotrophic culture climate, weather hardly, photoinduction is cultivated and in glass room, can be carried out under natural lighting or artificial lighting condition, thus, adopts method of the present invention can realize the continuous seepage of xenthophylls; (5) comprehensive utilization of the residue frond after separation and Extraction xenthophylls, can obtain extra product and economic benefit, reduces the comprehensive production cost of xenthophylls.
Described in summary, training mode of the present invention reasonably combines the respective advantage that heterotrophism and light autotrophy cultivate two kinds of modes, compared with other pattern, have that production efficiency is high, the array mode of culture systems is flexible and the advantage such as production cost is low, the advantage that heterotrophism pattern can obtain high-density algae liquid and photoinduction stage xenthophylls Rapid Accumulation can being given full play to, providing important technique means for solving the xenthophylls Industrialization coming from micro-algae.
Summary of the invention
One aspect of the present invention provides the method for xenthophylls in the micro-gonidium of a kind of Rapid Accumulation, the method comprises the Heterotrophic culture step of micro-algae, the step of photoinduction cultivation will be carried out after obtain micro-algae Heterotrophic culture liquid dilution, and optional frustule is gathered, the step of xenthophylls separation and Extraction.
The present invention provides a kind of xanthophyll production method coming from micro-algae on the other hand, and the method comprises the step of micro-algae heterotrophism, carry out the step of photoinduction cultivation, and frustule is gathered, the step of xenthophylls separation and Extraction after being diluted by the Heterotrophic culture liquid of micro-algae.
Method of the present invention can realize the Rapid Accumulation of xenthophylls in born of the same parents, substantially increases production efficiency, reduces production cost, and provides the xenthophylls of high-quality.
In an embodiment, described micro-algae is selected from: Chlorella pyrenoidesa (Chlorellapyrenoidosa), Chlorella vulgaris (Chlorella vulgaris), marine chlorella (Marine chlorella), Chlorella protothecoides (Chlorella protothecoides), chlorella ellipsoidea (Chlorella ellipsoidea), Chlorella zofingiensis, Chlorella regularis, Chlorella sorokina, Spongiococcumexcentricum and Chlamydomonas agloeformis.
In an embodiment, the step of described micro-algae Heterotrophic culture comprises: in bio-reactor, add the substratum that pH is 4.0 ~ 9.0, batch culture, fed batch cultivation, Semi-continuous cultivation or cultured continuously is carried out by the micro-algae algae kind of 0.1 ~ 30% access of working volume, culture temperature is 10 ~ 40 DEG C, control pH is less than 9.0, controls dissolved oxygen more than 1%.
In an embodiment, described photoinduction is cultivated and is comprised the algae liquid after by dilution and proceed in photoinduction device and carry out photoinduction, continuous illumination or intermittent illumination, and culture temperature is 5 ~ 50 DEG C, intensity of illumination is 0.1 ~ 150klx, and photoinduction culture cycle is 1 ~ 150 hour.
In an embodiment, Heterotrophic culture base is made up of nitrogenous source, organic carbon source and a small amount of inorganic salt, trace element and water; Photoinduction substratum is made up of nitrogenous source, inorganic salt and water.
In an embodiment, described heterotrophism step can be carried out in the bio-reactor of Heterotrophic culture at shaking flask, mechanical agitation type, air lift type or bubbling style, described photoinduction culturing step stands any can be used for such as bag and Pig bioreactor at the raceway pond of shaking flask or open type or circle pond, enclosed flat plate photobioreactor or duct type bioreactor or pillar bioreactor or film to carry out in the device that micro-algae light autotrophy cultivates, and light source is natural light or various artificial light.
In an embodiment, when product xenthophylls algae kind is Chlorella vulgaris, the substratum that heterotrophism uses consists of the following composition substantially: KNO
35 ~ 15 grams per liters, glucose 10 ~ 60 grams per liter, KH
2pO
40.3 ~ 0.9 grams per liter, Na
2hPO
412H
2o 1.0 ~ 10.0 grams per liter, MgSO
47H
2o 0.2 ~ 1.0 grams per liter, CaCl
20.05 ~ 0.3 grams per liter, FeSO
47H
2o 0.01 ~ 0.05 grams per liter, trace element 0.5 ~ 4ml and water, its medium trace element consist of H
3bO
35 ~ 15 grams per liters, ZnSO
47H
2o 5.0 ~ 10.0 grams per liter, MnCl
2h
2o1.0 ~ 2.0 grams per liter, (NH
4)
6mo
7o
244H
2o 0.5 ~ 1.5 grams per liter, CuSO
45H
2o 1.0 ~ 2.0 grams per liter and Co (NO
3)
26H
2o 0.1 ~ 0.9 grams per liter.
In an embodiment, when chlorella is Chlorella pyrenoidesa, the substratum that heterotrophism uses consists of the following composition substantially: glucose 10 ~ 60 grams per liter, urea 2 ~ 8 grams per liter, KH
2pO
41 ~ 2 grams per liter, Na
2hPO
412H
2o 1.0 ~ 10.0 grams per liter, MgSO
47H
2o 1 ~ 2 grams per liter, CaCl
20.05 ~ 0.1 grams per liter, trisodium citrate 0.1 ~ 2.0 grams per liter, Fe-EDTA solution 0.5 ~ 1mL, A5 solution 1 ~ 5mL and water; Wherein Fe-EDTA solution formula is FeSO
47H
2o 20 ~ 30 grams per liter and EDTA 20 ~ 40 grams per liter; A5 solution formula is H
3bO
32.5 ~ 4.0 grams per liters, MnCl
24H
2o 1.0 ~ 2.0 grams per liter, ZnSO
47H
2o0.1 ~ 0.6 grams per liter, CuSO
45H
2o 5 ~ 10 grams per liter and Na
2moO
40.01 ~ 0.05 grams per liter.
In an embodiment, adopt supercritical CO
2extraction process, organic solvent extraction or ultrasonic wave added solvent-extraction process extract xenthophylls.
In an embodiment, method of the present invention also comprises: mixed with other pigments by the frond after extraction xenthophylls and carry out spraying dry algae powder, or carry out separation and Extraction to the biologically active substance in frond.
In an embodiment, other pigments described comprise chlorophyll.In an embodiment, described biologically active substance comprises protein, chlorophyll and polysaccharide.
Accompanying drawing explanation
Fig. 1 and Fig. 2 shows in 5L bio-reactor/3L flat plate photobioreactor series system the process adopting heterotrophism-dilution-photoinduction series connection to cultivate Chlorella pyrenoidesa production xenthophylls.Wherein, Fig. 1 shows Chlorella pyrenoidesa in 5L bio-reactor Heterotrophic culture process; Fig. 2 shows Chlorella pyrenoidesa photoinduction culturing process in 3L flat plate photobioreactor.
Fig. 3 and Fig. 4 shows in 50L bio-reactor/10L column type bioreactor series system the process adopting heterotrophism-dilution-photoinduction series connection to cultivate Chlorella pyrenoidesa production xenthophylls.Wherein, Fig. 3 shows Chlorella pyrenoidesa in 50L bio-reactor Heterotrophic culture process; Fig. 4 shows Chlorella pyrenoidesa photoinduction culturing process in 10L column type bioreactor.
Fig. 5 and Fig. 6 shows in 50L bio-reactor/30L flat plate photobioreactor series system the process adopting heterotrophism-dilution-photoinduction series connection to cultivate Chlorella vulgaris production xenthophylls.Wherein, Fig. 5 shows Chlorella vulgaris Heterotrophic culture process in 50L bio-reactor; Fig. 6 show Chlorella vulgaris out of doors in 30L flat plate photobioreactor photoinduction cultivate.
Fig. 7 and Fig. 8 shows in 5L bio-reactor/3L flat bio-light bio-reactor series system the process adopting heterotrophism-dilution-photoinduction series connection to cultivate chlorella ellipsoidea production xenthophylls.Wherein, Fig. 7 shows chlorella ellipsoidea in 5L bio-reactor Heterotrophic culture process; Fig. 8 shows chlorella ellipsoidea photo induced processes in 3L flat plate photobioreactor.
Specific embodiments
The micro-algae being applicable to the application includes but not limited to Chlorella pyrenoidesa (Chlorella pyrenoidosa), Chlorella vulgaris (Chlorella vulgaris), marine chlorella (Marine chlorella), Chlorella protothecoides (Chlorella protothecoides), chlorella ellipsoidea (Chlorella ellipsoidea), Chlorellazofingiensis, Chlorella regularis, Chlorella sorokina, Spongiococcum excentricum and Chlamydomonas agloeformis.
In a preferred embodiment, the present invention adopts Chlorella pyrenoidesa, Chlorella vulgaris and chlorella ellipsoidea to produce xenthophylls.
1. the high Density Heterotrophic of micro-algae in bio-reactor
The object of this step is to obtain a large amount of frustule fast, for photoinduction stage Fast back-projection algorithm accumulation xenthophylls.
Various substratum well known in the art can be adopted to carry out micro-algae Heterotrophic culture.Usually, Heterotrophic culture base contains nitrogenous source, organic carbon source, a small amount of inorganic salt, trace element and water.
This kind of substratum comprises HA-SK substratum (Chinese patent ZL 200610024004.9), Endo substratum (Ogbonna J.C., Masui.H., Tanaka.H.Sequential heterotrophic:autotrophiccultivation-an efficient method of producing Chlorella biomass for health foodand animal feed.J.Appl.Phycol.1997,9,359 ~ 366) etc.
The present invention's HA-SK substratum used is substantially by KNO
3, glucose and a small amount of inorganic salt, trace element and water forms.In described technical scheme, described trace element should be selected from H
3bO
3, ZnSO
47H
2o, MnCl
2h
2o, (NH
4)
6mo
7o
244H
2o, CuSO
45H
2o and Co (NO
3)
26H
2one in O, multiple or whole.
Term used herein " substantially by ... composition " represent in composition of the present invention except containing main ingredient KNO
3, outside glucose and a small amount of inorganic salt, trace element and water, also can comprise the component that some fundamental characteristics for composition or new characteristic (can maintain micro-algae and reach higher level at shorter culture cycle inner cell density, activity substance content has a more substantial increase compared with conventional Heterotrophic culture simultaneously) do not affect in fact.Term used herein " by ... composition " represent that composition of the present invention is made up of pointed concrete component, there is no other components, but can with the impurity of content in usual scope.
In this substratum, each component of substratum can change within the specific limits and can not have very large materially affect to microalgae cell density and quality.Therefore, the consumption of these components should not by the strict restriction of embodiment.As known to those skilled in the art, in substratum, also can add a small amount of inorganic salt, such as magnesium sulfate, calcium chloride, ferrous sulfate and phosphoric acid salt etc., and a small amount of trace element is as Mn, Zn, B, I, M, Cu, Co etc.
In the present invention, preferably micro-component should be selected from H
3bO
3, ZnSO
47H
2o, MnCl
2h
2o, (NH
4)
6mo
7o
244H
2o, CuSO
45H
2o and Co (NO
3)
26H
2one or more in O.The consumption of inorganic salt and trace element can be determined according to Conventional wisdom.
HA-SK substratum of the present invention consists of the following composition substantially: KNO
35 ~ 15 grams per liters, glucose 10 ~ 60 grams per liter, KH
2pO
40.3 ~ 0.9 grams per liter, Na
2hPO
412H
2o 1.0 ~ 10.0 grams per liter, MgSO
47H
2o 0.2 ~ 1.0 grams per liter, CaCl
20.05 ~ 0.3 grams per liter, FeSO
47H
2o 0.01 ~ 0.05 grams per liter; Trace element 0.5 ~ 4ml, its medium trace element consist of H
3bO
35 ~ 15 grams per liters, ZnSO
47H
2o5.0 ~ 10.0 grams per liter, MnCl
2h
2o 1.0 ~ 2.0 grams per liter, (NH
4)
6mo
7o
244H
2o 0.5 ~ 1.5 grams per liter, CuSO
45H
2o 1.0 ~ 2.0 grams per liter and Co (NO
3)
26H
2o 0.1 ~ 0.9 grams per liter; Water.
The present invention's Endo substratum used consists of the following composition substantially: glucose 10 ~ 60 grams per liter, urea 2 ~ 8 grams per liter, KH
2pO
41 ~ 2 grams per liter, Na
2hPO
412H
2o 1.0 ~ 10.0 grams per liter, MgSO
47H
2o1 ~ 2 grams per liter, CaCl
20.05 ~ 0.1 grams per liter, trisodium citrate 0.1 ~ 2.0 grams per liter, Fe-EDTA solution 0.5 ~ 1mL, A5 solution 1 ~ 5mL; Wherein Fe-EDTA solution formula is FeSO
47H
2o 20 ~ 30 grams per liter and EDTA 20 ~ 40 grams per liter; A5 solution formula is H
3bO
32.5 ~ 4.0 grams per liters, MnCl
24H
2o 1.0 ~ 2.0 grams per liter, ZnSO
47H
2o 0.1 ~ 0.6 grams per liter, CuSO
45H
2o 5 ~ 10 grams per liter and Na
2moO
40.01 ~ 0.05 grams per liter; Water.
After according to above-mentioned formulated substratum, available conventional means as acid or alkali by as described in the pH of substratum be adjusted to 4.0 ~ 9.0, and at 115 ~ 120 DEG C autoclaving 15 ~ 20 minutes.Can adopt in batches, the various ways such as fed-batch, semicontinuous and cultured continuously implements Heterotrophic culture.
When Heterotrophic culture adopts fed-batch process, the corresponding substratum prepared is joined in bio-reactor, benefit adds water to working volume, usual coefficient is 0.6 ~ 0.8, then steam sterilizing (121 DEG C, maintain about 20 minutes), when temperature is down to 30 ~ 35 DEG C, start Heterotrophic culture by the micro-algae algae kind of 1 ~ 15% access of working volume.
After Heterotrophic culture for some time, need to carry out feed supplement when glucose in substratum has been consumed (being generally 27 ~ 45 hours), add carbon source (as glucose), (nitrogenous source e.g., cultivating Chlorella vulgaris is KNO to nitrogenous source
3, to cultivate the nitrogenous source of Chlorella pyrenoidesa be urea) and the nutritive salt such as inorganic salt, the nutritive salt added is the above-mentioned corresponding substratum after concentrating, and impels micro-algae continued growth.Can every feed supplement in 5 ~ 8 hours, the concentration of adding of glucose can be 15 ~ 25 grams per liters, and the concentration of adding of nitrogenous source solution can be 2 ~ 5 grams per liters.When after the nutritive salt adding certain number of times (being generally 4 ~ 7 times), when microalgae cell density reaches the highest, the Heterotrophic culture stage terminates.
No matter adopt which kind of training method, in culturing process, the culture condition that must control to be applicable to makes micro-algae normal growth.Usually, control temperature is 20 ~ 35 DEG C, such as 28 ~ 30 DEG C, and dissolved oxygen is not less than the air saturation concentration of 5%, and pH is not higher than 9.0.In a preferred embodiment, dissolved oxygen is not less than the air saturation concentration of 10%, and pH is not higher than 8.5.
Usually, when adopting batch culture, fed batch cultivation, at the end of the Heterotrophic culture stage, in bio-reactor, organic carbon source needs completely consumed complete.When adopting Semi-continuous cultivation mode, it is carry out when in bio-reactor, organic carbon source completely consumed is complete that band puts operation.
Heterotrophism can carry out in the bio-reactor of Heterotrophic culture at shaking flask, mechanical agitation type, air lift type, bubbling style etc.
2. the dilution of high density algae liquid
The object of this step is to make the micro-algae of product xenthophylls proceeding to photoinduction cultivation absorb luminous energy efficiently, improve optical energy utilization efficiency, reduce the mortality ratio of frustule simultaneously.Because light intensity is the decay in " time, sky, non-linear " in algae liquid, so under high-cell density, in reactor, frustule major part is in dark space, almost can't accept illumination, and such frustule is easy to death and can affects photoinduced efficiency.
The high-density algae liquid that Heterotrophic culture obtains should carry out dilution operation, with water with do not dilute highdensity algae liquid containing the substratum of organic carbon source, makes cell density maintain 0.1 ~ 10 grams per liter, adjustment pH to 5.0 ~ 8.0.In other embodiments, dilution algae liquid, makes cell density maintain 1 ~ 8 grams per liter.In a preferred embodiment, cell density is made to maintain 1.0 ~ 5.0 grams per liters.
Various known diluted medium can be adopted to dilute algae liquid.Usually, photoinduction substratum is made up of nitrogenous source, inorganic salt and water, relative to Heterotrophic culture base not containing organic carbon source.In preferred embodiments, the nitrogen concentration of described substratum is 2 ~ 10 grams per liters, is preferably 2 ~ 8 grams per liters.Described nitrogenous source can be identical or different with nitrogenous source used in Heterotrophic culture step.
In a preferred concrete scheme, the high-density frustule that Heterotrophic culture obtains should with not containing organic carbon source, nitrogen concentration be the initial medium of 2 ~ 10 grams per liters (such as, adopt when cultivating Chlorella vulgaris not containing the HA-SK substratum of glucose, adopt when cultivating Chlorella pyrenoidesa not containing the Endo substratum of glucose) suitably dilute.
The substratum that dilution adopts is without the need to autoclaving, and preparing rear adjustment pH to 5.0 ~ 8.0 can use.
In a specific embodiment, to Chlorella vulgaris, diluted medium (photoinduction substratum) consists of the following composition: KNO
31 ~ 8 grams per liter, MgSO
47H
2o, 0.5 ~ 1.0 grams per liter, CaCl
20.01 ~ 0.06 grams per liter, FeSO
47H
2o 0.01 ~ 0.06 grams per liter, EDTA 0.020 ~ 0.052 grams per liter.
To Chlorella pyrenoidesa, diluted medium (photoinduction substratum) consists of the following composition: urea 0.1 ~ 2.0 grams per liter, MgSO
47H
2o, 0.5 ~ 1.0 grams per liter, CaCl
20.01 ~ 0.06 grams per liter, FeSO
47H
2o0.01 ~ 0.06, EDTA 0.020 ~ 0.052 grams per liter, Trisodium Citrate 0.08 ~ 0.5 grams per liter.
3. photoinduction is cultivated
The object of this step allows produce the illumination that the micro-algae of xenthophylls accepts abundance, makes frustule rapid, high volume dynamic accumulation xenthophylls by photoinduction.
Gained diluent, after dilution, proceeds in photoinduction device and carries out photoinduction cultivation by high-density micro algae culturing liquid described above.Temperature controls at 5 ~ 50 DEG C, and intensity of illumination is 0.11 ~ 150klx, continuous illumination or intermittent illumination, and photoinduction culture cycle is 1 ~ 150 hour, and air flow is 0.1 ~ 2.0vvm.Wherein said bioreactor comprises all closed photo bioreactor (shaking flask, duct type, flat, pillar, film found bag and Pig etc.) and all Race-way photobioreactors (raceway pond, circle pond and the large basin of bubbling style etc.).
Usually, culture temperature can be controlled in the scope of 15 ~ 35 DEG C, such as 18 ~ 35 DEG C, 20 ~ 35 DEG C, 20 ~ 30 DEG C etc.Usually, intensity of illumination is 1 ~ 70klx, such as, 1 ~ 60,1 ~ 50,1 ~ 40,1 ~ 30,1 ~ 20,1 ~ 10klx etc., determined by the concrete condition of production.Usually, as caused algae liquid fully to mix by gas, then air flow is controlled is made as 0.15 ~ 2.0vvm, such as, 0.2 ~ 1.8,0.5 ~ 1.5,0.8 ~ 1.5,1.0 ~ 1.5vvm etc.In other embodiments, culture temperature controls at 10 ~ 50 DEG C, and intensity of illumination is 1 ~ 10klx, and air flow is 0.05 ~ 2.0vvm.
In other embodiments, photoinduction culture cycle is 8 ~ 100 hours, such as, according to the weather condition of reality, photoinduction culture cycle can be 8 ~ 90 hours, 8 ~ 80 hours, 8 ~ 60 hours, 8 ~ 48 hours, 8 ~ 24 hours not etc.; Or, photoinduction culture cycle can be 12 ~ 72 hours, 12 ~ 60 hours, 12 ~ 48 hours, 12 ~ 36 hours, 12 ~ 24 hours not etc. or 24 ~ 60 hours, 24 ~ 48 hours not etc.
In this application, " photoinduction culture cycle " includes whole photoinduction culturing process, and such as, the outdoor inducing culture cycle in cultivation time comprises the time not having illumination night.
In this application, " light application time " refers to use intensity of illumination described in the application micro-algae to be implemented to the time of photoinduction cultivation, and namely this time does not comprise the time not having illumination night.In certain embodiments, the light application time of photoinduction culturing step is 8 ~ 48 hours, and such as 8 ~ 36 hours, 8 ~ 24 hours, 8 ~ 18 hours, 8 ~ 12 hours, 12 ~ 36 hours, 12 ~ 24 hours are not etc., and any duration in above-mentioned scope.
Therefore, the photoinduction culturing step of the application also comprises light application time is photoinduction culturing step in 8 ~ 48 hours window.The mode of artificial lighting can be adopted to carry out photoinduction cultivation, the mode of natural lighting also can be utilized out of doors to carry out photoinduction cultivation.
In a specific embodiment, when nutrient solution Lutein concentration reaches the highest, terminate photoinduction and cultivate, results frustule carries out the separation and Extraction of xenthophylls or frustule of directly gathering carries out the preparation of algae powder.
4. frustule gather, xenthophylls separation and Extraction and frond comprehensive utilization
Photoinduction carries out centrifugal gathering to chlorella after cultivating and terminating, and obtains wet frond.The collecting method of frustule includes but not limited to high speed centrifugation, flocculation, the technology such as air supporting or filtration; Frustule wall-breaking method includes but not limited to the Wet-process wall breaking methods such as frond self-dissolving, high-pressure homogenization, enzymic hydrolysis, aqueous phase pyrolysis.
Traditional organic solvent extraction is adopted chlorella to be carried out to the extraction of xenthophylls.First organic solvent is joined in algae mud and extract, then carry out stirring centrifugal acquisition supernatant liquor and frond precipitation, concentrating under reduced pressure is carried out to supernatant liquor, stirring adds water, filters acquisition lutein crystal.
Other compositions in supernatant liquor progressively separation and Extraction can obtain lipid acid, chlorophyll etc., or directly all the components in supernatant liquor and frond are precipitated that mixed atomizing is dry obtains chlorella powder.
In one preferably scheme, adopt supercritical CO
2abstraction technique carries out the separation and Extraction of xenthophylls to chlorella.In a better scheme, Direct spraying drying after concentrated for the chlorella liquid obtained is obtained chlorella powder.
In the present invention, can the micro-algae cultivating gained be fully utilized, extract the various activeconstituents such as polyunsaturated fatty acid, protein, chlorophyll, polysaccharide wherein.The sequence of extraction of activeconstituents there is no particular restriction, but usually will meet this prerequisite of component damages that the step first extracted can not cause rear extraction.
Relate to frustule dry weight herein and lutein content measuring method is as follows:
Frustule dry weight measures: in micro-algae (as chlorella) culturing process, get nutrient solution V milliliter, centrifugal 10 minutes of 8000rpm, by the frond deionized water wash after centrifugal 3 times, is transferred to weighing bottle (W
1(gram)) in, dry to constant weight W in 105 DEG C of baking ovens
2(gram).Frond dry weight Cx can calculate according to following formula: Cx (grams per liter)=(W
2-W
1)/V/1000.
Xenthophylls measures: adopt high performance liquid chromatography (HPLC), concrete operation step is shown in document (ZhengYun Wu, Chun Lei Shi et al.Modeling of lutein production by heterotrophicChlorella in batch and fed-batch cultures.World Journal of Microbiology andBiotechnology, 2007,23:1233-1238).
Embodiment 1
Carry out steam sterilizing add following Heterotrophic culture base and water to 2.8L in 5L bio-reactor after, then access Chlorella pyrenoidesa when temperature drops to 30 DEG C, start Heterotrophic culture.
Heterotrophic culture condition: temperature is 30 ± 1 DEG C, air flow quantity is that 1vvm, pH are less than 8.0, controls dissolved oxygen more than 15%.
First time feed supplement after 53.9h after inoculation, carries out feed supplement every 5 ~ 8h afterwards, adds 4 times altogether, be cultured to 88.40h dry cell weight and reach 132.2g/L (see Fig. 1), and now Heterotrophic culture terminates to proceed to photoinduction cultivation.
High-density algae liquid after being terminated by Heterotrophic culture is diluted to 2.55g/L, and adds following photoinduction substratum, is transferred in 3L flat plate photobioreactor and carries out photoinduction cultivation.Photoinduction culture condition: temperature maintains 28 ~ 33 DEG C, air flow quantity is 1vvm, bilateral illumination, and every sidelight is by force 15klx.After 27h is cultivated in photoinduction, dry cell weight is reduced to 1.82g/L from 2.55g/L, and xenthophylls rises to 3.64mg/gDcw from 1.57mg/gDcw, and during photoinduction 27h, the productive rate of xenthophylls is 5.88mg/L/d; If the productive rate of xenthophylls can reach 12.1mg/L/d (for current micro-algae light autotrophy produces 2.5 times of xenthophylls maximum output 4.8mg/L/d) (see Fig. 2) during photoinduction 12h.
Heterotrophism and supplemented medium:
Glucose 60.0 grams of urea 8.0 grams of MgSO
47H
2o 2.0 grams
KH
2pO
41.1 grams of Na
2hPO
412H
2o 9.0 grams of CaCl
20.02 gram
Trisodium citrate 1.8 grams
Fe-EDTA solution 1.0ml trace element solution 4.5ml water 1000ml
Wherein Fe-EDTA solution formula is FeSO
47H
2o 15 grams per liter and EDTA1.4 grams per liter, trace element solution formula is H
3bO
32.11 grams per liters, MnCl
24H
2o 0.81 grams per liter, ZnSO
47H
2o 0.11 grams per liter, CuSO
45H
2o 10.0 grams per liter, Na
2moO
40.05 grams per liter.
Photoinduction substratum:
Urea 0.5 gram of MgSO
47H
2o 1.0 grams of trisodium citrates 0.05 gram
CaCl
20.01 gram of Fe-EDTA solution 0.4ml water 1000ml
Wherein Fe-EDTA solution formula is FeSO
47H
2o 15 grams per liter and EDTA1.4 grams per liter.
Embodiment 2
Add at 121 DEG C of sterilizing 20min after following Heterotrophic culture base and tap water to 25L in 50L bio-reactor, then when temperature is down to about 30 DEG C by 10% access Chlorella pyrenoidesa of working volume, start Heterotrophic culture.
Heterotrophic culture condition: temperature is 30 DEG C, air flow quantity be 1vvm, pH 6.0 ~ 8.0, control dissolved oxygen more than 15%.In culturing process, after glucose consumption is complete, carries out adding carbon source, when urea runs out of, add nitrogenous source.Add carbon source by 6 times, after adding nitrogenous source 3 times, reach 130.5g/L (see Fig. 3) at 98.89h algae cell density.
High-density algae after being terminated by Heterotrophic culture is diluted to 2.01g/L, and adds photoinduction substratum, is transferred in 10L cylinder bioreactor and carries out photoinduction.
Photoinduction culture condition: natural temperature, temperature is at 30 DEG C, and light intensity is at 3klx, and air flow quantity is 1vvm.After 30h is cultivated in photoinduction, dry cell weight is reduced to 1.66g/L from 2.01g/L, and xenthophylls rises to 3.47mg/gDcw from 2.55mg/gDcw, photoinduction 30h, and xenthophylls productive rate is 4.61mg/L/d; If only photoinduction 24h, the productive rate of xenthophylls reaches 5.73mg/L/d (see Fig. 4).Substratum is consistent with the substratum of embodiment 1.
Embodiment 3
Add at 121 DEG C of sterilizing 20min after following Heterotrophic culture base and tap water to 25L in 50L bio-reactor, then when temperature is down to about 30 DEG C by 13% access Chlorella vulgaris of working volume, start Heterotrophic culture.
Heterotrophic culture condition: temperature is 30 DEG C, air flow quantity is that 1vvm, pH are less than 9.0.In culturing process, after carbon source runs out of, carry out adding glucose, when nitrogenous source runs out of, add saltpetre.Add Carbon and nitrogen sources by 5 times, 58.20h algae cell density is up to 54.5g/L (see Fig. 5).
The high-density algae of Heterotrophic culture is diluted to about 3.2g/L, and adds photoinduction substratum, be transferred to 30L flat plate photobioreactor and carry out photoinduction cultivation out of doors.Photoinduction culture condition: natural temperature, natural lighting, air flow quantity is 1.0vvm.Photoinduction 28h, xenthophylls rises to 1.82mg/gDcw from initial 1.10mg/gDcw; During photoinduction 23h, lutein content is 1.67mg/gDcw, and xenthophylls productive rate reaches 5.23mg/L/d (see Fig. 6).
Heterotrophism and supplemented medium:
Glucose 60.0 grams of saltpetre 10.0 grams of MgSO
47H
2o 0.2 gram
KH
2pO
40.3 gram of Na
2hPO
412H
2o 8.8 grams of CaCl20.02 gram
Fe-EDTA solution 1.0ml trace element solution 3.5ml water 1000ml
Wherein Fe-EDTA solution formula is FeSO
47H
2o 15 grams per liter and EDTA1.4 grams per liter, trace element solution formula is H
3bO
32.86 grams per liters, MnCl
24H
2o 0.11 grams per liter, ZnSO
47H
2o 9.22 grams per liter, CuSO
45H
2o 1.00 grams per liter, (NH
4)
6mo
7o
244H
2o 0.1 grams per liter and Co (NO
3)
26H
2o0.9 grams per liter.
Photoinduction substratum:
Saltpetre 0.5 gram of MgSO
47H
2o 0.6 gram
CaCl
20.03 gram of Fe-EDTA solution 1.5ml water 1000ml
Wherein Fe-EDTA solution formula is FeSO
47H
2o 8 grams per liter and EDTA 10.4 grams per liter.
Embodiment 4
Carry out steam sterilizing add following Heterotrophic culture base and water to 2.8L in 5L bio-reactor after, then access chlorella ellipsoidea when temperature drops to 30 DEG C by 8% of working volume, start Heterotrophic culture.
Heterotrophic culture condition: temperature is 30 ± 1 DEG C, air flow quantity is that 1vvm, pH are less than 8.5, controls dissolved oxygen more than 5%.
At 66h after feed supplement twice, algae cell density is 53.0g/L (see Fig. 7), and now Heterotrophic culture terminates to proceed to photoinduction and cultivates.
High-density algae liquid after being terminated by Heterotrophic culture is diluted to 4.0g/L, and adds following photoinduction substratum, is transferred in 3L flat plate photobioreactor and carries out photoinduction cultivation.Photoinduction culture condition: temperature maintains 28 ~ 33 DEG C, air flow quantity is 1.0vvm, bilateral illumination, and every sidelight is by force 15klx.After 48h is cultivated in photoinduction, dry cell weight is reduced to 2.8g/L from 4.0g/L, and xenthophylls rises to 3.6mg/gDcw from 1.5mg/gDcw, and xenthophylls productive rate is 5.04mg/L/d; Photoinduction 24h, xenthophylls productive rate can reach 9.24mg/L/d (see Fig. 8).Substratum is consistent with the substratum of embodiment 3.
Although be described above object lesson of the present invention, having a bit is obvious to those skilled in the art, namely can make various changes the present invention and change under the premise without departing from the spirit and scope of the present invention.Therefore, claims cover all these variations within the scope of the present invention.
Claims (9)
1. the method for xenthophylls in the micro-gonidium of Rapid Accumulation, it is characterized in that, the method comprises the Heterotrophic culture step of micro-algae, the step of photoinduction cultivation will be carried out after obtain micro-algae Heterotrophic culture liquid dilution, and optional frustule is gathered, the step of xenthophylls separation and Extraction, wherein
The dilution of described algae liquid comprises that to be diluted to cell density with the algae liquid that heterotrophism obtains by substratum be 0.1 ~ 10 grams per liter, and described substratum does not contain organic carbon source, and its pH is 4.0 ~ 9.0;
Described micro-algae is selected from: Chlorella pyrenoidesa (Chlorella pyrenoidosa), Chlorella vulgaris (Chlorella vulgaris) and chlorella ellipsoidea (Chlorella ellipsoidea);
Described photoinduction is cultivated and is comprised the algae liquid after by dilution and proceed in photoinduction device and carry out photoinduction, and culture temperature is 5 ~ 50 DEG C, continuous illumination or intermittent illumination, and intensity of illumination is 0.1 ~ 150klx, and photoinduction culture cycle is 1 ~ 150 hour;
Described photoinduction substratum is made up of nitrogenous source, inorganic salt and water.
2. the method for claim 1, it is characterized in that, the step of described micro-algae Heterotrophic culture comprises: in bio-reactor, add the substratum that pH is 4.0 ~ 9.0, batch culture, fed batch cultivation, Semi-continuous cultivation or cultured continuously is carried out by the micro-algae algae kind of 0.1 ~ 30% access of working volume, culture temperature is 10 ~ 40 DEG C, control pH is less than 9.0, controls dissolved oxygen more than 1%.
3. the method according to any one of claim 1-2, is characterized in that, the dilution of described algae liquid comprises that to be diluted to cell density with the algae liquid that heterotrophism obtains by substratum be 1.0 ~ 5.0 grams per liters, and described substratum does not contain organic carbon source, and its pH is 4.0 ~ 9.0.
4. the method according to any one of claim 1-2, is characterized in that, described Heterotrophic culture base is made up of nitrogenous source, organic carbon source, inorganic salt, trace element and water.
5. the method according to any one of claim 1-2, it is characterized in that, described heterotrophism step can be carried out in the bio-reactor of Heterotrophic culture at shaking flask, mechanical agitation type, air lift type or bubbling style, described photoinduction culturing step shaking flask or open type raceway pond or circle pond, enclosed flat plate photobioreactor or duct type bioreactor or pillar bioreactor or found bag for the film that micro-algae light autotrophy is cultivated and Pig carries out, light source is natural light or artificial light.
6. the method for claim 1, is characterized in that, when chlorella is Chlorella vulgaris, the substratum that heterotrophism uses consists of the following composition: KNO
35 ~ 15 grams per liters, glucose 10 ~ 60 grams per liter, KH
2pO
40.3 ~ 0.9 grams per liter, Na
2hPO
412H
2o 1.0 ~ 10.0 grams per liter, MgSO
47H
2o 0.2 ~ 1.0 grams per liter, CaCl
20.05 ~ 0.3 grams per liter, FeSO
47H
2o 0.01 ~ 0.05 grams per liter, trace element 0.5 ~ 4ml and water, consisting of of its medium trace element: H
3bO
35 ~ 15 grams per liters, ZnSO
47H
2o 5.0 ~ 10.0 grams per liter, MnCl
2h
2o 1.0 ~ 2.0 grams per liter, (NH
4)
6mo
7o
244H
2o 0.5 ~ 1.5 grams per liter, CuSO
45H
2o1.0 ~ 2.0 grams per liter and Co (NO
3)
26H
2o 0.1 ~ 0.9 grams per liter.
7. the method for claim 1, is characterized in that, when chlorella is Chlorella pyrenoidesa, the substratum that heterotrophism uses consists of the following composition: glucose 10 ~ 60 grams per liter, urea 2 ~ 8 grams per liter, KH
2pO
41 ~ 2 grams per liter, Na
2hPO
412H
2o 1.0 ~ 10.0 grams per liter, MgSO
47H
2o 1 ~ 2 grams per liter, CaCl
20.05 ~ 0.1 grams per liter, trisodium citrate 0.1 ~ 2.0 grams per liter, Fe-EDTA solution 0.5 ~ 1mL, A5 solution 1 ~ 5mL and water;
Wherein Fe-EDTA solution formula is FeSO
47H
2o 20 ~ 30 grams per liter and EDTA 20 ~ 40 grams per liter;
A5 solution formula is H
3bO
32.5 ~ 4.0 grams per liters, MnCl
24H
2o 1.0 ~ 2.0 grams per liter, ZnSO
47H
2o 0.1 ~ 0.6 grams per liter, CuSO
45H
2o 5 ~ 10 grams per liter and Na
2moO
40.01 ~ 0.05 grams per liter.
8. the method according to any one of claim 1-2, is characterized in that, described method also comprises: mixed with other pigments by the frond after extraction xenthophylls and carry out spraying dry algae powder, or carry out separation and Extraction to the biologically active substance in frond.
9. the method according to any one of claim 1-2, is characterized in that, when micro-algae is to Chlorella vulgaris or chlorella ellipsoidea, photoinduction substratum consists of the following composition: KNO
31 ~ 8 grams per liter, MgSO
47H
2o, 0.5 ~ 1.0 grams per liter, CaCl
20.01 ~ 0.06 grams per liter, FeSO
47H
2o 0.01 ~ 0.06 grams per liter, EDTA 0.020 ~ 0.052 grams per liter;
When micro-algae is Chlorella pyrenoidesa, photoinduction substratum consists of the following composition: urea 0.1 ~ 2.0 grams per liter, MgSO
47H
2o, 0.5 ~ 1.0 grams per liter, CaCl
20.01 ~ 0.06 grams per liter, FeSO
47H
2o 0.01 ~ 0.06, EDTA 0.020 ~ 0.052 grams per liter, Trisodium Citrate 0.08 ~ 0.5 grams per liter.
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| WO2012065545A1 (en) * | 2010-11-16 | 2012-05-24 | 华东理工大学 | Microalgae culturing method for oil and lutein rapid accumulation |
| CN103103128A (en) * | 2011-11-10 | 2013-05-15 | 中国石油化工股份有限公司 | Method for high efficiency enrichment culture of microalgae |
| CN102517218B (en) * | 2011-12-28 | 2013-05-29 | 广东海洋大学 | Method for culturing microalgae on sea surface |
| CN103571906B (en) * | 2012-07-27 | 2018-12-11 | 上海泽元海洋生物技术有限公司 | A kind of new method efficiently producing astaxanthin using microalgae |
| CN104662162A (en) * | 2013-09-26 | 2015-05-27 | 华东理工大学 | Method using micro-algae for high-efficiency production of astaxanthin |
| CN111500464A (en) * | 2020-05-21 | 2020-08-07 | 福州大学 | Method for producing lutein by first mixotrophic-later autotrophic microalgae |
| CN114214202B (en) * | 2021-12-27 | 2024-02-13 | 广西源藻生物科技有限公司 | Method for co-producing lutein and protein by heterotrophic-photoinduction tandem culture of microalgae |
| CN116790382B (en) * | 2023-08-17 | 2023-11-03 | 藻辰(山东)生物工程有限公司 | High-yield lutein mutant chlorella, and preparation method and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1837351A (en) * | 2006-04-12 | 2006-09-27 | 华东理工大学 | Method for culturing chlorella with high-density and high-quality |
| CN1878872A (en) * | 2004-05-26 | 2006-12-13 | 雅马哈发动机株式会社 | Method of producing xanthophyll |
| CN101555454A (en) * | 2009-05-15 | 2009-10-14 | 华南理工大学 | Method for synchronously improving biomass and lutein of heterotrophic chlorella |
| CN102325874A (en) * | 2008-12-19 | 2012-01-18 | α-J研究有限合伙公司 | Optimization of algal product production through uncoupling cell proliferation and algal product production |
-
2010
- 2010-12-01 CN CN201010567920.3A patent/CN102094061B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1878872A (en) * | 2004-05-26 | 2006-12-13 | 雅马哈发动机株式会社 | Method of producing xanthophyll |
| CN1837351A (en) * | 2006-04-12 | 2006-09-27 | 华东理工大学 | Method for culturing chlorella with high-density and high-quality |
| CN102325874A (en) * | 2008-12-19 | 2012-01-18 | α-J研究有限合伙公司 | Optimization of algal product production through uncoupling cell proliferation and algal product production |
| CN101555454A (en) * | 2009-05-15 | 2009-10-14 | 华南理工大学 | Method for synchronously improving biomass and lutein of heterotrophic chlorella |
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