CN106167763B - The anaerobic culture device of liquid humid acid fertilizer bacterium - Google Patents
The anaerobic culture device of liquid humid acid fertilizer bacterium Download PDFInfo
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- CN106167763B CN106167763B CN201610776662.7A CN201610776662A CN106167763B CN 106167763 B CN106167763 B CN 106167763B CN 201610776662 A CN201610776662 A CN 201610776662A CN 106167763 B CN106167763 B CN 106167763B
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/44—Means for regulation, monitoring, measurement or control, e.g. flow regulation of volume or liquid level
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Abstract
The invention discloses a kind of anaerobic culture device of liquid humid acid fertilizer bacterium, reactor is set as IIIth area of Ith area of reactor, IIth area of reactor and reactor；By the combination of aeration tank and deoxygenation pond, by reduction-state AH2QDS is oxidized to AQDS, participates in electron transfer process again, realizes recycling for culture solution, reduces microbial inoculum cost of manufacture.It is simple to avoid means, the operating method such as topping up or manufacture vacuum environment compared with traditional anaerobic culture device by the present invention；And after each cultivation cycle, using bentonite adsorption microbial bacterial agent, microbial inoculum loss is avoided, guarantee is provided for efficiently using for follow-up microbial inoculum.
Description
Technical field
The present invention relates to the Anaerobic culturel of microbial bacterial agent culture apparatus more particularly to a kind of liquid humid acid fertilizer bacterium dresses
It sets.
Background technology
Traditional microbiological anaerobic culture device includes mainly two kinds of forms：When biochemical oxygen demand, i.e., by adding chemical drugs
Agent manufactures anaerobic environment using oxygen in consumption of chemical reaction device；Second is that using gas cylinder into reactor topping up
Or artificially manufacture vacuum environment.Liquid humid acid fertilizer bacterium is widely present in as a kind of anaerobic bacteria in river drift and soil, and
Environment greatly differs from each other with actual bed mud-overlying water system in traditional anaerobic culture device；Secondly, reactor is combined with gas cylinder,
Not only increase floor space, improves microbial inoculum cost of manufacture, and reactor aerobic state, Bu Nengbao are easily caused when every sub-sampling
Demonstrate,prove strictly anaerobic environment.
Liquid humid acid fertilizer bacterium, can be using organic matter as carbon source as a kind of microorganism with humic formula respiration capability
And electron donor, coupling energy are grown.Reduction-state humus can be restored further caused by humic formula respiration
Oxidation state species in environment, such as Fe (III), Mn (IV), Cr (VI), U (VI), nitroaromatic and polyhalo pollutant.
Therefore, humus respiration can influence the biogeochemical cycle of Carbon and nitrogen cycles and some trace metals in environment, and energy
Enough promote the detoxification of heavy metal and organic pollution, in-situ immobilization, dirt in the self-purification of water, contaminated soil and river drift
Water process etc. has broad application prospects, and at present for the pilot scale culture of liquid humid acid fertilizer bacterium still in blank rank
Section.
AQDS (electron transit mediator) is used as a kind of humus pattern object, has quinonyl structure similar with natural humus,
But its molecular weight is far smaller than the humus in soil and deposit, it is easier to be utilized by microorganism growth metabolism, 1mmol/
LAQDS can be coupled about 60 times of thalli growth.Therefore, the present invention designs a kind of Anaerobic culturel method, using AQDS culture solutions, and will
Culture solution recycles, and ensures the enrichment and growth that microbial inoculum is carried out under anaerobic environment.
Invention content
The purpose of the present invention, be in view of at present to the pilot scale culture of liquid humid acid fertilizer bacterium still in blank stage, for gram
The deficiency of the anaerobic culture device of the traditional liquid humid acid fertilizer bacterium of clothes, provides a kind of new clothes of liquid humid acid fertilizer bacterium efficiently concentrating culture
It sets.
The present invention is achieved by following technical solution.
A kind of anaerobic culture device of liquid humid acid fertilizer bacterium, including reactor, automatic liquid-level control device, water distributor and remove
Oxygen pond；It is characterized in that, the reactor is provided with IIIth area of Ith area of reactor, IIth area of reactor and reactor；
It is provided with unidirectional gas eduction unit 8 at the top of Ith area of reactor and activated sludge fills into mouth 9, the side in Ith area of reactor
Face is provided with culture solution and fills into a mouthful A10, automatic liquid-level control device A11 and valve A12, and the outlet end of valve A12 is provided with admittedly
Liquid/gas separator A13, solid-liquid separator A13 are connected with IIth area of reactor with aeration tank 19 respectively；
The side in IIth area of reactor is provided with culture solution and fills into a mouthful B14, automatic liquid-level control device B15 and valve B16, valve
The outlet end of door B16 is provided with solid-liquid separator B17, and solid-liquid separator B17 is connected with IIIth area of reactor with aeration tank 19 respectively
It is logical；
The other side in IIth area of reactor is provided with deoxygenation pond 3, and deoxygenation pond 3 is connected by centrifugal pump 4 with aeration tank 19；It removes
The top in oxygen pond 3 is provided with oxygen scavenger supply port 1 and automatic liquid-level control device 2, and deoxygenation pond 3 passes through multidirectional valve 6 and setting
Water distributor A5 in IIth area of reactor is connected with the water distributor B7 for being arranged in IIIth area of reactor；
The side in IIIth area of reactor is provided with automatic liquid-level control device C18；The bottom in IIIth area of reactor is provided with absorption
Area 21, adsorption zone 21 are connected by valve 20 with IIIth area of reactor；The side of adsorption zone 21 is provided with discharge gate 22, adsorption zone
21 bottom is provided with ultrafiltration membrane 23；The top of ultrafiltration membrane 23 is additionally provided with absorption carrier in adsorption zone 21, which is
Bentonite.
Ith area of the reactor, IIth area of reactor, IIIth area of reactor dischargeable capacity be respectively 0.2,0.2,0.4m3。
The gradient of the sections bottom partition board in Ith area of the reactor, IIth area of reactor is 3%~5%.
The deoxygenation pond 3,19 dischargeable capacity of aeration tank are 0.2m3, 21 dischargeable capacity of adsorption zone is 0.4m3。
The water distributor A5, water distributor B7 are the unilateral horizontal direction trepanning in lower section, opening diameter 50mm, and quantity is
20。
The absorption carrier is the bentonite through grinding and crossing 80 mesh sieve and high-temperature sterilization, and bentonite additive amount is adsorption zone
The 1/4 of dischargeable capacity；And the medicament KH through grinding and crossing 80 mesh sieve and high-temperature sterilization is added wherein2PO4、CaCl2、MgSO4And
NaHCO3, the quality proportioning of medicament is 5：2：1：1, the volume of medicament addition is the 5~10% of the volume of bentonite addition,
Medicament and bentonite are laid in after mixing above ultrafiltration membrane.
The present invention compensates for the blank of liquid humid acid fertilizer bacterium pilot scale culture device, and compared with traditional anaerobic culture device
Compared with avoiding topping up or manufacture means, the operating method such as vacuum environment be simple.In addition, passing through aeration tank and deoxygenation
The combination in pond, by reduction-state AH2QDS is oxidized to AQDS, participates in electron transfer process again, realizes the cycle profit of culture solution
With reducing microbial inoculum cost of manufacture.Meanwhile the absorption to microbial inoculum is realized using the stronger bentonite of adsorption capacity in adsorption zone,
Microbial inoculum loss is avoided, guarantee is provided for efficiently using for follow-up microbial inoculum.
Description of the drawings
Fig. 1 is liquid humid acid fertilizer bacterium anaerobic culture device schematic diagram.
Reference numeral is as follows：
1 --- oxygen scavenger supply port 2 --- automatic liquid-level control device
3 --- deoxygenation pond 4 --- centrifugal pumps
5 --- water distributor A 6 --- multidirectional valves
The unidirectional gas export mouths of 7 --- water distributor B 8 ---
Culture solution fills into a mouthful A to 9 --- activated sludge fills into mouth 10 ---
11 --- automatic liquid-level control device A 12 --- valve A
Culture solution fills into a mouthful B to 13 --- solid-liquid separator A 14 ---
15 --- automatic liquid-level control device B 16 --- valve B
17 --- solid-liquid separator B 18 --- automatic liquid-level control device C
19 --- aeration tank 20 --- valves
21 --- adsorption zone 22 --- discharge gates
23 --- ultrafiltration membrane 24 --- bentonites
Specific implementation mode
The present invention is prepared using conventional raw materials and common process method.
Bentonite is a kind of claystone, and quality loosely such as soil, can be dispersed into suspension or gel, therefore in aqueous medium
There is stronger adsorption capacity to liquid, organic substance, maximal absorptive capacity is up to 5 times and the weight of itself.
Bentonite be one kind using montmorillonite as the general designation of the nonmetallic minerals of main mineral constituent, according to interlayer cation
Difference is divided into as sodium bentonite, calcium-base bentonite, hydrogen-based bentonite and organobentonite.The adsorption capacity of sodium bentonite
It is substantially better than other type bentonites, therefore the preferred sodium bentonite of the present invention is as adsorbent, and carried out after being ground
It uses.
The operation principle of liquid humid acid fertilizer bacterium anaerobic culture device of the present invention is as follows：
The complete cultivation cycle of specific embodiment is 96h, including three phases.First cultivation stage is 48h, is being reacted
It is carried out in Ith area of device, dischargeable capacity 0.2m3；Second cultivation stage is for 24 hours, to be carried out in IIth area of reactor, dischargeable capacity is
0.2m3；Third cultivation stage is for 24 hours, to be carried out in IIIth area of reactor, dischargeable capacity 0.2m3；After each period culture,
There are about 0.3m3Thalline and culture solution enter adsorption zone, adsorbed using bentonite.
Culture solution is AQDS culture solutions, and main component is beer waste water or molasses containing waste water, and supplements AQDS into waste water and make
For electron acceptor, magnitude of recruitment is controlled in 40~60g/m3, injection nutrient solution volume is 0.1m3, therefore the control of AQDS magnitude of recruitments is 6g.
(1) first cultivation stage, incubation time 48h
Mouthful A10 is filled by culture solution and injects fresh medium to Ith area of reactor, when liquid level reaches automatic liquid level control
Stop when 11 setting height of device, i.e. I 1/2 volume of area of reactor processed.It takes sewage plant anaerobic activated sludge as inoculation mother liquor, connects
Kind amount is 0.08m3, activated sludge filling into Ith area of the entrance reactor of mouth 9 by cultivating.In incubation, anaerobe is with lemon
Lemon acid sodium is carbon source and electron donor, and AQDS is that electron acceptor carries out quinone breathing, transmittance process of the electronics in cell membrane respiratory chain
In, coupling generates energy and supports thalli growth.With the proliferation and metabolism of liquid humid acid fertilizer bacterium, AQDS receives electronics and is reduced to
AH2QDS, culture solution gradually become orange red by yellow.The CH for excluding to generate by unidirectional gas eduction unit 84、CO2And N2Deng
Gas.
After cultivating 48h, mouthful B14 is filled by culture solution and injects fresh medium to IIth area of reactor, when liquid level reaches
Stop when to 15 setting height of automatic liquid-level control device, i.e. II 1/2 volume of area of reactor.Open simultaneously valve A12, thalline and
Culture solution enters the second cultivation stage culture of progress of IIth area of reactor through solid-liquid separator A13, wherein thalline, and culture solution enters exposure
Valve A12 is closed in gas pond 19 when thalline and culture solution gradually empty.It is 0.6m to control aeration quantity in aeration tank3/ h, aeration
Time 1h, in aeration process, culture solution gradually becomes yellow by orange red, wherein the AH being reduced2QDS is gradually oxidized to
AQDS can be re-used as electron acceptor and participate in electron transmission, to realize recycling for culture solution.
In addition, after thalline in Ith area of reactor and culture solution gradually empty, mouthful A10 is filled into reactor I by culture solution
Fresh medium is injected in area, when liquid level reaches automatic liquid-level control device A11 setting heights, i.e. Ith area of reactor, 1/2 volume
When stop.Activated sludge enters Ith area of reactor, inoculum concentration 0.08m by filling into mouth 93, a new round is restarted in Ith area of reactor
The first stage of liquid humid acid fertilizer bacterium cultivates.
(2) second cultivation stages, incubation time are for 24 hours
The timing since thalline enters IIth area of reactor.
Culture solution in aeration tank 19 is pumped to through centrifugal pump 4 in deoxygenation pond 3, and aeration tank (19) are effective with deoxygenation pond 3
Volume is 0.2m3；Using sulphite as oxygen scavenger in deoxygenation pond 3, added amount of chemical 15g.Culture solution is in deoxygenation pond 3
2h is stopped, by automatic liquid-level control device 2 and changeover valve gate system 6, through water distributor A5, water distributor B7, there is 1/2 volume respectively
Culture solution enters IIth area of reactor and 1/2 volume culture solution enters IIIth area of reactor.Water distributor A5, water distributor B7 are that lower section is single
Side opening, and using Cyclic culture liquid from water distributor injection realize to lower section cultivate region hydraulic mixing, increase thalline with
The contact of culture solution, to improve phage surface mass-transfer efficiency.
After thalline culture for 24 hours, valve B16 is opened, thalline and culture solution pass through solid-liquid separator B17 in IIth area of reactor,
Thalline enters IIIth area of reactor and carries out third cultivation stage culture；Culture solution enters aeration tank 19, and control aeration quantity is 0.6m3/
H, aeration time 1h close valve B16 when thalline and culture solution gradually empty.
In addition, after thalline in IIth area of reactor and culture solution empty, mouthful B14 is filled into IIth area of reactor by culture solution
Fresh medium is injected, when liquid level reaches 15 setting height of automatic liquid-level control device, i.e. II 1/2 volume of area of reactor
Stop.
(3) third cultivation stage, incubation time are for 24 hours
The timing since thalline enters IIIth area of reactor.
Culture solution in aeration tank is pumped to deoxygenation pond 3 through centrifugal pump 4, after stopping 2h, by being controlled by automatic liquid level
Device 2 and changeover valve gate system 6, through water distributor B7, fully enter IIIth area of reactor.
After thalline culture for 24 hours, opens and be located at reactor bottom valve 20, thalline and culture solution enter adsorption zone 21, pass through
Automatic liquid-level control device 18 controls, and when liquid is flow to end, closes valve 20.
At this point, thalline has been subjected to 48h cultures in Ith area of reactor, valve A12, thalline and culture solution are opened through being separated by solid-liquid separation
Device A13, wherein thalline enter IIth area of reactor and carry out second stage culture, and culture solution enters aeration tank 19 and is aerated, when anti-
When the thalline and culture solution in Ith areas Ying Qi gradually empty, valve A12 is closed；It is 0.3~0.6m to control aeration quantity in aeration tank 193/
H, 0.5~1h of aeration time repeat step (2), (3), realize the continuous culture of liquid humid acid fertilizer bacterium.
In addition, after thalline in Ith area of reactor and culture solution gradually empty, mouthful A10 is filled into reactor I by culture solution
Fresh medium is injected in area, and liquid level is controlled by automatic liquid-level control device A11；Activated sludge fills into mouth by activated sludge
9 enter Ith area of reactor, and inoculum concentration is the 30~40% of I area's dischargeable capacity of reactor, and new round corruption is restarted in Ith area of reactor
Grow the first stage culture of matter reducing bacteria.
21 dischargeable capacity of adsorption zone is 0.4m3, absorption carrier is provided in adsorption zone 21, absorption carrier is through grinding 80
The bentonite of mesh sieve and high-temperature sterilization, additive amount 0.1m3；And it is added wherein through grinding 80 mesh sieve and high-temperature sterilization
Medicament KH2PO4、CaCl2、MgSO4、NaHCO3, mass ratio 5：2：1：1, additive amount is the 5% of bentonite amount.
Thalline and culture solution open discharge gate 22 and collect thalline, waste liquid is through adsorption zone lower end after adsorption zone 21 is adsorbed
Ultrafiltration membrane 23 is discharged.
Claims (6)
1. a kind of anaerobic culture device of liquid humid acid fertilizer bacterium, including reactor, automatic liquid-level control device, water distributor and deoxygenation
Pond；It is characterized in that, the reactor is provided with IIIth area of Ith area of reactor, IIth area of reactor and reactor；
It is provided with unidirectional gas eduction unit (8) at the top of Ith area of reactor and activated sludge fills into mouth (9), the side in Ith area of reactor
Face is provided with culture solution and fills into a mouthful A (10), automatic liquid-level control device A (11) and valve A (12), the outlet end of valve A (12)
It is provided with solid-liquid separator A (13), solid-liquid separator A (13) is connected with IIth area of reactor with aeration tank (19) respectively；
The side in IIth area of reactor is provided with culture solution and fills into a mouthful B (14), automatic liquid-level control device B (15) and valve B (16),
The outlet end of valve B (16) is provided with solid-liquid separator B (17), solid-liquid separator B (17) respectively with IIIth area of reactor and aeration
Pond (19) is connected；
The other side in IIth area of reactor is provided with deoxygenation pond (3), and deoxygenation pond (3) are connected by centrifugal pump (4) with aeration tank (19)
It is logical；Oxygen scavenger supply port (1) and automatic liquid-level control device (2) are provided at the top of deoxygenation pond (3), deoxygenation pond (3) pass through more
It is connected to valve (6) with the water distributor A (5) and the water distributor B (7) that is arranged in IIIth area of reactor for being arranged in IIth area of reactor；
The side in IIIth area of reactor is provided with automatic liquid-level control device C (18)；The bottom in IIIth area of reactor is provided with adsorption zone
(21), adsorption zone (21) is connected by valve (20) with IIIth area of reactor；The side of adsorption zone (21) is provided with discharge gate
(22), the bottom of adsorption zone (21) is provided with ultrafiltration membrane (23)；It is additionally provided with suction above adsorption zone (21) interior ultrafiltration membrane (23)
Appendix body, the absorption carrier are bentonite.
2. the anaerobic culture device of liquid humid acid fertilizer bacterium according to claim 1, which is characterized in that Ith area of the reactor,
IIth area of reactor, IIIth area of reactor dischargeable capacity be respectively 0.2,0.2,0.4m3。
3. the anaerobic culture device of liquid humid acid fertilizer bacterium according to claim 1, which is characterized in that Ith area of the reactor,
The gradient of the sections bottom partition board in IIth area of reactor is 3%~5%.
4. the anaerobic culture device of liquid humid acid fertilizer bacterium according to claim 1, which is characterized in that the deoxygenation pond (3),
Aeration tank (19) dischargeable capacity is 0.2m3, adsorption zone (21) dischargeable capacity is 0.4m3。
5. the anaerobic culture device of liquid humid acid fertilizer bacterium according to claim 1, which is characterized in that the water distributor A
(5), water distributor B (7) is the unilateral horizontal direction trepanning in lower section, opening diameter 50mm, quantity 20.
6. the anaerobic culture device of liquid humid acid fertilizer bacterium according to claim 1, which is characterized in that the absorption carrier is
Through grinding and crossing the bentonite of 80 mesh sieve and high-temperature sterilization, bentonite additive amount is the 1/4 of adsorption zone dischargeable capacity；And wherein
The medicament KH that addition sieves simultaneously high-temperature sterilization through grinding and crossing 80 mesh2PO4、CaCl2、MgSO4And NaHCO3, the quality proportioning of medicament is
5：2：1：1, the volume of medicament addition is the 5~10% of the volume of bentonite addition, and medicament is uniformly mixed with bentonite
After be laid in above ultrafiltration membrane.
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| CN111908609B (en) * | 2020-07-09 | 2021-12-10 | 北京工业大学 | Anaerobic denitrification device is reinforceed to short distance nitration coupling reduction state humus |
| WO2022160571A1 (en) * | 2021-01-29 | 2022-08-04 | 上海睿钰生物科技有限公司 | In vitro life culture system and method |
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| CN101307297A (en) * | 2007-12-21 | 2008-11-19 | 浙江大学 | Separating, purifying and identifying method with anaerobic humus respiration capability reducing bacterium |
| CN102443400B (en) * | 2011-10-22 | 2014-01-08 | 广东省生态环境与土壤研究所 | Ternary composite organic pollution repairing agent containing iron oxide, humus and reducing bacteria thereof, and preparation method of repairing agent |
| CN104458624B (en) * | 2014-12-19 | 2017-02-22 | 中国环境科学研究院 | Method for analyzing electron transfer capability of in-situ humus of soil |
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