CN105779268B - A kind of device and method of culture photosynthetic organism - Google Patents
A kind of device and method of culture photosynthetic organism Download PDFInfo
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- CN105779268B CN105779268B CN201410811144.5A CN201410811144A CN105779268B CN 105779268 B CN105779268 B CN 105779268B CN 201410811144 A CN201410811144 A CN 201410811144A CN 105779268 B CN105779268 B CN 105779268B
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Abstract
The present invention provides a kind of device and methods of culture photosynthetic organism, device therein includes the cultivation region for accommodating culture solution of top visible light, flow-disturbing mechanism is installed in the cultivation region, the flow-disturbing mechanism has can be along the inclined-plane of cultivation region bottom surface persistent movement；The lasting thrust obliquely that the flow-disturbing mechanism is used to generate the culture solution of the cultivation region bottom during its persistent movement by the inclined-plane pushes the culture solution of the cultivation region bottom to liquid level persistent movement.Device and method culture photosynthetic organism using the present invention can not only improve the growth efficiency of photosynthetic organism, and can save energy consumption.
Description
Technical field
The present invention relates to the devices and side of a kind of device and method of culture photosynthetic organism more particularly to a kind of culture microalgae
Method.
Background technology
Photosynthetic organism, which is one kind, to carry out photosynthetic biology, such as photosynthetic bacteria and microalgae using light.Microalgae is
The type that one kind is grown in water is various and is distributed extremely extensive rudimentary plant, it is the cell factory driven by sunlight, is led to
The efficient photosynthesis of microalgae cell is crossed, CO is absorbed2, the chemical energy of the carbohydrate such as fat or starch is converted light energy into,
And release O2.It is expected to reach simultaneously with chemicals using microalgae production bioenergy and substitutes fossil energy, reduces CO2The mesh such as discharge
's.
Photosynthetic organism is generally cultivated in bioreactor (or device).Bioreactor mainly has out at present
Put formula and two kinds closed.Either Race-way photobioreactor or closed photo bioreactor are required for react
Device is placed under light source and is cultivated, and light source can be sunlight, can also be artificial light source, in order to reduce toxigenic capacity, generally
Using sunlight in outside scenery.In the prior art, the either bioreactor of that form, is required to that liquid stream will be cultivated
(transmission) is moved up, is on the one hand to avoid photosynthetic organism " conglomeration ", influences its normal growth；On the other hand it is in order to more preferable
Biography light and mass transfer.
Although having numerous documents about chemical process mass transfer, the mass transfer and chemical process of photosynthetic organism incubation
Mass transfer it is different, and there is also the unexistent problem of chemical process in the incubation of photosynthetic organism, for example, mechanical force damage,
Pass light etc..
On the one hand, light can only propagate limited distance in water；On the other hand, mutually blocking between cell, also can shadow
The normal light of photosynthetic organism is rung, therefore when cultivating photosynthetic organism, it, all can be to the liquid of culture solution in order to ensure growth efficiency
Thickness is limited.For cultivating microalgae, in order to receive more sufficient illumination, the liquid thickness of culture solution is usually no more than 60cm,
Most common liquid thickness is 10cm~30cm.
In order to improve the light of photosynthetic organism, there is the method using Thin cell layer liquid culture, such as United States Patent (USP)
US5981271 discloses a kind of culture process and device for making culture solution form laminar flows, can greatly improve culture solution
Concentration and production efficiency.On the one hand, this method is not easily solved the loss problem of easy " escape " nutriment, such as CO2；It is another
Aspect establishes laminar flows and also results in higher energy consumption.
Chinese patent application CN102421887A discloses a kind of method of culture photosynthetic organism, including：Culturing room is provided,
Culture medium and photosynthetic organism are introduced to the culturing room, and is exposed under light, then flow control device is utilized to control gas
By culture medium, the air-flow drives the evaporation of the culture medium.It, must in order to avoid photosynthetic organism settles in actually cultivating
It must be passed through a large amount of gas, and this necessarily leads to increase and the explosive vaporization of culture solution of energy consumption, this is to reducing energy consumption and water consume
It is clearly unfavorable.
Chinese patent application CN102260629A discloses a kind of board-like bioreactor, including at least one runner,
Wherein, each runner includes：At least 2 pieces of overhead gages, are set on the inner wall of plane of illumination；At least 2 pieces of lower baffle plates, are set to unglazed
On the inner wall in face；Wherein, the length direction of overhead gage and lower baffle plate and the angle of culture solution flow direction are 20 °~70 °, and are pressed from both sides
Angular direction is opposite.The plate-type reactor has specific internal structure, and frustule can be realized in the light area of bioreactor
It shuttles between dark space, to improve the culture efficiency of microalgae.But it needs all cultivate liquid recycle streams when program operation
It moves up by the baffle in bioreactor, to significantly increase operation energy consumption.
Either energy consumption, material consumption are big or excessively complicated for the prior art above-mentioned, therefore are unsuitable for extensive, high efficiency
Culture photosynthetic organism, be especially not suitable for incubation for the purpose of obtaining bioenergy.
It is practical using pond or raceway pond when pilot scale culture microalgae.Raceway pond is exactly using paddle wheel driving culture
Liquid is flowed along the shallow pond of runway shape, and culture solution receives illumination during flowing.Due to the continuous proliferation of microalgae, frustule
Between the case where mutually blocking also constantly aggravate, therefore raceway pond is difficult to carry out High Density Cultivation, and the light of final culture solution is close
Angle value is generally less than 2.Pond equally exists above-mentioned problem, and asks there is also transmission, mass transfer, harvesting etc. are otherwise
Topic.
To sum up, in order to realize culture photosynthetic organism in large-scale, high-efficiency, there is an urgent need for researching and developing new culture technique, with
Preferably solve the problems, such as biography light, mass transfer and the transmission during culture photosynthetic organism.
The NOx in industrial waste gas is fixed using microalgae, on the one hand NOx can be eventually converted into useful microalgae biology
Matter；On the other hand the cost that culture microalgae can be reduced again, has both economy and the ambilateral significance of environment.Chemical engineering industry institute
The amount of NOx of generation is huge, so if fixing the NOx in industrial waste gas with microalgae, it is necessary to microalgae be made to fix the speed of NOx
Rate and the rate of industrial discharge NOx match, and reduce the floor space of micro algae culturing device to the greatest extent, with the life of current raceway pond
Efficiency is produced, this actual needs are also cannot be satisfied.In general, photoautotrophic efficiency is less than 30g.m-2.d-1, outdoor extensive
The efficiency of culture is generally below 10g.m-2.d-1, a large amount of soil can be occupied by carrying out industrial waste gas denitration with such efficiency,
Know and organic carbon source is added during microdisk electrode, the growth rate of microalgae can be greatly improved, however add organic carbon source pole
Easily incur algae solution to be contaminated by bacterial, seriously affect the normal growth of microalgae, even results in culture failure.And sterilization is used to grasp
Make, and inevitably results in cost and increase considerably.
Invention content
It is an object of the present invention to provide it is a kind of it is simple in structure, construction cost is low, culture efficiency higher and energy consumption smaller
Photosynthetic organism culture apparatus.It is a further object to provide a kind of methods using the device culture photosynthetic organism.
A further object of the present invention is to provide a kind of method for fixing industrial waste gas NOx using the device.
By background technology it is found that although the prior art such as raceway pond provides flowing for the growth of photosynthetic organism,
But production efficiency of these technologies in pilot scale culture is still relatively low, energy consumption is higher.Inventor has found by further investigation, leads to
It crosses pressure culture solution to flow in a particular manner, it will be able to further increase the culture efficiency of photosynthetic organism；Further research
It has also been found that only a small amount of EM bacterium need to be added in the algae solution of culture microalgae, it will be able to which the breeding for inhibiting harmful bacteria significantly carries
The growth rate of high microalgae, it is often more important that, after EM bacterium are added, even if can if not to culture environment progress sterilization treatment
It successfully carries out luminous energy and supports, thus greatly reduce the cost of culture.
The present inventor completes the present invention based on above-mentioned discovery, and main contents are as follows：
1. a kind of device of culture photosynthetic organism, includes the cultivation region for accommodating culture solution of top visible light, special
Sign is, flow-disturbing mechanism is equipped in the cultivation region, and the flow-disturbing mechanism has and can persistently be transported along the cultivation region bottom surface
Dynamic inclined-plane；The flow-disturbing mechanism is used for culture during its persistent movement to the cultivation region bottom by the inclined-plane
The lasting thrust obliquely that liquid generates pushes the culture solution of the cultivation region bottom to liquid level persistent movement；The inclined-plane
Moving region covers the wholly or largely region of the cultivation region bottom surface.
2. according to the device described in 1, which is characterized in that the cultivation region is cylindric, and the inclined-plane is that rectangle is tiltedly put down
Face simultaneously can persistently be rotated around cylindrical center's axis；The long side of the rectangle tapered plane is parallel to the bottom cylindrical face and along institute
State the radial setting of cylinder
3. according to the device described in 2, which is characterized in that the number on the inclined-plane is n, and is symmetrical with cylindrical center's axis
Axis is arranged by n times axial symmetry, n=3~20, preferably n=6~12；The angle of inclination of the inclined-plane with respect to the horizontal plane be 20 °~
80 °, preferably 30 °~60 °.
4. according to the device described in 2, which is characterized in that the flow-disturbing mechanism is by plectane and is fixed on rectangular on the plectane
Shape inclined plate is constituted, and the radius of the radius of the plectane and the cylinder is same as or slightly smaller than the radius of the cylinder.
5. according to any device of Claims 1 to 4, which is characterized in that the turbulence structure is by mechanical transmissioning machine
Structure or magnetic drive.
6. according to 1~5 any device, which is characterized in that automatic detection culture solution is arranged in the cultivation region
PH value and the aerator for adjusting aeration quantity accordingly.
7. a kind of method of culture photosynthetic organism, which is characterized in that in any device of claim 1~6 into
Row.
8. according to the method described in 7, which is characterized in that the photosynthetic organism is microalgae, preferably green alga or cyanobacteria, more preferably
Chlorella, scenedesmus, single needle algae or spirulina；Cultivation temperature is 15~40 DEG C, preferably 25~35 DEG C；Medium pH value be 6~
11, preferably 7~9；Light intensity is the lux of 1000 luxs~200000, the lux of preferably 5000 luxs~150000.
9. according to the method described in 8, which is characterized in that culture height of liquid layer is 0.1m~2m, the bevel altitude and training
The ratio of nutrient solution layer height is 0.01~0.5；Adjust the inclined-plane movement velocity make microalgae movement velocity be 5cm/s~
500cm/s, more preferably 40cm/s~400cm/s.
10. according to the method described in 8 or 9, which is characterized in that with NO3 -And/or NO2 -As nitrogen source.
11. a kind of method for fixing industrial waste gas NOx using microalgae, including：
(1) to industrial waste gas denitration the step of, to obtain salt made from earth containing a comparatively high percentage of sodium chloride and/or nitrosonium salts；
(2) the step of using the method culture microalgae of claim 10；In this step, the salt made from earth containing a comparatively high percentage of sodium chloride that is obtained with step (1)
And/or nitrosonium salts are as nitrogen source, and EM bacterium and organic carbon source are added in culture solution, the addition of EM bacterium is 1 × 106A/L trainings
Nutrient solution~9 × 108A/L culture solutions, preferably 1 × 107A/culture solution~5 × 10 L8A/L, by organic carbon source in culture solution
Concentration is controlled in 1g/L culture solutions~30g/L culture solutions, is preferably controlled in 2g/L culture solutions~10g/L culture solutions.
The present invention achieves following technique effect.
First, although providing various forms of flowings for culture photosynthetic organism in the prior art, these flowings
In, considerable part is unnecessary for " improving photosynthetic organism growth efficiency ".The present invention by force culture solution with
Specific mode flows, and can not only increase substantially the growth efficiency of photosynthetic organism, and can save energy consumption.
Second, the device of the invention is simple in structure, construction cost is low.
Third, whether photoautotrophy, or it is simultaneous foster, EM bacterium are added in algae solution, can effectively inhibit harmful thin
The breeding of bacterium increases substantially the growth rate of microalgae.This feature makes the present invention be more suitable for large-scale culturing micro-algae, especially
In simultaneous support, due to the sterilizing that need not carry out disinfection, make the present invention that there is the advantage of bigger.According to the present invention, microalgae with
High efficiency consumption is inorganic nitrogen-sourced, and the present invention is made to be quite suitable for the NOx in fixed industrial waste gas.
Other features and advantages of the present invention will be described in detail in subsequent specific embodiment part.
Description of the drawings
The sectional side view of Fig. 1 apparatus of the present invention and its vertical view of flow-disturbing mechanism.
The sectional side view of Fig. 2 comparative example devices and its vertical view of transmission mechanism.
The photosynthetic organism growth curve of Fig. 3 embodiments 1,2,3 and comparative example 1.
The photosynthetic organism growth curve of Fig. 4 embodiments 4,5,6,7 and comparative example 2.
The photosynthetic organism growth curve of Fig. 5 embodiments 8 and comparative example 3.
Reference sign
1 cultivation region, 2 driving mechanism
3 plectane, 4 inclined plate
5 side wall, 6 bottom wall
7 roof, 8 culture solution entrance
9 culture solutions export D direction of illuminations
Specific implementation mode
The specific implementation mode of the present invention is described in detail below.It should be understood that described herein specific
Embodiment is merely to illustrate and explain the present invention, and is not intended to restrict the invention.
In the present invention, such as without opposite explanation, the terms such as upper and lower or height are directed to for gravity direction.
The present invention provides a kind of devices of culture photosynthetic organism, include the training for accommodating culture solution of top visible light
Support area, which is characterized in that flow-disturbing mechanism is installed in the cultivation region, the flow-disturbing mechanism has can be along the cultivation region bottom
The inclined-plane of face persistent movement；The flow-disturbing mechanism is used to rely on the inclined-plane during its persistent movement to the cultivation region bottom
The lasting thrust obliquely that the culture solution in portion generates pushes the culture solution of the cultivation region bottom to liquid level persistent movement；Institute
The moving region for stating inclined-plane covers the wholly or largely region of the cultivation region bottom surface.
According to the present invention, " the top visible light " refer to the structure of described device or the material of selection can make it is described
The culture solution of cultivation region receives the irradiation of the sunlight at the top of it, for example the top of the cultivation region is opened wide or with light transmission material
Material manufacture.According to the present invention, the side wall of described device can use light transmission or lighttight material manufacture；It is preferred that with translucent material system
It makes so that the culture solution of the cultivation region can receive the irradiation of light in side.
According to the present invention, the inclined-plane can also be inclined lower concave curved surface either inclined plane.
According to the present invention, the movement velocity on the inclined-plane is preferably 20~500cm/s, more preferable 30~200cm/s.
According to the present invention, those skilled in the art can select according to actual needs, make the moving region on the inclined-plane
The base surface area of the cultivation region of the covering more than 50%, for example, more than 50% to 60%, 60%~70%, 70%~80%,
The base surface area of 80%~90%, 90%~95% or 95% or more the cultivation region.
According to the present invention, the floor space of the cultivation region is generally 1m2~500m2, preferably 1m2~100m2。
In the case of preferred, the cultivation region is cylindric, and the inclined-plane is rectangle tapered plane and can be around the cylinder
Central shaft persistently rotates；The long side of the rectangle tapered plane is parallel to the bottom cylindrical face and is set along the radial direction of the cylinder
It sets.In the case of preferred, the number on the inclined-plane is n, and is arranged by n times axial symmetry using cylindrical center's axis as symmetry axis, n
=3~20, preferably n=6~12.The angle of inclination of the inclined-plane with respect to the horizontal plane is 20 °~80 °, preferably 30 °~60 °；Institute
The height for stating inclined-plane is 0.05m~0.5m, preferably 0.1m~0.2m.
As a kind of specific implementation mode of above-mentioned preferable case, the flow-disturbing mechanism is by plectane and is fixed on the plectane
Rectangle inclined plate constitute, the radius of the radius of the plectane and the cylinder is same as or slightly smaller than the radius of the cylinder.
According to the present invention it is possible to drive the turbulence structure using any existing known mode, for example passed by machinery
Motivation structure or magnetic drive.
According to the present invention, automatic detection medium pH value can be set in the cultivation region and adjust the exposure of aeration quantity accordingly
Device of air.The present inventor is had found by a large number of experiments, for photoautotrophy or simultaneous foster training method, when microalgae is metabolized alkali metal
Nitrate, alkali metal nitrites salts, alkali carbonate, alkali metal hydrogencarbonate, alkali metal phosphate, alkali metal hydrogen phosphate
One of or it is arbitrary when combining, if not being passed through CO into algae solution in the incubation of microalgae2Or it is added without pH adjusting agent,
Then the pH value of algae solution can rise, especially when microalgae is metabolized alkali nitrates, alkali metal nitrites salts or combinations thereof, algae solution pH
Faster ascendant trend is presented in value.The pH value of general culture microalgae is 6~11, and pH when most common or culture efficiency highest is
Neutrality left and right, it is permitted beyond micro algae growth in order to avoid the pH value of culture solution when culture solution contains above-mentioned nutriment
Range more efficiently cultivates microalgae, and the present invention is preferably at least with containing CO2Gas provide part carbon for the incubation of microalgae
Source contains CO by control2Gas intake, can easily by the pH value of algae solution control in suitable range.
It, according to actual needs, can also be in addition, in order to preferably observe the growth conditions of condition of culture and photosynthetic organism
Dissolved oxygen probes, hygrosensor, Light-Intensity Detector, conductivity detector, photosynthetic organism concentration are provided in culture apparatus
The those skilled in the art such as detector it is thinkable for observe condition of culture and photosynthetic organism growth conditions various instrument
It is one or more in device, in order to observe and monitor parameters in real time.
The present invention also provides it is a kind of culture photosynthetic organism method, which is characterized in that in any device above-mentioned into
Row.
In the present invention, photosynthetic organism includes microalgae and photosynthetic bacteria, and the type of microalgae is not particularly limited, such as
It can select green alga or cyanobacteria.The present invention preferably cultivates oil-producing microalgae, and more preferably those are with larger industrial utilization
At least one of oil-producing engineering microalgae, such as chlorella, scenedesmus, spirulina, chrysophyceae and Phaeodactylum tricornutum.
Micro algae growth needs necessary condition, such as suitable temperature, sufficient illumination, enough water, CO2And nitrogen
The nutriments such as fertilizer, phosphate fertilizer, regulate and control that dissolved oxygen in algae solution, pH value is in suitable range etc..Although for different microalgaes,
These conditions are not quite similar, but these are all known in the art.
In general, cultivation temperature is 15~40 DEG C, preferably 25~35 DEG C；Medium pH value is 6~11, preferably 7
~9；Light intensity is the lux of 1000 luxs~200000, the lux of preferably 5000 luxs~150000.
It, can be according to the photosynthetic organism cultivated, using culture solution commonly used in the art, for micro algae growth for culture solution
The nutrients such as nitrogen, phosphorus, potassium needed for providing, such as BG11 culture solutions.
According to the Expenditure Levels of nutriment in the growth pattern of micro algae biomass and culture solution, need to be replenished in time not
The nutriment of foot.According to the present invention, any mode for adding nutriment is all available, for example segmentation is added or continuously mended
Add, as long as can control the amount of nutriment in suitable range.
According to the present invention, culture height of liquid layer is 0.1m~2m, and the ratio of the bevel altitude and culture height of liquid layer is
0.01~0.5.According to the present invention, both photosynthetic organism can be cultivated under existing culture height of liquid layer, it can also be higher than existing
Photosynthetic organism is cultivated under some culture height of liquid layer, for example cultivates photosynthetic organism under the culture height of liquid layer higher than raceway pond.
According to the present invention, adjusting the movement velocity on the inclined-plane makes the movement velocity of microalgae be 5cm/s~500cm/s, more
Preferably 40cm/s~400cm/s.
, according to the invention it is preferred to NO3 -And/or NO2 -As nitrogen source.
The present invention also provides a kind of methods for fixing industrial waste gas NOx using microalgae, including：
(1) to industrial waste gas denitration the step of, to obtain salt made from earth containing a comparatively high percentage of sodium chloride and/or nitrosonium salts；
(2) the step of using the method culture microalgae of claim 10；In this step, the salt made from earth containing a comparatively high percentage of sodium chloride that is obtained with step (1)
And/or nitrosonium salts are as nitrogen source, and EM bacterium and organic carbon source are added in culture solution, the addition of EM bacterium is 1 × 106A/L trainings
Nutrient solution~9 × 108A/L culture solutions, preferably 1 × 107A/culture solution~5 × 10 L8A/L, more preferably 1 × 107A/L trainings
Nutrient solution~1 × 108A/L；The concentration of organic carbon source in culture solution is controlled in 1g/L culture solutions~30g/L culture solutions, is preferably controlled
System is in 2g/L culture solutions~10g/L culture solutions.
Although can increase part toxigenic capacity using organic carbon source, culture efficiency also greatly improves, and makes following process
Process is simplified, so if sterile culture can be avoided, it will be able to be avoided consuming a large amount of steam and strictly be gone out to system
Bacterium is handled, to which toxigenic capacity be greatly reduced.According to the present invention, particularly preferably those simultaneous foster microalgaes, such as chlorella, grid
Algae, spirulina or single needle algae., it is surprising that in a manner of simultaneous support when these microalgaes of culture, as long as a certain number of EM are added
Bacterium, even if can be smoothed out if culture without sterilization, the growth rate of microalgae is greatly speeded up, even if water source contains greatly
It measures harmful bacteria and/or opens wide culture, be as a result also such；And when being added without EM bacterium, and support and would generally fail.
According to the present invention, available organic carbon source includes but not limited to sugar, organic acid, acylate, alcohol, cellulose hydrolysis
At least one of object and glucidtemns；Such as can be selected from glucose, fructose, acetic acid, sodium acetate, lactic acid, ethyl alcohol, methanol,
At least one of cellulose hydrolysate and cellulose hydrolysate, preferably selection is glucose.
The EM bacterium (Effective Microorganisms) belong to the prior art, mainly by belonging to photosynthetic bacteria
Group, lactobacillus, yeast flora, Gram positive actinomycetes group, fermentation system Filamentous flora tens kinds of microorganism groups at being
A kind of commercially available active bacteria formulation.The EM bacterium both can voluntarily prepare according to existing knowledge, can also be commercially available, and use
It is preceding to need to illustrate to ferment according to having knowledge or commercial preparations.
The present invention is described in more detail by taking a kind of specific embodiment as an example below in conjunction with attached drawing.
As shown in Figure 1, having the cultivation region 1 for accommodating culture solution of top visible light, training in the culture apparatus of the present invention
It supports in area 1 and is provided with flow-disturbing mechanism (plectane 3 and inclined plate 4) and its driving mechanism 2；The flow-disturbing mechanism is driven by driving mechanism 2,
It can be rotated in cultivation region 1, to guide the culture solution of cultivation region bottom (to be transported to cultivation region motion of melt surface to direction of illumination D
It is dynamic, movement degree of the hot housing liquid on direction of illumination).In the present invention, the effect of flow-disturbing mechanism is to drive cultivation region bottom
The culture solution in portion is different from existing static flow-disturbing component, the effect of these flow-disturbing components is all to cultivation region motion of melt surface
It is by hindering fluid flowing to form flow-disturbing.The flow-disturbing mechanism of the present invention is by driving the culture solution of cultivation region bottom to culture
Area's motion of melt surface, you can increase culture solution in the flow in illumination (sunlight) direction, reduce horizontal flow (perpendicular to direction of illumination
Flow), for large-scale culture photosynthetic organism, this can not only simplify device, improve efficiency, and can greatly save energy
Consumption.
As previously mentioned, flow-disturbing mechanism through the invention, can be established in cultivation region culture solution cultivation region bottom with
Reciprocating motion between the liquid level of cultivation region makes photosynthetic organism (such as microalgae) intermittently expose under light illumination.In present embodiment
In, the movement velocity of photosynthetic organism is preferably 5~500cm/s, more preferably 40~400cm/s.Under stronger illumination, light
The the movement velocity of symphysis object the high then, and culture effect is better, culture efficiency is higher.
The present invention flow-disturbing mechanism can there are many structure type, exemplarily only enumerate a kind of simple shape herein
Formula.As shown in Figure 1, the flow-disturbing mechanism includes plectane 3 and multiple rectangle inclined plates 4 for being installed on the plectane, driving mechanism 2
For driving flow-disturbing mechanism rotation in cultivation region 1, the radial setting of the long edge plectane 3 of rectangle inclined plate 4 adjacent oblique
Angle between plate is identical or essentially identical.As shown in Figure 1, culture solution is placed in a horizontal positioned cylindric pond, and
Flow-disturbing mechanism (plectane 3 and rectangle inclined plate 4) is positioned under the liquid level of culture solution, the bottom close to circle pond, and by certain
Mechanical structure is fixed.When the flow-disturbing mechanism rotates, the promotion of the culture solution through inclined plate 4 in circle bottom of pond portion is largely guided
Along direction of illumination D to motion of melt surface, and the culture solution for justifying pond liquid level is then moved along direction of illumination D to circle bottom of pond portion, passes through this
Kind mode, just establishes reciprocating motion of the culture solution between cultivation region bottom and cultivation region liquid level in cultivation region so that culture
Photosynthetic organism in liquid intermittently exposes under light illumination.
In the present embodiment, the radius in cylindric pond is 0.2~5m；The liquid thickness of culture solution be preferably 10cm~
60cm, more preferably 10cm~30cm；The height of inclined plate 4 and the ratio of culture solution liquid thickness are preferably 1/3~1/20, more preferably
1/3~1/10；Inclined plate 4 is preferably 20 °~80 ° relative to the angle of inclination of plectane 3 (plectane 3 is horizontally placed on round bottom of pond face),
More preferably 30 °~60 °.The slewing rate of the inclined plate is between 1rpm~200rpm, you can obtains preferable effect.Using
Above-mentioned mode and parameter, you can greatly improve culture efficiency.
In the present invention, culture apparatus can also be open, those skilled in the art can be according to tool either closed
The culture environment of body is selected.If using closed culture, to receive the irradiation of sunlight, the roof of culture apparatus should be used
Transparent material manufactures；In order to avoid photosynthetic organism is adherent, normal light is influenced, the roof should be with culture solution face interval one
Fixed distance.For example, the culture apparatus in Fig. 1 can be box cuboid, cylindric cultivation region is placed in the babinet, the case
The roof 7 of body is spaced apart at a certain distance with cultivation region liquid level.
In present embodiment, driving mechanism 2 is preferably motor.
Unless otherwise defined, all technical and scientific terms used herein all has those skilled in the art conventional
The meaning of understanding.In case of conflict, it is subject to the definition of this specification.
In the context of the present specification, other than the content clearly stated, any matters or item that do not mention are equal
It is directly applicable in those of known in the art without carrying out any change.Moreover, any embodiment described herein can be with
It is freely combined with one or more other embodiments described herein, the technical solution or technological thought being consequently formed are accordingly to be regarded as
A part for the original disclosure of the present invention or original description, and it is not considered as the new content for not disclosing or being expected herein,
Unless those skilled in the art think that the combination is apparent unreasonable.
All features disclosed in this invention can in any combination, these combinations should be understood in disclosed in this invention
Hold, unless those skilled in the art think that the combination is obviously unreasonable.Numerical point disclosed in this specification includes not only specific
Disclosed numerical point, further includes the endpoint of each numberical range, and the range that these numerical points arbitrarily combine is regarded as this hair
Bright published range, no matter whether separately disclosing these numerical value pair herein.
It is further illustrated the present invention below by embodiment.
Algae solution OD value (OD680Value) it measures：OD value spectrophotometric determination, is compared with distilled water, is measured
Light absorption value of the algae solution at wavelength 680nm, the index as microalgae concentration.
The measurement of solution nitrogen content：It is measured in aqueous solution using ICS3000 types ion chromatograph (Dionex companies of the U.S.)
NO3 -Content or NO2 -Content, instrument is equipped with E640 leacheates automatic generator, electric conductivity detector and the work of chameleon chromatography
It stands；IonPac AS11-HC types splitter (250mm × 4mm i.d.)；IonPac AG11 type guard columns (50mm × 4mm
i.d.)；Itself suppressor of ASRS-ULTRA anion.Leacheate：KOH solution；Flow velocity is 1mL/min；Eluent concentration：
30mmol/L；Sample size is 60 μ L；Column temperature is 30 DEG C；Inhibit electric current 100mA；External standard method peak area quantification.
Count of bacteria：Count of bacteria is carried out according to the following steps
1. sample washs：1ml samples are drawn, are washed 2-3 times with 1 × PBS；2. initial gross separation：According to algae and bacterium from
The difference of mental and physical efforts centrifuges 2min, initial gross separation algae with 1000rpm first (for bacterium in supernatant, algae is in precipitation)；If
When algae content is higher, repeat again；3. collecting supernatant, the amount of algae in supernatant is negligible at this time, 8000rpm centrifugations
5min abandons supernatant；4. precipitation is resuspended with 500ul bacterium rupture of membranes agent, 15min is reacted at room temperature；5.8000rpm centrifuges 5min, with 1 ×
PBS washs 2 bacterium solutions；6. 1 × PBS of 100ul, which are added, is resuspended thalline, 5ul PI dye liquor mother liquors are added, react at room temperature 30min；7.
Fluorescence microscopy microscopic observation bacterium simultaneously counts, and bacterial number is up to 1000 in 4 block plaids, when being more than 1000, dilution
Bacterium solution certain multiple counts again；8. calculation formula：
Bacterial density=count results/4 × extension rate × 4 × 10 in surveyed solution4A/ml
Main agents consumptive material：
| Agents useful for same consumptive material | Manufacturer |
| PI Viability Staining Solution | Four positive cypress Cat No.FXP002 |
| Rupture of membranes agent | Sharp that health Cat No.REK3004 |
| Phosphate buffer (10 × PBS, pH7.4, cell culture grade are sterile) | Sharp that health Cat No.REK3013 |
| Cell climbing sheet | NEST |
Key instrument：
| Instrument | Manufacturer |
| Tally | Shanghai precision instrument |
| Fluorescence microscope | Olympus BX-51 |
The culture medium of microalgae：Medium component is shown in Table 1~table 4.
1 culture medium BG11 of table
| Component | Composition, mg/L |
| K2HPO4·3H2O | 40 |
| NaNO3 | 1500 |
| Na2CO3 | 20 |
| MgSO4·7H2O | 75 |
| CaCl2·2H2O | 36 |
| Citric acid | 6 |
| Ferric citrate | 6 |
| EDETATE SODIUM | 1 |
| Micro- A5 | 1 |
2 trace element A5 of table
| Component | Composition, mg/L |
| H3BO3 | 2860 |
| MnCl2·4H2O | 1810 |
| ZnSO4·7H2O | 222 |
| CuSO4·5H2O | 79 |
| NaMoO4·5H2O | 390 |
| Co(NO3)2·6H2O | 50 |
3 Heterotrophic culture base of table
4 trace element of table
| Component | Composition, g/L |
| H3BO3 | 2.86 |
| MnCl2·4H2O | 0.11 |
| ZnSO4·7H2O | 9.22 |
| CuSO4·5H2O | 1.00 |
| (NH4)6Mo7O24·4H2O | 0.10 |
| Co(NO3)2·6H2O | 0.90 |
EM bacterium：Probiotics used in embodiment is the oasis Kang Yuan bio tech ltd production such as golden probiotics,
Illustrated into line activating processing, PH ＜ 4 by it using preceding.
Embodiment 1
The method that the present embodiment is used to illustrate the culture photosynthetic organism of the present invention.
Using photosynthetic organism culture apparatus culture chlorella as shown in Figure 1.Using BG11 culture mediums, nitrogenous fertilizer 0.3g/L
Urea, between 20~30 DEG C, algae initial concentration OD680It is 0.5, is passed through the CO of 2% (v/v)2Ventilation culture.
Cultivation region (circle pond) a diameter of 1m, a diameter of 0.9m of plectane, 4 number of inclined plate is 8, inclined plate length 0.45m, inclined plate and horizontal plane
Angle be 45 degree, inclined plate height be 3cm, algae solution liquid thickness be 20cm, start photosynthetic organism culture apparatus in motor make flow-disturbing
Mechanism (plectane and inclined plate) rotation (inclined plate angle with horizontal plane institute is identical as the algae solution direction of motion to direction), adjusts its rotation speed
Degree makes the movement velocity of microalgae be 60cm/s (being measured by Doppler anemometer).The OD of detection algae solution daily680Value, it is continuous to train
It is harvested after supporting 14 days, the growth curve of microalgae is shown in Fig. 3.
Embodiment 2
The method that the present embodiment is used to illustrate the culture photosynthetic organism of the present invention.
According to the method culture chlorella of embodiment 1, the difference is that only：The angle of inclined plate and horizontal plane is 60 degree,
Inclined plate height be 5cm, algae solution liquid thickness be 30cm, start photosynthetic organism culture apparatus in motor make flow-disturbing mechanism (plectane and tiltedly
Plate) rotation, adjusting its velocity of rotation makes the movement velocity of microalgae be 80cm/s (being measured by Doppler anemometer), and algae solution is being disturbed
It flows and moves up and down under the driving of mechanism.The growth curve of microalgae is shown in Fig. 3.
Embodiment 3
The method that the present embodiment is used to illustrate the culture photosynthetic organism of the present invention.
According to the method culture chlorella of embodiment 1, the difference is that only：The angle of inclined plate and horizontal plane is 30 degree,
Adjusting its velocity of rotation makes the movement velocity of microalgae be 40cm/s (being measured by Doppler anemometer).The growth curve of microalgae is shown in
Fig. 3.
Comparative example 1
According to the method culture chlorella of embodiment 1, the difference is that only：Do not start flow-disturbing mechanism, but uses paddle wheel
Algae solution flowing is driven, it is 60cm/s (being measured by Doppler anemometer) to make the movement velocity of microalgae.Fig. 2 is shown in specific difference,
Circle one paddle wheel of pond radially installed drives algae solution flowing with it, and the length of the paddle wheel is 90cm, does not start the flow-disturbing machine of the present invention
Structure, at this time inclined plate play spoiler (inclined plate angle with horizontal plane institute opposite with the algae solution direction of motion to direction).Microalgae
Growth curve is shown in Fig. 3.
Embodiment 4
It (is come from using BG11 culture mediums (adding nutritional ingredient by table 1, culture solution is without sterilization treatment) culture chlorella
Sinopec microalgae algae library, number Chlorella sp.RIPP-1), the glucose of 2g/L is added in incubation, controls temperature
Between 20~30 DEG C, it is passed through compressed air and CO2Culture, CO is passed through as algae solution PH ＞ 102, stop as algae solution PH ＜ 7.5
It is passed through CO2.Natural daylight culture, daylight intensity is used to reach as high as 60000 luxs, detect algae daily in incubation
The OD of liquid680Value, the growth curve of microalgae are shown in Fig. 4.Wherein, EM additive amounts are 3.6 × 106A/L algae solutions monitor in breeding process
The count of bacteria ＜ 8 × 10 of algae solution6A/mL algae solutions, continuous culture harvest after 14 days.
Embodiment 5
The present embodiment is differed only in embodiment 4：EM additive amounts are 1.8 × 107A/L algae solutions.It is supervised in breeding process
Survey the count of bacteria ＜ 1 × 10 of algae solution7A/mL algae solutions.The growth curve of microalgae is shown in Fig. 4.
Embodiment 6
The present embodiment is differed only in embodiment 4：EM additive amounts are 3.6 × 107A/L algae solutions.It is supervised in breeding process
Survey the count of bacteria ＜ 2 × 10 of algae solution7A/mL algae solutions.The growth curve of microalgae is shown in Fig. 4.
Embodiment 7
The present embodiment is differed only in embodiment 4：EM additive amounts are 7.2 × 107A/L algae solutions.It is supervised in breeding process
Survey the count of bacteria ＜ 5.8 × 10 of algae solution7A/mL algae solutions.The growth curve of microalgae is shown in Fig. 4.
Comparative example 2
This comparative example is differed only in embodiment 4：EM bacterium are not added.The count of bacteria of algae solution is monitored in breeding process
It has been up to 1.2 × 108A/mL algae solutions.The growth curve of microalgae is shown in Fig. 4.
As can be seen from Fig. 4, under the conditions of simultaneous support, addition EM bacterium promote the growth of microalgae.
Embodiment 8
BG11 culture mediums (adding nutritional ingredient by table 1, culture solution is without sterilization treatment) culture chlorella is used first
(coming from Sinopec microalgae algae library, number Chlorella sp.RIPP-1)；Work as OD680When value is 4, mended by 3 specified amount of table
Add a Heterotrophic culture base nutritional ingredient.Between 20~30 DEG C, it is passed through compressed air and CO2Culture, works as algae solution
It is passed through CO when PH ＞ 102, stop being passed through CO as algae solution PH ＜ 7.52.Natural daylight culture, daylight are used in incubation
Intensity reaches as high as 60000 luxs, adds the glucose of 2g/L, and press 2.9 × 107The amount of a/L algae solutions adds EM bacterium, often
The OD of its detection algae solution680Value；The glucose of 10g/L is added in culture again after 1 day, and presses 3.6 × 107A/L algae solutions add EM
Bacterium；Add glucose 10g/L again when culture was to the 5th day, in breeding process the count of bacteria of monitoring algae solution be up to 9.7 ×
106A/mL algae solutions, continuous culture harvest after 8 days, and last time stops being passed through CO after glucose is added2, terminate algae solution when cultivation
PH value is 8.6, is centrifugally separating to obtain algal gel and foster algae raffinate.The NO in algae raffinate is supported in analysis3 -With NO2 -10 μ g/ of total content ＜
g.The growth curve of microalgae is shown in Fig. 5.
Comparative example 3
This comparative example is differed only in embodiment 8：EM bacterium are not added.Monitor incubation in algae solution count of bacteria most
A height of 13.6 × 108A/mL algae solutions.The growth curve of microalgae is shown in Fig. 4.
As can be seen from Fig. 5, addition EM bacterium are greatly facilitated the growth of microalgae and consume rapidly inorganic nitrogen-sourced.
Claims (9)
1. a kind of device of culture microalgae, includes the cultivation region for accommodating culture solution of top visible light, which is characterized in that
Flow-disturbing mechanism is installed, the flow-disturbing mechanism has can be along the inclined-plane of cultivation region bottom surface persistent movement in the cultivation region；
What the flow-disturbing mechanism was used to generate the culture solution of the cultivation region bottom during its persistent movement by the inclined-plane
Lasting thrust obliquely pushes the culture solution of the cultivation region bottom to liquid level persistent movement；The moving region on the inclined-plane
The base surface area of the cultivation region of 80%~90%, 90%~95% or 95% or more covering；
The cultivation region is cylindric, and the inclined-plane is rectangle tapered plane and can persistently be rotated around cylindrical center's axis；Institute
The long side for stating rectangle tapered plane is parallel to the bottom cylindrical face and the radial direction setting along the cylinder；
The flow-disturbing mechanism is made of plectane and the rectangle inclined plate being fixed on the plectane, the radius of the plectane and the circle
The radius of column is same as or slightly smaller than the radius of the cylinder.
2. device described in accordance with the claim 1, which is characterized in that the number on the inclined-plane is n, and is with cylindrical center's axis
Symmetry axis is arranged by n times axial symmetry, n=3~20；The angle of inclination of the inclined-plane with respect to the horizontal plane is 20 °~80 °.
3. device described in accordance with the claim 1, which is characterized in that the turbulence structure is driven by mechanical transmission mechanism or magnetic force
It is dynamic.
4. device described in accordance with the claim 1, which is characterized in that automatic detection medium pH value is arranged in the cultivation region
And the aerator of aeration quantity is adjusted accordingly.
5. a kind of method of culture microalgae, which is characterized in that carried out in any device of Claims 1 to 4.
6. according to the method for claim 5, which is characterized in that the microalgae is green alga or cyanobacteria；Cultivation temperature be 15~
40℃；Medium pH value is 6~11；Light intensity is the lux of 1000 luxs~200000.
7. according to the method for claim 6, which is characterized in that culture height of liquid layer is 0.1m~2m, the bevel altitude
Ratio with culture height of liquid layer is 0.01~0.5；Adjusting the movement velocity on the inclined-plane makes the movement velocity of microalgae be 5cm/s
~500cm/s.
8. according to the method for claim 6, which is characterized in that with NO3 -And/or NO2 -As nitrogen source.
9. a kind of method for fixing industrial waste gas NOx using microalgae, including：
(1) to industrial waste gas denitration the step of, to obtain salt made from earth containing a comparatively high percentage of sodium chloride and/or nitrosonium salts；
(2) the step of using the method culture microalgae of claim 8；In this step, the salt made from earth containing a comparatively high percentage of sodium chloride that is obtained with step (1) and/or
EM bacterium and organic carbon source is added as nitrogen source in nitrosonium salts in culture solution, and the addition of EM bacterium is 1 × 106A/L culture solutions
~9 × 108A/L culture solutions control the concentration of organic carbon source in culture solution in 1g/L culture solutions~30g/L culture solutions.
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