CN108179100A - A kind of photosynthetic bacteria screening and culturing method - Google Patents
A kind of photosynthetic bacteria screening and culturing method Download PDFInfo
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- CN108179100A CN108179100A CN201810252837.3A CN201810252837A CN108179100A CN 108179100 A CN108179100 A CN 108179100A CN 201810252837 A CN201810252837 A CN 201810252837A CN 108179100 A CN108179100 A CN 108179100A
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Abstract
The invention belongs to photosynthetic bacteria culture technique field, specifically a kind of photosynthetic bacteria screening and culturing method, the bacteria screening cultural method includes the following steps：Sampling, unlatching culture systems, pregnant solution is prepared, enrichment, is detached, pure medium is prepared, isolates and purifies and produce in batches；The present invention manufactures a suitable culture environment for photosynthetic bacteria, is conducive to a large amount of enrichments of photosynthetic bacteria, can shorten the screening duration of photosynthetic bacteria, improve the screening precision of photosynthetic bacteria, is conducive to screening and the scale evaluation of photosynthetic bacteria.
Description
Technical field
The invention belongs to photosynthetic bacteria culture technique field, specifically a kind of photosynthetic bacteria screening and culturing method.
Background technology
Photosynthetic bacteria be occur on the earth earliest, generally existing in nature, the protokaryon with original luminous energy synthetic system
Biology, be it is a kind of using light as the energy, can under anaerobism illumination or aerobic dark condition using in nature organic matter, vulcanize
Object, ammonia etc. carry out photosynthetic microorganism as hydrogen donor and carbon source.Photosynthetic bacteria is distributed widely in the soil of nature, water
Field, marsh, lake, Jiang Hai etc. are distributed mainly on the anoxic zone that light can be transmitted in aquatic environment.
In aquaculture, noxious materials, the realization such as nitrite, sulfide that photosynthetic bacteria can degrade in water body are filled
When bait, purify water, prevent disease, as functions such as feed addictives；Photosynthetic bacteria is adaptable, can restrain oneself high concentration
Organic wastewater, have to poisonous substances such as phenol, cyanogen it is certain endure and capacity of decomposition, there is stronger decomposition and inversion ability.But photosynthetic bacteria
Growth mainly by temperature, illumination, pH and influence of oxygen content.To realize the large-scale culture of photosynthetic bacteria, at present, photosynthetic bacteria
Culture medium is mainly placed on addition strain in installation for fermenting by 1. and cultivated by culture, and this training method is to required training
Foster base is of high cost, equipment investment is big；2. using plastic barrel, culture medium and strain are put into plastic barrel, light is provided using sunlight
Energy and thermal energy, but this mode is influenced by external environment and the depositional of photosynthetic bacteria, causes product stability poor, it is difficult to
Realize large-scale culture.Meanwhile need to obtain strain before photosynthetic bacteria is cultivated, by cultivating strain come mass propgation light
Close bacterium, then being one for the strain that photosynthetic bacteria is filtered out before mass propgation photosynthetic bacteria will solve the problems, such as.
In consideration of it, a kind of photosynthetic bacteria screening and culturing method of the present invention, is mainly used for the screening to photosynthetic bacteria
And culture, the screening precision of photosynthetic bacteria and culture scale can be improved.
Invention content
In order to make up for the deficiencies of the prior art, the present invention proposes a kind of photosynthetic bacteria screening and culturing method, and the present invention is main
The screening and culture to photosynthetic bacteria are used for, improves the screening precision of photosynthetic bacteria and culture scale, makes photosynthetic bacteria can be with
Volume production.The phase that the present invention passes through incubator, controller one, stirring module, environment adjustment module, nutrition case and gas Switching Module
Mutual cooperating can give photosynthetic bacteria one suitable growing environment, and pass through the effect of lighting module so that photosynthetic thin
The culture of bacterium is compensated automatically under conditions of natural light underutilization by artificial lighting, is conducive to the screening of photosynthetic bacteria
And scale evaluation.
The technical solution adopted by the present invention to solve the technical problems is：A kind of photosynthetic bacteria screening and culturing method, this is thin
Bacterium screening and culturing method uses following culture systems, which includes lighting module, incubator, controller one, stirring mould
Block, environment adjustment module, nutrition case and gas Switching Module, the incubator are used to cultivate photosynthetic bacteria；The lighting module
Positioned at incubator side, lighting module is used for photosynthetic bacteria photosynthesis；Circulation pipe and cycle are provided in the lighting module
The both ends of pipe are connected with incubator；Infusion pump one is provided on the circulation pipe, infusion pump one is used to convey in circulation pipe
Liquid；The stirring module is installed on incubator upper end, and stirring module is used to stir the photosynthetic bacteria in incubator with nutrient solution
It mixes uniformly；The controller one is located at stirring module upper end, and controller one stirs module, gas exchanges mould for adjusting and controlling
Block, environment adjustment module and nutrition case；The environment adjustment module is arranged on incubator, and environment adjustment module is trained for adjusting
Support the environment in case；The nutrition case is located at stirring module side, and nutrition case is connected with incubator, and nutrition case is for temporary battalion
Nutrient solution and culture solution is conveyed into incubator；The gas exchanges module is located at incubator side and gas exchanges module is with cultivating
Case is connected, and gas exchanges module is used to replacing gas in incubator and removes oxygen in gas；The upper end of the incubator
Bacterium solution inlet port is provided with, the lower end of incubator is provided with bacterium solution conveying end；
The bacteria screening cultural method includes the following steps：
A. it samples：Muddy water is taken from Reservoir Sediment, low temperature is taken back；
B. culture systems are opened：After sampling, culture systems are carried out with sterilization treatment, and utilize the control of controller one,
It is 28 DEG C ± 1 DEG C that the temperature in incubator is adjusted by environment adjustment module, is updated in incubator by gas exchanges module
Gas, and the oxygen in incubator is removed；
C. pregnant solution is prepared：After completing step b, the brine containing 2% salt is taken, and add in：NH4Cl、CH3COONa、
MgSO47H2O, yeast extract and peptone adjust pH to 7.0, and 121 DEG C sterilize 30 minutes；After sterilizing add in NaHCO3 solution and
KH2PO4 solution；And pregnant solution is put into nutrition case；
D. it is enriched with：The muddy water in step a is taken to be put into incubator from bacterium solution inlet port, using the control of controller one,
The pregnant solution in nutrition case is made to enter in incubator；Pregnant solution in nutrition case is introduced in incubator；It will by stirring module
Pregnant solution is uniformly mixing to obtain mixed bacteria liquid with muddy water, and the PH that the mixed bacteria liquid in incubator is adjusted by environment adjustment module is
7.0, mixed bacteria liquid is conveyed by delivery pump one by circulation pipe and is entered in lighting module；Lighting module is adjusted, makes lighting module
Illumination is carried out to mixed bacteria liquid, the standard that intensity of illumination is made to be not less than 34 μ Em-2s-1, Light To Dark Ratio 15h:9h, culture 7
After it, the enrichment bacterium solution after culture is taken out from bacterium solution conveying end；
E. isolation and purification culture basigamy system：After step d to be done, the brine containing 2% salt and addition are taken：NH4Cl、
CH3COONa, MgSO47H2O, yeast extract and agar, it is 7.0 to adjust pH value, is sterilized 30 minutes at 121 DEG C；It is added in after sterilizing
NaHCO3 and K2HPO4；
F. it isolates and purifies：In an aseptic environment, the enrichment bacterium solution in Step d is pipetted in sterile petri dish, is poured into
Culture medium described in being cooled to 50-55 DEG C of step d, slowly shakes up, and solidification is stood, and be sealed with film, in 28 DEG C ± 1 DEG C
Quiescent culture, the standard that intensity of illumination is made to be not less than 34 μ Em-2s-1, illumination cultivation in 36 hours incubators pass through sight
It examines, picking red single bacterium colony, repetition carries out scribing line separation in culture medium described in d, obtains Rhodopseudomonas photosynthetic bacteria；
G. it produces in batches：The Rhodopseudomonas photosynthetic bacteria obtained in f is taken, the operation of step b, c is repeated, by Step d
In muddy water be changed to the Rhodopseudomonas photosynthetic bacteria of acquisition and repeat Step d, you can volume production Rhodopseudomonas is photosynthetic
Bacterium.
The lighting module includes annular lamp tube group, annular reflective mirror, annular black ink shading group, tusche supply mains, control
Device two processed, valve one, valve two, black ink storage tank, black ink storage tank and infusion pump two, the annular lamp tube group is surrounds
Multiple first fluorescent tubes of one circle；The annular reflective mirror is located on the inside of annular lamp tube group, for the light sent out to annular lamp tube group
It is reflected；The circulation pipe cycle is wrapped on annular light group outer wall；The annular black ink shading group is forms a circle
Multiple black inks be in charge of, annular black ink shading group is set on the outside of circulation pipe, and annular black ink shading group is used for following
Light on the outside of endless tube is blocked；The tusche supply mains is located at annular black ink shading group lower end, tusche supply mains with it is every
A black ink is in charge of unicom；The photosensitive sensor is distributed on annular black ink shading group outer wall, and photosensitive sensor is used to examine
Survey the light intensity outside annular black ink shading group；The controller two is located at annular lamp tube group lower end, and controller two passes through
Photosensitive sensor input signal controls opening for annular light tube group, valve one, valve two, infusion pump two and black ink storage tank
It closes；The valve one is located at black ink and is in charge of bottom end, and valve one is used to that black ink to be controlled to be in charge of interior black ink flow direction；The valve
Door two is located at tusche supply mains bottom end, and valve two is used to that the black ink in tusche supply mains to be controlled to flow to；The infusion pump two
In two lower end of valve, infusion pump two is used to the black ink of black ink storage tank being transported to during black ink is in charge of.In use, it opens
Infusion pump one, controller one, controller two, valve one, valve two, photosensitive sensor, black ink storage tank and infusion pump two, it is defeated
Mixed bacteria liquid in incubator is extracted and conveyed to circulation pipe and mixed bacteria liquid being made to be recycled always in circulation pipe by liquid pump one
Flowing, at this point, the intensity of illumination that photosensitive sensor sensing is extraneous, and signal is transmitted to controller two, pass through the control of controller two
System, when extraneous intensity of illumination is sufficiently large；Annular lamp tube group and infusion pump two are closed；Valve one, valve two are opened, tusche moisture
Black ink in pipe falls under the effect of gravity, and black ink is flowed back into black ink storage tank；When extraneous intensity of illumination is to cycle
When intensity of illumination on front side of pipe is enough and to intensity of illumination deficiency on rear side of circulation pipe, pass through the control of controller two, circular lamp
The first fluorescent tube on front side of pipe group is closed, and the first fluorescent tube of rear side is opened, and the valve one and valve two of front side are opened, and infusion pump two closes
Close, the black ink on front side of annular black ink shading group be in charge of in black ink whereabouts be back in black ink storage tank after, it is extraneous
Light can direct projection on front side of circulation pipe, the valve one of front side is closed, and the valve one of rear side is opened, by the effect of infusion pump two,
By the black ink in black ink storage tank be transported to rear side black ink be in charge of in, at this point, ambient can not direct projection to recycle
On rear side of pipe, the first fluorescent tube on front side of annular lamp tube group can be to carrying out illumination on rear side of circulation pipe, the valve one of rear side is closed；It realizes
Intensity of illumination in circulation pipe is conducive to photosynthetic bacteria and fully carries out photosynthesis, Fast Growth always not less than standard value.
The incubator includes housing, heating insulation layer and clock, and the heating insulation layer is attached in inner walls, adds
Hot insulating layer is used for incubator inside heating and thermal insulation；The clock is located on housing exterior walls, and clock is used for the note of incubation time
When.In use, temperature sensor detects the temperature in incubator and feeds back to controller one, controller one controls and adjusts heating
Insulating layer is to incubator inside heating and thermal insulation, the time of culture is determined by watching the clock on housing exterior walls.
The stirring module includes motor one, hollow agitating shaft, axle sleeve and the first spherical shell, and the motor one is fixed on culture
Case upper end；The hollow agitating shaft is connect with motor one；The axle sleeve is located at one lower end of motor, is provided in first in axle sleeve
Chamber；The hollow agitating shaft is adapted with axle sleeve；The upper end of the hollow agitating shaft is provided with the second spherical shell, is set on the second spherical shell
It is equipped with multiple first through hole and the second spherical shell is located in first inner chamber；Second inner chamber, the first ball are provided in first spherical shell
Multiple second through-holes, first inner chamber and second inner chamber unicom are equipped on shell, multi-disc stirring is equipped on the outer wall of the first spherical shell
Blade is also equipped with multiple second fluorescent tubes on the outer wall of the first spherical shell.In use, under the control of controller one, one turn of motor
Dynamic, motor one drives hollow agitating shaft to rotate, and hollow agitating shaft drives the rotation of the first spherical shell, and stirring blade follows the first spherical shell to turn
It is dynamic, meanwhile, nutrient solution is stored in nutrition case, controller one controls nutrition case to pour into nutrient solution into the first inner chamber of axle sleeve,
Nutrient solution in axle sleeve enters in hollow pipe and flows in the second spherical shell, and nutrient solution is thrown in incubator by the rotation of the second spherical shell,
Stirring blade rotates, and nutrient solution is made uniformly to be mixed with mixed bacteria liquid, meanwhile, the rotation of stirring blade in incubator but also be heated
Uniformly, hot-spot phenomenon will not be generated, the rotation of stirring blade will not generate local PH but also the PH in incubator is uniform
It is worth excessively high, temperature in incubator is with PH suitable for so that the enrichment culture of photosynthetic bacteria or batch are smooth into producing；And
Second fluorescent tube lights, and photosynthetic bacteria can be made to carry out photosynthesis mixed bacteria liquid illumination.
The environment adjustment module includes temperature sensor, PH detector and PH regulating boxs, the temperature sensor setting
In one end of stirring blade, temperature sensor is connect with controller one, and temperature sensor is used to sense the temperature in incubator；Institute
The other end that PH detector is arranged on stirring blade is stated, PH detector is connect with controller one, and PH detector is cultivated for detecting
PH value in case；The PH regulating boxs are located at incubator side；It is provided with the first pipe between the PH regulating boxs and axle sleeve, first
The first inner chamber of valve three, the first pipe unicom PH regulating boxs and axle sleeve is provided on pipe；The first inner chamber of the axle sleeve and nutrition
The second pipe is connected between case, valve four, first inner chamber and the nutrition case of the second pipe unicom axle sleeve are provided on the second pipe.It uses
When, temperature sensor and PH detector by the signal transmission detected to controller one, under the control and conciliation of controller one,
Heating insulation layer regulates and controls the temperature in incubator, adjusts liquid equipped with PH in PH regulating boxs, PH regulating boxs are by the first pipe
PH is conveyed into the first inner chamber of axle sleeve and adjusts liquid, PH adjusts liquid and enters the second spherical shell, and be flowed into the first ball from the second spherical shell
In shell, motor one drives the rotation of the first spherical shell, and PH adjusting liquid is thrown in incubator by the first spherical shell makes PH adjust liquid adjusting culture
The environment in incubator is adjusted in pH value in case.
The gas exchanges module includes pneumatic filter, deoxygenation machine and snorkel, and the deoxygenation machine is located at gas filtration
Between device and incubator, the snorkel unicom incubator and deoxygenation machine, and snorkel also unicom deoxygenation machine and pneumatic filter.
In use, under the control of controller one, pneumatic filter can be filtered to entering the gas in incubator, make to enter training
The gas supported in case is pure, avoids pollution incubator, because photosynthetic bacteria does not like oxygen, by deoxygenation machine to entering in incubator
Gas carries out deoxygenation to produce the environment suitable for photosynthetic bacterium growth.
The beneficial effects of the invention are as follows：
1. a kind of photosynthetic bacteria screening and culturing method of the present invention, the present invention is by sampling, opening culture systems, richness
Liquid collecting preparation, enrichment, isolation and purification culture basigamy system, the combination for isolating and purifying and producing in batches etc. cultural methods, to photosynthetic thin
Bacterium manufactures a suitable culture environment, is conducive to a large amount of enrichments of photosynthetic bacteria, can shorten the screening duration of photosynthetic bacteria, carry
The screening precision of high photosynthetic bacteria.
2. the mutual knot of a kind of photosynthetic bacteria screening and culturing method of the present invention, the lighting module and incubator
It closes, makes illumination compensation of the lighting module to mixed bacteria liquid in incubator；So that the culture of photosynthetic bacteria is in natural light underutilization
Under conditions of compensated automatically by artificial lighting, improve the photosynthesis of photosynthetic bacteria, be conducive to the screening of photosynthetic bacteria
And scale evaluation.
3. a kind of photosynthetic bacteria screening and culturing method of the present invention, the incubator, controller one, stirring module,
Environment adjustment module, nutrition case and gas Switching Module be combined with each other, and nutrition case, environment adjustment module provide suitable for incubator
Suitable nutrient solution and the growing environment suitable for photosynthetic bacterium growth, stirring module are distributed culture solution mixing in incubator, make light
It is balanced to close nutrition needed for bacterium, meanwhile, stirring module makes temperature in incubator and PH uniform, will not generate hot-spot or office
Portion's pH value is excessively high and inhibits the culture of photosynthetic bacteria, is conducive to the Scale Growth of photosynthetic bacteria.
4. a kind of photosynthetic bacteria screening and culturing method of the present invention, the incubator and gas Switching Module is mutual
With reference to, controlled the oxygen in incubator, the growing environment for making photosynthetic bacteria is suitable, be conducive to photosynthetic bacteria enrichment and
Scale evaluation.
Description of the drawings
The invention will be further described below in conjunction with the accompanying drawings.
Fig. 1 is flow chart of the method for the present invention；
Fig. 2 is the overall structure diagram of the present invention；
Fig. 3 is A-A sectional views in Fig. 2；
Fig. 4 is the stirring modular structure schematic diagram of the present invention；
In figure：Lighting module 1, circulation pipe 11, infusion pump 1, annular lamp tube group 12, annular reflective mirror 13, annular are black
Ink shading group 14, tusche supply mains 141, valve 1, valve 2 143, controller 2 15, photosensitive sensor 16, black ink
Storage tank 17, infusion pump 2 18, incubator 2, bacterium solution inlet port 21, bacterium solution liquid outlet 22, housing 23, heating insulation layer 24, when
Clock 25, controller 1, stirring module 4, motor 1, hollow agitating shaft 42, axle sleeve 43, the first spherical shell 44, stirring blade 441,
Second fluorescent tube 45, environment adjustment module 5, temperature sensor 51, PH detector 52, PH regulating boxs 53, valve 3 531, nutrition case
6th, valve 4 61, gas exchanges module 7, pneumatic filter 71, deoxygenation machine 72 and snorkel 73.
Specific embodiment
In order to be easy to understand the technical means, the creative features, the aims and the efficiencies achieved by the present invention, tie below
Specific embodiment is closed, the present invention is further explained.
As shown in Figures 1 to 4, a kind of photosynthetic bacteria screening and culturing method, the bacteria screening cultural method is using following training
The system of supporting, the culture systems include lighting module 1, incubator 2, controller 1, stirring module 4, environment adjustment module 5, nutrition
Case 6 and gas Switching Module 7, the incubator 2 are used to cultivate photosynthetic bacteria；The lighting module 1 is located at 2 side of incubator,
Lighting module 1 is used for photosynthetic bacteria photosynthesis；The both ends that circulation pipe 11 and circulation pipe 11 are provided in the lighting module 1 are equal
It is connected with incubator 2；Infusion pump 1 is provided on the circulation pipe 11, infusion pump 1 is used to convey in circulation pipe 11
Liquid；The stirring module 4 is installed on 2 upper end of incubator, and stirring module 4 is used for the photosynthetic bacteria in incubator 2 and nutrition
Liquid stirs evenly；The controller 1 is located at 4 upper end of stirring module, and controller 1 stirs module 4, gas for adjusting and controlling
Body Switching Module 7, environment adjustment module 5 and nutrition case 6；The environment adjustment module 5 is arranged on incubator 2, and environment is adjusted
Module 5 is used to adjust the environment in incubator 2；The nutrition case 6 is located at stirring 4 side of module, and nutrition case 6 and incubator 2
It connects, nutrition case 6 is used to keep in nutrient solution and convey culture solution into incubator 2；The gas exchanges module 7 is located at incubator
2 sides and gas exchanges module 7 is connected with incubator 2, gas exchanges module 7 is for replacing gas and removing in incubator 2
Oxygen in gas；The upper end of the incubator 2 is provided with bacterium solution inlet port 21, and the lower end of incubator 2 is provided with bacterium solution and goes out liquid
Mouth 22；
The bacteria screening cultural method includes the following steps：
A. it samples：Muddy water is taken from Reservoir Sediment, low temperature is taken back；
B. culture systems are opened：After sampling, culture systems are carried out with sterilization treatment, and utilize the control of controller 1,
It is 28 DEG C ± 1 DEG C that the temperature in incubator 2 is adjusted by environment adjustment module 5, updates incubator 2 by gas exchanges module 7
Interior gas, and the oxygen in incubator 2 is removed；
C. pregnant solution is prepared：After completing step b, the brine containing 2% salt is taken, and add in：NH4Cl、CH3COONa、
MgSO47H2O, yeast extract and peptone adjust pH to 7.0, and 121 DEG C sterilize 30 minutes；After sterilizing add in NaHCO3 solution and
Pregnant solution is obtained after KH2PO4 solution；It and will be in pregnant solution input nutrition case 6；
D. it is enriched with：The muddy water in step a is taken to be put into incubator 2 from bacterium solution inlet port 21, utilizes the control of controller 1
System introduces the pregnant solution in nutrition case 6 in incubator 2；It is uniformly mixing to obtain with muddy water by stirring module 4 by pregnant solution mixed
Bacterium solution is closed, the PH that the mixed bacteria liquid in incubator 2 is adjusted by environment adjustment module 5 is 7.0, passes through conveying by circulation pipe 11
One conveying mixed bacteria liquid of pump enters in lighting module 1；Lighting module 1 is adjusted, lighting module 1 is made to carry out illumination to mixed bacteria liquid,
The standard that intensity of illumination is made to be not less than 34 μ Em-2s-1, Light To Dark Ratio 15h:9h；After culture 7 days, from bacterium solution conveying end 22
Take out the enrichment bacterium solution after culture；
E. isolation and purification culture basigamy system：After step d to be done, the brine containing 2% salt and addition are taken：NH4Cl、
CH3COONa, MgSO47H2O, yeast extract and agar, it is 7.0 to adjust pH value, is sterilized 30 minutes at 121 DEG C；It is added in after sterilizing
NaHCO3 and K2HPO4；
F. it isolates and purifies：In an aseptic environment, the enrichment bacterium solution in Step d is pipetted in sterile petri dish, pours into cooling
It to culture medium described in 50-55 DEG C of step d, slowly shakes up, stands solidification, and be sealed with film, in 28 DEG C of ± 1 DEG C of standings
It cultivates, 36 μ Em-2s-1 of intensity of illumination, illumination cultivation in 36 hours incubators, passes through observation, picking red single bacterium colony, weight
It is multiple that scribing line separation is carried out in culture medium described in d, obtain Rhodopseudomonas photosynthetic bacteria；
G. it produces in batches：The Rhodopseudomonas photosynthetic bacteria obtained in f is taken, the operation of step b, c is repeated, by Step d
In muddy water be changed to the Rhodopseudomonas photosynthetic bacteria of acquisition and repeat Step d, you can volume production Rhodopseudomonas is photosynthetic
Bacterium.
The lighting module 1 includes annular lamp tube group 12, annular reflective mirror 13, annular black ink shading group 14, black ink
Manifold 141, valve 1, valve 2 143, controller 2 15, photosensitive sensor 16, black ink storage tank 17 and infusion pump two
18, the annular lamp tube group 12 is multiple first fluorescent tubes to form a circle；The annular reflective mirror 13 is located at annular lamp tube group 12
Inside, the light for being sent out to annular lamp tube group 12 reflect；The cycle of circulation pipe 11 is wrapped in outside annular lamp tube group 12
On wall；The annular black ink shading group 14 is that the multiple black inks to form a circle are in charge of, and annular black ink shading group 14 is arranged
In 11 outside of circulation pipe, and annular black ink shading group 14 is used to block the light in 11 outside of circulation pipe；The tusche
Supply mains 141 is located at annular 14 lower end of black ink shading group, and tusche supply mains 141 is in charge of unicom with each black ink；The light
Dependent sensor 16 is distributed on annular 14 outer wall of black ink shading group, and photosensitive sensor 16 is used to detect annular black ink shading group
Light intensity outside 14；The controller 2 15 is located at 12 lower end of annular lamp tube group, and controller 2 15 passes through photosensitive sensor
16 input signals control annular light tube group 12, valve 1, valve 2 143, infusion pump 2 18 and black ink storage tank 17
It opens and closes；The valve 1 is located at black ink and is in charge of bottom end, and valve 1 is used to that black ink to be controlled to be in charge of interior tusche flow
To；The valve 2 143 is located at 141 bottom end of tusche supply mains, and valve 2 143 is used to control the tusche in tusche supply mains 141
Flow to；The infusion pump 2 18 is located at 2 143 lower end of valve, and infusion pump 2 18 is used for the black ink of black ink storage tank 17
It is transported to during black ink is in charge of.In use, open infusion pump 1, controller 1, controller 2 15, valve 1, valve
2 143, photosensitive sensor 16, black ink storage tank 17 and infusion pump 2 18, infusion pump 1 is by the Mixed Microbes in incubator 2
Liquid is extracted and is conveyed to circulation pipe 11 and mixed bacteria liquid being made to be circulated always in circulation pipe 11, at this point, photosensitive sensor
The extraneous intensity of illumination of 16 sensings, and signal is transmitted to controller 2 15, by the control of controller 2 15, when ambient light is according to strong
When spending sufficiently large；Annular lamp tube group 12 and infusion pump 2 18 are closed；Valve 1, valve 2 143 are opened, and black ink is in charge of interior
Black ink fall under the effect of gravity, black ink is flowed back into black ink storage tank 17；When extraneous intensity of illumination is to circulation pipe
When the intensity of illumination of 11 front sides is enough and to the intensity of illumination deficiency of the rear side of circulation pipe 11, pass through the control of controller 2 15, ring
First fluorescent tube of 12 front side of shape light tube group is closed, and the first fluorescent tube of rear side is opened, the valve 1 of front side and 2 143 dozens, valve
It opens, infusion pump 2 18 is closed, and the black ink of annular 14 front side of black ink shading group is in charge of interior black ink whereabouts and is back to tusche
After in water storage tank 17, ambient can direct projection to 11 front side of circulation pipe, the closing of valve 1 of front side, the valve one of rear side
142 open, and by the effect of infusion pump 2 18, the black ink that the black ink in black ink storage tank 17 is transported to rear side is in charge of
It is interior, at this point, ambient can not direct projection to the rear side of circulation pipe 11, the first fluorescent tube of 12 front side of annular lamp tube group can be to circulation pipe
11 rear sides carry out illumination, and the valve 1 of rear side is closed；Realize that the intensity of illumination in circulation pipe 11 is not less than standard value always,
Be conducive to photosynthetic bacteria and fully carry out photosynthesis, Fast Growth.
The incubator 2 includes housing 23, heating insulation layer 24 and clock 25, and the heating insulation layer 24 is attached at housing
On 23 inner walls, heating insulation layer 24 is used for 2 inside heating and thermal insulation of incubator；The clock 25 is located on 23 outer wall of housing, when
Clock 25 clocks for incubation time.In use, temperature sensor 51 detects the temperature in incubator 2 and feeds back to controller one
3, the control of controller 1 and adjusting heating insulation layer 24 are to 2 inside heating and thermal insulation of incubator, by watching on 23 outer wall of housing
Clock 24 come determine culture time.
The stirring module 4 includes motor 1, hollow agitating shaft 42,43 and first spherical shell 44 of axle sleeve, the motor one
41 are fixed on 2 upper end of incubator；The hollow agitating shaft 42 is connect with motor 1；The axle sleeve 43 is located under motor 1
It holds, first inner chamber is provided in axle sleeve 43；The hollow agitating shaft 42 is adapted with axle sleeve 43；The hollow agitating shaft 42 it is upper
End is provided with the second spherical shell, and multiple first through hole are provided on the second spherical shell and the second spherical shell is located in first inner chamber；Described
Second inner chamber is provided in one spherical shell 44, multiple second through-holes are equipped on the first spherical shell 44, first inner chamber joins with second inner chamber
It is logical, multi-disc stirring blade 441 is equipped on the outer wall of the first spherical shell 44, multiple second are also equipped on the outer wall of the first spherical shell 44
Fluorescent tube 45.In use, under the control of controller 1, motor 1 rotates, and motor 1 drives hollow agitating shaft 42 to rotate,
Hollow agitating shaft 42 drives the first spherical shell 44 to rotate, and stirring blade 441 follows the first spherical shell 44 to rotate, meanwhile, it is stored up in nutrition case 6
There is a nutrient solution, controller 1 controls nutrition case 6 to pour into nutrient solution into the first inner chamber of axle sleeve 43, the nutrition in axle sleeve 43
Liquid enters in hollow pipe and flows in the second spherical shell, and nutrient solution is transferred in the first spherical shell 44 by the second spherical shell, the first spherical shell 44
Nutrient solution is thrown in incubator 2 by rotation, and stirring blade 441 rotates, and nutrient solution is made uniformly to be mixed with mixed bacteria liquid, meanwhile, it stirs
The rotation of blade 441 is mixed but also being heated evenly in incubator 2, hot-spot phenomenon, the rotation of stirring blade 441 will not be generated
But also the PH in incubator 2 is uniform, it will not generate that local pH is excessively high, the temperature in incubator 2 is with PH suitable for so that photosynthetic
The enrichment culture or batch of bacterium are smooth into producing；And the second fluorescent tube 45 lights, and can make light to mixed bacteria liquid illumination
It closes bacterium and carries out photosynthesis.
The environment adjustment module 5 includes temperature sensor 51, PH detector 52 and PH regulating boxs 53, the temperature sensing
Device 51 is arranged on one end of stirring blade 441, and temperature sensor 51 is connect with controller 1, and temperature sensor 51 is used to sense
Temperature in incubator 2；The PH detector 52 is arranged on the other end of stirring blade 441, PH detector 52 and controller 1
Connection, PH detector 52 are used to detect the pH value in incubator 2；The PH regulating boxs 53 are located at 2 side of incubator；The PH tune
The first pipe is provided between section case 53 and axle sleeve 43, valve 3 531,53 He of the first pipe unicom PH regulating boxs are provided on the first pipe
The first inner chamber of axle sleeve 43；The second pipe is connected between the first inner chamber of the axle sleeve 43 and nutrition case 6, is provided on the second pipe
Valve 4 61, first inner chamber and the nutrition case 6 of the second pipe unicom axle sleeve 43.In use, temperature sensor 51 and PH detector 52
By the signal transmission detected to controller 1, under the control and conciliation of controller 1, heating insulation layer 24 is to incubator 2
Interior temperature is regulated and controled, and liquid is adjusted equipped with PH in PH regulating boxs 53, and PH regulating boxs 53 pass through the first pipe to the first of axle sleeve 43
PH is conveyed in inner cavity and adjusts liquid, PH adjusts liquid and enters the second spherical shell, and be flowed into the first spherical shell 44 from the second spherical shell, motor one
41 the first spherical shells 44 of drive rotate, and PH adjusting liquid is thrown in incubator 2 by the first spherical shell 44 makes PH adjust in liquid adjusting incubator 2
PH value, the environment in incubator 2 is adjusted.
The gas exchanges module 7 includes pneumatic filter 71, deoxygenation machine 72 and snorkel 73, and the deoxygenation machine 72 is located at
Between pneumatic filter 71 and incubator 2；The 73 unicom incubator 2 of snorkel and deoxygenation machine 72, and the also unicom of snorkel 73
Deoxygenation machine 72 and pneumatic filter 71.In use, under the control of controller 1, pneumatic filter 71 can be cultivated entering
Gas in case 2 is filtered, and makes the gas entered in incubator 2 pure, pollution incubator 2 is avoided, because photosynthetic bacteria does not like
Oxygen carries out deoxygenation to produce the environment suitable for photosynthetic bacterium growth by deoxygenation machine 72 to the gas entered in incubator 2.
Concrete operations flow is as follows：
During work, under the control of controller 1, pneumatic filter 71 can carry out the gas entered in incubator 2
Filtering makes the gas entered in incubator 2 pure, pollution incubator 2 is avoided, by deoxygenation machine 72 to entering in incubator 2
Gas carries out deoxygenation；Under the control of controller 1, motor 1 rotates, and motor 1 drives hollow agitating shaft 42 to rotate, empty
Heart agitating shaft 42 drives the first spherical shell 44 to rotate, and stirring blade 441 follows the first spherical shell 44 to rotate, meanwhile, controller 1 controls
Nutrition case 6 pours into nutrient solution into the first inner chamber of axle sleeve 43, and the nutrient solution in axle sleeve 43 enters in hollow pipe and flows to second
In spherical shell, nutrient solution is transferred in the first spherical shell 44 by the second spherical shell, and the first spherical shell 44 is rotated is thrown to incubator 2 by nutrient solution
In, stirring blade 441 rotates, and nutrient solution is made uniformly to be mixed with mixed bacteria liquid, under the control and conciliation of controller 1, heating
Insulating layer 24 regulates and controls the temperature in incubator 2, and liquid is adjusted equipped with PH in PH regulating boxs 53, and PH regulating boxs 53 pass through first
Pipe conveys PH into the first inner chamber of axle sleeve 43 and adjusts liquid, and PH adjusts liquid and enters the second spherical shell, and be flowed into the from the second spherical shell
In one spherical shell 44, motor 1 drives the first spherical shell 44 to rotate, and PH adjusting liquid is thrown in incubator 2 by the first spherical shell 44 makes PH tune
The pH value in liquid adjusting incubator 2 is saved, the environment in incubator 2 is adjusted；Meanwhile the rotation of stirring blade 441, also make
It obtains and is heated evenly in incubator 2, hot-spot phenomenon will not be generated, but also the PH in incubator 2 is uniform, part will not be generated
PH value is excessively high, after the second fluorescent tube 45 is opened, photosynthetic bacteria can be made to carry out photosynthesis mixed bacteria liquid illumination；Pass through manufacture
Go out a suitable environment and so that the enrichment culture of photosynthetic bacteria or batch are smooth into producing.
After the control environment for producing suitable photosynthetic bacteria survival, at this point, opening infusion pump 1, controller 1, control
Device 2 15 processed, valve 1, valve 2 143, photosensitive sensor 16, black ink storage tank 17 and infusion pump 2 18, infusion pump one
Mixed bacteria liquid in incubator 2 is extracted and conveyed to circulation pipe 11 and mixed bacteria liquid being made to be followed always in circulation pipe 11 by 111
Circulation moves, at this point, photosensitive sensor 16 senses extraneous intensity of illumination, and signal is transmitted to controller 2 15, passes through controller
2 15 control, when extraneous intensity of illumination is sufficiently large；Annular lamp tube group 12 and infusion pump 2 18 are closed；Valve 1, valve
2 143 open, and black ink is in charge of interior black ink and falls under the effect of gravity, and black ink is flowed back into black ink storage tank 17；
When extraneous intensity of illumination to the intensity of illumination of the front side of circulation pipe 11 enough and to the intensity of illumination deficiency of 11 rear side of circulation pipe when, lead to
The control of controller 2 15 is crossed, the first fluorescent tube of 12 front side of annular lamp tube group is closed, and the first fluorescent tube of rear side is opened, the valve of front side
Door 1 and valve 2 143 are opened, and infusion pump 2 18 is closed, and the black ink on front side of annular black ink shading group 14 is in charge of interior
After black ink whereabouts is back in black ink storage tank 17, ambient can be on front side of direct projection to circulation pipe 11, the valve one of front side
142 close, and the valve 1 of rear side is opened, by the effect of infusion pump 2 18, the black ink in black ink storage tank 17 is defeated
Be sent to rear side black ink be in charge of in, at this point, ambient can not direct projection to the rear side of circulation pipe 11, on front side of annular lamp tube group 12
First fluorescent tube can carry out 11 rear side of circulation pipe illumination, and the valve 1 of rear side is closed；Realize that the illumination in circulation pipe 11 is strong
Degree not less than standard value, is conducive to photosynthetic bacteria and fully carries out photosynthesis, Fast Growth, Batch Culture photosynthetic bacteria always.
The basic principles, main features and advantages of the invention have been shown and described above.The technical staff of the industry should
Understand, the present invention is not limited to the above embodiments, and the above embodiments and description only describe the originals of the present invention
Reason, without departing from the spirit and scope of the present invention, various changes and improvements may be made to the invention, these changes and improvements
It all fall within the protetion scope of the claimed invention.The claimed scope of the invention is by appended claims and its equivalent circle
It is fixed.
Claims (6)
- A kind of 1. photosynthetic bacteria screening and culturing method, it is characterised in that：The bacteria screening cultural method uses following culture systems, The culture systems include lighting module (1), incubator (2), controller one (3), stirring module (4), environment adjustment module (5), Nutrition case (6) and gas Switching Module (7), the incubator (2) is for cultivating photosynthetic bacteria；The lighting module (1) is located at Incubator (2) side, lighting module (1) is for photosynthetic bacteria photosynthesis；Circulation pipe is provided in the lighting module (1) (11) and the both ends of circulation pipe (11) are connected with incubator (2)；Infusion pump one (111) is provided on the circulation pipe (11), Infusion pump one (111) is for conveying the liquid in circulation pipe (11)；The stirring module (4) is installed on incubator (2) upper end, stirs Module (4) is mixed for the photosynthetic bacteria in incubator (2) to be stirred evenly with nutrient solution；The controller one (3) is positioned at stirring Module (4) upper end, controller one (3) stir module (4), gas exchanges module (7), environment adjustment module for adjusting and controlling (5) and nutrition case (6)；The environment adjustment module (5) is arranged on incubator (2), and environment adjustment module (5) is trained for adjusting Support the environment in case (2)；The nutrition case (6) is positioned at stirring module (4) side, and nutrition case (6) is connected with incubator (2), Nutrition case (6) is for keeping in nutrient solution and to conveying culture solution in incubator (2)；The gas exchanges module (7) is positioned at culture Case (2) side and gas exchanges module (7) and incubator (2) are connected, and gas exchanges module (7) is interior for replacing incubator (2) Oxygen in gas and removing gas；The upper end of the incubator (2) is provided with bacterium solution inlet port (21), incubator (2) Lower end is provided with bacterium solution liquid outlet (22)；The bacteria screening cultural method includes the following steps：A. it samples：Muddy water is taken from Reservoir Sediment, low temperature is taken back；B. culture systems are opened：After sampling, culture systems are carried out with sterilization treatment, and using the control of controller one (3), lead to It is 28 DEG C ± 1 DEG C to cross the temperature that environment adjustment module (5) is adjusted in incubator (2), is updated and cultivated by gas exchanges module (7) Gas in case (2), and the oxygen in incubator (2) is removed；C. pregnant solution is prepared：After completing step b, the brine containing 2% salt is taken, and add in：NH4Cl、CH3COONa、MgSO4· 7H2O, yeast extract and peptone adjust pH to 7.0, and 121 DEG C sterilize 30 minutes；NaHCO3 solution is added in after sterilizing and KH2PO4 is molten Pregnant solution is obtained after liquid；It and will be in pregnant solution input nutrition case (6)；D. it is enriched with：The muddy water in step a is taken to be put into incubator (2) from bacterium solution inlet port (21), utilizes controller one (3) Control introduces the pregnant solution in nutrition case (6) in incubator (2)；Pregnant solution and muddy water are stirred by stirring module (4) Even to obtain mixed bacteria liquid, the PH that the mixed bacteria liquid in incubator (2) is adjusted by environment adjustment module (5) is 7.0, passes through cycle Pipe (11) conveys mixed bacteria liquid by delivery pump one and enters in lighting module (1)；Lighting module (1) is adjusted, makes lighting module (1) Illumination is carried out to mixed bacteria liquid, the standard that intensity of illumination is made to be not less than 34 μ Em-2s-1, Light To Dark Ratio 15h:9h；Culture 7 After it, the enrichment bacterium solution after culture is taken out from bacterium solution conveying end (22)；E. isolation and purification culture basigamy system：After step d to be done, the brine containing 2% salt and addition are taken：NH4Cl、 CH3COONa, MgSO47H2O, yeast extract and agar, it is 7.0 to adjust pH value, is sterilized 30 minutes at 121 DEG C；It is added in after sterilizing NaHCO3 and K2HPO4；F. it isolates and purifies：In an aseptic environment, the enrichment bacterium solution in Step d is pipetted in sterile petri dish, is poured into and is cooled to 50- Culture medium described in 55 DEG C of step d, slowly shakes up, and stands solidification, and is sealed with film, in 28 DEG C of ± 1 DEG C of quiescent cultures, 36 μ Em-2s-1 of intensity of illumination, illumination cultivation in 36 hours incubators pass through observation, and picking red single bacterium colony is repeated in d Scribing line separation is carried out in the culture medium, obtains Rhodopseudomonas photosynthetic bacteria；G. it produces in batches：The Rhodopseudomonas photosynthetic bacteria obtained in f is taken, repeats the operation of step b, c, it will be in Step d The Rhodopseudomonas photosynthetic bacteria that muddy water is changed to acquisition repeats Step d, you can volume production Rhodopseudomonas photosynthetic bacteria.
- 2. a kind of photosynthetic bacteria screening and culturing method according to claim 1, it is characterised in that：The lighting module (1) Including annular lamp tube group (12), annular reflective mirror (13), annular black ink shading group (14), tusche supply mains (141), valve one (142), valve two (143), controller two (15), photosensitive sensor (16), black ink storage tank (17) and infusion pump two (18), The annular lamp tube group (12) is multiple first fluorescent tubes to form a circle；The annular reflective mirror (13) is positioned at annular lamp tube group (12) inside, the light for being sent out to annular lamp tube group (12) reflect；Circulation pipe (11) cycle is wrapped in circular lamp On pipe group (12) outer wall；The annular black ink shading group (14) is that the multiple black inks to form a circle are in charge of, annular black ink Shading group (14) is set on the outside of circulation pipe (11), and annular black ink shading group (14) is for the light on the outside of circulation pipe (11) Line is blocked；The tusche supply mains (141) positioned at annular black ink shading group (14) lower end, tusche supply mains (141) with Each black ink is in charge of unicom；The photosensitive sensor (16) is distributed on annular black ink shading group (14) outer wall, photosensitive biography Sensor (16) is for detecting the external light intensity of annular black ink shading group (14)；The controller two (15) is positioned at circular lamp Pipe group (12) lower end, controller two (15) control annular light tube group (12), valve by photosensitive sensor (16) input signal One (142), the keying of valve two (143), infusion pump two (18) and black ink storage tank (17)；The valve one (142) is located at Black ink is in charge of bottom end, and valve one (142) flows to for black ink to be controlled to be in charge of interior black ink；Described two (143) position of valve In tusche supply mains (141) bottom end, valve two (143) is for controlling the black ink in tusche supply mains (141) to flow to；It is described defeated Liquid pump two (18) is positioned at valve two (143) lower end, and infusion pump two (18) is for the black ink of black ink storage tank (17) to be conveyed In being in charge of to black ink.
- 3. a kind of photosynthetic bacteria screening and culturing method according to claim 1, it is characterised in that：Incubator (2) packet Housing (23), heating insulation layer (24) and clock (25) are included, the heating insulation layer (24) is attached on housing (23) inner wall, adds Hot insulating layer (24) is for heating and thermal insulation internal to incubator (2)；The clock (25) is on housing (23) outer wall, clock (25) clocking for incubation time.
- 4. a kind of photosynthetic bacteria screening and culturing method according to claim 1, it is characterised in that：The stirring module (4) Including motor one (41), hollow agitating shaft (42), axle sleeve (43) and the first spherical shell (44), the motor one (41) is fixed on culture Case (2) upper end；The hollow agitating shaft (42) connect with motor one (41)；The axle sleeve (43) positioned at motor one (41) lower end, First inner chamber is provided in axle sleeve (43)；The hollow agitating shaft (42) is adapted with axle sleeve (43)；The hollow agitating shaft (42) upper end is provided with the second spherical shell, and multiple first through hole are provided on the second spherical shell and the second spherical shell is located at first inner chamber In；Second inner chamber is provided in first spherical shell (44), multiple second through-holes, first inner chamber are equipped on the first spherical shell (44) With second inner chamber unicom, multi-disc stirring blade (441), the outer wall of the first spherical shell (44) are equipped on the outer wall of the first spherical shell (44) On be also equipped with multiple second fluorescent tubes (45).
- 5. a kind of photosynthetic bacteria screening and culturing method according to claim 4, it is characterised in that：The environment adjustment module (5) including temperature sensor (51), PH detector (52) and PH regulating boxs (53), the temperature sensor (51) is arranged on stirring One end of blade (441), temperature sensor (51) are connect with controller one (3), and temperature sensor (51) is for sensing incubator (2) temperature in；The PH detector (52) is arranged on the other end of stirring blade (441), PH detector (52) and controller One (3) connect, and PH detector (52) is for detecting the pH value in incubator (2)；The PH regulating boxs (53) are positioned at incubator (2) Side；The first pipe is provided between the PH regulating boxs (53) and axle sleeve (43), valve three (531) is provided on the first pipe, the The first inner chamber of one pipe unicom PH regulating boxs (53) and axle sleeve (43)；The first inner chamber of the axle sleeve (43) and nutrition case (6) it Between be connected with the second pipe, be provided with valve four (61) on the second pipe, first inner chamber and the nutrition case of the second pipe unicom axle sleeve (43) (6)。
- 6. a kind of photosynthetic bacteria screening and culturing method according to claim 4, it is characterised in that：The gas exchanges module (7) including pneumatic filter (71), deoxygenation machine (72) and snorkel (73), the deoxygenation machine (72) is positioned at pneumatic filter (71) Between incubator (2)；Snorkel (73) the unicom incubator (2) and deoxygenation machine (72), and snorkel (73) also unicom is removed Oxygen machine (72) and pneumatic filter (71).
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| CN114350509A (en) * | 2022-03-17 | 2022-04-15 | 南方海洋科学与工程广东省实验室(广州) | High-pressure environment biological enrichment and spraying type solid separation culture device |
| CN114350508A (en) * | 2022-03-17 | 2022-04-15 | 南方海洋科学与工程广东省实验室(广州) | High-pressure environment marine microorganism enrichment culture and gravity type separation device |
| WO2023173494A1 (en) * | 2022-03-17 | 2023-09-21 | 南方海洋科学与工程广东省实验室(广州) | High-pressure environment biological enrichment and spraying-type solid isolation culture apparatus |
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