CN108823097A - A method of high poly- PHA Mixed Microbes are enriched with using sulfate-containing organic wastewater acclimation sludge - Google Patents
A method of high poly- PHA Mixed Microbes are enriched with using sulfate-containing organic wastewater acclimation sludge Download PDFInfo
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Abstract
The invention discloses a kind of method for being enriched with high poly- PHA Mixed Microbes using sulfate-containing organic wastewater acclimation sludge, the method includes：The water inlet of anaerobic baffled reactor uses artificial synthesized Organic Wastewater Containing Sulfate, and sugar is cooked carbon source, and sulfate is Na2SO4、(NH4)2SO4And MgSO4·7H2The mixture of O, uses Na2CO3Solution adjusts water inlet pH value to 7 ± 0.5.Keep 33 ± 1 DEG C of temperature during reactor start-up, hydraulic detention time for 24 hours, COD/SO4 2‑Ratio 20：1；Influent COD is stepped up 3000mg/L, SO by 1000mg/L4 2‑Concentration is stepped up 150mg/L by 50mg/L；Reactor runs the starting of completion in 70 days.Reactor the 1st is controlled in operational process, the water flow upflow velocity of 2 compartments is 0.36m/h；Influent COD maintains COD4000mg/L constant after increasing to 4000mg/L by 3000mg/L；Adjust SO4 2‑Concentration gradually increases to 1000mg/L by 320mg/L, makes COD/SO4 2‑Ratio is in 12.5-4.0 range；Using Na2CO3Solution adjust reactor the 1st, 2 compartment aqueous pH value respectively to 4.7 ± 3 and 5.2 ± 3.The present invention may be implemented efficiently to produce PHA and utilization of wastewater resource, and PHA production cost is greatly reduced.
Description
Technical field
The invention belongs to environmental protection and field of biotechnology, and in particular to a kind of to select richness using sulfate-containing organic wastewater anaerobic fermentation
Collect the method with the mixed bacterial of poly-hydroxy fatty acid accumulation ability.
Background technique
Poly-hydroxy fatty acid（PHA）It is one kind polyester intracellular synthesized under unbalanced growth conditions by microorganism, PHA is not only
Possess thermoplasticity similar with petrochemicals such as polyethylene, polypropylene and mechanical property, and have biodegradability,
The features such as biocompatibility.Biological production PHA generallys use two-step method, and the first step realizes that the selection to PHA accumulation flora is enriched with,
Second step generates PHA using these microbial fermentation substrates.In two-step process, the bacterium of the high poly- PHA ability of first step enrichment
Can group be a step of most critical, using the yield for directly affecting subsequent fermentation production PHA and use cheap waste the bottom of as
Object produces PHA.That is, if the first step using sulfate-containing organic wastewater as the high poly- PHA flora of substrate rich, with this
The population that mode filters out can use sulfate-containing organic wastewater to produce PHA as raw material when second step ferments.
Summary of the invention
The present invention provides one kind using sulfate-containing organic wastewater as substrate, and the method for the high poly- PHA mixed bacterial of anaerobic fermentation enrichment is real
PHA and utilization of wastewater resource are now efficiently produced, PHA production cost is greatly reduced.
The method for being enriched with high poly- PHA Mixed Microbes using sulfate-containing organic wastewater acclimation sludge mainly includes following content：
（1）Using anaerobic baffled reactors more than two compartments, preferably five compartment anaerobic baffled reactors.
（2）Reactor water inlet uses artificial synthesized Organic Wastewater Containing Sulfate, and carbon source therein is sugar, and sulfate is
Na2SO4、(NH4)2SO4And MgSO4·7H2The mixture of O, and use Na2CO3PH value of waste water is adjusted to 7.0 or so.
（3）The start-up course of reactor：Kept for 33 ± 1 DEG C of temperature, hydraulic detention time（HRT）For for 24 hours, COD/SO4 2-Than
Value is 20：1；Influent COD is stepped up 3000mg/L, SO by 1000mg/L4 2-Concentration is stepped up 150mg/ by 50mg/L
L；After reactor is run 70 days, reactor start-up success.Preferred optimum state at this time is：Liquid in reactor the 1st, 2 compartments
PH value be respectively 5.0 and 5.9, the mean concentration of VFA is respectively 14.5mmol/L and 11.2mmol/L, and PHA accounts for mud dry weight
4.7%.
（4）The running-course control of reactor the 1st, the 2nd compartment：Whole service process hydraulic detention time（HRT）For
For 24 hours, reactor water flow upflow velocity is 0.36m/h；Influent COD increases to 4000mg/L by 3000mg/L, then maintains
COD4000mg/L is constant；SO4 2-Concentration gradually increases to 1000mg/L by 320mg/L, makes COD/SO4 2-Ratio is in 12.5-4.0 model
It encloses；Using Na2CO3The pH value that solution adjusts liquid in the 1st, 2 compartments is respectively 4.7 ± 3 and 5.2 ± 3.
（5）Seed sludge when reactor start-up can be derived from sugar refinery IC reactor, can also be derived from containing for other sources
Sulfate organic sludge, seed sludge volume number are preferably the 1/3 of reactor dischargeable capacity.
Detailed description of the invention
Fig. 1 shows reactor the 1st of the present invention, the PHA content in 2 compartment sludge.
Fig. 2 a, 2b are to work as COD/SO respectively4 2-Reactor the 1st, 2 compartment sludge TEM figure when ratio is 12.5.
Fig. 3 a, 3b are to work as COD/SO respectively4 2-Reactor the 1st, 2 compartment sludge TEM figure when ratio is 9.3.
Fig. 4 a, 4b are to work as COD/SO respectively4 2-Reactor the 1st, 2 compartment sludge TEM figure when ratio is 4.0.
Specific embodiment
The detailed description of one step is made to the present invention below with reference to embodiment, convenient for better understanding to the present invention.
1. the reactor assembly of sludge acclimatization：Using the anaerobic baffled reactor of five compartments（ABR）, total measurement (volume) 77L,
Dischargeable capacity is 61.6L.Reactor is divided into 5 compartments, and each compartment is made of upper and lower flow chamber, and upper and lower flow chamber volume ratio is 3：1.
The dog-ear that baffle plate bottom margin leads to upstream room is that upward 45 DEG C of deflector makes convenient for sending water to the center of upstream room
Muddy water is sufficiently mixed contact, maintains higher sludge concentration.The each upstream room top of reactor is mounted with three phase separator, top
Air guide port is set.Reactor controls flow of inlet water by peristaltic pump, is heated at reflux water using return pipe encapsulation reaction device outer wall and protects
Warm device controls temperature of reactor（33±1℃）.It should be noted that the enrichment of PHA Mixed Microbes of the present invention pertains only to the of ABR
1 and the 2nd compartment.
2. seed sludge and the water inlet of reactor：Seed sludge when reactor start-up is derived from the IC reactor in certain sugar refinery
（Internal Circulation）, the upstream room of each compartment, seed sludge volume are uniformly added into after nutrient solution impregnates 3 days
Number is the 1/3 of reactor dischargeable capacity.Water inlet is artificial synthesized Organic Wastewater Containing Sulfate：Using sucrose as carbon source, nitrogen source is
NH4HCO3, phosphorus source KH2PO4, COD:N:P = 200：5：1；Sulfate is Na2SO4、(NH4)2SO4And MgSO4·7H2O's is mixed
Object is closed, and uses Na2CO3Adjusting pH value is 7.0 or so.
3. the characterization of the high poly- PHA Mixed Microbes of domestication enrichment in reactor：Relate generally to the 1st and the 2nd compartment, including water phase
Middle COD removal rate, SO4 2-Removal rate, VFA concentration and component, PHA content in sludge phase, transmission electron microscope（TEM）Observation
The form of PHA and its producing strains.
4. the starting of ABR reactor：When reactor start-up, holding HRT is 30h, COD/SO4 2-Ratio is 20：1.Water inlet
COD is stepped up 3000mg/L, SO by 1000mg/L4 2-Concentration is stepped up 150mg/L by 50mg/L.Reactor operation
To after 70 days, COD removal rate is up to 89% or more, SO4 2-For removal rate up to 95% or more, total gas production is 4.1 L/h, and methane production is
3.3 L/h are discharged VFA in 4.0mmol/L hereinafter, reactor start-up success.At this point, the pH value of the 1st, 2 compartments is respectively 5.0 and
5.9, VFA mean concentration is respectively 14.5mmol/L and 11.2mmol/L, and PHA accounts for the 4.7% of mud dry weight.
5. the control parameter of the 1st, 2 compartments enrichment PHA Mixed Microbes in ABR reactor operational process：Regulation enrichment PHA is mixed
The key parameter of combined bacteria is reactor influent COD/SO4 2-Ratio and COD and SO4 2-Concentration, the 1st, the upflow velocity and pH of 2 compartments
Value.
（1）Influent COD increases to 4000mg/L by 3000mg/L first, then maintains COD4000mg/L constant, SO4 2-Concentration
1000mg/L is gradually increased to by 320mg/L, makes COD/SO4 2-Ratio is in 12.5-4.0 range.
（2）Whole service process HRT is for 24 hours, to make reactor water conservancy upflow velocity 0.36m/h.
（3）Using Na2CO3The pH value that solution adjusts the 1st, 2 compartments is respectively 4.7 ± 3 and 5.2 ± 3.
Under aforesaid operations Parameter Conditions, as influent COD/SO4 2-Ratio is respectively 12.5 (4 g COD/0.32 g
SO4 2-)、9.3 (4 g COD/0.43 g SO4 2-) and 4.0 (4 g COD/1 g SO4 2-) when, ABR reactor the 1st and the 2nd every
The COD and SO of room4 2-Removal rate, VFAs concentration and component see the table below 1.As it can be seen from table 1 the COD removal rate of the 1st compartment exists
Between 20-24%, the COD removal rate of the 2nd compartment is less than 10%；And SO4 2-It is largely removed in the 1st compartment, removal rate exists
Between 90.6-67.3%, remaining a small amount of SO4 2-Further removed in the 2nd compartment.With COD/SO4 2-Ratio is down to by 12.5
4.0, SO4 2-Removal gradually moved back by the 1st compartment to the 2nd compartment.1st, total VFA of 2 compartments（Including acetic acid, propionic acid, butyric acid
With lactic acid etc.）In 19.4-26.3 mmoll-1Between.Work as COD/SO4 2- When ratio is 12.5 and 9.3, the 2nd compartment acetic acid accounts for master
It leads, works as COD/SO4 2- When ratio is reduced to 4.0, the 2nd compartment acetic acid and butyric acid account for leading.In the present invention, the 1st, the production of 2 compartments
Sour bacterium（APB）By the sucrose inversion in water inlet be VFAs (such as acetic acid, propionic acid, butyric acid and lactic acid), these VFAs further by
Sulfate reducing bacteria（SRB）Using while sulphate reducing generate more small molecule fatty acid, show acetic acid account for it is leading.Just
Just it is that these VFAs (such as acetic acid, propionic acid, butyric acid and lactic acid) for PHA producing strains provide carbon source abundant, has promoted high poly-
The enrichment of PHA Mixed Microbes.In different influent COD/SO4 2-Under the conditions of ratio, the PHA content in the 1st, 2 compartment sludge is shown in Fig. 1.By
Fig. 1 knows, COD/SO4 2-When being 12.5, the 1st, 2 compartment PHA account for the 37.6% and 19.4% of mud dry weight respectively；COD/SO4 2-It is 9.3
When, the 1st, 2 compartment PHA account for the 26.5% and 35.0% of mud dry weight respectively；COD/SO4 2-When being 4.0, the 1st, 2 compartment PHA difference
Account for the 15.0% and 29.3% of mud dry weight.To sum up, COD/SO4 2- When ratio is 12.5 and 9.3, the PHA content in sludge is higher,
Work as COD/SO4 2- Ratio it is near 4.0 when, PHA content has downward trend；And with influent COD/SO4 2- Reduction, that is, SO of ratio4 2-
The increase of concentration, high-content of the PHA in sludge are gradually moved back by the 1st compartment to the 2nd compartment.
6. the form of PHA and PHA producing strains in sulfate-containing organic wastewater system for anaerobic treatment.
Influent COD/SO4 2-When ratio is respectively 12.5,9.3 and 4.0, the transmission electron microscope of the 1st, 2 compartment sludge
（TEM）Observation result is shown in Fig. 2, Fig. 3 and Fig. 4 respectively.By the form and somatic cells of the visible PHA producing strains of Fig. 2, Fig. 3 and Fig. 4
The graininess PHA inside largely assembled；Show the dirt that PHA producing strains individual is big, graininess PHA aggregation is more compared with the result of Fig. 1
Mud PHA content is also high.
Claims (4)
1. a kind of method for being enriched with high poly- PHA Mixed Microbes using sulfate-containing organic wastewater acclimation sludge, the method include at least：
（1）The reactor of use is the multi-compartment anaerobic baffled reactor of at least two compartments or more；
（2）The water inlet of reactor uses artificial synthesized Organic Wastewater Containing Sulfate, in composition：Using sugar as carbon source, sulfate
It is Na2SO4、(NH4)2SO4And MgSO4·7H2The mixture of O, and use Na2CO3Solution adjusts water inlet pH value to 7 ± 0.5；
（3）The start-up course of reactor：Kept for 33 ± 1 DEG C of temperature, hydraulic detention time is COD/SO for 24 hours4 2-Ratio is 20：1；
Influent COD is stepped up 3000mg/L, SO by 1000mg/L4 2-Concentration is stepped up 150mg/L by 50mg/L；Reactor
Starting is completed after operation 70 days；
（4）The running-course control of reactor the 1st, the 2nd compartment：Whole service process hydraulic detention time is reactor water for 24 hours
Stream upflow velocity is 0.36m/h；Influent COD increases to 4000mg/L by 3000mg/L, then maintains COD4000mg/L constant；It adjusts
Save SO4 2-Concentration gradually increases to 1000mg/L by 320mg/L, makes COD/SO4 2-Ratio is in 12.5-4.0 range；Using Na2CO3It is molten
Liquid adjusts the aqueous pH value of reactor the 1st, the 2nd compartment respectively to 4.7 ± 3 and 5.2 ± 3.
2. according to the method described in claim 1, for step（3）In reactor the 1st, the 2nd compartment aqueous, pH value point
It Kong Zhi not be controlled respectively in 5.0 and 5.9, the mean concentration of VFA and account for mud dry weight in 14.5mmol/L and 11.2mmol/L, PHA
4.7%.
3. according to the method described in claim 2, wherein reactor start-up when seed sludge be derived from sugar refinery IC reactor, be inoculated with
Amount of sludge is the 1/3 of reactor dischargeable capacity.
4. method according to claim 1,2 or 3, reactor therein is five compartment anaerobic baffled reactors.
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| CN104150591A (en) * | 2014-07-31 | 2014-11-19 | 桂林电子科技大学 | Method for enriching methanosarcina and methanothrix during sulfate organic wastewater treatment |
| US20160251474A1 (en) * | 2015-02-27 | 2016-09-01 | Afyren | Process for producing polyhydroxyalkanoates from precursors obtained by anaerobic fermentation from fermentable biomass |
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