CN103710273B - Paecilomyces variot bainier F1-23 and method for processing methanal-containing industrial wastewater by using same - Google Patents
Paecilomyces variot bainier F1-23 and method for processing methanal-containing industrial wastewater by using same Download PDFInfo
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Abstract
The invention belongs to field of microorganisms and application of the microorganisms, and particularly relates to paecilomyces variot bainier F1-23 and a method for processing methanal-containing industrial wastewater by using the paecilomyces variot bainier F1-23. The paecilomyces variot bainier F1-23 CCTCC No.M2013471 or a carrier fixed with the paecilomyces variot bainier F1-23 CCTCC No.M2013471 is inoculated into the methanal-containing industrial wastewater for degrading the methanal in the methanal-containing industrial wastewater after mixing and vibrating or fully contacting; after degradation, the content of methanal in the industrial wastewater is less than 6.0g/L. The method solves the problem that the existing microorganism only degrades the industrial wastewater with low concentration of methanal, and the paecilomyces variot bainier F1-23 has the advantages of having high capacity in degrading methanal, having good heredity, and degrading high concentration methanal-containing industrial wastewater through thallus fixation and circular process.
Description
Technical field
The invention belongs to microorganism and application, refer to a kind of paecilomyces varioti bacterium F1-23 especially and utilize its process containing the method for formaldehyde industrial wastewater.
Background technology
Formaldehyde (HCHO), for having pungency and asphyxiant colourless gas under normal temperature, commodity are its aqueous solution, one of ten kinds of chemical that output is the highest in the world.Its aqueous solution is generally used as sterilization, anticorrosion and fumigant in agricultural, forestry, livestock industry, biology and medicine.Formaldehyde is harmful, mainly because formaldehyde with the protein bound in human body, can change protein interior structure and make it solidify, thus having lethality.At present, except the method for formaldehyde mainly contains chemical reaction method, physical adsorption techniques, ozone anion technology, nano photo catalyzed oxidation, plasma technology etc.; These methods all exist degraded not thoroughly, long, high in cost of production shortcoming (the Saeed M. of degradation time, 2005.World Journal of Microbiology and Biotechnology.21:1299-1301.) microorganism rule with the probiotics in ecotope for material degradation formaldehyde, cost is low, effective, and non-secondary pollution.
The beginning of this century, microbiological deterioration formaldehyde is adopted just to be paid close attention to day by day widely.An oxidation of formaldehyde approach (Barber RD. is extensively there is in microorganism, 1998.Journal of Molecular Biology.280(5): 775-784.), act on key enzyme formaldehyde dehydrogenase (the formaldehyde dehydrogenase of this approach, FADH, EC1.2.1.1) be in medium Xin Lian alcoholdehydrogenase family one, be present in (Barber RD., 1996.Journal of Bacteriology.178(5) in most protokaryon and all eukaryotic microorganisms: 1386-1393.).Formaldehyde removing toxic substances for microorganism has vital role, is also the key link of current microorganisms degradation of formaldehyde.Currently reported Formaldehyde degradation bacterium major part is Rhodopseudomonas (Pseudomonas sp.) (Saeed M., 2005.World Journal of Microbiology and Biotechnology.21:1299-1301; Adroer N., 1990.Applied Microbiology and Biotechnology.33:217-220; Xie Wenjuan, waits .2011. microbiology to circulate a notice of .38 (11): 1626-1631; The .2010. environmental science .31(10 such as Xu Yun): 2481-2486; The .2011. environmental engineering journal .5(11 such as Li Zhangliang): 2547-2551.), the concentration that these Institute of Micro-biology can degrade is lower, be generally 0.1-0.8g/L, and the concentration of formaldehyde content of trade effluent is generally 3g/L, this just needs the bacterial strain that seed selection tolerable concentration is higher, degradation capability is stronger.
Traditional microorganism method for artificially breeding has selection by mutation, cross-breeding (comprising protoplast fusion) and genetic engineering breeding.Wherein selection by mutation is the most extensive, adopt physics or chemical factor process microorganism cells colony, impel hereditary thing (DNA) structure in wherein a few cell to change, thus cause microorganism hereditary proterties to change, from colony, then filter out the process of excellent mutant strain.Obtained the mutant strain of part degrading high concentration formaldehyde by existing breeding method, and these methods will become the Critical policies of strain improvement.
At present; report that more is that formaldehyde waste water is by aerobic (Hidalgo A.; 2002.Applied Microbiology and Biotechnology.58(2): 260-263.) or anaerobism mode (Mingbo QU., 1997.Biotechnology and Bioengineering.55(5-6): 727-736; Omil F., 1991.Enzyme Microbiol.24(5-6): 255-262.) carry out biological degradation, the main active sludge PARA FORMALDEHYDE PRILLS(91,95) waste water that adopts carries out processing (Wang Zhihai etc., 2009. Chinese water supply and drainage .25(1): 55-57; Xu Zhong impartial .2008. environmental engineering journal .2(9): 1174-1176; Eiroa M., 2005.Bioresource Technology.96(17): 1914-1918.), and almost do not have about the report of using microbe immobilization technology degradation of formaldehyde waste water.Dispose of sewage compared with other Treatment process with immobilized microorganism technology, have and improve the removal efficiency of pollutent, alleviate the burden of follow-up sludge disposal, be conducive to the mud-water separation of precipitation process, the features such as, good stability strong to the ability to bear of toxic substance.Therefore, immobilized microorganism technology has become the study hotspot (in king difficult to understand etc., 2004. University Of Chongqing journal .27(3) of domestic and international field of waste water treatment: 125-129; Zhang Lei etc., 2009. Hubei University of Education journal .26(2): 39-42.).
Summary of the invention
An object of the present invention is to provide a kind of paecilomyces varioti bacterium F1-23.
Two of object of the present invention is to provide the method utilizing paecilomyces varioti bacterium F1-23 process containing formaldehyde industrial wastewater.
Overall technology design of the present invention is:
A kind of paecilomyces varioti bacterium F1-23(paecilomyces variotii F1-23), its deposit number is CCTCC No.M2013471.
Utilize the process of paecilomyces varioti bacterium containing the method for formaldehyde industrial wastewater, paecilomyces varioti bacterium F1-23 CCTCC No.M2013471 maybe will be fixed with the carrier access of paecilomyces varioti bacterium F1-23CCTCC No.M2013471 containing in formaldehyde industrial wastewater, and after mixing oscillation treatment or fully contact, degraded is containing the formaldehyde in formaldehyde industrial wastewater; In described trade effluent, the content of formaldehyde is lower than 6.0g/L.
Concentration of formaldehyde is too high, also can suppress the degradation property of paecilomyces varioti bacterium F1-23CCTCC No.M2013471, and therefore the concentration of formaldehyde solution can not higher than 6.0g/L.
Mentioned microorganism submits China typical culture collection center (being called for short CCTCC) preservation on October 13rd, 2013, and its deposit number is CCTCC No.M2013471, and the address of this depositary institution is positioned at Wuhan City, Hubei Province Wuhan University.
The morphological specificity of mentioned microorganism is: chemoheterotrophy; Addicted to temperature; Become acid.Cultivate 5d for 25 DEG C, bacterium colony is put down thin, coarse, slightly in particulate state; Outer rim is white, and inside is brownish black; Dry without transudate, there is irregular radiation fold rill on surface.Diaphragm is not had in mycelia; Sporangiospore (4-5) × (2-3) μm, oval, in cystic structures.
Concrete technical conceive of the present invention also has:
The solid medium of paecilomyces varioti bacterium F1-23CCTCC No.M2013471 adopts the component of following unit mass to form:
Glucose 10-30; Extractum carnis 2-4; KCl0.5; MgSO
47H
2o1; FeSO
47H
2o0.1; Agar 18-20; Water 1000; PH=7.2-7.5;
The liquid nutrient medium of paecilomyces varioti bacterium F1-23CCTCC No.M2013471 adopts the component of following unit mass number to form:
Glucose 10-30; Extractum carnis 2-4; KCl0.5; MgSO
47H
2o1; FeSO
47H
2o0.1; Water 1000; PH=7.2-7.5.
Described carry out degradation of formaldehyde process containing formaldehyde industrial wastewater before, also comprise a pre-treatment step, this pre-treatment step is for reducing the concentration of COD, the BOD in trade effluent and metal ion.
The described pre-treatment containing formaldehyde industrial wastewater can adopt multiple existing techniques in realizing, consider the cost of application and time, and the waste water after active sludge treatment is little on the impact of paecilomyces varioti bacterium F1-23CCTCC No.M2013471, and pre-treatment step preferably adopts following technique to realize:
A, collection active sludge
Active sludge is gathered from sewage work;
B, mixing shaking culture
Active sludge is mixed with the trade effluent containing formaldehyde, carries out shaking culture;
C, standing sedimentation
By the product standing sedimentation after shaking culture in step B, recycling precipitate recycled, supernatant liquor is used for the degradation treatment of formaldehyde.
Described carrier refers to the porous material thalline of paecilomyces varioti bacterium CCTCC No.M2013471 to height adsorptive power.Described carrier preferably adopts but is not limited to tubular fibre, kaolin, sintered glass, gac, silica gel.
Described paecilomyces varioti bacterium CCTCC No.M2013471 fixedly comprising the steps: on carrier
A, by carrier, the liquid nutrient medium be placed in containing paecilomyces varioti bacterium CCTCC No.M2013471 is cultivated;
B, paecilomyces varioti bacterium CCTCC No.M2013471 are adsorbed in immobilization carrier completing thalline.
The measuring method of degradation of formaldehyde ability is as follows, utilizes this method as the evaluation basis of screening paecilomyces varioti bacterium F1-23CCTCC No.M2013471.
The mensuration of formaldehyde, formaldehyde reacts with methyl ethyl diketone and ammonium ion and generates yellow substance under pH=5.5-7.0 condition, has absorption peak at wavelength 414nm, more quantitative with standard series.
(1) demarcation (using anhydrous sodium carbonate standardization) of sulfuric acid standard titration solution: when configuring the sulfuric acid standard titration solution of 0.5mol/L, get the vitriol oil 72ml of 98%, slowly injects water along the walls of beaker filling distilled water; Treat that solution temperature is down to room temperature, then adding distil water is settled to 1L, shakes up.When demarcating concentration, take the benchmark anhydrous sodium carbonate 1.1g dried to constant weight through 60 DEG C, put in 250ml iodine flask, adding distil water 50ml makes dissolving.Methylate is red-and tetrabromo-mcresolsulfonphthalein mixing indicating liquid 1-2 drips, with the sulfuric acid titrating solution titration of preparation.When solution becomes red-purple from green, boil 2min.Be cooled to room temperature, continue to be titrated to solution when becoming mulberry from green, the consumption of record sulfuric acid titrating solution.The mean value getting three parallel determinations is net result, and the error between Duplicate Samples must not be greater than 0.1%.
The sulfuric acid titrating solution concentration c represented with molar mass, is calculated as follows:
c（mol/L）=m/（0.1060×γ）
In formula: m---anhydrous sodium carbonate quality, g;
γ---sulfuric acid titrating solution volume, ml;
0.1060---1mol/L sulfuric acid titrating solution 1ml is equivalent to 0.1060g anhydrous sodium carbonate.
(2) demarcation of formaldehyde standardized solution: formaldehyde and excessive neutral sodium sulfite solution reaction, generates sodium hydroxide, using thymolphthalein as indicator, with the titration of sulfuric acid titrand.During mensuration, and add 50ml sodium sulfite solution in 250ml Erlenmeyer flask and 2-3 drips thymolphthalein indicating liquid, be neutralized to blueness with sulfuric acid standard titration solution and just disappear.Take 1.3-1.5g formaldehyde standardized solution with Subtraction method, be accurate to 0.0002g, put into above-mentioned Erlenmeyer flask, being neutralized to that blueness just disappeared with sulfuric acid standard titration solution is terminal.The arithmetical av getting three parallel determinations is measurement result.The error of three parallel determinations must not higher than 0.1%.
Represent that formaldehyde content x is calculated as follows with mass percent:
x=γ×c×2×100×0.3003/m=γ×c×6.006/m
In formula: γ---titration consumes the volume of sulfuric acid standard titration solution, ml;
C---the reagent concentration of sulfuric acid standard titration solution, mol/L;
The quality of m---sample, g;
0.03003---with 1.00ml sulfuric acid standard titration solution ［ c(1/2H
2sO
4)=1.000mol/L ］ suitable formaldehyde quality (g).
(3) mensuration of formaldehyde typical curve: get clean tube 6, get 0,0.05,0.10,0.15,0.20 and the above-mentioned formaldehyde reference liquid of 0.25ml in test tube, add distilled water to 5ml, add methyl ethyl diketone 1ml, boil 15min in 55 DEG C of water-baths, the reaction times needs strict control.With blank tube suppressed zero, measure light absorption value A in 414nm place, take concentration as X-coordinate, A value is ordinate zou, draws out typical curve.
When detecting bacterial classification, thalline access in inclined-plane is equipped with in the 250ml Erlenmeyer flask of 50ml assay medium, 28 DEG C, 180rpm cultivates 5d.Filtering fermentation liquor, filtrate is according to standard curve determination residual formaldehyde content.
Substantive distinguishing features acquired by the present invention and significant technical progress are:
1, the paecilomyces varioti bacterium F1-23CCTCC No.M2013471 in the present invention is improved on HCHO-resistance and degradation capability, and be divided into and improve 22.2% and 23.8%, genetic stability is excellent.
2, the present invention adopts the carrier possessing stronger adsorptive power to fix tubular fibre and fixes paecilomyces varioti bacterium F1-23CCTCC No.M2013471, can (2h) degradation of formaldehyde (3.725g/L) at short notice, this concentration is higher than the formaldehyde content (3%) of common industrial waste water.For the trade effluent of high-concentration formaldehyde content, can be solved by the mode of circular treatment.
3, in addition, the present invention uses microbiological deterioration as degradation of formaldehyde major way, can not produce secondary pollution in operational process, can reuse, save production cost, and is conducive to mass-producing and expands.
Described in the present invention in submission on October 13rd, 2013 China typical culture collection center (being called for short CCTCC) preservation, its deposit number is CCTCC No.M2013471.
Embodiment
Below in conjunction with embodiment, the present invention is described further; but should not be construed as limitation of the invention; the content that protection scope of the present invention is recorded with claim is as the criterion, any according to the equivalent technical elements replacement done by specification sheets, does not all depart from protection scope of the present invention.
Embodiment 1
A kind of paecilomyces varioti bacterium F1-23(paecilomyces variotii F1-23), its deposit number is CCTCC No.M2013471.
Utilize the process of paecilomyces varioti bacterium containing the method for formaldehyde industrial wastewater, paecilomyces varioti bacterium F1-23CCTCC No.M2013471 maybe will be fixed with the carrier access of paecilomyces varioti bacterium F1-23CCTCC No.M2013471 containing in formaldehyde industrial wastewater, and after mixing oscillation treatment or fully contact, degraded is containing the formaldehyde in formaldehyde industrial wastewater; In described trade effluent, the content of formaldehyde is lower than 6.0g/L.
The solid medium of paecilomyces varioti bacterium F1-23CCTCC No.M2013471 adopts the component of following unit mass to form:
Glucose 10-30; Extractum carnis 2-4; KCl0.5; MgSO
47H
2o1; FeSO
47H
2o0.1; Agar 18-20; Water 1000; PH=7.2-7.5;
The liquid nutrient medium of paecilomyces varioti bacterium F1-23CCTCC No.M2013471 adopts the component of following unit mass number to form:
Glucose 10-30; Extractum carnis 2-4; KCl0.5; MgSO
47H
2o1; FeSO
47H
2o0.1; Water 1000; PH=7.2-7.5.
Described carry out degradation of formaldehyde process containing formaldehyde industrial wastewater before, also comprise a pre-treatment step, this pre-treatment step is for reducing the concentration of COD, the BOD in trade effluent and metal ion.
The described pre-treatment containing formaldehyde industrial wastewater can adopt multiple existing techniques in realizing, consider the cost of application and time, and the waste water after active sludge treatment is little on the impact of paecilomyces varioti bacterium F1-23CCTCC No.M2013471, and pre-treatment step preferably adopts following technique to realize:
A, collection active sludge
Active sludge is gathered from sewage work;
B, mixing shaking culture
Active sludge is mixed according to the ratio that mass ratio is 1:2, at 25 DEG C, 150rpm shaking culture 4-6 hour with the trade effluent containing formaldehyde;
C, standing sedimentation
By the product standing sedimentation after shaking culture in step B, recycling precipitate recycled, supernatant liquor is used for the degradation treatment of formaldehyde.
It is as follows that the trade effluent different to following formaldehyde content containing the liquid nutrient medium of paecilomyces varioti bacterium F1-23CCTCC No.M2013471 adopting aforesaid method to prepare carries out result:
1, concentration of formaldehyde 1.12g/L, thalline inoculum size 10%, mixing vibration 12h, formaldehyde is thoroughly degraded.
2, concentration of formaldehyde 2.24g/L, thalline inoculum size 10%, mixing vibration 36h, formaldehyde is thoroughly degraded.
3, concentration of formaldehyde 3.73g/L, thalline inoculum size 10%, mixing vibration 36h, formaldehyde is thoroughly degraded.
4, concentration of formaldehyde 5.60g/L, thalline inoculum size 10%, mixing vibration 36h, Degradation Formaldehyde rate 69%; Thalline inoculum size 20%, mixing vibration 36h, thorough degradation of formaldehyde.
Embodiment 2
In the present embodiment bacterial classification, bacterial classification cultivation and containing the pre-treatment of formaldehyde industrial wastewater as embodiment 1, the tubular fibre (aperture is 0.1 μm) of 10 long 10cm is restrainted into a bundle, insert in same liquid nutrient medium, 3-5 days is cultivated under 25 DEG C, 180 revs/min conditions, cause paecilomyces varioti bacterium F1-23CCTCC No.M2013471 to adsorb on the hollow fibers, complete immobilization process.In gnotobasis, chopping is attached with paecilomyces varioti bacterium F1-23CCTCC No.M2013471(biomass and is about 0.1g) tubular fibre, be filled in the pillar (ψ=13mm, L=10cm) of hollow; Pillar is injected with the formaldehyde solution that concentration is 3.725g/L by 0.45ml/min flow velocity, when liquid flows out pillar, hardly containing formaldehyde by peristaltic pump.
Adopt said apparatus process containing after the trade effluent 2h of formaldehyde, use washed with de-ionized water pillar, can again recycle afterwards.
Meanwhile, test containing the formaldehyde solution of concentration of formaldehyde lower than 3.725g/L, flowed through this device, all can formaldehyde thoroughly in degraded solutions in 2h, and can Reusability for several times.
For the paper mill effluent that dustiness is higher, its formaldehyde content is up to 5g/L, and with this waste water of said apparatus process, in 2h, Degradation Formaldehyde is 70%, and the Formaldehyde In Wastewarter namely flowed out is 1.5g/L; Again wastewater streams is carried out secondary treatment through device, its formaldehyde of can thoroughly degrading.This reusable advantage is that other are removed not available for carbaldehyde Method.
There is substantial connection the time (i.e. immobilization time) of thalline absorption with kind of carrier, and design parameter is as shown in the table.
For proving that paecilomyces varioti bacterium F1-23CCTCC No.M2013471 is fixed on carrier and is directly inoculated in the trade effluent containing formaldehyde whether have difference, applicant carried out following experiment.
To in its liquid nutrient medium, under 25 DEG C, 180 revs/min conditions, cultivate 3-5 days from picking paecilomyces varioti bacterium F1-23CCTCC No.M2013471 solid medium, obtain mycelium pellet; Getting 5ml liquid medium (mycelium pellet containing some amount) is in the formaldehyde solution of 3.725g/L to concentration of formaldehyde; And the tubular fibre (aperture is 0.1 μm) of 10 long 10cm is restrainted into a bundle, insert in same liquid nutrient medium, under 25 DEG C, 180 revs/min conditions, cultivate 3-5 days, cause paecilomyces varioti bacterium F1-23CCTCCNo.M2013471 to adsorb on the hollow fibers, complete immobilization process.
Immobilized paecilomyces varioti bacterium F1-23CCTCC No.M2013471 can thorough degradation of formaldehyde concentration be the formaldehyde solution of 3.725g/L in 2 hours, showed immobilization way to the original Degradation Formaldehyde performance of microorganism without impact.
Claims (5)
1. a paecilomyces varioti bacterium F1-23 (
paecilomyces variotiif1-23), it is characterized in that its deposit number is CCTCC No.M2013471.
2. utilize paecilomyces varioti bacterium F1-23 described in claim 1 (
paecilomyces variotiif1-23) process is containing the method for formaldehyde industrial wastewater, it is characterized in that paecilomyces varioti bacterium F1-23 maybe will to be fixed with the carrier access of paecilomyces varioti bacterium F1-23 containing in formaldehyde industrial wastewater, after mixing oscillation treatment or fully contact, degraded is containing the formaldehyde in formaldehyde industrial wastewater; In described trade effluent, the content of formaldehyde is lower than 6.0g/L, described carry out degradation of formaldehyde process containing formaldehyde industrial wastewater before, also comprise a pre-treatment step, this pre-treatment step is for reducing the concentration of COD, the BOD in trade effluent and metal ion.
3. according to claim 2 utilize paecilomyces varioti bacterium F1-23 (
paecilomyces variotiif1-23) process is containing the method for formaldehyde industrial wastewater, it is characterized in that described pre-treatment step comprises the steps:
A, collection active sludge
Active sludge is gathered from sewage work;
B, mixing shaking culture
Active sludge is mixed with the trade effluent containing formaldehyde, carries out shaking culture;
C, standing sedimentation
By the product standing sedimentation after shaking culture in step B, recycling precipitate recycled, supernatant liquor is used for the degradation treatment of formaldehyde.
4. according to claim 2 utilize paecilomyces varioti bacterium F1-23 (
paecilomyces variotiif1-23) process is containing the method for formaldehyde industrial wastewater, it is characterized in that described carrier is selected from the one in tubular fibre, kaolin, sintered glass, gac, silica gel.
5. according to claim 2 utilize paecilomyces varioti bacterium F1-23 (
paecilomyces variotiif1-23) process is containing the method for formaldehyde industrial wastewater, it is characterized in that described paecilomyces varioti bacterium F1-23 fixedly comprising the steps: on carrier
A, by carrier, the liquid nutrient medium be placed in containing paecilomyces varioti bacterium F1-23 is cultivated;
B, paecilomyces varioti bacterium F1-23 are adsorbed in immobilization carrier completing thalline.
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| CN102021120A (en) * | 2010-08-13 | 2011-04-20 | 广东省生态环境与土壤研究所 | Paecilonyces variotii strain and application thereof |
| RU2448107C2 (en) * | 2010-06-02 | 2012-04-20 | Учреждение Российской Академии Наук Институт Нефтехимии И Катализа Ран | Method of producing 3,3'-bi-1,5,3-dithiazepinane and use thereof as fungicidally active agent |
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| CN102021120A (en) * | 2010-08-13 | 2011-04-20 | 广东省生态环境与土壤研究所 | Paecilonyces variotii strain and application thereof |
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