CN110129401A - A kind of sewage ferments the method for wooden carbon source synthesis PHBV - Google Patents
A kind of sewage ferments the method for wooden carbon source synthesis PHBV Download PDFInfo
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- CN110129401A CN110129401A CN201910493083.5A CN201910493083A CN110129401A CN 110129401 A CN110129401 A CN 110129401A CN 201910493083 A CN201910493083 A CN 201910493083A CN 110129401 A CN110129401 A CN 110129401A
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
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- C12P7/00—Preparation of oxygen-containing organic compounds
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Abstract
It ferments the method for wooden carbon source synthesis PHBV the invention discloses a kind of sewage, comprising the following steps: 1) by being hydrolyzed, being digested with high temperature and pressure hydrothermal pretreatment method to Woody Biomass；2) with wood hydrolysis-enzymolysis liquid (reduced sugar) for carbon source, volatile fatty acid (VFAs) is generated to its anaerobic fermentation using the sewage containing activated sludge；3) using VFAs carbon source, by aerobic dynamic be fed method the sewage containing activated sludge is tamed, enrichment can synthesizing polyhydroxyalkanoateby (PHAs) microorganism；4) by adjusting the factors such as VFAs component (even number carbonic acid and odd number carbonic acid) ratio, carbon source concentration (with VFAs densimeter), pH value and C/N ratio, the PHBV with different HB/HV ratios is synthesized.The present invention realizes the efficient higher value application of waste wood resource, turns waste into wealth；Secondly, sewage tunning is converted to the environmental-friendly plastic PHAs with good utility value, this has Great significance for environmental protection and resources sustainable development.
Description
Technical field
The invention belongs to the efficient high-value-use of biomass resource, biodegradable plastics and field of solid waste treatment,
It is related to a kind of method of wooden carbon source synthesis poly 3-hydroxy butyrate 3- hydroxyl pentanoate copolymer (PHBV) of sewage fermentation.
Background technique
Plastics are indispensable materials in people's daily life, and the density having due to plastic product is small, intensity is high, again
It measures that light, corrosion-resistant, processing and forming is convenient, electrical insulation capability is excellent and the good characteristics such as cheap is developed rapidly.
However, Fossil fuel consumption and Global Environmental Problems caused by petroleum-based plastics attract wide attention, this makes people to benefit
Great interest is produced with the recyclable organism base product that natural resources synthesizes.Polyhydroxyalkanoate (PHAs) is that one kind has
The environmentally friendly high molecule plastic of biological degradability and biocompatibility, it is similar with the physical property of conventional plastic.Wherein gather
3-hydroxybutyrate ester/3- hydroxyl valerate (PHBV) structural unit is by 3-hydroxybutyrate ester (3HB) and 3- hydroxyl valerate
(3HV) composition, the two determine the brittleness and toughness of final products respectively, and the ratio by adjusting the two, which can get, has dissimilarity
The PHBV of energy, meets the needs of different, expands its application field.However the method for synthesis PHBV mainly uses pure bacterium method at present,
The organic acid using glucose or manually prepared requires sterilization in production process, production cost is almost tradition as carbon source
4-9 times of plastics limits its industrialized production.
Currently, the cheap carbon source of synthesis PHBV mainly has saccharic carbon source, methanol and organic acid carbon source.Wherein saccharic carbon source packet
Include glucose, sucrose, starch etc.；Organic acid carbon source includes acetic acid, propionic acid, butyric acid or mixed acid.It is synthesized by carbon source of methanol
When PHBV, since the content of methanol bacterium accumulation PHB is not high, PHB cost recovery is big, so that the molecular weight of PHB is smaller；And with saccharic
When for carbon source, synthesizing the content of PHBV compared with organic acid is the low of carbon source.So synthesizing PHBV with important by carbon source of mixed acid
Research significance.
Wooden Biomass resource is from a wealth of sources as a kind of reproducible resource, including agricultural stalk class waste, forest
And forest products processing waste etc., but utilization rate and the rate of recovery are low, therefore it is imperative to carry out resource utilization to it.There is research to send out
Existing, cellulose and hemicellulose hydrolyzable in Wooden Biomass resource produce the carbon of acid as sludge anaerobic at small molecule carbohydrate
Source can be used as Mixed Microbes in use, volatile fatty acid (VFAs) wide variety, including acetic acid, propionic acid, butyric acid etc. generated
Effective substrate of kind synthesis PHAs.
Therefore, the present invention generates mixed small molecule carbohydrate using the Wooden Biomass hydrolysis of recycling, with carbohydrate hydrolyzate
Activated sludge mixed bacterial, which is fed, in conjunction with anerobic sowage production VFAs synthesizes PHBV.The present invention is provided using discarded Wooden Biomass
Domestication mixed bacterial synthesizes PHBV behind source and sewage fermentation, replaces industrial expensive substrate using cheap carbon source, drops significantly
The low production cost of PHBV, to promote the marketization of PHAs product to play an important role.Meanwhile it can by changing synthesis condition
The ratio for regulating and controlling PHB and PHV in PHAs makes the PHBV of synthesis have different performances, can meet the needs of different, expand it and answer
Use range.
Summary of the invention
The present invention in order to overcome the deficiencies of the prior art, provides a kind of wooden carbon source synthesis poly 3-hydroxy butyrate of sewage fermentation
The method of 3- hydroxyl pentanoate copolymer (PHBV).
The present invention is the technical problem solved in background technique, the wooden carbon source the technical solution adopted is that a kind of sewage ferments
The method for synthesizing poly 3-hydroxy butyrate 3- hydroxyl pentanoate copolymer (PHBV), key step are as follows:
1) wood powder and ultrapure water are reacted through high-temperature high-pressure reaction kettle, is diluted after cooling, added into the hydrolyzate after dilution
Enter enzymolysis liquid, is filtered after sterilizing spare；
2) filtrate obtained by step 1) is added in sewage, nitrogen source, phosphorus source and microelement domestication, the sewage after domestication is added
Fermentation and acid obtains carbon source VFAs；
3) domestication enrichment is carried out to the mixed bacterial of polyhydroxyalkanoate in sewage (PHAs):
(1) prepare wastewater influent liquid: the VFAs obtained using step 3) as carbon source, control carbon source concentration for 1000mg/L (with
COD meter), nitrogen concentration 50mg/L, phosphorus source concentration is 10mg/L, and every 1L water inlet includes 100mg/L MgSO4It is micro with 1mg/L
Element liquid；
(2) sewage is added to tame in 2L container and be cultivated, the method for operation of acclimation period be water inlet → aeration+stirring →
Precipitating → water outlet, cycle of operation 12h；
(3) the precipitating phase time is 1h in the cycle of operation, and inflow and outflow is 500ml；
4) PHAs is synthesized:
(1) sewage after domestication is added in 500mL container, the initial MLSS of sewage is 2000mg/L, synthesis cycle
The method of operation is water inlet → aeration+stirring → precipitating → water outlet, cycle of operation 12h；
(2) prepare wastewater influent liquid: using VFAs obtained in step 3) as carbon source, carbon source concentration (by VFAs densimeter) is
800-1000mg/L, nitrogen concentration 100-300mg/L, phosphorus source concentration are 20-60mg/L, and every 1L water inlet includes 100mg/L
MgSO4With 1mg/L liquid microelement；
(3) when synthesizing PHAs, control VFAs (even number carbonic acid and odd number carbonic acid) ratio is 88:12-49:51, carbon source concentration
(with VFAs densimeter) is 800-1000mg/L, and pH value 7-9, C/N ratio is 100:3-100:7.
Optimum condition in the step 4)-(3) are as follows: VFAs component ratio be 63:37, carbon source concentration 1000mg/L, no
PH is controlled, C/N ratio is 100:5, and the PHAs yield synthesized at this time reaches maximum value.
Wood powder is that the wood wastes of industrial or agricultural pulverize and sieve acquisition in the step 1), industrial waste such as timber
The industrial wood wastes such as processing factory, Furniture Factory and waste wood etc., agricultural wastes such as stalk, bagasse etc..
Sewage is respectively the dirt of sewage and aerobic activated sludge containing anaerobic activated sludge in the step 2) and step 3)
Water.
The flora of sewage includes bacillus, Chromobacterium, Nocardia, deformation in the step 2) and step 3)
Bacillus, Shigella, Salmonella, Escherichia, pseudomonas, Blastocystis, actinomyces, bacillus flavus
One of category is a variety of.
Liquid microelement group becomes 40mg/L CaCl in the step 2) and step 3)2, 3mg/L FeCl3·3H2O,
0.24mg/L ZnSO4·7H2O, 0.3mg/L CoCl2·6H2O, 0.3mg/L MnCl2·4H2O, 0.3mg/L H3BO3,
0.06mg/L CuSO4·5H2O, 0.06mg/L KI.
The preferred NH of nitrogen source in the step 2) and step 3)4Cl, the preferred KH of phosphorus source2PO4。
Nutrient preferred proportion in water inlet in the step 3)-(2) are as follows: C:N:P=100:5:1.
Water inlet pH value is 7 ± 0.2 in the step 2), and nitrogen content abundance being passed through keeps anaerobism shape to remove oxygen
State.
Water inlet pH value is 7 ± 0.2 in the step 3)-(2), and the amount of oxygen being passed through is sufficient.
The VFAs component synthesized in the step 2) is mainly the mixed acid of acetic acid, propionic acid and butyric acid.
The PHAs ingredient synthesized in the step 4) is PHBV.
PHB the and PHV ratio synthesized in the step 4)-(3) is controllable, can get the PHBV of different performance, to expand
Its big application range.
" water inlet " refers to the initial water inlet in each period in reaction time in the step 2) and step 3), and " water outlet " refers to
Water outlet after each reaction time end cycle.
It further include continuously monitoring the content that PHAs is synthesized in the microorganism in the step 4).
The factors such as VFAs component ratio, carbon source concentration, pH value and the C/N ratio being added to the container are adjusted in the step 4),
The ratio of adjustable PHB and PHV can meet the requirement of different performance, expand its application range.
In PHAs synthesis process, measurement is sampled to sewage after the 6th end cycle, after mixed liquor is centrifuged,
It takes and is deposited in -50 DEG C of freeze-dryings, the yield and ingredient of gas Chromatographic Determination PHAs are used through being esterified.
The yield of PHAs and composition measurement specific steps are as follows:
It draws 8mL sewage and is centrifuged 10min at 10000rpm, remove portion's thallus and be freeze-dried under the conditions of -50 DEG C for 24 hours,
It weighs and records after drying.
The preparation of PHA esterification reaction solution: accurately weighing 1.0g benzoic acid (analysis is pure), mixed with 100mL methanol (chromatographically pure)
It closes, adds the 10mL concentrated sulfuric acid (excellent pure grade), esterification reaction solution is made to being completely dissolved in ultrasonic 5min.
Sample preparation procedure: 2mL chloroform, 1mL esterification is added in esterification pipe in the thallus after weighing a certain amount of drying
Liquid, 100 DEG C of reaction 4h are added 1mL high purity water, acutely shake, stand in 4 DEG C after being cooled to room temperature.When lower layer's organic phase is clarified
When, lower layer's organic phase 1.0-1.5mL, which is drawn, with 1mL injection needle is transferred in gas chromatographic sample introduction bottle, it can sample introduction.
Chromatographic condition:
Injection port: 200 DEG C of temperature, carrier gas N2, pressure 10.33psi, flow 40mL/min, split ratio 25:1.
Post case: temperature programming, keeps 2min by 70 DEG C of initial temperature, 140 DEG C then are raised to the speed of 7 DEG C/min, is protected
1min is held, is increased to 220 DEG C again with the speed of 10 DEG C/min, keeps 1min.
Detector: fid detector, 250 DEG C of temperature, H2Flow 40mL/min, air mass flow 450mL/min.
The PHAs ingredient of synthesis is PHBV.
The composition and content of PHAs is calculated according to the ratio between mark product and the interior peak area for being marked on chromatogram.By to difference
The comparative analysis of experimental group PHAs yield and dry cell weight can determine the optimum synthesis condition of mixed bacteria.
Compared with prior art, the present invention the performance of its distinguishing feature is as follows:
1, the present invention is using wooden hydrolysate and the volatile fatty acid of sewage fermenting and producing as the carbon source of mixed bacterial, not only
Discarded wooden resource is rationally utilized, and sewage is made to realize the developing direction of minimizing and recycling, realizes PHAs original
The low cost of material；
2, PHAs is synthesized using sewage, wherein being PHAs synthesis bacterium containing a large amount of mixed bacterial, does not need stringent go out
Bacterium equipment and gnotobasis, considerably reduce manufacturing technique requirent and product cost；
3, compared with studying the saccharic carbon source used mostly at present, PHAs yield can be improved in the present invention, and manually prepares
Mixed organic acid carbon source is compared, and the present invention uses renewable resource cheap carbon source abundant；
4, by controlling different synthesis conditions, it can get the PHAs of different PHB and PHV components, be PHAs product multiplicity
Metaplasia, which produces, provides simple controllable technology pattern.
5, during mixed bacterial synthesis PHAs, by adjusting VFAs component ratio and C/N ratio, it can be improved PHAs's
Yield, maximum output condition are that VFAs component ratio is 63:37, and carbon source concentration (with VFAs densimeter) 1000mg/L does not adjust pH,
C/N ratio is 100:5, and maximum output reaches 1431.5mg/L, and PHAs accounts for dry cell weight 24.54% at this time；In C/N than unbalanced or control
Under the conditions of pH processed, it is unfavorable for the increase of its yield.
Detailed description of the invention
Fig. 1 is total figure: (two peaks before 6.89min are HB and HV).
Fig. 2 is HB component.
Fig. 3 is HV component.
Specific embodiment
The present invention is described in detail below in conjunction with the drawings and specific embodiments.The embodiment of the present invention is in order to more
So that those skilled in the art is more fully understood the present invention well, any limitation is not made to the present invention.
1) 40-80 mesh wood powder and ultrapure water are uniformly mixed with 1:4 ratio and are placed in 200 DEG C of high-temperature high-pressure reaction kettles 40 points
Clock is diluted to 1L after cooling and adjusts pH to 4.8；45 DEG C of magnetic force of 3mL CTec2 enzymolysis liquid are added in hydrolyzate after dilution to stir
48h is mixed, is filtered after sterilizing spare.
2) filtrate obtained by step 1) is added in sewage, nitrogen source, phosphorus source and microelement is added, keep 32 DEG C or so progress
Domestication in 30 days by a definite date；Sewage after domestication adds the fermentation and acid in 500mL container, and control carbon source concentration is 6g/L, pH value
It is 8, fermentation temperature is 35 DEG C, fermentation time 12h, adjusts a pH to initial value every 2h in acid process.
3) domestication enrichment is carried out to the mixed bacterial of polyhydroxyalkanoate in sewage (PHAs):
(1) prepare wastewater influent liquid: for the VFAs obtained using step 3) as carbon source, control carbon source concentration is 1000mg/L, nitrogen
Source concentration is 50mg/L, and phosphorus source concentration is 10mg/L, and every 1L water inlet includes 100mg/L MgSO4With 1mg/L liquid microelement；
(2) sewage is added to tame in 2L container and be cultivated, the method for operation of acclimation period be water inlet → aeration+stirring →
Precipitating → water outlet, cycle of operation 12h；
(3) the precipitating phase time is 1h in the cycle of operation, and inflow and outflow is 500ml；
4) PHAs is synthesized:
(1) sewage after domestication is added in 500mL container, the initial MLSS of sewage is 2000mg/L, synthesis cycle
The method of operation is water inlet → aeration+stirring → precipitating → water outlet, cycle of operation 12h；
(2) prepare wastewater influent liquid: using VFAs obtained in step 3) as carbon source, carbon source concentration (by VFAs densimeter) is
800-1000mg/L, nitrogen concentration 100-300mg/L, phosphorus source concentration are 20-60mg/L, and every 1L water inlet includes 100mg/L
MgSO4With 1mg/L liquid microelement；
(3) when synthesizing PHAs, control VFAs (even number carbonic acid and odd number carbonic acid) ratio is 88:12-49:51, carbon source concentration
(pressing VFAs densimeter) is 800-1000mg/L, and pH value 7-9, C/N ratio is 100:3-100:7.Above-mentioned steps 2) and step 3)
The middle preferred NH of nitrogen source4Cl, the preferred KH of phosphorus source2PO4。
Each embodiment technological condition of table 1 and result
Embodiment 1
Sewage after taming is added in 500mL container, the initial MLSS of sewage is 2000mg/L, accumulates the operation side in period
Formula is water inlet → aeration+stirring → precipitating → water outlet, cycle of operation 12h.Water inlet is VFAs, and carbon source concentration is (with VFAs concentration
Meter) it is 900mg/L, nitrogen concentration 200mg/L, phosphorus source concentration is 40mg/L, and every 1L water inlet includes 100mg/L MgSO4With
1mg/L liquid microelement, VFAs component (even number carbonic acid and odd number carbonic acid) ratio are 88:12, and C/N ratio is 100:5.Synthesis process
In do not adjust pH.Measurement is sampled to activated sludge in the 6th cultivation cycle.After mixed liquor is centrifuged, takes and be deposited in -50 DEG C
Freeze-drying, is esterified, and the yield and ingredient of gas chromatography measurement PHAs are used.
Embodiment 2
Sewage after taming is added in 500mL container, the initial MLSS of sewage is 2000mg/L, accumulates the operation side in period
Formula is water inlet → aeration+stirring → precipitating → water outlet, cycle of operation 12h.Water inlet is VFAs, and carbon source concentration is (with VFAs concentration
Meter) it is 900mg/L, nitrogen concentration 200mg/L, phosphorus source concentration is 40mg/L, and every 1L water inlet includes 100mg/L MgSO4With
1mg/L liquid microelement, VFAs component (even number carbonic acid and odd number carbonic acid) ratio are 63:37, and C/N ratio is 100:5.Synthesis process
In do not adjust pH.Measurement is sampled to activated sludge in the 6th cultivation cycle.After mixed liquor is centrifuged, takes and be deposited in -50 DEG C
Freeze-drying, is esterified, and the yield and ingredient of gas chromatography measurement PHAs are used.
Embodiment 3
Sewage after taming is added in 500mL container, the initial MLSS of sewage is 2000mg/L, accumulates the operation side in period
Formula is water inlet → aeration+stirring → precipitating → water outlet, cycle of operation 12h.Water inlet is VFAs, and carbon source concentration is (with VFAs concentration
Meter) it is 900mg/L, nitrogen concentration 200mg/L, phosphorus source concentration is 40mg/L, and every 1L water inlet includes 100mg/L MgSO4With
1mg/L liquid microelement, VFAs component (even number carbonic acid and odd number carbonic acid) ratio are 49:51, and C/N ratio is 100:5.Synthesis process
In do not adjust pH.Measurement is sampled to activated sludge in the 6th cultivation cycle.After mixed liquor is centrifuged, takes and be deposited in -50 DEG C
Freeze-drying, is esterified, and the yield and ingredient of gas chromatography measurement PHAs are used.
Embodiment 4
Sewage after taming is added in 500mL container, the initial MLSS of sewage is 2000mg/L, accumulates the operation side in period
Formula is water inlet → aeration+stirring → precipitating → water outlet, cycle of operation 12h.Water inlet is VFAs, and carbon source concentration is (with VFAs concentration
Meter) it is 900mg/L, nitrogen concentration 200mg/L, phosphorus source concentration is 40mg/L, and every 1L water inlet includes 100mg/L MgSO4With
1mg/L liquid microelement, VFAs component (even number carbonic acid and odd number carbonic acid) ratio are 63:37, and C/N ratio is 100:5.Each period
The pH that mixed liquor is adjusted after water inlet is 7.Measurement is sampled to activated sludge in the 6th cultivation cycle.After mixed liquor is centrifuged,
It takes and is deposited in -50 DEG C of freeze-dryings, be esterified, use the yield and ingredient of gas chromatography measurement PHAs.
Embodiment 5
Sewage after taming is added in 500mL container, the initial MLSS of sewage is 2000mg/L, accumulates the operation side in period
Formula is water inlet → aeration+stirring → precipitating → water outlet, cycle of operation 12h.Water inlet is VFAs, and carbon source concentration is (with VFAs concentration
Meter) it is 900mg/L, nitrogen concentration 200mg/L, phosphorus source concentration is 40mg/L, and every 1L water inlet includes 100mg/L MgSO4With
1mg/L liquid microelement, VFAs component (even number carbonic acid and odd number carbonic acid) ratio are 63:37, and C/N ratio is 100:5.Each period
The pH that mixed liquor is adjusted after water inlet is 9.Measurement is sampled to activated sludge in the 6th cultivation cycle.After mixed liquor is centrifuged,
It takes and is deposited in -50 DEG C of freeze-dryings, be esterified, use the yield and ingredient of gas chromatography measurement PHAs.
Embodiment 6
Sewage after taming is added in 500mL container, the initial MLSS of sewage is 2000mg/L, accumulates the operation side in period
Formula is water inlet → aeration+stirring → precipitating → water outlet, cycle of operation 12h.Water inlet is VFAs, and carbon source concentration is (with VFAs concentration
Meter) it is 800mg/L, nitrogen concentration 150mg/L, phosphorus source concentration is 30mg/L, and every 1L water inlet includes 100mg/L MgSO4With
1mg/L liquid microelement, VFAs component (even number carbonic acid and odd number carbonic acid) ratio are 63:37, and C/N ratio is 100:5.Synthesis process
In do not adjust pH.Measurement is sampled to activated sludge in the 6th cultivation cycle.After mixed liquor is centrifuged, takes and be deposited in -50 DEG C
Freeze-drying, is esterified, and the yield and ingredient of gas chromatography measurement PHAs are used.
Embodiment 7
Sewage after taming is added in 500mL container, the initial MLSS of sewage is 2000mg/L, accumulates the operation side in period
Formula is water inlet → aeration+stirring → precipitating → water outlet, cycle of operation 12h.Water inlet is VFAs, and carbon source concentration is (with VFAs concentration
Meter) it is 1000mg/L, nitrogen concentration 300mg/L, phosphorus source concentration is 60mg/L, and every 1L water inlet includes 100mg/L MgSO4With
1mg/L liquid microelement, VFAs component (even number carbonic acid and odd number carbonic acid) ratio are 63:37, and C/N ratio is 100:5.Synthesis process
In do not adjust pH.Measurement is sampled to activated sludge in the 6th cultivation cycle.After mixed liquor is centrifuged, takes and be deposited in -50 DEG C
Freeze-drying, is esterified, and the yield and ingredient of gas chromatography measurement PHAs are used.
Embodiment 8
Sewage after taming is added in 500mL container, the initial MLSS of sewage is 2000mg/L, accumulates the operation side in period
Formula is water inlet → aeration+stirring → precipitating → water outlet, cycle of operation 12h.Water inlet is VFAs, and carbon source concentration is (with VFAs concentration
Meter) it is 900mg/L, nitrogen concentration 200mg/L, phosphorus source concentration is 40mg/L, and every 1L water inlet includes 100mg/L MgSO4With
1mg/L liquid microelement, VFAs component (even number carbonic acid and odd number carbonic acid) ratio are 63:37, and C/N ratio is 100:3.Synthesis process
In do not adjust pH.Measurement is sampled to activated sludge in the 6th cultivation cycle.After mixed liquor is centrifuged, takes and be deposited in -50 DEG C
Freeze-drying, is esterified, and the yield and ingredient of gas chromatography measurement PHAs are used.
Embodiment 9
Sewage after taming is added in 500mL container, the initial MLSS of sewage is 2000mg/L, accumulates the operation side in period
Formula is water inlet → aeration+stirring → precipitating → water outlet, cycle of operation 12h.Water inlet is VFAs, and carbon source concentration is (with VFAs concentration
Meter) it is 900mg/L, nitrogen concentration 200mg/L, phosphorus source concentration is 40mg/L, and every 1L water inlet includes 100mg/L MgSO4With
1mg/L liquid microelement, VFAs component (even number carbonic acid and odd number carbonic acid) ratio are 63:37, and C/N ratio is 100:6.Synthesis process
In do not adjust pH.Measurement is sampled to activated sludge in the 6th cultivation cycle.After mixed liquor is centrifuged, takes and be deposited in -50 DEG C
Freeze-drying, is esterified, and the yield and ingredient of gas chromatography measurement PHAs are used.
As shown in Figure 1, the mass spectrogram of PHAs, two peaks before 6.89min are HB and HV.Fig. 2 is HB component, and Fig. 3 is HV
Component.
Although above in conjunction with attached drawing, invention has been described, and the invention is not limited to above-mentioned specific implementations
Mode, the above mentioned embodiment is only schematical, rather than restrictive, and those skilled in the art are at this
Under the enlightenment of invention, without deviating from the spirit of the invention, many variations can also be made, these belong to of the invention
Within protection.
Claims (9)
- A kind of method of wooden carbon source synthesis PHBV 1. sewage ferments, which is characterized in that key step is as follows:1) wood powder and ultrapure water are reacted through high-temperature high-pressure reaction kettle, is diluted after cooling, enzyme is added into the hydrolyzate after dilution Liquid is solved, is filtered after sterilizing spare；2) filtrate obtained by step 1) is added in sewage, nitrogen source, phosphorus source and microelement domestication, the sewage fermentation after domestication is added Acid is produced, carbon source VFAs is obtained；3) domestication enrichment is carried out to the mixed bacterial of polyhydroxyalkanoate in sewage:(1) wastewater influent liquid: the carbon source VFAs obtained with step 2) is prepared, control carbon source concentration is 1000mg/L (in terms of COD), Nitrogen concentration is 50mg/L, and phosphorus source concentration is 10mg/L, and every 1L water inlet includes 100mg/L MgSO4With 1mg/L liquid microelement；(2) sewage is added to tame in 2L container and is cultivated, the method for operation of acclimation period is water inlet → aeration+stirring → precipitating → water outlet, cycle of operation 12h；(3) the precipitating phase time is 1h in the cycle of operation, and inflow and outflow is 500ml；4) PHAs is synthesized:(1) sewage after domestication is added in 500mL container, the initial MLSS of sewage is 2000mg/L, the operation of synthesis cycle Mode is water inlet → aeration+stirring → precipitating → water outlet, cycle of operation 12h；(2) prepare wastewater influent liquid: using VFAs obtained in step 3) as carbon source, carbon source concentration (pressing VFAs densimeter) is 800- 1000mg/L, nitrogen concentration 100-300mg/L, phosphorus source concentration are 20-60mg/L, and every 1L water inlet includes 100mg/L MgSO4 With 1mg/L liquid microelement；(3) synthesize PHAs when, control VFAs in the ratio of even number carbonic acid and odd number carbonic acid be 88:12-49:51, carbon source concentration (with VFAs densimeter) it is 800-1000mg/L, pH value 7-9, C/N ratio is 100:3-100:7.
- The method of wooden carbon source synthesis PHBV 2. a kind of sewage according to claim 1 ferments, which is characterized in that the step It is rapid 1) in wood powder be that the wood wastes of industrial or agricultural pulverize and sieve acquisition.
- The method of wooden carbon source synthesis PHBV 3. a kind of sewage according to claim 1 ferments, which is characterized in that the step It is rapid that 2) and in step 3) sewage is respectively the sewage of sewage and aerobic activated sludge containing anaerobic activated sludge.
- The method of wooden carbon source synthesis PHBV 4. a kind of sewage according to claim 1 ferments, which is characterized in that described The flora of sewage includes bacillus, Chromobacterium, Nocardia, Proteus, will he in step 2) and step 3) One of Bordetella, Salmonella, Escherichia, pseudomonas, Blastocystis, actinomyces, Flavobacterium Or it is a variety of.
- The method of wooden carbon source synthesis PHBV 5. a kind of sewage according to claim 1 ferments, which is characterized in that described Liquid microelement group becomes 40mg/L CaCl in step 2) and step 3)2, 3mg/L FeCl3·3H2O, 0.24mg/L ZnSO4·7H2O, 0.3mg/L CoCl2·6H2O, 0.3mg/L MnCl2·4H2O, 0.3mg/L H3BO3, 0.06mg/L CuSO4·5H2O, 0.06mg/L KI.
- The method of wooden carbon source synthesis PHBV 6. a kind of sewage according to claim 1 ferments, which is characterized in that described The preferred NH of nitrogen source in step 2) and step 3)4Cl, the preferred KH of phosphorus source2PO4。
- The method of wooden carbon source synthesis PHBV 7. a kind of sewage according to claim 1 ferments, which is characterized in that described Nutrient preferred proportion in water inlet in step 3)-(2) are as follows: C:N:P=100:5:1.
- The method of wooden carbon source synthesis PHBV 8. a kind of sewage according to claim 1 ferments, which is characterized in that described Water inlet pH value is 7 ± 0.2 in step 2), and nitrogen content abundance being passed through keeps anaerobic state, the step to remove oxygen 3) water inlet pH value is 7 ± 0.2 in-(2), and the amount of oxygen being passed through is sufficient.
- The method of wooden carbon source synthesis PHBV 9. a kind of sewage according to claim 1 ferments, which is characterized in that described Synthesis VFAs component is mainly the mixed acid of acetic acid, propionic acid and butyric acid in step 2).
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