CN107760605A - A kind of research method that tofu wastewater is purified using microdisk electrode - Google Patents
A kind of research method that tofu wastewater is purified using microdisk electrode Download PDFInfo
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Abstract
The present invention discloses a kind of research method that tofu wastewater is purified using microdisk electrode, and key step includes：1) microdisk electrode；2) tofu wastewater is handled；3) batch cultivation is tested；4) fed-batch culture experiment.Microalgae C.pyrenoidosa of the present invention average biomass productivity ratio reaches 0.64gL‑1d‑1, average lipid content is 37.00 ± 9.34%, and lipid production rate is up to 0.40gL‑1d‑1, higher than the lipid production rate of batch cultivation, so as to learn that fed-batch culture is best suitable for tofu wastewater culture C.pyrenoidosa, the double action with synchronous water reuse and with the production of cost-benefit biomass.
Description
Technical field
The present invention relates to microalgae biologic treating technique field, and in particular to a kind of to purify tofu wastewater using microdisk electrode
Research method.
Background technology
One ton of soybean processing is produced into about 7-10 ton waste water into bean curd, the COD (COD) of waste water is 10-20g/L.
Research to tofu wastewater (SPW) processing all concentrates on anaerobic filter AF (anaerobic filters), and upflow type anaerobic is dirty
The technique such as mud fluid bed (UASB), sequencing batch reactor (SBR).Although Anaerobic Treatment is suitable for the high-concentration waste water of energy regenerating
Processing, but method has several deficiencies, and Anaerobic Treatment can not remove biological nitrogen and phosphorus, and need constantly regulate processing procedure
The basicity of middle liquid.Aerobic needs to meet that the requirement of COD bioconversions, denitrogenation dephosphorizing target and sludge condensation will
Ask, and process not only wastes organic carbon and nutrients resource, can also produce CO2And sludge.One optimal processing procedure should
The pollutant of the production of useful organisms, while organics removal, nitrogen and phosphorus can be utilized.
Tofu wastewater (SPW) includes monose, oligosaccharide, vitamin, organic acid, amino acid, lipid, lactalbumin, different Huang
Ketone, saponin, P, Ca, Fe and other nutriments[1-2].Some researchs, which have been inquired into, utilizes membrane technology recovery oligosaccharides, protein
Or isoflavones, but this method can only reclaim some compositions in waste water, and leave the waste water that may need further to handle.
Some researchers explore the microdisk electrode mode being combined with wastewater treatment.These microalgaes can not only remove
Pollutant, and caused lipid, can be converted to biodiesel[3-5].Because SPW contains obvious available nutriment
Typically without poisonous and harmful, it is impossible to suppress micro algae growth, the grease micro-algae culture medium in such medium can reduce microalgae life
The cost of thing diesel oil.
Microalgae has growth rate high, strong adaptability is (i.e. for other can be converted into the agriculture plant of biomass product
Make still to grow under conditions of extremely arduous), and will not be with other product competitions, land occupation area is few, oil-producing
Measure the advantages of high and trigger people's concern.In bioreactor, carbon dioxide can be fixed using solar energy by microalgae and made
For carbon source, machine compound (i.e. glucose and acetate) is changed into energy with autotrophy heterotrophism or mixed culturing method and is used for biology
Amount growth.Mixed culture operation has several advantages, including delustring demand, higher micro algae growth, cost-benefit biomass
Harvest, and degradation of substrates, under mixing condition, microalgae can be by breathing and photosynthesis, while absorbs inorganic and organic bottom
Thing, this is autotrophy and heterotrophic growth sum.
Micro algae growth and wastewater treatment are linked together, that is, make use of the nutriment enriched in tofu wastewater, and
Biomass productivity is promoted, has reached the effect of double optimization.The training method of microalgae has 4 kinds in bioreactor,
Respectively batch cultivation, fed-batch culture, Semi-continuous cultivation, continuous culture.Batch cultivation refers to disposable in the reactor
After putting into nutrient solution and inoculation microalgae liquid and aseptically cultivating certain time, disposable nutrient solution of releasing is post-processed
Cultural method.Fed-batch culture refers to a certain amount of nutrient solution loading reactor is aseptically first inoculated with into microalgae
Liquid, to be cultivated, cell is constantly grown, product is constantly formed, and with the continuous consumption of nutriment during this, as being
New nutritional ingredient is supplemented in system is metabolized cell further growth, and product is taken out after whole culture terminates.
The content of the invention
The defects of it is an object of the invention to overcome above-mentioned background technology to exist, there is provided one kind utilizes microdisk electrode purification beans
The research method of rotten waste water.
Technical scheme:A kind of research method that tofu wastewater is purified using microdisk electrode, is comprised the following steps：
1), microdisk electrode：C.pyrenoidosa is inoculated with sterile conditions, 100mL is used in 250mL conical flasks
Autoclaved SE culture mediums are cultivated C.pyrenoidosa, are subsequently placed in light incubator and are cultivated；
Microdisk electrode condition：Light intensity=25-30 μm of ol photon m-2S-1, Light To Dark Ratio=14：10th, temperature=25 ± 1
DEG C, 24h is cultivated, and batch (-type) shakes in the training period；
2), tofu wastewater pre-processes：Counterbalance cell SW1 and anaerobic hydrolysis reactor SW2 tofu wastewater is collected respectively, in height
Centrifuged in fast refrigerated centrifuge, adjust the pH 6-7 of supernatant, in 100-121 DEG C of autoclaving 20 minutes, and in 1-4 DEG C of guarantor
Deposit, obtain SW1 and SW2 wastewater mediums；
3), batch cultivation is tested：Using 3 500ml conical flasks as bioreactor P1, P2 and P3, add respectively
Enter SW1, SW2 wastewater medium and SE culture mediums, C.pyrenoidosa inoculation algae solution trainings are then added into P1, P2 and P3
Support, mixed solution with magnetic stirring apparatus；
In light intensity=27 μm ol photon m-2S-1, Light To Dark Ratio=14：10th, cultivated at temperature=25 ± 1 DEG C
120h；
4), fed-batch culture experiment：Using 1 500 milliliters of conical flask as bioreactor, add
100mL C.pyrenoidosa inoculums and 50ml SW2 culture mediums, are mixed mixed solution with magnetic stirring apparatus；In light intensity
=27 μm of ol photon m-2S-1, Light To Dark Ratio=14：10th, culture 24h is carried out at temperature=25 ± 1 DEG C；
In ensuing four days, cultivated daily in the same time to the SW2 culture mediums of microalgae addition various dose,
The dosage of four days is respectively 75mL, 100mL, 0mL, 50mL.
Step 2) the high speed refrigerated centrifuge is centrifuged 10 minutes with 10355rpm rotating speed.
The step 3) adds 100mL C.pyrenoidosa inoculation algae solutions and 100mL SW1 waste water cultures in P1
The distilled water of base and 70mL；
100mL C.pyrenoidosa inoculation algae solutions and 100mL SW2 wastewater mediums and 70mL are added in P2
Distilled water；In P3,170ml SE culture mediums and 100ml inoculum C are added.
Compared with prior art, the present invention has the advantage that：
1), result of study of the present invention shows：After 120h fed-batch culture, microalgae removes 77.8 respectively ±
5.7% soluble chemical oxygen demand (SCODcr), 88.8 ± 1% total nitrogen (TN), 89.1 ± 0.6% ammonium nitrogen (NH4 +-
N) and 70.3 ± 11.4% total phosphorus (TP).C.pyrenoidosa average biomass productivity ratio reaches 0.64gL simultaneously-1d-1, average lipid content is 37.00 ± 9.34%, and lipid production rate is up to 0.40gL-1d-1, higher than the lipid production of batch cultivation
Rate, so as to learn that fed-batch culture is best suitable for tofu wastewater culture C.pyrenoidosa.
2), together with the present invention couples wastewater treatment with biomass production, it is not necessary to it is extra to supplement the nutrients, tool can be turned into
There are synchronous water reuse and the dual strategy with the production of cost-benefit biomass.
Brief description of the drawings
Fig. 1 is C.pyrenoidosa batch and fed-batch culture growth schematic diagram in tofu wastewater.
Fig. 2 is SCODCr schematic diagrames in C.pyrenoidosa fed-batch culture processed soybeans processing waste waters：
SW1 represents the supernatant of counterbalance cell centrifugation tofu wastewater；
SW2 represents the supernatant of the centrifugation tofu wastewater from anaerobic hydrolysis reactor；
R1 represents the SCODCr clearances of SW2 culture medium fed-batch cultures；
R2 represents the SCODCr clearances of SW2 culture medium batch cultivations；
R3 represents the SCODCr clearances of SW1 culture medium batch cultivations.
Fig. 3 is TN, TP clearance schematic diagram in C.pyrenoidosa fed-batch cultures：
(a) figure is changed over time for TN；
(b) figure is changed over time for TP.
Fig. 4 is VFAs, SC clearance schematic diagram in C.pyrenoidosa fed-batch cultures.
Embodiment
Below by specific embodiments and the drawings, the present invention is further illustrated.Embodiments of the invention are in order to more
Those skilled in the art is more fully understood the present invention well, any limitation is not made to the present invention.
A kind of research method that tofu wastewater is purified using microdisk electrode of the present invention, is comprised the following steps：
1), the culture of microalgae：C.pyrenoidosa is inoculated with sterile conditions, used in 250mL conical flasks
The autoclaved SE culture mediums of 100mL are cultivated C.pyrenoidosa, are subsequently placed in light incubator and are cultivated, SE
The composition of culture medium：NaNO3 0.25(g/L)、K2HPO4.3H2O 0.075(g/L)、MgSO4.7H2O 0.075(g/L)、
CaCl2.2H2O 0.025(g/L)、KH2PO4 0.175(g/L)、NaCL0.025(g/L)、Soilextract 40(mL/L)、
FeCl3·6H2O 0.005(g/L)、Fe-EDTA 1(mL/L)、A5solution 1(mL/L)、distilled water 958
(mL/L)；
Wherein A5solution collocation method is：H is dissolved in 100ml deionized water3BO3 286mg、ZnSO4·
7H2O 22mg、MnCl2·4H2O 181mg、CuSO4·5H2O7.9mg、(NH4)6Mo7O24·4H2O 3.9mg；Fe-EDTA's matches somebody with somebody
Put method：A liquid Na2EDTA 1g, distilled water 50ml, B liquid FeCl3·6H2O81mg、HCl(0.1N)(0.1mol/
L) 50ml, A liquid, B liquid is respectively configured, after being dissolved after being sufficiently stirred, by A liquid, B liquid is mixed evenly.
C.pyrenoidosa microalgaes are a kind of green algas, the condition of culture of microalgae：Light intensity=27 μm ol photon m-2S-1、
Light To Dark Ratio=14：10th, temperature=25 ± 1 DEG C, and batch (-type) shakes in the training period；
2), tofu wastewater pre-processes：In order to obtain C.pyrenoidosa wastewater medium, counterbalance cell is collected respectively
(SW1) and anaerobic hydrolysis reactor (SW2) tofu wastewater, COD=13215mg/L, TN=267.1mg/L of tofu wastewater,
NH4 +- N=52.1mg/L, TP=56.3mg/L are Na=1387mg/L, Cu=0.55mg/L, Mg=also including content
173.5mg/L, Mn=0.21mg/L salt, contained nutriment is higher than SE culture medium respective concentrations, in high speed freezing centrifuge
In with 10,355 rotating speed, centrifuge 10 minutes, take supernatant, adjust the pH 6.5 of supernatant, divide in 121 DEG C of autoclavings 20
Clock, and in 1 DEG C of preservation；Centrifuged supernatant from counterbalance cell (SW1) and anaerobic hydrolysis reactor (SW2) contains 10160 respectively
With 8087mg/L COD；
3), batch cultivation is tested：Using three 500ml conical flasks as bioreactor (P1, P2, P3), respectively
SW1, SW2 wastewater medium and SE culture mediums are added, C.pyrenoidosa inoculation algae solutions are then added into P1, P2, P3,
Solution is mixed with magnetic stirring apparatus；
Respectively into P1 and P2 add 100mL C.pyrenoidosa inoculation algae solution, then respectively add 100mL SW1,
SW2 wastewater mediums, finally add 70mL distilled water；
In P3, the inoculum C.pyrenoidosa inoculations algae solution of the SE culture mediums and 100ml that add 170ml compares；
In light intensity=27 μm ol photon m-2S-1, Light To Dark Ratio=14：10th, cultivated at temperature=25 ± 1 DEG C
120h；
4), fed-batch culture experiment：One 500 milliliters of conical flask adds 100mL's as bioreactor
The SW2 culture mediums of C.pyrenoidosa inoculums and 50ml, mixed solution is mixed with magnetic stirring apparatus；In the μ of light intensity=27
mol photon m-2S-1, Light To Dark Ratio=14：10th, culture 24h is carried out at temperature=25 ± 1 DEG C；In ensuing four days, often
It the same time to microalgae addition various dose SW2 cultivated, four days addition microalgae dosage be respectively 75mL,
100mL、0mL、50mL。
Data Detection is carried out below：
(1), biomass：
Collect algae suspension 1mL measure C.pyrenoidosa biomass, under conditions of being 5074 turns, 4 DEG C in rotating speed from
The heart 10 minutes, by particle drying to constant weight at 65 DEG C.Each three parts of sample, blank sample is also with identical method
Reason, the sample without algae is as control.
Above-mentioned batch cultivation and feed batch fermentation, the biomass schematic diagram of 120 hours, as shown in Figure 1.
Analysis chart 1, it can be seen that the biomass of 120h C.pyrenoidosa microalgaes is cultivated in SW1 and SW2 culture mediums
Respectively 2.09 ± 0.13 and 2.15 ± 0.21g/L.On the contrary, the life of the C.pyrenoidosa microalgaes using SE medium cultures
Object amount is only up to 0.66 ± 0.03g/L.
(2), water quality detection：
To measure water quality, microdisk electrode centrifuges (10355 turns, 4 DEG C, 10 minutes), and supernatant is fine by 0.45 μm of acetic acid
Tie up filter membrane.Filtrate measure analysis soluble chemical oxygen demand (SCODcr), total nitrogen (TN), total phosphorus (TP), COD according to
GB 11914-89 standards detect (State Environmental Protection Administration China, 2002)[6].Detected using elemental analyser
Carbon (C), hydrogen (H), oxygen (O), the content of nitrogen (N) in C.pyrenoidosa.By the use of glucose as standard, pass through phend-sulphuric acid
Measure carbohydrate[7]。
As shown in Fig. 2 in fed-batch incubation, after adding SPW, SCODCr concentration in fed-batch incubation
High value is increased to, but concentration quickly falls to a relatively low concentration, because C.pyrenoidosa absorption and having
The consumption of machine compound.In batches with the SCOD of fed-batch cultureCrRemoval efficiency gradually increases.In fed-batch experiments, use
The SCOD of the microalgae of SW2 culturesCrReach 77.8 ± 5.7%.
As shown in figure 3, after SPW waste water is added, TN and TP progressively increase to a higher concentration, but because
Absorption and digestion of the microalgae to nutrients make concentration quickly reduce.Terminate in the mixotrophism phase, TN and TP concentration reduce respectively
To 19.1 ± 0.7 and 11.7 ± 4.7mg L-1.It is relative, TN and TP clearance is respectively 88.8 ± 1.0% and 70.3 ±
11.4%.
(3) Volatile fatty acid contents (VFAs) in biomass, are analyzed, filtrate is collected in 1.5 milliliters of gas-chromatographies
(GC) bottle, it is about 4 to add 3% phosphorus acid for adjusting pH.Using equipped with flame ionization detector (FID) and analytical column
CPWAX52CB (30m × 0.53mm × 1 μm) gas chromatograph determines VFA (C2-C5) concentration, sample size 1mL.Note
Emitter and FID temperature are respectively 200 and 220 DEG C.Using nitrogen as carrier gas, flow velocity is 50 ml/mins.GC column ovens
Program setting is 110 DEG C, starts to be kept for 2 minutes, then rises to 220 DEG C with 10 DEG C/min of speed, and kept for 2 points at 220 DEG C
Clock.
As shown in Figure 4, it is shown that during fed-batch culture in pretreated SPW VFAs removal effect.By
After 120h culture, VFA in SPWSConcentration drop to 162.4 ± 68.6mgL-1, corresponding clearance is 96.7 ± 1.4%, beans
Carbohydrate (SC) concentration is 59.8 ± 12.1mg L in rotten waste water-1, corresponding clearance is 41.1 ± 1.9%.
In fed-batch culture, microalgae C.pyrenoidosa can effectively remove the nutrients of tofu wastewater, and produce
Raw very big biomass productivity.
It should be appreciated that embodiment and example discussed herein simply to illustrate that, to those skilled in the art
For, it can be improved or be converted, and all these modifications and variations should all belong to the protection of appended claims of the present invention
Scope.
Pertinent literature：
[1].Lopes Barbosa,A.C.,Lajolo,F.M.,Genovese,M.I.,2006.Influence of
temperature,pH and ionic strength on the production of isoflavone-rich soy
protein isolates.Food Chem.98,757–766.
[2].Tang,C.-H.,Ma,C.-Y.,2009.Effect of high pressure treatment on
aggregation and structural properties of soy protein isolate.LWT–Food
Sci.Technol.42,606–611.
[3].Li,Q.,Du,W.,Liu,D.,2008a.Perspectives of microbial oils for
biodiesel production.Appl.Microbiol.Biotechnol.80,749–756.
[4].Bhatnagar,A.,Bhatnagar,M.,Chinnasamy,S.,Das,K.,2010.Chlorella
minutissima a promising fuel alga for cultivation in municipal
wastewaters.Appl.Biochem.Biotechnol.161,523–536
[5].Perez-Garcia,O.,Escalante,F.M.E.,de-Bashan,L.E.,Bashan,Y.,
2011.Heterotrophic cultures of microalgae:metabolism and potential
products.Water Res.45,11–36.
[6].State Environmental Protection Administration of China,2002.Water
andWastewater Analyzing Methods,fourth ed.China Environmental Science.
[7].Herbert,D.,Philipps,P.J.,Strange,R.E.,1971.Methods Enzymol.5B,
265–277.
Claims (3)
1. a kind of research method that tofu wastewater is purified using microdisk electrode, it is characterised in that comprise the following steps：
1), microdisk electrode：C.pyrenoidosa is inoculated with sterile conditions, 100mL high pressures are used in 250mL conical flasks
The SE culture mediums of sterilizing are cultivated C.pyrenoidosa, are subsequently placed in light incubator and are cultivated；
Microdisk electrode condition：Light intensity=25-30 μm of ol photon m-2S-1, Light To Dark Ratio=14：10th, temperature=25 ± 1 DEG C, culture
24h, and batch (-type) shakes in the training period；
2), tofu wastewater pre-processes：Counterbalance cell SW1 and anaerobic hydrolysis reactor SW2 tofu wastewater is collected respectively, in high speed cold
Freeze in centrifuge and centrifuge, adjust the pH 6-7 of supernatant, in 100-121 DEG C of autoclaving 20 minutes, and in 1-4 DEG C of preservation, obtain
Obtain SW1 and SW2 wastewater mediums；
3), batch cultivation is tested：Using 3 500ml conical flasks as bioreactor P1, P2 and P3, it is separately added into
SW1, SW2 wastewater medium and SE culture mediums, C.pyrenoidosa inoculation algae solution cultures are then added into P1, P2 and P3,
Solution is mixed with magnetic stirring apparatus；
In light intensity=27 μm ol photon m-2S-1, Light To Dark Ratio=14：10th, culture 120h is carried out at temperature=25 ± 1 DEG C；
4), fed-batch culture experiment：Using 1 500 milliliters of conical flasks as bioreactor, add 100mL's
The SW2 culture mediums of C.pyrenoidosa inoculums and 50ml, mixed solution is mixed with magnetic stirring apparatus；In the μ of light intensity=27
mol photon m-2S-1, Light To Dark Ratio=14：10th, culture 24h is carried out at temperature=25 ± 1 DEG C；
In ensuing four days, cultivated daily in the same time to the SW2 culture mediums of microalgae addition various dose, four days
Dosage be respectively 75mL, 100mL, 0mL, 50mL.
2. according to the method for claim 1, it is characterised in that step 2) the high speed refrigerated centrifuge is with 10355rpm
Rotating speed, centrifuge 10 minutes.
3. according to the method for claim 1, it is characterised in that the step 3) adds 100mL's in P1
C.pyrenoidosa is inoculated with algae solution and 100mL SW1 wastewater mediums and 70mL distilled water；
100mL C.pyrenoidosa inoculation algae solutions and 100mL SW2 wastewater mediums and 70mL distillation is added in P2
Water；In P3,170ml SE culture mediums and 100ml inoculum C are added.Pyrenoidosa inoculations algae solution compares experiment.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108410920A (en) * | 2018-05-10 | 2018-08-17 | 天津大学 | The optimization method of polysaccharide and grease is produced using high concentration tofu wastewater culture chlorella L166 |
| CN108424857A (en) * | 2018-04-16 | 2018-08-21 | 天津大学 | A kind of research method of simultaneous foster pattern scenedesmus culture for waste water fume treatment |
| CN108641965A (en) * | 2018-04-16 | 2018-10-12 | 天津大学 | A kind of optimization method of and foster pattern scenedesmus culture |
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| CN104357327A (en) * | 2014-07-31 | 2015-02-18 | 深圳市深港产学研环保工程技术股份有限公司 | Method for culturing microalgae on large scale by bean product wastewater |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108424857A (en) * | 2018-04-16 | 2018-08-21 | 天津大学 | A kind of research method of simultaneous foster pattern scenedesmus culture for waste water fume treatment |
| CN108641965A (en) * | 2018-04-16 | 2018-10-12 | 天津大学 | A kind of optimization method of and foster pattern scenedesmus culture |
| CN108410920A (en) * | 2018-05-10 | 2018-08-17 | 天津大学 | The optimization method of polysaccharide and grease is produced using high concentration tofu wastewater culture chlorella L166 |
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