CN102115776A - Microalgae screening method and system thereof - Google Patents
Microalgae screening method and system thereof Download PDFInfo
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- CN102115776A CN102115776A CN2010100012068A CN201010001206A CN102115776A CN 102115776 A CN102115776 A CN 102115776A CN 2010100012068 A CN2010100012068 A CN 2010100012068A CN 201010001206 A CN201010001206 A CN 201010001206A CN 102115776 A CN102115776 A CN 102115776A
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Abstract
The invention discloses a microalgae screening method and a system thereof. The method comprises the following steps: microalgae strains to be screened are respectively placed in a plurality of photo bioreactors in a microalgae culture device to be cultured; a controllable gas outlet pipe and at least a constant-temperature water tank on a gas distribution unit in the microalgae culture device provide photosynthetic reaction gas at a set flow rate and a set culture temperature for the microalgae strains in each photo bioreactor; a lamp bracket placed on the side surface of each photo bioreactor provides even illumination at a set intensity for the microalgae strains; and performance detection equipment acquires the microalgae strains cultured in each photo bioreactor and conducts biological performance index detection, thereby screening out the microalgae strains with the best biological performance indexes. The biological performance indexes of different and identical types of microalgae strains under different culture conditions can be accurately evaluated, compared and screened, thereby achieving efficient, rapid and batched screening of the microalgae strains.
Description
Technical field
The present invention relates to biological technical field, refer to a kind of little algae screening method and system that the strain of little algae algae and growth conditions thereof are screened of realizing especially.
Background technology
Little algae is meant the unicellular of some microcosmic or many cells colony or thread algae, and as chlorella, salt algae, spirulina etc., great majority are planktonic algaes, and biomass is big, distribution is wide.Little algae can obtain biomass fuel by pyrolysis, is the important recyclable organism energy; Many little algaes can accumulate a large amount of greases in cell, high-content can reach more than 70% of dry cell weight, and the method by biorefinery is rich in from these extracts grease greasy microalgae cell, be used to prepare edible oil and biofuel; Also containing nutritive ingredients such as multivitamin, carotenoid, protein, polyunsaturated fatty acid in the microalgae cell, is the important source material source of biological medicine, functional food and cosmetics production; Little algae also can be used in the industrial processes such as wastewater treatment, to reduce environmental pollution.
Little algae with simple cell structure, with respect to more effectively conversion solar can (the photosynthesis transformation efficiency can reach more than 10%) with the Lu Sheng higher plant, and reproduction speed is very fast, and yield per unit is the several times of Lu Sheng higher plant.Owing to these reasons, many oil-containing micro-algaes are compared with high Lu Sheng oil crops, and they can produce 20 times of greases more than the amount in the unit surface.Therefore, little algae is the most potential liquid biofuel raw materials for production---greasy important source.
Determining of the evaluation of good little algae algae kind, screening and algae strain optimal growth condition; be the essence work of micro-algae large-scale cultivation and little algae product development; and the determining and must compare by the biological property to the various algae strains of the growth under various growth conditionss of the optimal growth condition of the evaluation of good little algae algae kind, screening and algae strain, as the biochemical composition composition in the speed of growth, the cell, to the resistance of environment etc.The principal element that influences algae strain growth and cellular constituent is illumination, temperature, salinity, basicity (pH) etc., therefore, in the evaluation and screening experiment of the strain of little algae algae, when cultivating the little algae algae kind of to be evaluated and screening, need to guarantee culture condition such as illumination, temperature and ventilatory effect, come the reasonable algae strain of screenability from a plurality of algae strains, and the optimal culture condition of definite algae strain.
In the prior art, mainly by triangular flask being placed growth cabinet or using the illumination shaking table to carry out the cultivation and the screening of the strain of little algae algae.Wherein, the triangular flask of cultivating the strain of little algae algae is placed growth cabinet, feed photosynthetic response gas, carry out little algae when cultivating, because growth cabinet can provide the temperature of comparison homogeneous and controllable, coordinate mutually with culture temperature with ventilatory effect but be difficult to regulate illumination.Realize the cultivation of little algae algae strain by the illumination shaking table, shaking table can regulating and controlling temperature and illumination, simultaneously can be by shaking the accelerating gas exchange, improve ventilatory effect, but its control to ventilatory effect is not very good, can not regulate accurately as required, and its illumination is also inhomogeneous, can't reaches the illumination demand of micro algae growth.
And the shared space of triangular flask is bigger, the difficult screening that realizes the strain of a large amount of little algae algae, and shaking table once can only be cultivated a kind of algae strain, and the screening that realize the strain of a large amount of little algae algae is difficulty relatively also.
Publication number is to have mentioned a kind of scheme of Dunaliella salina (Dunaliella prava) being carried out the exploration of optimum growh (cultivation) condition by the small light bio-reactor in the international patent application of WO2006020177, be diluted to unified initial growth condition to little algae every day, illumination condition is changed to 2000 μ Em-2s-1 by 150 μ Em-2s-1, the 50 μ Em-2s-1 that raise every day, the whole optimization time needs 40 days.This method also can only be explored a kind of little algae interior growing state of short period of time under given conditions at every turn, can't monitor the situation of the growth and the metabolite accumulation in algae strain whole growth cycle under given conditions.And this heuristic approach is consuming time very long, can not be used for cultivation and screening of cultivating the different mutant strains of different algae strains or same algae strain in enormous quantities simultaneously.
As seen, triangular flask of the prior art adds stable homogeneous and the coordination that growth cabinet and illumination shaking table all are difficult to accomplish various culture condition, can not well provide some little algae to same kind to carry out different culture condition required in the process of optimal culture condition screening and to the culture condition of stable and consistent required in the process that different types of little algae is estimated and screens under the same culture conditions.And, can not disposablely carry out screening and culturing so that carry out the evaluation and test of biological property index at a large amount of algae strains because it takes up room greatly, needs the lot of manpower and material resources input.Therefore, little algae of the prior art is cultivated and screening system all can not well be used for the cultivation and the screening of a large amount of algae strains, can't be efficiently, mass ground carries out the biological property index that the algae strain was screened and evaluated and tested accurately to little algae.
Summary of the invention
The embodiment of the invention provides a kind of little algae screening method and system, can not be efficiently in order to solve in the prior art, fast, mass carries out the problem that little algae is screened and accurately evaluates and tests algae strain biological property index.
A kind of little algae screening method comprises:
The micro algae culturing liquid that little algae algae strain to be screened places some bioreactors of micro algae culturing device to hold is respectively cultivated;
At least one Water Tank with Temp.-controlled of controlled escape pipe on the gas distribution member that links to each other with the inlet mouth of each bioreactor in the described micro algae culturing device and the described bioreactor of arrangement is for the little algae algae strain in each bioreactor provides the photosynthetic response gas of setting flow and the culture temperature of setting; Be placed on the adjustable lamp bracket in described bioreactor side and position, the even illumination of setting intensity is provided for the strain of described little algae algae;
Performance detection apparatus obtains in each bioreactor the strain of cultured microalgae algae and carries out the biological property index and detect, and realizes filtering out little algae algae strain of biological property index the best.
A kind of little algae screening system comprises: micro algae culturing device, little algae performance detection apparatus;
Described micro algae culturing device, be used for cultivating some little algae algae strain to be screened, for the little algae algae strain in each bioreactor provides the photosynthetic response gas of setting flow, the culture temperature of setting and the even illumination of setting intensity by some bioreactors;
Described little algae performance detection apparatus is used for obtaining the strain of cultured microalgae algae from described micro algae culturing device, and the biological property index of little algae algae strain of obtaining is detected; Realization filters out described little algae algae strain of biological property index the best according to the biological property index that detection obtains.
The micro algae culturing liquid that little algae algae strain to be screened places some bioreactors of micro algae culturing device to hold is respectively cultivated by little algae screening method and system that the embodiment of the invention provides; At least one Water Tank with Temp.-controlled of controlled escape pipe on the gas distribution member that links to each other with the inlet mouth of each bioreactor in the described micro algae culturing device and the described bioreactor of arrangement is for the little algae algae strain in each bioreactor provides the photosynthetic response gas of setting flow and the culture temperature of setting; Be placed on the adjustable lamp bracket in described bioreactor side and position, the even illumination of setting intensity is provided for the strain of described little algae algae; Performance detection apparatus obtains in each bioreactor the strain of cultured microalgae algae and carries out the biological property index and detect, and realizes filtering out little algae algae strain of biological property index the best.This method can realize from different little algae algae kinds good algae kind of screening and the optimal culture condition of exploring a certain little algae algae kind, can require to provide identical or different separately culture condition according to different screenings for each bioreactor, realize intensity of illumination, the stable and consistent of culture temperature and ventilatory effect, and can compare monitoring to the biological property of the different growth phases of little algae, improve accuracy and comparability that little algae screening and the speed of estimating and efficient and biological property detect greatly, realized the efficient of little algae, fast, the mass screening.
Description of drawings
Fig. 1 is the structural representation of micro algae culturing device in the embodiment of the invention;
Fig. 2 is the structural representation of bioreactor in the embodiment of the invention;
Fig. 3 is the structural representation of Water Tank with Temp.-controlled in the embodiment of the invention;
Fig. 4 is the structural representation of gas distributor in the embodiment of the invention;
Fig. 5 is the structural representation of lamp bracket in the embodiment of the invention;
Fig. 6 is the structural representation of reactor frame in the embodiment of the invention;
Fig. 7 is the structural representation of little algae screening system in the embodiment of the invention;
Fig. 8 is the schema of little algae screening method in the embodiment of the invention;
Fig. 9 is at the schema of little algae screening method of same algae kind in the embodiment of the invention one;
Figure 10 is at the schema of little algae screening method of different algae kinds in the embodiment of the invention two.
Embodiment
Little algae screening method and system that the embodiment of the invention provides, can the pilot-gas flow by adopting, the micro algae culturing device of intensity of illumination and culture temperature, the little algae algae strain that is embodied as identical type provides different culture condition so that filter out little algae algae strain of biological property index the best and determine its pairing culture condition, and is embodied as the strain of different types of little algae algae and provides identical culture condition so that filter out little algae algae strain of biological property index the best.
The micro algae culturing device that the embodiment of the invention provides is illustrated in figure 1 as the structural representation of micro algae culturing device, comprising: some bioreactors 1, at least one Water Tank with Temp.-controlled 2, gas distribution member 3 and at least one lamp bracket 4.Only illustrated a Water Tank with Temp.-controlled 2 and a lamp bracket 4 among Fig. 1, in the practical application, a Water Tank with Temp.-controlled 2 can dispose more than one lamp bracket 4, and when the quantity of Water Tank with Temp.-controlled 2 increased, the quantity of lamp bracket 4 generally also can increase thereupon.
Water Tank with Temp.-controlled 2 is used to settle bioreactor 1, and the culture temperature of setting is provided for the strain of little algae algae.
Gas distribution member 3 is provided with some controlled escape pipes that can the pilot-gas flow, and each controlled escape pipe is connected with the inlet mouth of a bioreactor 1, and being used to be respectively the bioreactor that is connected provides the photosynthetic response gas of setting flow.
The structure of above-mentioned bioreactor 1 is an enclosed bioreactor as shown in Figure 2, comprises glass pillar reaction vessel 101, the glass capillary 102 as inlet pipe, the glass capillary 103 as escape pipe, plug 104.Wherein, glass pillar reaction vessel 101 is used to hold micro algae culturing liquid and cultivates little algae algae strain to be screened as the reaction vessels of cultivating little algae, wherein, seal with plug 104 at glass pillar reaction vessel 101 tops, glass capillary 102 and 103 inserts in the glass pillar reaction vessel 101 by the through hole on the plug 104, the derivation of the residual gas that produces after the importing of realization photosynthetic response gas and the photosynthetic response.
In addition, glass capillary 103 bloats bubble in the plug 104 by inserting to photosynthetic response gas that bioreactor 1 provides by controlled escape pipe, enters in the micro algae culturing liquid.This mode can play the effect of effectively washing away with the internal surface of clear light bio-reactor simultaneously, has avoided the strain of little algae algae in the accumulation of reactor internal surface and/or adhere to.
Except can directly blasting the gas by glass capillary, can also use small-sized, a simple gas distributor or scatterer, realize bioreactor inner better ventilation and stirring effect.
The mechanism of above-mentioned Water Tank with Temp.-controlled 2 is provided with water-in 201 and water outlet 202 as shown in Figure 3 on the Water Tank with Temp.-controlled 2.Water-in 201 is used to allow the recirculated water (can be high-temperature water or water at low temperature) of design temperature to enter Water Tank with Temp.-controlled 2.Water Tank with Temp.-controlled 2 is used to allow the bioreactor 1 of recirculated water and arrangement to carry out heat exchange.Water outlet 202 is used for the recirculated water (water at low temperature that obtains after the high-temperature water heat exchange or water at low temperature carry out obtain after the heat exchange high-temperature water) that carries out after the heat exchange is discharged Water Tank with Temp.-controlled 2.Bioreactor 1 is immersed in the water of design temperature of Water Tank with Temp.-controlled 2, can provide recirculated water for Water Tank with Temp.-controlled 2 by water circulating pump and circulation line, guarantees that water temperature is constant.By temperature control device, for example: Arrangement for heating or cooling water in receptacle for live fish, the temperature that the recirculated water that is provided is provided.
Also can not use Water Tank with Temp.-controlled 2, bioreactor 1 is exposed in the air, regulate and control the temperature of bioreactor 1 by the temperature of bioreactor place environment space.
Above-mentioned gas allocation units 3 specifically comprise: at least one mixed gas tank 31 and at least one gas distributor 32.Only illustrated a gas distributor 32 among the figure.
Some controlled escape pipes are set on the gas distributor 32, provide the photosynthetic response gas of setting flow for the bioreactor that is connected by controlled escape pipe.Controlled escape pipe comprises: the flowrate control valve that is provided with on outlet pipe and each outlet pipe, and by the gas flow on the flowrate control valve control outlet pipe.
For example: the structure of above-mentioned gas divider 32 comprises inlet mouth 301, gas distributing chamber 302 that is communicated with mixed gas tank 31 and the controlled escape pipe of being made up of the flowrate control valve 303 that is provided with on outlet pipe 304 and each outlet pipe as shown in Figure 4.Thereby realize that mixed photosynthetic response gas distributes to each outlet pipe 304 in gas distributing chamber, for the bioreactor 1 that is connected with each breather line 304 provides the photosynthetic response gas of required flow separately.
Gas distribution member 3 also comprises: the air compressor 33 and the first flow meter 34 that is connected, and the carbon dioxide steel cylinder 35 and second under meter 36.Wherein, air compressor 33 is used to mixed gas tank 31 that pressurized air is provided; First flow meter 34 links to each other with air compressor 33, is used to monitor the flow that air compressor 33 offers the air that mixes gas tank 31.Carbon dioxide steel cylinder 35 is used to mixed gas tank 31 that carbon dioxide is provided; Second under meter 36 links to each other with carbon dioxide steel cylinder 35, is used to monitor the flow of carbon dioxide steel cylinder 35 for the carbon dioxide that mixes gas tank 31 and provide.
That is to say that provide air, carbon dioxide steel cylinder 35 that carbon dioxide is provided by air compressor 33, its body of carbonic acid gas can be taken from industrial tail gas or waste gas, therefore, little algae is cultivated and can realize that waste gas reclaims environment purification.A certain proportion of air and carbon dioxide are mixed, by the volume ratio of first flow meter 34 and second under meter, 36 control airs and carbon dioxide.Mixing the gas flow of each bioreactor of back feeding controls by flowrate control valve.
The structure of above-mentioned lamp bracket 4 comprises some fluorescent tubes that are arranged in parallel or other piped light sources such as LED fluorescent tube as shown in Figure 5.Realize the light radiation intensity uniformity of light source on each bioreactor by evenly gather fluorescent tube or LED fluorescent tube.The photoirradiation intensity higher as needs can all be arranged lamp bracket at the bilateral of bioreactor.
Above-mentioned micro algae culturing device can also comprise reactor frame 5 as shown in Figure 6.Reactor frame 5 can be placed on Water Tank with Temp.-controlled 2, is used to place bioreactor 1, realizes bioreactor 1 is placed in the Water Tank with Temp.-controlled 2.Reactor frame 2 shown in Fig. 6 is the synthetic glass support bodys with three-decker, wherein go up some holes that are used to insert bioreactor 1 are set on the two-layer dividing plate, after bioreactor inserts, support by the dividing plate of bottom and the hole of last two-layer dividing plate correspondence position, realize vertically placing.
Use above-mentioned micro algae culturing device can make up a kind of little algae screening system, as shown in Figure 7, comprising: above-mentioned micro algae culturing device 10 and little algae performance detection apparatus 20.
Wherein, the structure of micro algae culturing device 10 and function are as mentioned above.
Little algae performance detection apparatus 20 is used for obtaining the strain of cultured microalgae algae from micro algae culturing device, and the biological property index of little algae algae strain of obtaining is detected.Then, the mode by artificial screening or device software screening according to the biological property index that detection obtains, filters out little algae algae strain of biological property index the best.
In above-mentioned little algae screening system, can select different little algae performance detection apparatus 20 for use according to different detection indexs and detection method.Wherein, little algae performance detection apparatus can be selected any one or a few performance detection apparatus of the prior art for use, for example: comprise at least a in the following equipment:
Be used to measure the spectrophotometer of algae strain optical density(OD), the precision balance that is used to measure algae strain dry weight, the chlorophyll content that is used to measure the algae strain and other pigment contents spectrophotometer or high performance liquid chromatograph, be used to measure Nitrogen evaporator, precision balance and the gas chromatograph of the fat content of algae strain.Detect the corresponding relation of index, detection method and selected little algae performance detection apparatus 20, as shown in table 1 below:
Table 1
Wherein, optical density(OD) is the detection index that characterizes the cell quantity of little algae algae strain, and dry weight is the detection index that characterizes the biomass of little algae algae strain.
Some bioreactors of above-mentioned micro algae culturing device 10 can be used to cultivate little algae algae strain of same kind, to obtain its optimal culture condition.
Accordingly, the Water Tank with Temp.-controlled of micro algae culturing device 10 is set to a plurality of, specifically be used to bioreactor that different culture temperature is provided, the controlled escape pipe of gas distribution member then specifically is used to each bioreactor that the photosynthetic response gas of different flow or the different proportionings of same traffic is provided.
Accordingly, little algae performance detection apparatus 20 is used for carrying out the biological property index from little algae algae strain that micro algae culturing device 10 obtains the same kind of cultivation and detects; So that follow-up the screening, filter out little algae algae strain of biological property the best in little algae algae strain of the same kind of being cultivated, and determine the pairing culture condition of little algae algae strain of this biological property the best; Wherein, culture condition comprises the ventilation situation of culture temperature, intensity of illumination and photosynthetic response gas.
Some bioreactors of above-mentioned micro algae culturing device 10 can be used to cultivate the strain of different types of little algae algae, with the performance of each the algae strain of evaluation under this culture condition, thereby obtain the optimum algae kind of setting under the culture condition.
Accordingly, the Water Tank with Temp.-controlled of micro algae culturing device 10 specifically is used to bioreactor that identical culture temperature is provided; The controlled escape pipe of gas distribution member specifically is used to each bioreactor that the photosynthetic response gas of same traffic is provided.
Accordingly, little algae performance detection apparatus 20 is used for carrying out the biological property index from different types of little algae algae strain that micro algae culturing device 10 obtains cultivation and detects; So that follow-up the screening filters out under the same culture conditions that is provided, little algae algae strain of biological property the best in the strain of different types of little algae algae, and the algae kind of little algae algae strain of definite this biological property the best.
Above-mentioned little algae screening system also comprises: industrialization culture apparatus 40.
When micro algae culturing device is used to cultivate little algae algae strain of same kind and provides different culture condition for it, after determining the pairing culture condition of little algae algae strain of biological property the best, industrialization culture apparatus 40 is used to adopt the pairing culture condition of little algae algae strain of biological property the best of determining and the pairing algae kind of little algae algae strain of same kind to carry out little algae industrialization cultivation.
When micro algae culturing device is used to cultivate the strain of different types of little algae algae and provides identical culture condition for it, after determining the pairing algae kind of little algae algae strain of biological property the best, industrialization culture apparatus 40 is used to adopt the pairing algae kind of little algae algae strain of the culture condition determined and biological property the best to carry out little algae industrialization and cultivates.
Utilize flow process that above-mentioned little algae screening system realizes the screening of little algae shown in 8 figure, comprise the following steps:
Step S11: the micro algae culturing liquid that little algae algae strain that will be to be screened places some bioreactors of micro algae culturing device to hold is respectively cultivated.
Can bioreactor be placed in the Water Tank with Temp.-controlled by being placed on the reactor frame in the Water Tank with Temp.-controlled.
Step S12: micro algae culturing device provides Incubation Condition for the strain of little algae algae.
Micro algae culturing device can provide required Incubation Condition for the little algae algae strain in each bioreactor, wherein:
(1) with gas distribution member that the inlet mouth of each bioreactor links to each other on controlled escape pipe the photosynthetic response gas of setting flow is provided for the little algae algae strain in each bioreactor.Specifically comprise:
Air and carbon dioxide that two inlet mouths of mixed gas tank that will be from gas distribution member enter the setting volume ratio of mixing gas tank mix, and obtain photosynthetic response gas; By the controlled escape pipe pilot-gas flow on the gas distributor in the gas distribution member, provide the photosynthetic response gas of setting flow for the bioreactor that is connected.
Promptly control the ventilation situation of photosynthetic response gas in each bioreactor, comprise ventilation flow rate and bubble size etc. by controlled escape pipe.The photosynthetic response gas that feeds is the suitable photosynthetic air of little algae or air and carbon dioxide gas mixture.The proportioning of general air and carbon dioxide is: 0.5%-40%.
(2) temperature of at least one Water Tank with Temp.-controlled of arrangement bioreactor is for the little algae algae strain in each bioreactor provides corresponding culture temperature.Specifically comprise:
To Water Tank with Temp.-controlled, inject the recirculated water (high-temperature water or water at low temperature) of design temperature from the water-in of Water Tank with Temp.-controlled, recirculated water carries out getting rid of from the water outlet of Water Tank with Temp.-controlled after the heat exchange with the bioreactor of settling in Water Tank with Temp.-controlled, is embodied as the culture temperature that little algae algae strain in each bioreactor provides setting.
Generally according to the characteristic of the algae kind of the little algae algae strain in the bioreactor and the culture condition of required concern, determine the temperature of recirculated water, then, the recirculated water of design temperature enters from the water-in 201 of Water Tank with Temp.-controlled, discharge by water outlet 202, guarantee the temperature of bioreactor.
The temperature that generally needs to keep the micro algae culturing liquid in the bioreactor is in 1 5-37 ℃ scope, and is preferable, and the temperature that keeps the micro algae culturing liquid in the bioreactor is in 1 5-25 ℃.Water in the Water Tank with Temp.-controlled has certain attenuation for intensity of illumination, but is not fairly obvious through measurement, so its decay to intensity of illumination is little to the influence of micro algae growth.
(3) be placed on the lateral lamp bracket of bioreactor, the even illumination of setting intensity is provided for the strain of little algae algae.
The lamp bracket position is adjustable, the quantity and spacing of fluorescent tube is also adjustable on the lamp bracket, thereby realizes being adjusted to the purpose that little algae algae strain in the bioreactor provides the illumination of varying strength.
Step S13: performance detection apparatus obtains in each bioreactor the strain of cultured microalgae algae and carries out the biological property index and detect.
The biological property index comprises at least a in the following index: the chlorophyll content of algae strain optical density(OD), algae strain dry weight, algae strain and the fat content of other pigment contents and algae strain etc.; Can select for use equipment given in the above-mentioned table 1 to carry out the biological property index detects.
Step S14: the little algae algae strain that filters out biological property index the best.
Ventilation situation among the above-mentioned steps S12 in each photosynthetic reactor is all adjustable separately, and a large amount of O that can produce when taking photosynthesis out of and carry out
2, the process of accelerating gas exchange promotes photosynthetic carrying out.Because what bioreactor adopted among the application is the short column reactor of optical path, can make the little algae algae liquid in the bioreactor can thorough mixing by the control of aeration condition, make the strain of little algae algae can enter light incidence section and dark space in the short period of time and realize being alternately distributed, thereby can utilize luminous energy to carry out photosynthesis fully.That is to say by enough gas-liquid contact times being provided and providing little algae the required enough dark and the residence time, guarantee photosyntheticly efficiently to carry out, specifically can provide enough duration of contact by the length and the ventilation speed of selecting reactor, need simultaneously to consider that multiple factors such as the volume of the composition of absorption rate, air inlet of the speed of growth, the carbon of the algae kind of being cultivated and flow and liquid and height are to the influence of duration of contact, so that reach required mass transfer level in the middle of gas and liquid medium.By the cross-sectional area of selecting reactor, the abundant turbulence of nutrient solution when guaranteeing ventilation for little algae provides the required dark and the residence time, and is avoided little algae sedimentation and is deposited on the bottom of reactor.
That is to say, when the cross-sectional area (or saying internal diameter) of the glass capillary of selecting reactor and ventilation, need to consider the gas flow that controlled escape pipe provided, required turbulence effect, the size of bubble and the size of gas distributor etc. of gas distributor, so that select suitable cross-sectional area or say internal diameter.If the internal diameter of the glass capillary of ventilation is excessive, then can cause bubble very big, carbonic acid gas is descended from the mass transfer level that gas phase enters liquid phase, assimilation effect also descends thereupon; If the internal diameter of the glass capillary of ventilation is too small, then can cause bubble less, may make the strain of little algae algae under capillary effect, leave liquid medium with bubble floating, very unfavorable for the growth of little algae algae strain.
Screening to little algae biological property index, mainly be divided into two kinds of situations: the one, provide different culture condition at little algae algae strain of same kind, filter out wherein little algae algae strain of biological property index the best, with the optimal culture condition of little algae algae strain of obtaining selected a certain kind.The 2nd, at the strain of different types of little algae algae, identical culture condition is provided, filter out wherein little algae algae strain of biological property index the best,, realize obtaining the purpose of good algae kind with biological property the best of little algae algae strain which kind under the culture condition that is provided is provided.Adopt two specific embodiments to be elaborated at both of these case below.
Embodiment one
Little algae screening method that the embodiment of the invention one provides is explored growth (cultivation) condition of its best at little algae algae strain of same kind, its flow process as shown in Figure 9, performing step is as follows:
Step S101: little algae algae strain of cultivating same kind in the micro algae culturing liquid that holds in the some bioreactors in micro algae culturing device.
For example: the diameter of the glass column reactor of selecting for use is 40mm, highly is 600mm, and the volume 750ml of whole reactor, reactor are the borosilicate glass material.In each bioreactor of micro algae culturing device, hold little algae algae strain of identical algae kind.
Step S102: micro algae culturing device provides the culture condition that has nothing in common with each other for the little algae algae strain in each bioreactor.
Promptly, feed the photosynthetic response gas of the different proportionings of different flow or same traffic respectively at little algae algae strain in each bioreactor, and controller bubble size.Bioreactor is placed in a plurality of Water Tanks with Temp.-controlled that different culture temperature are provided obtains different culture temperature.
Can also provide different intensities of illumination by the position of adjusting lamp bracket and the quantity and spacing of top fluorescent tube thereof.Above-mentioned intensity of illumination, culture temperature, ventilation situation can be chosen according to orthogonal test.
For example, use five cover bioreactors to cultivate grape algae (Botryococcus braunii), nutrient solution, promptly substratum is Modified Bold 3N Medium, the inoculum density of little algae algae strain is provided with five gradients.CO in the photosynthetic response gas
2Content be 1%-25%, and five gradients are set.At this moment, then need respectively by a plurality of gas distribution member, the gas distributor of photosynthetic response gas by separately inserted needs prescribed concentration CO
2The glass capillary of photosynthetic response gas in.The culture temperature scope is 19-31 ℃, and five gradients also are set, by five Water Tank with Temp.-controlled regulation and control.
Place the bioreactor of five different vaccination concentration like this in each Water Tank with Temp.-controlled, and the needed CO of each bioreactor
2The photosynthetic response gas that concentration is different is provided by different gas distribution member.Promptly realized selecting three factors, five horizontal quadrature test design, so that effectively explore the growth conditions of the best of selected little algae algae kind.
The component of above-mentioned substratum Modified Bold 3N Medium is as shown in table 3.
Table 3
Step S103: the detection of biological property index is carried out in little algae algae strain of obtaining the same kind of cultivating in each bioreactor.
The little algae algae strain that can obtain different growth phases detects, to observe the upgrowth situation of each different steps.
Step S104: determine little algae algae strain of biological property index the best.Specifically comprise:
Filter out little algae algae strain of biological property the best in little algae algae strain of same kind, and determine the pairing culture condition of little algae algae strain of this biological property the best.Wherein, culture condition comprises the ventilation situation of culture temperature, intensity of illumination and photosynthetic response gas.
In for example above-mentioned orthogonal test, determine upgrowth situation the little algae algae strain under multiple different culture condition, i.e. the best algae strain of biological property index.
Step S105: the pairing culture condition of little algae algae strain of determining biological property index the best.
For example: determine temperature, intensity of illumination of the photosynthetic response gas flow that feeds in the bioreactor of little algae algae strain of biological property the best in the above-mentioned orthogonal test, place Water Tank with Temp.-controlled etc.
Embodiment two
Little algae screening method that the embodiment of the invention two provides, at the strain of different types of little algae algae, give the culture condition of stable and consistent, explore biological property the best in which algae kind of specific growth (cultivation) condition, its flow process as shown in figure 10, performing step is as follows:
Step S201: cultivate the strain of some different types of little algae algaes in the micro algae culturing liquid that holds in the some bioreactors in micro algae culturing device.
For example: the diameter 40mm of the glass column reactor of selecting for use 101, height 600mm, the volume 750ml of whole reactor, reactor are the borosilicate glass material.In each bioreactor of micro algae culturing device, hold little algae algae strain of different algae kinds.For example: little algae algae strain of Chlorococcum (Chlorococcum sp.) and two kinds of different algae kinds of chlorella (Chlorella vulgaris), substratum is BG11, each algae strain is provided with three parallel samples.Wherein, substratum is that the component of BGll is as shown in table 3.
Table 3
Step S202: micro algae culturing device provides identical culture condition for the different types of little algae algae strain in each bioreactor.
Promptly, feed the photosynthetic response gas of same traffic respectively, and controller bubble size is consistent at little algae algae strain in each bioreactor.Bioreactor is placed at least one Water Tank with Temp.-controlled that identical culture temperature is provided obtains identical culture temperature.Provide identical intensity of illumination by the position of adjusting lamp bracket and the quantity and spacing of top fluorescent tube thereof.
For example: the energy-saving sight lamp of 20 36W of horizontally-arranged that gathers on the lamp bracket provides illumination.Feeding contains 1%-2%CO
2Photosynthetic response gas.
Step S203: obtain different types of little algae algae strain of cultivating in each bioreactor and carry out the detection of biological property index.
Measure the biological property index such as dry weight, optical density(OD) of different growth phases, be used to estimate the biological properties such as the speed of growth, biomass and cellular constituent of different algae strains.Can be according to the period measurement of setting, for example: survey once every day.
Step S204: determine little algae algae strain of biological property index the best.Specifically comprise:
Little algae algae strain of biological property the best in the strain of different types of little algae algae is provided under the same culture conditions that is provided.
Step S205: the pairing algae kind of little algae algae strain of determining biological property index the best.
For example: determine the little algae algae kind in the bioreactor of little algae algae strain of biological property the best, and the temperature of the photosynthetic response gas flow that feeds, place Water Tank with Temp.-controlled, intensity of illumination etc.
Filter out little algae algae strain of biological property the best by above-mentioned screening process after, then can carry out the industrialization cultivation according to this pairing culture condition of little algae algae strain that filters out and affiliated little algae algae kind.Comprising: to the pairing culture condition of little algae algae strain of biological property the best in little algae algae strain of the same kind determined among the embodiment one, adopt the culture condition of determining that the pairing algae kind of this little algae algae strain is carried out little algae industrialization and cultivate.And, adopt culture condition of setting and the algae kind of being determined to carry out little algae industrialization and cultivate the affiliated algae kind of little algae algae strain of biological property the best in the little algae algae strain not of the same race of the culture condition of the setting determined among the embodiment two.
Can also carry out pilot scale before industrialization is cultivated amplifies, checking uses the culture condition of determining that selected algae kind is carried out the feasibility that industrialization is cultivated, that is to say, according to biological property evaluation to the strain of little algae algae, the algae kind of the excellent property that filters out under the identical culture condition or the optimal culture condition of exploring at same algae kind, adopt corresponding culture condition to carry out the pilot scale amplification culture the good algae kind that filters out, because the culture condition of pilot scale amplification culture and large-scale technical grade reactor are in illumination, everyways such as ventilation are more approaching, therefore, little algae of suitable pilot scale amplification culture also can be good at adapting to this culture environment of technical grade reactor.Wherein the technical grade reactor comprises that optical paths such as tubular reactor, plate-type reactor are relatively short, the large-scale preferably reactor of ventilatory effect.
Little algae screening method and system that the embodiment of the invention provides, the micro algae culturing device of identical or different culture condition can be provided respectively for a plurality of bioreactors simultaneously by design, solved in the prior art and can not provide different grown cultures conditions respectively for the strain of some different types of little algae algaes, to explore the problem of good algae strain, simultaneously also solved in the prior art and can not provide different culture condition respectively, to determine the problem of its optimal growth condition for some algae strains of identical algae kind.Temperature control, ventilatory effect control, intensity of illumination control by micro algae culturing device, intensity of illumination, culture temperature and ventilatory effect etc. corresponding, stable and consistent can be provided according to demand, the experiment that above-mentioned little algae screening method is finished one-period needs the time generally only to need 7-10 days, promptly can finish in a short time a large amount of little algae screening mutant strains, therefrom filter out the good little algae algae strain of biological property and determine its only culture condition, shorten the cycle of little algae screening and culturing greatly, realized little algae screening efficient, quick, mass.Be convenient to simultaneously observe under the different culture condition, the upgrowth situation of different algae strains, and rationally contrast, obtain the biological property of little each growth phase of algae easily.
Above-mentioned little algae screening method and system are not only applicable to the cultivation of little algae and carry out the algae strain according to biological property and estimate and screen, and are equally applicable to other photosynthetic organism (for example: have photosynthetic bacterium etc.) evaluation and screening.
The above; only be the preferable embodiment of the present invention; but protection scope of the present invention is not limited thereto; anyly be familiar with those skilled in the art in the technical scope that the present invention discloses; the variation that can expect easily, replace or be applied to other similar devices, all should be encompassed within protection scope of the present invention.Therefore, protection scope of the present invention should be as the criterion with the protection domain of claims.
Claims (18)
1. a little algae screening method is characterized in that, comprising:
The micro algae culturing liquid that little algae algae strain to be screened places some bioreactors of micro algae culturing device to hold is respectively cultivated;
At least one Water Tank with Temp.-controlled of controlled escape pipe on the gas distribution member that links to each other with the inlet mouth of each bioreactor in the described micro algae culturing device and the described bioreactor of arrangement is for the little algae algae strain in each bioreactor provides the photosynthetic response gas of setting flow and the culture temperature of setting; Be placed on the adjustable lamp bracket in described bioreactor side and position, the even illumination of setting intensity is provided for the strain of described little algae algae;
Performance detection apparatus obtains in each bioreactor the strain of cultured microalgae algae and carries out the biological property index and detect, and realizes filtering out little algae algae strain of biological property index the best.
2. the method for claim 1 is characterized in that, gas distribution member specifically comprises for the strain of little algae algae provides the photosynthetic response gas of setting flow:
The air and the carbon dioxide that will enter the setting volume ratio of described mixed gas tank from two inlet mouths of the mixed gas tank the described gas distribution member mix, and obtain described photosynthetic response gas;
By the controlled escape pipe pilot-gas flow on the gas distributor in the described gas distribution member, provide the photosynthetic response gas of setting flow for the described bioreactor that is connected.
3. method as claimed in claim 2 is characterized in that, counts described mixed gas tank by air compressor in the described gas distribution member and the first flow that is connected the pressurized air of setting flow is provided; And
Count described mixed gas tank by carbon dioxide steel cylinder in the described gas distribution member and second flow that is connected the carbon dioxide of setting flow is provided.
4. as the arbitrary described method of claim 1-3, it is characterized in that, to described Water Tank with Temp.-controlled, inject the recirculated water of design temperature from the water-in of described Water Tank with Temp.-controlled, described recirculated water carries out getting rid of from the water outlet of described Water Tank with Temp.-controlled after the heat exchange with the bioreactor of settling in described Water Tank with Temp.-controlled, is embodied as the culture temperature that little algae algae strain in each bioreactor provides setting.
5. the method for claim 1, it is characterized in that, when little algae algae strain described to be screened is the little algae algae strain of same kind, described bioreactor is placed in a plurality of Water Tanks with Temp.-controlled that different culture temperature are provided, and the controlled escape pipe of described gas distribution member provides the photosynthetic response gas of setting flow for each bioreactor.
6. method as claimed in claim 5 is characterized in that, the described described little algae algae strain that filters out biological property index the best specifically comprises:
Filter out little algae algae strain of biological property the best in little algae algae strain of described same kind, and determine the algae kind and the pairing culture condition of little algae algae strain of this biological property the best; Described culture condition comprises the ventilation situation of culture temperature, intensity of illumination and photosynthetic response gas.
7. the method for claim 1, it is characterized in that, when little algae algae strain described to be screened is different types of little algae algae strain, described bioreactor is placed at least one Water Tank with Temp.-controlled that identical culture temperature is provided, and the controlled escape pipe of described gas distribution member provides the photosynthetic response gas of same traffic for each bioreactor.
8. method as claimed in claim 7 is characterized in that, the described described little algae algae strain that filters out biological property index the best specifically comprises:
Little algae algae strain of biological property the best in the strain of described different types of little algae algae, and the algae kind and the pairing culture condition of little algae algae strain of definite this biological property the best are provided under the same culture conditions that is provided; Described culture condition comprises the ventilation situation of culture temperature, intensity of illumination and photosynthetic response gas.
9. as claim 6 or 8 described methods, it is characterized in that, adopt the pairing algae kind of little algae algae strain and the pairing culture condition of described biological property the best of determining, carry out little algae industrialization and cultivate.
10. a little algae screening system is characterized in that, comprising: micro algae culturing device and little algae performance detection apparatus;
Described micro algae culturing device, be used for cultivating some little algae algae strain to be screened, for the little algae algae strain in each bioreactor provides the photosynthetic response gas of setting flow, the culture temperature of setting and the even illumination of setting intensity by some bioreactors;
Described little algae performance detection apparatus is used for obtaining the strain of cultured microalgae algae from described micro algae culturing device, and the biological property index of little algae algae strain of obtaining is detected; Realization filters out described little algae algae strain of biological property index the best according to the biological property index that detection obtains.
11. system as claimed in claim 10 is characterized in that, described micro algae culturing device specifically comprises: some bioreactors, gas distribution member, at least one Water Tank with Temp.-controlled and at least one lamp bracket;
Described bioreactor is placed in the described Water Tank with Temp.-controlled, is used to hold micro algae culturing liquid and cultivates little algae algae strain to be screened;
Described gas distribution member is provided with some controlled escape pipes that can the pilot-gas flow, each controlled escape pipe is connected with the inlet mouth of a described bioreactor, and being used to be respectively the bioreactor that is connected provides the photosynthetic response gas of setting flow;
Described Water Tank with Temp.-controlled is used to settle described bioreactor, and the culture temperature of setting is provided for the strain of described little algae algae;
Described lamp bracket is placed on the side of described Water Tank with Temp.-controlled, and described lamp bracket position is adjustable, is used to the little algae algae strain in the described bioreactor that the even illumination of setting intensity is provided.
12. system as claimed in claim 11 is characterized in that, described micro algae culturing device also comprises: be placed on the reactor frame in the described Water Tank with Temp.-controlled;
Described reactor frame is used to place described bioreactor, and described bioreactor is placed in the described Water Tank with Temp.-controlled.
13. device as claimed in claim 11 is characterized in that, described gas distribution member specifically comprises: mix gas tank and at least one gas distributor
Described mixed gas tank, the air and the carbon dioxide that are used for the setting volume ratio that will feed from two inlet mouths that are provided with mix, and obtain photosynthetic response gas, and offer described gas distributor;
Some described controlled escape pipes are set on the described gas distributor, provide the photosynthetic response gas of setting flow for the described bioreactor that is connected by described controlled escape pipe.
14. device as claimed in claim 13 is characterized in that, described controlled escape pipe comprises: the flowrate control valve that is provided with on outlet pipe and each outlet pipe;
Described flowrate control valve is used to control the gas flow on the described outlet pipe.
15. device as claimed in claim 13 is characterized in that, described gas distribution member also comprises: the air compressor and the first flow meter that is connected, and the carbon dioxide steel cylinder and second under meter;
Described air compressor is used to described mixed gas tank that pressurized air is provided;
Described first flow meter links to each other with described air compressor, is used to monitor the flow that described air compressor offers the air of described mixed gas tank;
Described carbon dioxide steel cylinder is used to described mixed gas tank that carbon dioxide is provided;
Described second under meter links to each other with described carbon dioxide steel cylinder, is used to monitor the flow of the carbonic acid gas that described carbon dioxide steel cylinder provides for described mixed gas tank.
16. as the arbitrary described device of claim 11-15, it is characterized in that, water-in and water outlet be set on the described Water Tank with Temp.-controlled;
Described water-in is used to allow the recirculated water of design temperature to enter described Water Tank with Temp.-controlled;
Described Water Tank with Temp.-controlled is used to allow the recirculated water of described design temperature and the bioreactor of arrangement to carry out heat exchange;
Described water outlet is used for the recirculated water after the heat exchange is discharged described Water Tank with Temp.-controlled.
17. system as claimed in claim 11 is characterized in that, described some bioreactors are used to cultivate little algae algae strain of same kind; Accordingly,
Described Water Tank with Temp.-controlled is a plurality of, specifically is used to described bioreactor that different culture temperature is provided; The controlled escape pipe of described gas distribution member specifically is used to each bioreactor that the photosynthetic response gas of different flow or the different proportionings of same traffic is provided; Accordingly,
Described little algae performance detection apparatus is used for carrying out the biological property index from little algae algae strain that described micro algae culturing device obtains the same kind of cultivation and detects; Realization filters out little algae algae strain of biological property the best in little algae algae strain of described same kind, and determines the algae kind and the pairing culture condition of little algae algae strain of this biological property the best; Described culture condition comprises the ventilation situation of culture temperature, intensity of illumination and photosynthetic response gas.
18. system as claimed in claim 11 is characterized in that, described some bioreactors are used to cultivate the strain of different types of little algae algae; Accordingly,
Described Water Tank with Temp.-controlled specifically is used to described bioreactor that identical culture temperature is provided; The controlled escape pipe of described gas distribution member specifically is used to each bioreactor that the photosynthetic response gas of setting flow is provided; Accordingly,
Described little algae performance detection apparatus is used for carrying out the biological property index from different types of little algae algae strain that described micro algae culturing device obtains cultivation and detects; Realization filters out under the same culture conditions that is provided, little algae algae strain of biological property the best in the strain of described different types of little algae algae, and the algae kind and the pairing culture condition of little algae algae strain of definite this biological property the best; Described culture condition comprises the ventilation situation of culture temperature, intensity of illumination and photosynthetic response gas.
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