CN106167763A - 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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- CN106167763A CN106167763A CN201610776662.7A CN201610776662A CN106167763A CN 106167763 A CN106167763 A CN 106167763A CN 201610776662 A CN201610776662 A CN 201610776662A CN 106167763 A CN106167763 A CN 106167763A
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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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Abstract
The invention discloses the anaerobic culture device of a kind of liquid humid acid fertilizer bacterium, reactor is set to reactor I district, reactor II district and reactor III district；By Aeration tank and the combination in deoxygenation pond, by reduction-state AH2QDS is oxidized to AQDS, again participates in electron transfer process, it is achieved that recycling of culture fluid, reduces microbial inoculum cost of manufacture.The present invention is compared with tradition anaerobic culture device, it is to avoid topping up or manufacture the means such as vacuum environment, and operational approach is simple；And after each cultivation cycle terminates, use bentonite adsorption microbial bacterial agent, it is to avoid and microbial inoculum loss, provide guarantee for effectively utilizing of follow-up microbial inoculum.
Description
Technical field
The present invention, about microbial bacterial agent culture apparatus, particularly relates to the Anaerobic culturel dress of a kind of liquid humid acid fertilizer bacterium
Put.
Background technology
Traditional microbiological anaerobic culture device mainly includes two kinds of forms: one is biochemical oxygen demand, i.e. by adding chemical drugs
Agent, utilizes oxygen in consumption of chemical reaction device, manufactures anaerobic environment；Two is to use gas cylinder topping up in reactor
Or artificially manufacture vacuum environment.Liquid humid acid fertilizer bacterium, as a kind of anaerobe, is widely present in river drift and soil, and
In tradition anaerobic culture device, environment greatly differs from each other with actual bed mud-overlying water system；Secondly, reactor is combined with gas cylinder,
Not only increase floor space, improve microbial inoculum cost of manufacture, and easily cause reactor aerobic state during every sub-sampling, it is impossible to protect
Card strictly anaerobic environment.
Liquid humid acid fertilizer bacterium has the microorganism of humic formula respiration capability as a class, it is possible to use organic as carbon source
And electron donor, coupling energy grows.Reduction-state humus produced by humic formula Repiration can reduce further
Oxidation state species in environment, such as Fe (III), Mn (IV), Cr (VI), U (VI), nitroaromatic and polyhalo pollutant.
Therefore, humus respiration can affect Carbon and nitrogen cycles and the biogeochemical cycle of some trace metals in environment, and energy
Enough promote heavy metal and the detoxification of organic pollution, in in-situ immobilization, the dirt of the self-purification of water, contaminated soil and river drift
The aspects such as water process have broad application prospects, and the current pilot scale culture for liquid humid acid fertilizer bacterium is still in blank rank
Section.
AQDS (electron transit mediator), as a kind of humus pattern thing, has the quinonyl structure similar to natural humus,
But its molecular weight is far smaller than the humus in soil and deposit, it is easier to utilized by growth of microorganism metabolism, 1mmol/
LAQDS can coupling thalli growth about 60 times.Therefore, the present invention designs a kind of Anaerobic culturel method, uses AQDS culture fluid, and will
Culture fluid recycles, it is ensured that carry out the enrichment and growth of microbial inoculum under anaerobic environment.
Summary of the invention
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 the blank stage, for gram
The deficiency of the anaerobic culture device of clothes tradition liquid humid acid fertilizer bacterium, it is provided that the new clothes that a kind of liquid humid acid fertilizer bacterium efficiently concentrating is cultivated
Put.
The present invention is achieved by following technical solution.
The anaerobic culture device of a kind of liquid humid acid fertilizer bacterium, including reactor, automatic liquid-level control device, water distributor with remove
Oxygen pond；It is characterized in that, described reactor is provided with reactor I district, reactor II district and reactor III district；
The top in reactor I district is provided with unidirectional gas eduction unit 8 and activated sludge fills into mouth 9, the side in reactor I district
Face is provided with culture fluid and fills into a mouthful A10, automatic liquid-level control device A11 and valve A12, and the port of export of valve A12 is provided with solid
Liquid/gas separator A13, solid-liquid separator A13 are connected with Aeration tank 19 with reactor II district respectively；
The side in reactor II district is provided with culture fluid and fills into a mouthful B14, automatic liquid-level control device B15 and valve B16, valve
The port of export of door B16 is provided with solid-liquid separator B17, solid-liquid separator B17 and is connected with Aeration tank 19 with reactor III district respectively
Logical；
The opposite side in reactor II district is provided with deoxygenation pond 3, and deoxygenation pond 3 is connected with Aeration tank 19 by centrifugal pump 4；Remove
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 is by multidirectional valve 6 and setting
Water distributor A5 in reactor II district and the water distributor B7 being arranged on reactor III district is connected；
The side in reactor III district is provided with automatic liquid-level control device C18；The bottom in reactor III district is provided with absorption
District 21, adsorption zone 21 is connected with reactor III district by valve 20；The side of adsorption zone 21 is provided with discharge gate 22, adsorption zone
The bottom of 21 is provided with ultrafilter membrane 23；In adsorption zone 21 ultrafilter membrane 23 be arranged above absorption carrier, this absorption carrier is
Bentonite.
Described reactor I district, reactor II district, the dischargeable capacity in reactor III district be respectively 0.2,0.2,0.4m3。
Described reactor I district, the gradient of sections bottom dividing plate in reactor II district are 3%～5%.
Described deoxygenation pond 3, Aeration tank 19 dischargeable capacity are 0.2m3, adsorption zone 21 dischargeable capacity is 0.4m3。
Described water distributor A5, water distributor B7 are the unilateral horizontal direction perforate in lower section, and opening diameter is 50mm, and quantity is
20。
Described absorption carrier is that bentonite addition is adsorption zone through grinding and crossing 80 mesh sieves the bentonite of high temperature sterilize
The 1/4 of dischargeable capacity；And add wherein through grinding and crossing 80 mesh sieves the medicament KH of high temperature sterilize2PO4、CaCl2、MgSO4And
NaHCO3, the quality proportioning of medicament is 5:2:1:1, and the volume that medicament adds is the 5～10% of the volume that bentonite adds,
It is laid in above ultrafilter membrane after medicament is mixed homogeneously with bentonite.
The present invention compensate for the blank of liquid humid acid fertilizer bacterium pilot scale culture device, and compared with tradition anaerobic culture device
Relatively, it is to avoid topping up or manufacture the means such as vacuum environment, operational approach is simple.It addition, by Aeration tank and deoxygenation
The combination in pond, by reduction-state AH2QDS is oxidized to AQDS, again participates in electron transfer process, it is achieved that the circulation profit of culture fluid
With, reduce microbial inoculum cost of manufacture.Meanwhile, the bentonite that absorbability is stronger is used to realize the absorption to microbial inoculum in adsorption zone,
Avoid microbial inoculum to run off, provide guarantee for effectively utilizing of follow-up microbial inoculum.
Accompanying drawing explanation
Fig. 1 is liquid humid acid fertilizer bacterium anaerobic culture device schematic diagram.
Reference is as follows:
1 oxygen scavenger supply port 2 automatic liquid-level control device
3 deoxygenation pond 4 centrifugal pumps
The 5 multidirectional valves of water distributor A 6
7 water distributor B 8 unidirectional gas export mouth
9 activated sludge fill into mouth 10 culture fluid and fill into a mouthful A
11 automatic liquid-level control device A 12 valve A
13 solid-liquid separator A 14 culture fluid fill into a mouthful B
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 ultrafilter membrane 24 bentonite
Detailed description of the invention
The present invention uses conventional raw materials and common process method to be prepared.
Bentonite is a kind of claystone, and quality is loose as native, can be dispersed into suspension or gel, therefore in aqueous medium
Liquid, organic substance are had stronger absorbability, maximal absorptive capacity up to 5 times with the weight of self.
Bentonite is the general designation of the class nonmetallic minerals with montmorillonite as main mineral constituent, according to interlayer cation
Difference, is divided into again sodium bentonite, calcium-base bentonite, hydrogen-based bentonite and organobentonite.The absorbability of sodium bentonite
It is substantially better than other kind bentonite, therefore the preferred sodium bentonite of the present invention is as adsorbent, and carry out after being ground
Use.
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, in reaction
Device I is carried out in district, and dischargeable capacity is 0.2m3；Second cultivation stage is 24h, carries out in reactor II district, and dischargeable capacity is
0.2m3；3rd cultivation stage is 24h, carries out in reactor III district, and dischargeable capacity is 0.2m3；Each cycle cultivates after terminating,
There are about 0.3m3Thalline and culture fluid enter adsorption zone, use bentonite adsorb.
Culture fluid is AQDS culture fluid, is mainly composed of beer waste water or molasses containing waste water, and supplements AQDS work in waste water
For electron acceptor, magnitude of recruitment controls 40～60g/m3, injecting culture fluid volume is 0.1m3, therefore AQDS magnitude of recruitment controls as 6g.
(1) first cultivation stage, incubation time is 48h
Fill into mouthful A10 by culture fluid and inject fresh medium to reactor I district, when liquid level reaches automatic liquid level control
Device 11 setting height processed, i.e. stops during reactor I district's 1/2 volume.Take Sewage Plant anaerobic activated sludge as inoculation mother solution, connect
Amount of planting is for 0.08m3, activated sludge is by filling into cultivation mouth 9 enters reactor I district.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, and electronics is at the transmittance process of cell membrane respiratory chain
In, coupling produces energy and supports thalli growth.Along with the proliferation and metabolism of liquid humid acid fertilizer bacterium, AQDS accepts electronics and is reduced to
AH2QDS, culture fluid is gradually become orange red by yellow.The CH produced is got rid of by unidirectional gas eduction unit 84、CO2And N2Deng
Gas.
After cultivating 48h, fill into mouthful B14 by culture fluid and inject fresh medium to reactor II district, when liquid level reaches
To automatic liquid-level control device 15 setting height, i.e. stop during reactor II district's 1/2 volume.Open valve A12 simultaneously, thalline and
Culture fluid is through solid-liquid separator A13, and wherein thalline entrance reactor II district carries out the second cultivation stage cultivation, and culture fluid enters and exposes
Gas pond 19, when thalline and culture fluid gradually empty, closes valve A12.In control Aeration tank, aeration rate is 0.6m3/ h, aeration
Time 1h, in aeration process, culture fluid is gradually become yellow by orange red, the AH being wherein reduced2QDS is gradually oxidized to
AQDS, can participate in electron transmission as electron acceptor again, it is achieved thereby that the recycling of culture fluid.
Additionally, after thalline and culture fluid gradually empty in reactor I district, fill into mouthful A10 to reactor I by culture fluid
Fresh medium is injected in district, when liquid level reaches automatic liquid-level control device A11 setting height, i.e. reactor I district 1/2 volume
Time stop.Activated sludge is entered reactor I district by filling into mouth 9, and inoculum concentration is 0.08m3, a new round is restarted in reactor I district
The first stage of liquid humid acid fertilizer bacterium cultivates.
(2) second cultivation stages, incubation time is 24h
Enter reactor II district from thalline and start timing.
Culture fluid in Aeration tank 19 is during pump 4 is pumped to deoxygenation pond 3 by centrifugation, and Aeration tank (19) is effective with deoxygenation pond 3
Volume is 0.2m3；Using sulphite as oxygen scavenger in deoxygenation pond 3, added amount of chemical is 15g.Culture fluid is in deoxygenation pond 3
Stop 2h, by automatic liquid-level control device 2 and changeover valve gate control 6, through water distributor A5, water distributor B7, have 1/2 volume respectively
Culture fluid enters reactor II district and 1/2 volume culture fluid enters reactor III district.It is single that water distributor A5, water distributor B7 are lower section
STH, and utilize Cyclic culture liquid to realize hydraulic mixing to cultivation region, lower section from the injection of water distributor, increase thalline with
The contact of culture fluid, thus improve phage surface mass-transfer efficiency.
After yeast culture 24h, opening valve B16, in reactor II district, thalline and culture fluid are through solid-liquid separator B17,
Thalline enters reactor III district and carries out the 3rd cultivation stage cultivation；Culture fluid enters Aeration tank 19, and control aeration rate is 0.6m3/
H, aeration time 1h, when thalline and culture fluid gradually empty, close valve B16.
Additionally, after thalline and culture fluid empty in reactor II district, fill into mouthful B14 to reactor II district by culture fluid
Inject fresh medium, when liquid level reaches automatic liquid-level control device 15 setting height, i.e. during reactor II district's 1/2 volume
Stop.
(3) the 3rd cultivation stages, incubation time is 24h
Enter reactor III district from thalline and start timing.
Culture fluid in Aeration tank pump 4 by centrifugation is pumped to deoxygenation pond 3, after stopping 2h, by being controlled by automatic liquid level
Device 2 and changeover valve gate control 6, through water distributor B7, fully enter reactor III district.
After yeast culture 24h, opening and be positioned at reactor bottom valve 20, thalline and culture fluid enter adsorption zone 21, pass through
Automatic liquid-level control device 18 controls, and when liquid flows to end, closes valve 20.
Now, in reactor I district, thalline is cultivated through 48h, opens valve A12, thalline and culture fluid through solid-liquid separation
Device A13, wherein thalline entrance reactor II district carries out second stage cultivation, and culture fluid enters Aeration tank 19 and carries out aeration, when instead
When the thalline in Ying Qi I district and culture fluid gradually empty, close valve A12；In control Aeration tank 19, aeration rate is 0.3～0.6m3/
H, aeration time 0.5～1h, repeats step (2), (3), it is achieved the continuous cultivation of liquid humid acid fertilizer bacterium.
Additionally, after thalline and culture fluid gradually empty in reactor I district, fill into mouthful A10 to reactor I by culture fluid
Fresh medium is injected in district, controls liquid level by automatic liquid-level control device A11；Activated sludge is filled into mouth by activated sludge
9 enter reactor I district, and inoculum concentration is the 30～40% of reactor I district dischargeable capacity, and it is rotten that a new round is restarted in reactor I district
The first stage growing matter reducing bacteria cultivates.
Adsorption zone 21 dischargeable capacity is 0.4m3, it being provided with absorption carrier in adsorption zone 21, absorption carrier was for through grinding 80
Mesh sieve the bentonite of high temperature sterilize, addition is 0.1m3；And add wherein through grinding 80 mesh sieves high temperature sterilize
Medicament KH2PO4、CaCl2、MgSO4、NaHCO3, mass ratio is 5:2:1:1, and addition is the 5% of bentonite amount.
Thalline and culture fluid, after adsorption zone 21 is adsorbed, is opened discharge gate 22 and are collected thalline, and waste liquid is through lower end, adsorption zone
Ultrafilter membrane 23 is discharged.
Claims (6)
1. an anaerobic culture device for liquid humid acid fertilizer bacterium, including reactor, automatic liquid-level control device, water distributor and deoxygenation
Pond；It is characterized in that, described reactor is provided with reactor I district, reactor II district and reactor III district；
The top in reactor I district is provided with unidirectional gas eduction unit (8) and activated sludge fills into mouth (9), the side in reactor I district
Face is provided with culture fluid and fills into a mouthful A (10), automatic liquid-level control device A (11) and valve A (12), the port of export of valve A (12)
It is provided with solid-liquid separator A (13), solid-liquid separator A (13) to be connected with Aeration tank (19) with reactor II district respectively；
The side in reactor II district is provided with culture fluid and fills into a mouthful B (14), automatic liquid-level control device B (15) and valve B (16),
The port of export of valve B (16) be provided with solid-liquid separator B (17), solid-liquid separator B (17) respectively with reactor III district and aeration
Pond (19) is connected；
The opposite side in reactor II district is provided with deoxygenation pond (3), and deoxygenation pond (3) are connected with Aeration tank (19) by centrifugal pump (4)
Logical；The top in deoxygenation pond (3) is provided with oxygen scavenger supply port (1) and automatic liquid-level control device (2), and deoxygenation pond (3) are by many
It is connected with the water distributor A (5) being arranged on reactor II district and the water distributor B (7) being arranged on reactor III district to valve (6)；
The side in reactor III district is provided with automatic liquid-level control device C (18)；The bottom in reactor III district is provided with adsorption zone
(21), adsorption zone (21) are connected with reactor III district by valve (20)；The side of adsorption zone (21) is provided with discharge gate
(22), the bottom of adsorption zone (21) is provided with ultrafilter membrane (23)；Adsorption zone (21) interior ultrafilter membrane (23) be arranged above inhale
Appendix body, this absorption carrier is bentonite.
The anaerobic culture device of liquid humid acid fertilizer bacterium the most according to claim 1, it is characterised in that described reactor I district,
Reactor II district, the dischargeable capacity in reactor III district be respectively 0.2,0.2,0.4m3。
The anaerobic culture device of liquid humid acid fertilizer bacterium the most according to claim 1, it is characterised in that described reactor I district,
The gradient of the sections bottom dividing plate in reactor II district is 3%～5%.
The anaerobic culture device of liquid humid acid fertilizer bacterium the most according to claim 1, it is characterised in that described deoxygenation pond (3),
Aeration tank (19) dischargeable capacity is 0.2m3, adsorption zone (21) dischargeable capacity is 0.4m3。
The anaerobic culture device of liquid humid acid fertilizer bacterium the most according to claim 1, it is characterised in that described water distributor A
(5), water distributor B (7) be the unilateral horizontal direction perforate in lower section, opening diameter is 50mm, and quantity is 20.
The anaerobic culture device of liquid humid acid fertilizer bacterium the most according to claim 1, it is characterised in that described absorption carrier is
Through grinding and crossing 80 mesh sieves the bentonite of high temperature sterilize, bentonite addition is the 1/4 of adsorption zone dischargeable capacity；And wherein
Add through grinding and crossing 80 mesh sieves the medicament KH of high temperature sterilize2PO4、CaCl2、MgSO4And NaHCO3, the quality proportioning of medicament is
5:2:1:1, the volume that medicament adds is the 5～10% of the volume that bentonite adds, is mixed homogeneously with bentonite by medicament
After be laid in above ultrafilter membrane.
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Cited By (2)
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| CN111908609A (en) * | 2020-07-09 | 2020-11-10 | 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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