CN102021126A - Salt-resistant degrading bacterium for treating high-concentration persistent organic wastewater and application thereof - Google Patents

Salt-resistant degrading bacterium for treating high-concentration persistent organic wastewater and application thereof Download PDF

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CN102021126A
CN102021126A CN 201010180648 CN201010180648A CN102021126A CN 102021126 A CN102021126 A CN 102021126A CN 201010180648 CN201010180648 CN 201010180648 CN 201010180648 A CN201010180648 A CN 201010180648A CN 102021126 A CN102021126 A CN 102021126A
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cgmcc
sewage
concentration
acidovorax facilis
reactor
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CN102021126B (en
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张忠智
马庆霞
罗一菁
乔玮
关迎春
王祖佑
陈怡�
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China University of Petroleum Beijing CUPB
China National Petroleum Corp
Petrochina Lanzhou Petrochemical Co
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China University of Petroleum Beijing CUPB
China National Petroleum Corp
Petrochina Lanzhou Petrochemical Co
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Abstract

The invention discloses a salt-resistant degrading bacterium for treating high-concentration persistent organic wastewater and application thereof. The salt-resistant degrading bacterium is acidovorans with a preservation number of CGMCC (China General Microbiological Culture Collectio Center) No.3706, CGMCC No.3707 or CGMCC No.3708. The invention also provides the applications of the acidovorans in the treatment of high-concentration persistent organic wastewater and a method for treating high-concentration persistent organic wastewater with the acidovorans. The acidovorans is a salt-resistant microbial strain for efficiently degrading high-concentration chemical wastewater, has the characteristics of strong adaptability for high-concentration chemical wastewater, good biological stability and the like, can be used for treating wastewater under the conditions of high mineralization and high salt content, and can effectively reduce COD (Chemical Oxygen Demand) of wastewater.

Description

Be used for salt tolerant degradation bacteria and application thereof that high-concentration difficult-degradation organic sewage is handled
Technical field
The invention relates to the degradation bacteria and the application thereof that are used for sewage disposal, particularly about be used for salt tolerant degradation bacteria that high-concentration difficult-degradation organic sewage handles, this salt tolerant degradation bacteria is in the application of handling high-concentration difficult-degradation organic sewage and utilize this salt tolerant degradation bacteria to handle the method for high-concentration difficult-degradation organic sewage, belongs to technical field of sewage.
Background technology
Be the high concentrated organic wastewater of difficult degradation greatly in the trade effluent.The organic concentration (in COD) that so-called " high density " is meant waste water more than 2000mg/L, have in addition up to every liter several ten thousand to tens0000 milligrams; So-called " difficult degradation " is meant the lower (BOD of biodegradability of waste water 5/ COD value general all below 0.3 in addition lower), be difficult to biological degradation.Therefore, in the industry generally with COD concentration greater than 2000mg/L, BOD 5/ COD value is lower than 0.3 organic waste water unification and is called " high concentration hard-degraded organic waste water (sewage) ".
The chemical industrial organic wastewater water-quality constituents is complicated, mainly contains petroleum-type, volatile phenol, sulfide, suspended substance, cyanogen, aromatic amine, nitrogen heterocyclic and polycyclic aromatic hydrocarbon compounds etc., belongs to high concentration hard-degraded organic waste water.Pollutent in the wastewater from chemical industry according to chemical composition can be divided into: organism (phenols, organophosphorus etc., mainly from industries such as basic organic, synthetic materials, agricultural chemicals, dyestuffs), inorganics (various metals and nonmetal, as mercury, lead, arsenic, chromium, soda acid salt, prussiate and sulfide etc., mainly from industries such as inorganic salt, nitrogenous fertilizer, phosphate fertilizer, sulphur nitric acid and soda ash) and the mixture (mainly from industries such as atmosphere alkali, sensitive materials and coating) of organism and inorganics.The pollutent of these difficult degradations has four fundamental characteristics: 1. extended residual: in case be discharged in the environment, be difficult to be decomposed, can retain several years or longer time in surrounding mediums such as water body, soil and bed mud; 2. biological accumulation: generally have the feature of low water solubility, fat-solubility, can in the biological fatty tissue of majority, accumulate; 3. half volatile: can move in the atmospheric environment medium and long distance; 4. high toxicity: humans and animals is generally had toxic action, can cause organism endocrine regulation, reproduction and immunity function imbalance, have in addition cause serious disease such as cancer.
At present, domestic and international treatment process to wastewater of chemical industry with high concentration mainly contains physics method, chemical method and biological process etc.The method of traditional processing high-concentration difficult-degradation organic sewage mainly adopts physical-chemical technology, relevant sewage treatment technology process mostly is " two-phase method " of " three-stage process " and " coagulation-filtration " of " oil removal-coagulation-filtration ", these technology are to the suspended state in the waste water, part emulsification attitude and be partly dissolved macromolecular cpd removal effect is preferably arranged, but relatively poor for the removal effect of most of solvability organic pollutants such as all kinds of phenols and derivative thereof etc.The COD that this part pollutent brings, BOD 5Reaching other pollutent indexs such as ammonia nitrogen, sulfide, volatile phenol etc. is the major cause that causes that outer draining exceeds standard, and the compliance rate of outer draining only is about 50%.Adopt physical-chemical art breading high-concentration difficult-degradation organic sewage, also need carry out second-stage treatment technology to guarantee the wastewater from chemical industry emission compliance usually with biological method.
Biological process is present most widely used a kind of method for processing organic wastewater, mainly comprises active sludge, biomembrance process, aerobic-anaerobic process etc.It mainly is the metabolism that utilizes microorganism, and the cohesion by microorganism, absorption, oxygenolysis etc. are used for the organism in the degradation of sewage, have applied range, big, the low cost and other advantages of treatment capacity.Organic microorganism is generally various degradation bacteria in the described degradation of sewage, research report about organic degradation bacteria in the degradation of sewage is a lot of at present, the kind of described degradation bacteria is also a lot, but the degradation bacteria that really can handle high-concentration hardly-degradable chemical industry organic sewage under high salinity, high salinity condition seldom.For example, people such as Lv Ronghu, Fu Qiang is in " seed selection of high-concentration phenol degradation bacteria and fall the phenol performance " (environmental science, the 26th the 5th phase of volume, the phenol bacterium falls in the advantage that has disclosed the high phenol concentration of degrading that is screened in September, 2005), yet, do not mention in the document and describedly fall the phenol bacterium and whether can handle high salinity, high salinity sewage, in fact, the tolerance of salinity of these bacterium is not high, the highest 5000~6000mg/L that only is about of salinity of its actual treatable sewage.
Summary of the invention
One object of the present invention is to provide the degradation bacteria that can handle the chemical industry organic sewage under high salinity, high salinity condition.
Another object of the present invention provides a kind of preparation that contains acidovorax facilis of the present invention, preserves and uses with convenient.
Another object of the present invention provides described degradation bacteria or the application of bacteria preparation in handling high-concentration difficult-degradation organic sewage.
Another object of the present invention provides the method for utilizing described degradation bacteria or bacteria preparation to handle high-concentration difficult-degradation organic sewage.
The present invention has filtered out 3 strain bacterial strains from the high-concentration chemical industry sewage water sample of petrochemical plant, respectively called after JF-2, JF-3, JF-6.Described JF-2 bacterial strain has been preserved in (address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, China Committee for Culture Collection of Microorganisms common micro-organisms center on 04 02nd, 2010, Institute of Microorganism, Academia Sinica), deposit number: CGMCC No.3706; Described JF-3 bacterial strain has been preserved in (address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, China Committee for Culture Collection of Microorganisms common micro-organisms center on 04 02nd, 2010, Institute of Microorganism, Academia Sinica), deposit number: CGMCC No.3707; Described JF-6 bacterial strain has been preserved in (address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, China Committee for Culture Collection of Microorganisms common micro-organisms center on 04 02nd, 2010, Institute of Microorganism, Academia Sinica), deposit number: CGMCC No.3708.
The bacterial strain JF-2 that the present invention filtered out, JF-3, JF-6, be acidovorax facilis (Acidovoraxsp.) through evaluation, can be used for handling high-concentration difficult-degradation organic sewage, can in the environment of COD 5000~6000mg/L, survive, and the tolerance of salinity can reach 3.5%, can handle the organic sewage of salinity up to 25000~35000mg/L.
The present invention also provides a kind of acidovorax facilis preparation, and containing deposit number of the present invention in this acidovorax facilis preparation is the acidovorax facilis of CGMCC No.3706, CGMCC No.3707 and/or CGMCC No.3708.This acidovorax facilis preparation can be solid-state bacteria preparation, also can be liquid bacteria preparation, preserves and uses with convenient.
The present invention also provides described acidovorax facilis JF-2, JF-3, the JF-6 preparation of acidovorax facilis of the present invention (or contain) application in handling high-concentration difficult-degradation organic sewage.
Utilizing acidovorax facilis JF-2 of the present invention, JF-3, the JF-6 preparation of acidovorax facilis of the present invention (or contain) when handling high-concentration difficult-degradation organic sewage, the pH value 6.0~10.0 of the preferred control high-concentration difficult-degradation organic sewage of handling; Described treatment temp is 20 ℃~40 ℃.
Utilizing acidovorax facilis JF-2 of the present invention, JF-3, the JF-6 preparation of acidovorax facilis of the present invention (or contain) when handling high-concentration difficult-degradation organic sewage, controlling preferably that phenolic comp ' ds pollution concentration≤1200mg/L for example can be 1000mg/L~1200mg/L in the described high-concentration difficult-degradation organic sewage; Described phenolic comp ' ds pollution can be to comprise phenol, p-tolyl aldehyde, dibutyl phthalate and/or 2,6 ditertiary butyl p cresol.
When handling high-concentration difficult-degradation organic sewage, saltiness≤3.5% of preferably controlling in the described high-concentration difficult-degradation organic sewage for example can be 1.5%~3.5% utilizing acidovorax facilis JF-2 of the present invention, JF-3, the JF-6 preparation of acidovorax facilis of the present invention (or contain); Salinity≤35000mg/L for example can be 10000mg/L~35000mg/L, perhaps 20000mg/L~35000mg/L, perhaps 25000mg/L~35000mg/L.
Utilizing acidovorax facilis JF-2 of the present invention, JF-3, the JF-6 preparation of acidovorax facilis of the present invention (or contain) when handling high-concentration difficult-degradation organic sewage, can be in processing sewage the complex biological nutritional formulation, to satisfy the growth needs of bacterial classification of the present invention.
The present invention also provides the method for utilizing described acidovorax facilis JF-2, JF-3, the JF-6 preparation of acidovorax facilis of the present invention (or contain) to handle high-concentration difficult-degradation organic sewage.Among the present invention, handle high-concentration difficult-degradation organic sewage and carried out the laboratory lab scale, and be that the pilot scale processing has been carried out on the basis with the determined best degradation condition of lab scale to using described acidovorax facilis, determined reasonably can industrial applications sewage treatment process.Wherein, adopt complex biological nutritional formulation and biological reinforced measure, wastewater of chemical industry with high concentration is handled, make it reach the water inlet index of conventional biosystem.Concrete, the method for utilizing described acidovorax facilis JF-2, JF-3, JF-6 to handle high-concentration difficult-degradation organic sewage of the present invention comprises step: treatment sewage is carried out acidifying, and sewage after the acidifying is diluted; With in the biological reactor of described sewage transport to, acidovorax facilis CGMCC No.3706 of the present invention, CGMCC No.3707 and/or CGMCC No.3708 (described acidovorax facilis can be carrier with the active sludge) have been added in the described bio-reactor in reactor then; Temperature is 20 ℃~40 ℃ in the controlling reactor, adopts acidication-contact oxidation technology, by aeration, sewage is purified.
According to specific embodiments of the present invention, in the sewage water treatment method of the present invention, can be the pH value 6~10 of adjusting treatment sewage, preferred pH value 6.5~8.5, most preferably the pH value about 8; Sewage after the acidifying is diluted, COD is about 3500~6000mg/L in may command dilution back water inlet, can add simultaneously the required nutritive substance of acidovorax facilis growth (the conventional acidovorax facilis needed nutritive substance of growing gets final product, for example: organic nitrogen source, sal epsom, potassium primary phosphate and some metal ion etc.).According to specific embodiments of the present invention, before described treatment sewage enters reactor, phenolic comp ' ds pollution concentration≤1200mg/L wherein, saltiness≤3.5%, salinity≤35000mg/L.Behind bio-reactor,, under acidovorax facilis JF-2 of the present invention, JF-3 in reactor in the active sludge and/or the effect of JF-6, sewage is purified sewage transport by aeration.
In a specific embodiments of the present invention, be to have added acidovorax facilis preparation of the present invention in the described bio-reactor, this acidovorax facilis preparation is for being cultured to concentration 10 with acidovorax facilis CGMCC No.3706, CGMCC No.3707 and/or CGMCC No.3708 in enrichment medium (the conventional liquid substratum preparation of acidovorax facilis in can be according to prior art) 8~10 9Cells/ml and making, the dosage of this acidovorax facilis preparation in described bio-reactor is 4%~8% (V/V) of sewage quantity in the bio-reactor.Preferably, contain acidovorax facilis CGMCC No.3706, CGMCC No.3707 and CGMCC No.3708 in the described bio-reactor simultaneously, the ratio of adding of three kinds of bacterium is 0.5~2: 0.5~2: 0.5~2 (in the initial cell number of the bacterial classification that adds).
Acidovorax facilis JF-2 of the present invention, JF-3, JF-6 and utilize this acidovorax facilis to carry out the method for sewage disposal, have the following advantages and effect: acidovorax facilis JF-2 of the present invention, JF-3, JF-6 are salt tolerant efficient degradation wastewater of chemical industry with high concentration microbial strainss, have characteristics such as strong to wastewater of chemical industry with high concentration adaptability, that biologically stable is good.They not only can reduce the COD of sewage effectively, its sharpest edges are can (salinity can reach 25000~35000mg/L), sewage is handled under higher salts contg (for example saltiness 1.5%~3.5%) condition, still has the good phenol performance of falling in high salinity.In one embodiment of the invention, utilize this bacterial classification to carry out biochemical field experiment, device operation 160 days, microorganism growth is good, system's stable water outlet.
Description of drawings
Fig. 1 is a phenolic comp ' ds pollution degradation rate chart under the three strain salt tolerant bacterium condition of different pH;
Fig. 2 is a phenolic comp ' ds pollution degradation rate chart under the three strain salt tolerant bacterium condition of different temperatures;
Fig. 3 is a phenolic comp ' ds pollution degradation rate chart under the different saltiness conditions of three strain salt tolerant bacterium;
Fig. 4 is a phenolic comp ' ds pollution degradation rate chart under the different Pollutant levels conditions of three strain salt tolerant bacterium;
Fig. 5 is that salt tolerant bacterium of the present invention is handled high-concentration chemical industry sewage device synoptic diagram;
Fig. 6 is a COD of sewage Changing Pattern chart before and after the effect of three strain salt tolerant bacterium.
The microorganism that is used for patented procedure preserves:
(1) salt tolerant bacterium JF-2 of the present invention
Preservation date: on 04 02nd, 2010;
Depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center (CGMCC);
Deposit number: CGMCC No.3706;
Classification name: acidovorax facilis (Acidovorax sp.).
(2) salt tolerant bacterium JF-3 of the present invention
Preservation date: on 04 02nd, 2010;
Depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center (CGMCC);
Deposit number: CGMCC No.3707;
Classification name: acidovorax facilis (Acidovorax sp.).
(3) salt tolerant bacterium JF-6 of the present invention
Preservation date: on 04 02nd, 2010;
Depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center (CGMCC);
Deposit number: CGMCC No.3708;
Classification name: acidovorax facilis (Acidovorax sp.).
Embodiment
Further describe the characteristics of salt tolerant bacterium of the present invention below by specific embodiment, and utilize this salt tolerant bacterium to handle the method for high-concentration hardly-degradable sewage and the technique effect that is had, but therefore the present invention is not subjected to any restriction.
Separation, evaluation and the preservation of embodiment 1, salt tolerant bacterium JF-2 of the present invention, JF-3, JF-6
1, artificial screening of microorganism and domestication
Get petrochemical plant high-concentration chemical industry sewage water sample (the Suizhong SZ-361 crude Treatment factory crude oil pollution water sample of gathering on December 12nd, 2008) 250mL as original bacterium source, method for domesticating carries out artificial screening and domestication to adopt conventional pursuing to measure in batches, concrete operations comprise: getting the original bacterium of 0.2mL source, to coat with 100mg/L phenol be in the Agar Plating of sole carbon source, every kind of original bacterium source do three parallel, 30 ℃, (getting the original bacterium of 0.2mL source simultaneously coats in the complete nutrition Agar Plating in the 48h cultivation, each makees three parallel samples, 30 ℃, 48h cultivates, in contrast); And successively the bacterium liquid of being cultivated is transferred into 200,300,400mg/L ... 1000mg/L's is in the screening culture medium of sole carbon source with the phenols, reaches the concentration 1000mg/L that requires up to phenol concentration; Get the bacterium liquid of final acquisition, get earlier in the sterilized water that 1mL joins 9mL, make 10 -1Diluent, again from 10 -1Get equally in the diluent in the sterilized water that 1mL joins 9mL, be prepared into 10 -2Diluent, by that analogy, the bacterium liquid of final acquisition is done a series of gradient dilutions, and gets 0.2mL respectively and coat the Agar Plating that phenol concentration is 1000mg/L, cultivate 48h for 30 ℃; Nearly hundred strains of picking list bacterium colony at random, the purifying of on slant medium, ruling, 30 ℃, 48h cultivates.To inoculate with 1000mg/L be in the substratum of sole carbon source with the mixed phenol through enrichment culture to get naturalized strain, and 30 ℃, 150rpm cultivates 48h, and the phenol rate falls in mensuration, chooses the bacterial strain of phenol degrading rate maximum.
Among the present invention,, isolate the different bacterial strain of 3 strains through above-mentioned cultivation, domestication, difference called after JF-2, JF-3, JF-6 is used for the high-concentration chemical industry treatment of Organic Wastewater.
2, strain identification
According to " common bacteria system identification handbook ", the isolated 3 strain bacterial strain JF-2 of the present invention, JF-3, JF-6 are identified, mainly identify genus from bacterial strain individual morphology feature, colony characteristics, staining reaction, biochemical reactions etc., content comprises morphologic observation, gramstaining, the reaction of hydrogen peroxide catalase, oxydase experiment, glucose oxidase fermenting experiment, methyl red experiment etc., qualification result such as following table 1.
Table 1
The mensuration project JF-2 JF-3 JF-6
Gramstaining Negative Negative Negative
Mobility Do not have Do not have Do not have
Aerobism Amphimicrobian Amphimicrobian Amphimicrobian
The reaction of hydrogen peroxide catalase Positive Positive Positive
Peroxidase test Positive Positive Positive
The glucose oxidase fermenting experiment Fermented type Produce the alkali type Fermented type
The methyl red experiment Positive Negative Negative
Mierocrystalline cellulose decomposes experiment Negative Negative Negative
Colony colour Oyster white Oyster white Oyster white
Colonial morphology Translucent, circle, protuberance is smooth, glossy Opaque, half particulate state projection, more moistening Opaque, little protuberance, full edge is smooth, glossy
Thalli morphology Shaft-like Shaft-like Shaft-like
In addition, according to conventional strain identification method, extract the genomic dna of bacterial strain JF-2, JF-3, JF-6 respectively, the design primer carries out pcr amplification, amplified production detects through agarose gel electrophoresis, selects ideal PCR product to deliver to the living worker's order-checking in Shanghai and carries out dna sequencing.The pcr amplification product sequencing result of JF-2, JF-3, JF-6 is respectively shown in SEQ ID No.:1 (JF-2), SEQ ID No.:2 (JF-3) and SEQ ID No.:3 (JF-6).Product sequencing result submission NCBI retrieves with homology with BLAST and compares, and utilizes software building systematic evolution trees such as SepnConverter, Clustalx1.83, Phylip3.63 and TreeView.By the 16SrDNA sequential analysis, determine that bacterial strain JF-2, JF-3 and JF-6 are Acidovorax (Acidovorax) (three strain bacterium corresponding representative bacterial classification in Genbank sees table 2).
Table 2
Bacterial strain Base (bp) Bacterial classification/the genus of similar sequences among the Genbank Homology (%)
JF-2 407 NC?008782.1 Acidovorax?sp.JS42 100%
JF-3 442 NC?011992.1 Acidovorax?ebreus?TPSY 99%
JF-6 417 NC?008782.1 Acidovorax?sp.JS42 98%
Bacterial strain JF-2 of the present invention, JF-3, JF-6 can adopt following store method:
(1) short-term is preserved: above-mentioned bacterial strains is gone up line in slant medium (phenol concentration is 500mg/L), and through 30 ℃, after 48h cultivates, 4 ℃ of preservations.
(2) prolonged preservation:
Can adopt the glycerine freeze preservation: scrape from fresh slant medium and get a few ring bacterium, be transferred to-70 ℃ of freezing preservations are housed in the 1.5mL 30% sterile glycerol glycerine pipe.
Or adopt skimmed milk freezing method: scrape from fresh slant medium and get a few ring bacterium, be transferred in the glycerine pipe that the sterilization skimmed milk is housed-70 ℃ of freezing preservations.
Embodiment 2, salt tolerant bacterium JF-2, JF-3, JF-6 organic experiment in the degradation of sewage under different condition
Plant daughter bacteria liquid: (with reference to the prior art preparation, essentially consist comprises: phenol 1g/L, (NH in enrichment medium with bacterial classification 4) 2SO 41.5g/L, KH 2PO 40.5g/L, K 2HPO 40.5g/L, NaCl 1.0g/L, MnSO 4H 2The O trace, FeSO 47H 2O trace etc.) be cultured to concentration 10 8-10 9Cells/ml as seed liquor, carries out the experimental study of present embodiment and the pilot plant test research of subsequent embodiment.
Simulated sewage prescription: KH 2PO 40.5g, K 2HPO 40.5g, MgSO 47H 2O 0.2g, CaCl 20.1g, NaCl 0.2g, MnSO 4H 2The O trace, FeSO 47H 2The O trace, NH 4NO 31.0g, distilled water 1L, according to the experiment needs, the adding phenolic comp ' ds pollution is some.Described phenolic comp ' ds pollution: (the mixing quality ratio is about 1: 1: 1 to the mixture of phenol, p-tolyl aldehyde, dibutyl phthalate, 2,6 ditertiary butyl p cresol: 1).
Pollutant levels adopt the amino antipyrine spectrophotometry of 4-, under the aseptic condition, get the nutrient solution of cultivating behind the 24h, in the centrifugal 10min of 4000r/min, get supernatant liquor and measure residual contaminants content, calculate degradation rate.
Experiment 1, the organic experiment in the degradation of sewage under different pH of salt tolerant bacterium
Adopt the 100mL triangular flask, the simulated sewage (phenolic comp ' ds pollution concentration is 1000mg/L in the simulated sewage nutrient solution) that adds 50mL in each triangular flask, drop into the seed liquor of three strain bacterium respectively, the dosage of three strain bacterium liquid is 6% (V/V), cultivate in 30 ℃ of vibrations (150r/min), the pH of simulated sewage is respectively 3.0,4.0,5.0,6.0,7.0,8.0,9.0,10.0, measures the changing value of pollutent in the simulated sewage of salt tolerant bacterium effect front and back, calculates degradation rate.
Experimental result sees also shown in Figure 1, shows that this three strains bacterium after phenolic comp ' ds pollution concentration is to cultivate 24h under the 1000mg/L condition of different pH, all degrading to pollutent in varying degrees, and its best pH is a meta-alkalescence.
Experiment 2, the organic experiment in the degradation of sewage under differing temps of salt tolerant bacterium
Adopt the 100mL triangular flask, the simulated sewage (phenolic comp ' ds pollution concentration is 1000mg/L in the simulated sewage nutrient solution) that adds 50mL in each triangular flask, drop into the seed liquor of three strain bacterium respectively, the dosage of three strain bacterium liquid is 6% (V/V), vibration (150r/min) is cultivated, culture temperature has adopted 20 ℃, 25 ℃, 30 ℃, 35 ℃, 40 ℃, 45 ℃, 50 ℃ respectively, simulated sewage pH is 7.0, other parameter and experimental technique are with experiment 1, by measuring the changing value of pollutent in the simulated sewage of salt tolerant bacterium effect front and back, calculate degradation rate.
Experimental result sees also shown in Figure 2, shows that this three strains bacterium after phenolic comp ' ds pollution concentration is to cultivate 24h under the 1000mg/L condition of different temperatures, all degrading to pollutent in varying degrees, and its optimum temps is 30-40 ℃.
The organic experiment in the different saltiness sewage of degrading of experiment 3, salt tolerant bacterium
Adopt the 100mL triangular flask, the simulated sewage (phenolic comp ' ds pollution concentration is 1000mg/L in the simulated sewage nutrient solution) that adds 50mL in each triangular flask, drop into the seed liquor of three strain bacterium respectively, the dosage of three strain bacterium liquid is 6% (V/V), cultivate in 30 ℃ of vibrations (150r/min), saltiness has adopted 0.5% respectively, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4% (involved saltiness all is to regulate by the sodium-chlor that adds different amounts among the present invention), simulated sewage pH is 7.0, other parameter and experimental technique are with experiment 1, by measuring the changing value of pollutent in the simulated sewage of salt tolerant bacterium effect front and back, calculate degradation rate.
Experimental result sees also shown in Figure 3, shows that this three strains bacterium after phenolic comp ' ds pollution concentration is to cultivate 24h under the different saltiness conditions of 1000mg/L, all degrading to pollutent in varying degrees, and its maximum salt tolerant amount is 3.5%.
The organic experiment in the different saltiness sewage of degrading of experiment 4, salt tolerant bacterium
Adopt the 100mL triangular flask, (phenolic comp ' ds pollution concentration is for having adopted 500mg/L, 800mg/L, 1000mg/L, 1200mg/L, 1500mg/L respectively in the simulated sewage nutrient solution to add the simulated sewage of 50mL in each triangular flask, drop into three strain bacterium respectively, the dosage of three strain bacterium is 6% (V/V), cultivate in 30 ℃ of vibrations (150r/min), simulated sewage pH is 7.0, other parameter and experimental technique are with experiment 1, by measuring the changing value of pollutent in the simulated sewage of salt tolerant bacterium effect front and back, calculate degradation rate.
Experimental result sees also shown in Figure 4, shows that this three strains bacterium all has higher degradation rate to pollutent, but along with constantly the increasing of Pollutant levels, its degradation rate descends gradually, and its best phenolic comp ' ds pollution concentration is 1000mg/L.
Embodiment 3, utilize organic pilot plant test in salt tolerant bacterium JF-2, JF-3, the JF-6 degradation of sewage
In the present embodiment, utilize salt tolerant bacterium JF-2 provided by the invention, JF-3, JF-6 to handle wastewater of chemical industry with high concentration, concrete grammar is:
(1) wastewater from chemical industry to high density carries out acidifying in acidification pool 1, control pH value 6~10, and preferred pH value is about 8, and the organic phase of generation and aqueous phase separation after the acidifying can be used as resource product and reclaims; (2) in order to satisfy the basic demand of microorganism to saltiness, the chemical engineering sewage after the acidifying dilutes (about extension rate 10) again, adds nutritive substance simultaneously; (3) chemical engineering sewage and nutrient mixing liquid are transported in the high-efficient biological reactor 3 by volume pump 2, reactor is by the temperature in heating unit and the temperature control instrument 5 control bio-reactors, make it keep about 30 ℃, adopt acidication-contact oxidation technology, utilize air pump 4 in reactor, to inflate, pass through aeration, active sludge is suspended state, and fully contact with waste water, by effects such as sludge absorption and microbiological deteriorations, waste water is purified thus; (4) purify back waste water and separate in secondary sedimentation basins 6 with active sludge, a mud part of separating after concentrating is returned the aeration tank, and to guarantee to keep in the aeration tank active sludge of higher concentration, all the other are discharged by system for excess sludge.The supernatant liquid sampling analysis is measured the COD value; Effluent quality COD reaches national emission request less than 500mg/L (COD of water outlet is difference with the difference of the COD of import, increases the residence time, and the COD of outlet is even lower, and the rate of removing of COD can bring up to 90%).
In the present embodiment, (each bacterial classification bacterium liquid blending ratio is 1: 1: 1 to add 3 high-efficiency degradation bacterium in the reactor, total dosage is 6% (V: V)), makes it form high-concentration activated sludge (dominant strain in this mud is 3 strain salt tolerant bacterium of the present invention) by the sludge carrier in the reactor.Main processing parameter sees table 3 (data in the form are more conservative) in its treating processes.
Test run on January 9 in 2008 begins to carry out continuous 160 days Experiment Data Records after 7 days, although during 45 ℃ of overtemperatures appearred reaching several times, system stability is good, shock resistance is strong, has recovered normal operation soon, the running of 160 days continous-stables.
Main technologic parameters during table 3 wastewater of chemical industry with high concentration is handled
In test, after former water diluted, the COD value was 3513.25mg/L, and adding nutrition (not carbonaceous sources nutrition) back COD value is 5702.5mg/L.COD of sewage goes out water removal rate can reach 87%, and sewage salinity is not less than 20000mg/L (Theoretical Calculation), surveys about 25000mg/L.Fig. 6 shown during this water inlet, effluent COD concentration contrast, this has proved absolutely that biosystem has formed that anti-high salinity, degraded are poisonous, the advantage group system of the extreme microorganism of high concentration organic contaminant.
Sequence table
<110〉China National Petroleum Corporation (CNPC)
CNPC's Lanzhou Petrochemical Company
Chinese Petroleum Univ.
 
<120〉be used for salt tolerant degradation bacteria and the application thereof that high-concentration difficult-degradation organic sewage is handled
 
<130>GAI10CN0452C
 
<160>3
 
<170>PatentIn?version?3.5
 
<210>1
<211>407
<212>DNA
<213〉acidovorax facilis JF-2 (Acidovorax sp.)
 
<400>1
ctcggatcta?gcgtctgaga?tggattcgtg?cccgaaaggg?aacctgcaca?caggtgctgc 60
atggctgtcg?tcagctcgtg?tcgtgagatg?ttgggttaag?tcccgcaacg?agcgcaaccc 120
ttgccattag?ttgctacgaa?agggcactct?aatgggactg?ccggtgacaa?accggaggaa 180
ggtggggatg?acgtcaagtc?ctcatggccc?ttataggtgg?ggctacacac?gtcatacaat 240
ggctggtaca?gagggttgcc?aacccgcgag?ggggagctaa?tcccataaag?ccagtcgtag 300
tccggatcgc?agtctgcaac?tcgactgcgt?gaagtcggaa?tcgctagtaa?tcgcggatca 360
gaatgtcgcg?gtgaatacgt?tcccgggtct?tgtacacacc?gcccgta 407
 
<210>2
<211>442
<212>DNA
<213〉acidovorax facilis JF-3 (Acidovorax sp.)
 
<400>2
tttaacgcga?agaaccttac?ccacctttga?catggcagga?agtttccaga?gatggattcg 60
tgcccgaaag?ggaacctgca?cacaggtgct?gcatggctgt?cgtcagctcg?tgtcgtgaga 120
tgttgggtta?agtcccgcaa?cgagcgcaac?ccttgccatt?agttgctacg?aaagggcact 180
ctaatgggac?tgccggtgac?aaaccggagg?aaggtgggga?tgacgtcaag?tcctcatggc 240
ccttataggt?ggggctacac?acgtcataca?atggctggta?cagagggttg?ccaacccgcg 300
agggggagct?aatcccataa?agccagtcgt?agtccggatc?gcagtctgca?actcgactgc 360
gtgaagtcgg?aatcgctagt?aatcgcggat?cagaatgtcg?cggtgaatac?gttcccgggt 420
cttgtacaca?ccgcccgtaa?aa 442
 
<210>3
<211>417
<212>DNA
<213〉acidovorax facilis JF-6 (Acidovorax sp.)
 
<400>3
cgtactgata?ttattactgc?gaattctgat?cgcgattact?agcgattccg?acttcacgca 60
gtcgagttgc?agactgcgat?ccggactacg?actggcttta?tgggattagc?tccccctcgc 120
gggttggcaa?ccctctgtac?cagccattgt?atgacgtgtg?tagccccacc?tataagggcc 180
atgaggactt?gacgtcatcc?ccaccttcct?ccggtttgtc?accggcagtc?ccattagagt 240
gccctttcgt?agcaactaat?ggcaagggtt?gcgctcgttg?cgggacttaa?cccaacatct 300
cacgacacga?gctgacgaca?gccatgcagc?acctgtgtgc?aggttccctt?tcgggcacga 360
atccatctct?ggaaacttcc?tgccatgtca?aaggtgggta?aggttcttcg?cgttaaa 417

Claims (10)

1. deposit number is the acidovorax facilis of CGMCC No.3706, CGMCC No.3707 or CGMCC No.3708.
2. acidovorax facilis preparation, containing deposit number in this acidovorax facilis preparation is the acidovorax facilis of CGMCC No.3706, CGMCC No.3707 and/or CGMCC No.3708, this acidovorax facilis preparation is solid-state or liquid bacteria preparation.
3. the application of the described acidovorax facilis preparation of described acidovorax facilis of claim 1 or claim 2 in handling high-concentration difficult-degradation organic sewage.
4. application according to claim 3, wherein, the pH value 6.0~10.0 of described high-concentration difficult-degradation organic sewage; Described treatment temp is 20 ℃~40 ℃.
5. application according to claim 3, wherein, phenolic comp ' ds pollution concentration≤1200mg/L in the described high-concentration difficult-degradation organic sewage; Described phenolic comp ' ds pollution comprises phenol, p-tolyl aldehyde, dibutyl phthalate and/or 2,6 ditertiary butyl p cresol.
6. application according to claim 3, wherein, the saltiness in the described high-concentration difficult-degradation organic sewage≤3.5%; Salinity≤35000mg/L.
7. utilize the described acidovorax facilis preparation of described acidovorax facilis of claim 1 or claim 2 to handle the method for high-concentration difficult-degradation organic sewage, the method comprising the steps of:
Treatment sewage is carried out acidifying, and sewage after the acidifying is diluted; Then with in the biological reactor of described sewage transport to, add have the right requirement 1 described acidovorax facilis CGMCC No.3706, CGMCC No.3707 and/or CGMCC No.3708 in the described bio-reactor, perhaps contain the described acidovorax facilis preparation of claim 2; Temperature is 20 ℃~40 ℃ in the controlling reactor, adopts acidication-contact oxidation technology, by aeration, sewage is purified.
8. method according to claim 7, wherein, before described treatment sewage enters reactor, pH value 6.0~10.0, phenolic comp ' ds pollution concentration≤1200mg/L, saltiness≤3.5%, salinity≤35000mg/L.
9. according to claim 7 or 8 described methods, wherein, be to add the requirement 2 described acidovorax facilis preparations of having the right in the described bio-reactor, this acidovorax facilis preparation is for to be cultured to concentration 10 with acidovorax facilis CGMCC No.3706, CGMCC No.3707 and/or CGMCC No.3708 in enrichment medium 8~10 9Cells/ml and making, the dosage of this acidovorax facilis preparation in described bio-reactor is 4%~8% (V/V) of sewage quantity in the bio-reactor.
10. method according to claim 9, wherein, described bio-reactor contains acidovorax facilis CGMCC No.3706, CGMCC No.3707 and CGMCC No.3708, and the ratio of adding of three kinds of bacterium is 0.5~2: 0.5~2: 0.5~2.
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Cited By (3)

* Cited by examiner, † Cited by third party
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CN103468598A (en) * 2012-11-07 2013-12-25 上海大学 Humic acid degrading strain, and screening method and application method thereof
CN105219685A (en) * 2015-11-10 2016-01-06 武汉大学 A kind of dephenolize halophilic bacterium and application thereof
CN110467274A (en) * 2019-08-27 2019-11-19 广东省生物工程研究所(广州甘蔗糖业研究所) The new opplication of acidovorax facilis LS-1 and kit and its application for repairing arsenic pollution

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103468598A (en) * 2012-11-07 2013-12-25 上海大学 Humic acid degrading strain, and screening method and application method thereof
CN103468598B (en) * 2012-11-07 2015-07-29 上海大学 A kind of humic acid degrading bacterial strain and screening and application method thereof
CN105219685A (en) * 2015-11-10 2016-01-06 武汉大学 A kind of dephenolize halophilic bacterium and application thereof
CN105219685B (en) * 2015-11-10 2018-11-13 武汉大学 A kind of dephenolize Halophiles and its application
CN110467274A (en) * 2019-08-27 2019-11-19 广东省生物工程研究所(广州甘蔗糖业研究所) The new opplication of acidovorax facilis LS-1 and kit and its application for repairing arsenic pollution

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