CN1176206C - Pseudomonads GT 241-1 for degradating chloro-aromatic compound and aromatic compound and its application - Google Patents
Pseudomonads GT 241-1 for degradating chloro-aromatic compound and aromatic compound and its application Download PDFInfo
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- CN1176206C CN1176206C CNB001312847A CN00131284A CN1176206C CN 1176206 C CN1176206 C CN 1176206C CN B001312847 A CNB001312847 A CN B001312847A CN 00131284 A CN00131284 A CN 00131284A CN 1176206 C CN1176206 C CN 1176206C
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Abstract
The present invention belongs to microorganism strains of toxic substances in waste water treatment and a purpose of the microorganism strains for degrading toxic waste water and other residual substances. The present invention is mainly characterized in that pseudomonas strains GT241-1 which are specially used for degrading a chloro-aromatic compound and an aromatic compound are domesticated, sieved and selected. The microorganism strains can fully degrade 2, 4-dichlorophenol, 2, 4-D, benzoic acid, salicylic acid and salt thereof, which are difficult to degrade at present. Compared with the prior art, the microorganism strains have specificity and strong adaptability, which is convenient for proliferation, utilization and development.
Description
Technical field
The invention belongs to the microorganism strains of toxic substance in improvement waste water or the environment and its purposes, relevant with a kind of pseudomonas of seed selection with its application purpose technical field.
Background technology
Phenolic compound is that the important pollutent, particularly chlorinated phenol of coal, oil and coking industry discharging is widely used as wood preservation, rust-preventive agent, mycocide and general agrochemical.The solubleness of this compounds in water is big, is a big class of poisonous foreign compound.Repair the attention that causes society to the processing that contains chlorophenol waste water and to the environment that is polluted by chlorophenol.2,4-two chlorophenols are used for production Wood preservation agent Pentachlorophenol and weedicide 2 in a large number, the 4-dichlorphenoxyacetic acid (2,4-D) with 2,4,5-trichlorophenoxyacetic acid (2,4,5-T), these two kinds of compounds have toxicity (the S.L.Seuferer et al of height, Pestic.Biochem.Physiol.15:213-221,1979).2,4-two chlorophenols are released to environment by direct discharging, and chlorinated aromatic compound 2,4 dichlorophenoxyacetic acid and 2,4 also produces 2 in the microbiological deterioration process of 5-trichlorophenoxyacetic acid, and 4-two chlorophenols also are released to environment.Country such as the U.S., China is 2, and 4-two chlorophenols are listed objectionable impurities or priority pollutant " Black List " in the water in.China sewage comprehensive discharge standard GB 8978-1996 regulation, in pollutant discharging unit's sewage effluent of building later on January 1st, 1,998 2,4-two chlorophenols must be lower than 1.0mg/L.
The toxicity of chlorophenol increases with the increase of chlorination degree, and the toxicity of chlorophenol is that also it can only be by the microbiological degradation of minority in addition.Up to the present, the isolated Pseudomonas B of people
13Degradable 2-chlorophenol, 3-chlorophenol and 4-chlorophenol (Schmidt, E., M.Hellwing and H.J.Knackmuss.Appl.Environ.Microbiol.46:1038-1044 (1983)), Azotobacter sp bacterium and Pseudomonas pickettii degradable 2,4,6-trichlorophenol (Deng-Yu Li, Jurgen eberspacher et al., Appl.Environ.Microbiol., 57:1920-1928 (1991); Hohzoh Kiyohara, Takashi Hatta et al., Appl.Environ.Microbiol., 58:1276-1283 (1992)), Pseudomonas cepacia AC1100 degradable Pentachlorophenol (Kams, J.S., J.J.Kilbane, S.Duttagupta and A.M.Chakrabry., Appl.Environ.Microbiol., 46:1176-1181 (1993)).
Studies show that in a large number under aerobic condition, the lower aromatic compound of some chloro degree is than readily biodegradable, degradation rate is also very fast; And higher aromatic compound of some chloro degree such as Pentachlorophenol and biphnyl polychloride are just than difficult for biological degradation (B.E.Haigler et al, Appl.Environ.Microbiol.55 (2): 372-379 (1989)).Under anaerobic, 2, the degradation speed of 4-two chlorophenols is slow, and it is not thorough to degrade, as reports such as GERT-WIELAND KOHRINGD, under anaerobic 2,4-two chlorophenol dechlorinations and change into 4-chlorophenol (GERT-WIELAND KOHRINGD etc., Appl.Environ.Microbiol, 55:348-353 (1989); GERT-WIELAND KOHRINGD et al, Appl.Environ.Microbiol.55:2735-2737 (1989)).
2,4-D (2,4-dichlorphenoxyacetic acid) is from finding so far surplus in the of existing 50 year that it is mainly used in cereal Tanaka and removes broadleaved herb.When having sunlight or UV-light to exist, 2,4-D can be decomposed to form the intermediate product oxyhydroquinone, and then continues to be oxidized to class humic acids (Tang remove silly etc. write chemistry of pesticide, press of Nankai University, 1998).2, when 4-D is degraded by microorganisms, produce 2,4-two chlorophenols further are degraded again.
Phenylformic acid and Whitfield's ointment (salicylic acid) reach and all belong to aromatic compound, all than difficult for biological degradation.
File an application day by the end of the present invention and end, do not see relevant programmed screening seed selection degradable 2 as yet, 4-two chlorophenols, 2, the report of the pseudomonad strain of 4-D, phenylformic acid and salt thereof and Whitfield's ointment and salt thereof and its purposes aspect.
Summary of the invention
The objective of the invention is to, overcome the defective of prior art, by the domestication, aromatic compound in separation screening aerobic degradation waste water or the environment and chlorinated aromatic compound, particularly degrade 2, the bacterial strain of 4-two chlorophenols is used for processing and contains 2,4-two chlorophenols, 2, the waste water of chlorinated aromatic compound such as 4-D and phenylformic acid and aromatic compounds such as salt and Whitfield's ointment and salt thereof thereof, and being used for by 2, the environment that chlorinated aromatic compound such as 4-two chlorophenols and aromatic compound pollute is repaired.
The present invention is achieved through the following technical solutions:
Residual chlorinated aromatic compound and aromatic compound in degrading waste water or the environment, particularly 2,4-two chlorophenols, 2, (2,4-D), the bacterial strain of phenylformic acid and salt and Whitfield's ointment and salt thereof, described bacterial strain is a kind of pseudomonads GT 241-1-1 to the 4-dichlorphenoxyacetic acid, (Pseudomonas sp), be deposited in CCTCC, deposit number CCTCC No.M200039, preservation date on December 6th, 2000.
Utilize aforesaid pseudomonas, it is characterized in that in degraded 2, the application in 4-two chlorophenols, 2,4 dichlorophenoxyacetic acid, phenylformic acid and salt thereof and Whitfield's ointment (salicylic acid) and the salt thereof.
Below the invention will be further described:
The characteristic of the bacterial strain that used strains separation screening method, degradability measuring method and the institute's separation screening of the present invention obtains is as follows:
One, used substratum during strains separation screening method, degradability are measured:
1, inorganic salt liquid culture medium prescription: K
2HPO
41.73 gram, KH
2PO
40.68 gram, NH
4NO
31.0 gram, MgSO
47H
2O 0.1 gram, MnSO
4H
2O 0.03 gram, FeSO
47H
2O 0.03 gram, CaCl
22H
2O 0.02 gram, water 1000ml, pH7.4.Inorganic salt solid medium: for the inorganic salt liquid substratum adds agar 1.6%.
2, LA culture medium prescription: tryptone 10 grams, yeast extract 5 grams, sodium-chlor 10 grams, agar 15～20 grams, 1000 milliliters in water, pH7.2.
Two, the separating screening method of bacterial strain GT241-1:
1, soil sample and domestication:
From Zhejiang, the production 2 on ground such as Jiangsu, Shanghai, Henan, 4-two chlorophenols and related products 2, the sewage draining exit soil sampling in the chemical plant of 4-D etc. is mixed soil sample.Mixed soil sample is positioned over 20～30 ℃, suitable moisture and good aeration condition is provided, and often a certain amount of 2, of the domestication of 4-two chlorophenols so that wherein microorganism is carried out to wherein adding.After processing in three months, promptly begin from this soil sample, to separate 2 4-two chlorophenol degradation bacteria strains.
2,2, the separation screening of 4-two chlorophenol degradation bacteria strains:
Adopt and add 2, the selectivity cultural method of 4-two chlorophenols separates 2,4-two chlorophenol degradation bacteria strains.
In 100 milliliters of inorganic salt liquid substratum, add through about 10 grams of the earth of domestication and 2,10 milligram (2 of 4-two chlorophenol, 4-two chlorophenols are mixed with the storage liquid of 4 mg/ml earlier, in the process for preparation earlier with NaOH with 2, the dissolving of 4-two chlorophenols, add HCl again with solution furnishing neutrality), put 30 ℃ of shakings and cultivate a week.Get the 10ml nutrient solution and add in 90 milliliters of inorganic salt liquid substratum, add 2,12 milligrams of 4-two chlorophenols, put 30 ℃ again shaking cultivate a week.Proceed third and fourth selectivity of taking turns afterwards as stated above and cultivate, increase by 2, the working concentration of 4-two chlorophenols adds 2 respectively, 15,20 milligrams of 4-two chlorophenols.Get 0.1 milliliter of nutrient solution and coat and add 2, the inorganic salt solid medium flat board of 4-two chlorophenols 80～200 mg/litre is put 28～30 ℃ of cultivations, grows bacterium colony in 4～7 days.Bacterium colony is streak culture in adding 2, and the LA substratum of 4-dioxy phenol 100～200 mg/litre carries out purifying.By measuring the purifying bacterial strain to 2, the degradation capability of 4-two chlorophenols is done further checking.
Three, the biological characteristics of bacterial strain GT241-1:
Bacterial strain GT241-1 is a bacillus, and flagellum is extremely living, Gram-negative, and glucose oxidase produces acid, the oxidase test positive, the catalase test positive, the nitrate reduction positive.Growth temperature range is 15-38 ℃, and optimum growth temperature is 25-30 ℃.The pH scope of growing in the LB substratum is pH5.5-9.5.Utilize 2, the concentration range of 4-two chlorophenols: be below the 500mg/L in the inorganic salt liquid substratum; Be below the 1000mg/L on the LA flat board.
Bacterial strain GT241-1 has resistance to penbritin, tsiklomitsin, erythromycin and Hg, can on the penbritin, tsiklomitsin, erythromycin and the Hg LA flat board that add 100mg/L, 30mg/L, 50mg/L and 60mg/L, grow respectively, but it is to the kantlex non-resistant.
Four, bacterial strain GT241-1 is to 2, the degradation property and the testing method of 4-two chlorophenols etc.:
1, to 2, degraded of 4-two chlorophenols and utilization:
(1) degradability is measured: bacterial strain GT241-1 is after spreading cultivation on the LA flat board, be inoculated in and add 80～120 mg/litre analytically pure 2, the inorganic salt liquid substratum of 4-two chlorophenols, centrifugal after cultivating 2～4 days, measure the light absorption value (light absorption value reduces with the prolongation of incubation time) of 286nm place clear liquid, the reference standard curve determines wherein 2, the content of 4-two chlorophenols.
(2) utilize 2,4-two chlorophenols are sole carbon source growth test method: with inorganic salt solid medium moist heat sterilization (121 ℃) 30 minutes, treat to add when temperature is reduced to 50 ℃ of left and right sides 20～800mg/L 2,4-two chlorophenols shake up the back and fall dull and stereotyped.Picking examination lawn, streak inoculation are put 28 ℃ and were cultivated 4～7 days in the solid medium planar surface.
(3) to 2,4-two chlorophenols are degraded and utilized characteristic: bacterial strain GT241-1 is to 2, and the suitableeest degradation temperature of 4-two chlorophenols is 25-30 ℃, degradative phase: be 18-48h under 25-30 ℃ of temperature; Under 15 ℃ of temperature, 48h begins degraded.Can be at bacterial strain GT241-1 under the suitableeest degradation temperature with 2 of 90mg/L in 48 hours, 4-two chlorophenols degraded 91%.Degradation solution is analyzed with ethyl acetate extraction, concentrated laggard capable chromatograph-mass spectrometer coupling (GC/MS), confirmed that bacterial strain GT241-1 can be with 2,4-two chlorophenols are degraded fully, do not accumulate mesostate in degradation process.
Bacterial strain GT241-1 can be 2, and 4-two chlorophenols are to grow on the inorganic salt solid medium of sole carbon source.Utilize 2, the scope of 4-two chlorophenols is 20～800mg/L.Grow bacterium colony in 28 ℃ of cultivations after 2 days, single bacterium colony size is about 0.1～0.4mm.
2, to 2, the utilization (degraded) of 4-D, phenylformic acid or its salt, Whitfield's ointment or its salt:
(1) utilize 2,4-D, phenylformic acid or its salt, Whitfield's ointment or its salt are sole carbon source growth test method:
With inorganic salt solid medium moist heat sterilization (121 ℃) 30 minutes, treat that temperature adds 2 of 100～2000mg/L, 100～8000mg/L and 100～2000mg/L respectively when reducing to 50 ℃ of left and right sides, 4-D, phenylformic acid or its salt and Whitfield's ointment or its salt shake up the back and fall dull and stereotyped.Bacterial strain GT241-1 is adding 2 of 80～200mg/L, and after the LA media surface enlarged culturing of 4-two chlorophenols and the activation, streak inoculation is put 28 ℃ and cultivated 2～7 days in this solid medium planar surface.
(2) utilize situation; Bacterial strain GT241-1 can utilize 2, and 4-D, phenylformic acid and salt thereof, Whitfield's ointment and salt thereof are the sole carbon source growth.Utilize concentration to be respectively 100～1500mg/L, 100～8000mg/L and 100～1200mg/L.When interpolation 100～6000mg/L Sodium Benzoate is the sole carbon source growth, cultivates and grew plentiful bacterium colony in 1 day; Adding 100～1500mg/L 2, when 4-D, sodium salicylate are the sole carbon source growth, are cultivating and grew bacterium colony in 2～4 days, single colony diameter is 0.1～0.4mm.
Accompanying drawing and explanation thereof
Fig. 1: be pseudomonads GT 241-1 of the present invention-1 pairs 2, the degraded typical curve of 4-two chlorophenols.Ordinate zou represents 2 among the figure, the concentration of 4-two chlorophenols; The light absorption value that the X-coordinate representative records under 286nm.
Fig. 2: be pseudomonads GT 241-1 of the present invention-1 pairs 2, the degradation curve of 4-two chlorophenols.The light absorption value that the ordinate zou representative records under 286nm among the figure; X-coordinate is represented degradation time; Ck represents contrast.
Good effect of the present invention is:
2,4-Dichlorophenol is a kind of poisonous, difficult degradation chlorinated aromatic compound, is that efforts at environmental protection person faces to the processing that contains chlorophenol waste water Important topic. The degraded 2 of separation screening of the present invention, existing stronger aerobic the falling of the pseudomonad strain GT241-1 of 4-Dichlorophenol Separate the ability of 2,4-Dichlorophenol, can also degrade other aromatic such as benzoic acid and salt thereof and salicylic acid and salt thereof and chloro fragrance Compound is such as 2,4-D, its stable performance. Bacterial strain GT241-1 takes a sample from the sewage draining exit of producing 2,4-Dichlorophenol and Related product, And process is tamed and purifies and separates is come out, toxic environment there is certain adaptive capacity, be expected in containing the chlorophenol wastewater treatment, send out Wave important function.
Embodiment
The separation screening of embodiment one, bacterial strain GT241-1
(1) soil sample collection and domestication
From Zhejiang, the production 2 on ground such as Jiangsu, Shanghai, Henan, each soil sampling 500 gram such as the sewage draining exit in the chemical plant of 4-two chlorophenols and related products 2,4 dichlorophenoxyacetic acid etc. are with it mixing.Mixed soil sample is positioned over 20～30 ℃, suitable moisture and good aeration condition is provided, and often a certain amount of 2 to wherein adding, and 4-two chlorophenols are to tame microorganism wherein.After the some months processing, promptly begin from this soil sample, to separate 2 4-two chlorophenol degradation bacteria strains.The residue soil sample continues to preserve as stated above and tame.
(2) strains separation screening
Separation screening inorganic salt liquid substratum: K
2HPO
41.73 gram, KH
2PO
40.68 gram, NH
4NO
31.0 gram, MgSO
47H
2O 0.1 gram, MnSO
4H
2O 0.03 gram, FeSO
47H
2O 0.03 gram, CaCl
22H
2O 0.02 gram, water 1000ml, pH7.4.Inorganic salt solid culture based formulas: for above-mentioned inorganic salt liquid substratum adds agar 1.6% again.LA culture medium prescription: tryptone 10 grams, yeast extract 5 grams, sodium-chlor 10 grams, agar 15～20 grams, 1000 milliliters in water, pH7.2.All adopted moist heat sterilization (121 ℃) 30 minutes after the above substratum preparation.
The domestication of bacterial strain selectivity is cultivated and is separated: add in 100 milliliters of inorganic salt liquid substratum through about 10 grams of the earth of domestication and 2,10 milligram (2 of 4-two chlorophenol, 4-two chlorophenols are mixed with the storage liquid of 4 mg/ml earlier, in the process for preparation earlier with NaOH with 2, the dissolving of 4-two chlorophenols, add HCl again with solution furnishing neutrality), put 30 ℃ of shakings and cultivate a week.Get 10ml nutrient solution (supernatant liquid) and add in 90 milliliters of inorganic salt liquid substratum, add 2,12 milligrams of 4-two chlorophenols, a week is cultivated in 30 ℃ of shakings.Proceed third and fourth selectivity of taking turns afterwards as stated above and cultivate, increase by 2, the working concentration of 4-two chlorophenols adds 2 respectively, 15,20 milligrams of 4-two chlorophenols.Get 0.1 milliliter of nutrient solution and coat and add 2, the inorganic salt solid medium flat board of 4-two chlorophenols 80～200 mg/litre is put 28～30 ℃ of cultivations, grows bacterium colony in 4～7 days.
The bacterial strain purifying: with LA substratum heat fused, treat to add 2 when temperature is reduced to 50 ℃ of left and right sides, 4-two chlorophenols 100 mg/litre shake up the back and fall dull and stereotyped.The bacterium colony that picking grows on inorganic salt solid medium flat board, streak inoculation are put 28 ℃ in its surface, cultivate 24～48 hours.Picking list bacterium colony, purifying one to three time once more as stated above, until be defined as behind the bacterium colony homogeneous that grows at last, the microscopy purebred till.
2,4-two chlorophenol degradation capabilities are measured: it is K that degradability is measured with the inorganic salt liquid culture medium prescription
2HPO
41.73 gram, KH
2PO
40.68 gram, NH
4NO
31.0 gram, MgSO
47H
2O 0.1 gram, MnSO
4H
2O 0.03 gram, FeSO
47H
2The O0.03 gram, CaCl
22H
2O 0.02 gram, water 1000ml, pH7.4.Moist heat sterilization (121 ℃) 30 minutes.2,4-two chlorophenols add before inoculation.With transfering loop picking 2～3 ring purifying bacterial strains, be inoculated in above-mentioned interpolation 2, in the inorganic salt liquid substratum of 4-two chlorophenols, put 28 ℃ of shakings and cultivated 2～7 days.With medium centrifugal, measure the light absorption value of 286nm place clear liquid, determine 2 according to typical curve, the content of 4-two chlorophenols.Typical curve is seen shown in Figure 1.
Embodiment two, slant strains and cultivation thereof:
Culture medium prescription: tryptone 10 grams, yeast extract 5 grams, sodium-chlor 10 grams, agar 15～20 grams, 1000 milliliters in water, pH7.2.Substratum packing test tube (18 * 180mm), loading amount 10ml, tampon beyond the Great Wall, moist heat sterilization (121 ℃) 30 minutes.When treating that temperature is reduced to 50 ℃ of left and right sides, add 2 in every test tube, 1～1.5 milligram of 4-two chlorophenol are put into the inclined-plane after shaking up.
Adopt aseptic technique inoculation GT241-1 bacterial strain in the test tube slant, cultivated 24～48 hours for 28 ℃, the visual inspection lawn is plentiful.Putting 4 ℃ can preserve three months to six months.
Embodiment three, bacterial strain GT241-1 be with 2, and 4-two chlorophenols are the growth on the inorganic salt solid medium of sole carbon source
Inorganic salt solid culture based formulas: K
2HPO
41.73 gram, KH
2PO
40.68 gram, NH
4NO
31.0 gram, MgSO
47H
2O 0.1 gram, MnSO
4H
2O 0.03 gram, FeSO
47H
2O 0.03 gram, CaCl
22H
2O 0.02 gram, agar 15～20 grams, water 1000ml, pH7.4.Substratum moist heat sterilization (121 ℃) 30 minutes.With 2 of 20-1000mg/L, 4-two chlorophenols, are treated to add 2 when temperature is reduced to 50 ℃ of left and right sides with above-mentioned solid medium heat fused in adding before the flat board, and 4-two chlorophenols shake up flat board of back.Get the test tube slant bacterial classification, streak inoculation is put 28 ℃ and was cultivated 4～7 days in the solid medium planar surface.
Bacterial strain GT241-1 can be with 2, and 4-two chlorophenols are the growth on the inorganic salt solid medium of sole carbon source.It utilizes 2, and the concentration range of 4-two chlorophenols is 20～800mg/L, is lower than 20mg/L and is higher than 800mg/L all can not grow.Utilize 2, the suitable concentration range of 4-two chlorophenols is 60～300mg/L, and the optimal concentration scope is 80～200mg/L.Under optimum concentration, bacterial strain GT241-1 bacterium colony occurs two days later in 28 ℃ of cultivations; Cultivate after 4～7 days, single bacterium colony size is about 0.1～0.4mm.
Embodiment four, bacterial strain GT241-1 under liquid condition to 2, the degraded of 4-two chlorophenols:
(1) bacterial strain GT241-1 is in LA media surface enlarged culturing
LA culture medium prescription: tryptone 10 grams, yeast extract 5 grams, sodium-chlor 10 grams, agar 15～20 grams, 1000 milliliters in water, pH7.2.Substratum moist heat sterilization (121 ℃) 30 minutes.With above-mentioned solid medium heat fused, to treat to add 2 when temperature is reduced to 50 ℃ of left and right sides, 4-two chlorophenols shake up the back and fall dull and stereotyped.Get the test tube slant bacterial classification, streak inoculation is put 28 ℃ and was cultivated 24～48 hours in solid LA culture medium flat plate surface.
(2) to 2, the degraded of 4-two chlorophenols:
Degraded test medium prescription: K
2HPO
41.73 gram, KH
2PO
40.68 gram, NH
4NO
31.0 gram, MgSO
47H
2O 0.1 gram, MnSO
4H
2O 0.03 gram, FeSO
47H
2O 0.03 gram, CaCl
22H
2O 0.02 gram, water 1000ml, pH7.4.Moist heat sterilization (121 ℃) 30 minutes.With 2 of 20-800mg/L, 4-two chlorophenols add before inoculation.With transfering loop picking 2～3 ring test tube slant bacterial classifications, be inoculated in above-mentioned interpolation 2, in the degraded test medium of 4-two chlorophenols, put 28 ℃ of shakings and cultivated 2 in the culturing process in the sampling and measuring nutrient solution, 4-two chlorophenol content 4～7 days.Method is: with medium centrifugal, measure the light absorption value (light absorption value reduces with the prolongation of incubation time) of 286nm place clear liquid, determine 2 according to typical curve, the content of 4-two chlorophenols.
Under above-mentioned liquid condition, 2 below the bacterial strain GT241-1 degradable 500mg/L, 4-two chlorophenols.The degradation temperature scope is 15-38 ℃, and the suitableeest degradation temperature is 25-30 ℃.Under the suitableeest degradation temperature, it can be with 2 of 90mg/L in 48hr, 4-two chlorophenols degraded 91%.Typical degradation curve is seen shown in Figure 2.Chromatogram one mass spectrometry (GC/MS) analysis revealed, this bacterial strain can be with 2, and 4-two chlorophenols are thoroughly degraded, and do not accumulate mesostate.
Embodiment five, bacterial strain GT241-1 be to 2, the utilization of 4-D, Sodium Benzoate and sodium salicylate (degraded):
(1) bacterial strain GT241-1 is adding 2, the LA media surface enlarged culturing and the activation of 4-two chlorophenols:
LA culture medium prescription: tryptone 10 grams, yeast extract 5 grams, sodium-chlor 10 grams, agar 15～20 grams, 1000 milliliters in water, pH7.2.Substratum moist heat sterilization (121 ℃) 30 minutes.With above-mentioned solid medium heat fused, to treat to add 2 when temperature is reduced to 50 ℃ of left and right sides, 4-two chlorophenols shake up the back and fall dull and stereotyped.Get the test tube slant bacterial classification, streak inoculation is put 28 ℃ and was cultivated 24～48 hours in this solid medium planar surface.
(2) to 2, the utilization of 4-D, Sodium Benzoate and sodium salicylate (degraded):
Inorganic salt solid culture based formulas: K
2HPO
41.73 gram, KH
2PO
40.68 gram, NH
4NO
31.0 gram, MgSO
47H
2O 0.1 gram, MnSO
4H
2O 0.03 gram, FeSO
47H
2O 0.03 gram, CaCl
22H
2O 0.02 gram, agar 15～20 grams, water 1000ml, pH7.4.Substratum moist heat sterilization (121 ℃) 30 minutes.With above-mentioned solid medium heat fused, treat to add respectively when temperature is reduced to 50 ℃ of left and right sides 2 of 100～2000mg/L, 100～8000mg/L and 100～2000mg/L, 4-D, Sodium Benzoate and sodium salicylate shake up the back and fall dull and stereotyped as sole carbon source.Streak inoculation is in adding 2, and the LA media surface enlarged culturing of 4-two chlorophenols and activatory bacterial strain GT241-1 put 28 ℃ and cultivated 2～7 days in this solid medium planar surface.
Bacterial strain GT241-1 can utilize 2, and 4-D, Sodium Benzoate and sodium salicylate are the sole carbon source growth.Utilize concentration to be respectively 100～1500mg/L, 100～8000mg/L and 100～1200mg/L.When the interpolation Sodium Benzoate is the sole carbon source growth, cultivates and grew plentiful bacterium colony in 1 day; Adding 2, when 4-D, sodium salicylate are the sole carbon source growth, are cultivating and grew bacterium colony in 2～4 days, single colony diameter is 0.1～0.4mm.
Claims (2)
1, residual chlorinated aromatic compound and aromatic compound in a kind of degrading waste water or the environment, particularly 2,4-two chlorophenols, 2, the bacterial strain of 4-dichlorphenoxyacetic acid, phenylformic acid and salt thereof and Whitfield's ointment and salt thereof, it is characterized in that, described bacterial strain is a kind of pseudomonas (Pseudomonas sp) GT241-1, CCTCC NO.M200039.
2, the described bacterial strain of claim 1 is in degraded 2, the application in 4-two chlorophenols, 2,4 dichlorophenoxyacetic acid, phenylformic acid and salt thereof and Whitfield's ointment and the salt thereof.
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| CN100391867C (en) * | 2005-11-04 | 2008-06-04 | 中国地质大学（武汉） | Application of chlorine resisting strain No.3 |
| CN102732440B (en) * | 2011-04-12 | 2014-04-23 | 华中农业大学 | Pseudomonas putida T2-2 able to reduce nitrosamines in flue-cured tobacco leaves and application thereof |
| CN108611290A (en) * | 2016-12-13 | 2018-10-02 | 香港城市大学深圳研究院 | The screening technique and used medium of left drug degradation bacteria in sewage |
| CN107941938B (en) * | 2017-11-19 | 2018-11-16 | 浙江大学 | Bacterium polycyclic aromatic hydrocarbon metabolite high efficiency extraction and degradation pathway determination method |
| CN108823122B (en) * | 2018-06-05 | 2022-03-11 | 常州大学 | 2,4-dichlorophenol degrading bacterium and degrading method |
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