CN101935622A - Pseudomonas strain and application thereof - Google Patents
Pseudomonas strain and application thereof Download PDFInfo
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- CN101935622A CN101935622A CN2010101084561A CN201010108456A CN101935622A CN 101935622 A CN101935622 A CN 101935622A CN 2010101084561 A CN2010101084561 A CN 2010101084561A CN 201010108456 A CN201010108456 A CN 201010108456A CN 101935622 A CN101935622 A CN 101935622A
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Abstract
The invention discloses a pseudomonas strain and application thereof. The strain provided by the invention is pseudomonas sp. SG-1 CGMCC No.3556. The microbial inoculum provided by the invention contains the active ingredient of pseudomonas sp. SG-1 CGMCC No.3556. The strain provided by the invention has a wider degradation spectrum and reaches the degradation rates to various toxic compounds over 75 percent. The microbial inoculum of the invention has the advantages of low production use cost, convenient use, wider degradation spectrum and good removal effect. The strain and the microbial inoculum provided by the invention are suitable for the biological treatment of Cr-contained wastewater generated in a tanning and dyeing process and wastewater generated in other petrochemical processes and can ensure that the toxic substance and Cr (VI) contents in the wastewater accord with the wastewater emission standard. The invention successfully solves the problem of poor toxic substance removal effect in a Cr-contained organic wastewater treatment process, reduces the workload in the production and use processes, decreases the production and use costs and has important meanings to protect the ecological environment and keep the body health of people.
Description
Technical field
The present invention relates to a pseudomonas bacterial strain and an application thereof.
Background technology
Aniline, phenol, cresols, para-chlorophenol, 1-naphthols, naphthalene and derivative thereof are important chemical material, discover that these compounds all have bigger harm to human and environmental organism.Chromium (VI) is the important composition of many wastewater from chemical industry, and chromium (VI) has tangible toxic action to human body; Simultaneously, the existence of chromium (VI) has inhibition significantly to the biological removal effect of organic pollutant in the waste water, and general biological treating is difficult to make the water outlet qualified discharge of the waste water that contains mentioned component.Microorganism is because of having powerful degradation function and stronger adaptability, and often is used as the important tool that environmental pollution is repaired.
Summary of the invention
The purpose of this invention is to provide a pseudomonas bacterial strain and an application thereof.
Pseudomonas provided by the invention (Pseudomonas sp.), name is called SG-1, belong to Rhodopseudomonas (Pseudomonas), separation is in wastewater treatment equipment, be preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center on December 29th, 2009 and (be called for short CGMCC, the address is: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City), preserving number is CGMCC No.3556.
The present invention also protects a kind of microbial inoculum, and its activeconstituents is pseudomonas (Pseudomonassp.) SG-1CGMCC No.3556.
The application in the degraded toxic substance of described bacterial strain and/or described microbial inoculum also belongs to protection scope of the present invention; Described toxic substance is at least a in phenol, ortho-methyl phenol, m-methyl phenol, p-methyl phenol, para-chlorophenol, naphthalene, 1-naphthols and the aniline.
The application in removing chromium (VI) of described bacterial strain and/or described microbial inoculum also belongs to protection scope of the present invention.
Described chromium (VI) claims sexavalent chrome again.
The present invention also protects a kind of method of the toxic substance of degrading, and is with described bacterial strain and/or described microbial inoculum degraded toxic substance; Described toxic substance is at least a in phenol, ortho-methyl phenol, m-methyl phenol, p-methyl phenol, para-chlorophenol, naphthalene, 1-naphthols and the aniline.
The present invention also protects described bacterial strain and/or the application of described microbial inoculum in wastewater treatment.
Described waste water specifically can be in chromate waste water that process hides and dyeing process produce and other petrochemical process and produces waste water etc.
The present invention also protects a kind of method of handling waste water, is described bacterial strain and/or described microbial inoculum are added in the described waste water.
Described waste water specifically can be in chromate waste water that process hides and dyeing process produce and other petrochemical process and produces waste water etc.
The technology of producing microbial inoculum is: inclined-plane kind-shake bottle a kinds-seeding tank-production jar-product (packing formulation is liquid bacterial agent or solid absorption microbial inoculum).
Prepare described microbial inoculum with described bacterial strain and specifically can adopt following steps:
1, bacterial strain SG-1 (original seed) is activated on culture dish, and measure degradation property, be inoculated on the test tube slant standby.
2, the test tube kind is inoculated in 1000ml and shakes in the bottle (containing the 200ml broth culture), constant-temperature shaking culture is prepared the inoculation seeding tank to logarithmic phase.
3, preparation fermention medium (glucose 0.8%, (NH
4)
2SO
41%, K
2HPO
40.2%, MgSO
40.05%, NaCl 0.01%, CaCO
30.3%, yeast extract paste 0.02%, pH value 7.2-7.5), the 400L fermention medium is added the 500L seeding tank, 121 ℃ of high pressure moist heat sterilizations, after being cooled to 33 ℃, will shaking bottle bacterial classification and inoculate into seeding tank, be cultured to logarithmic phase by the inoculum size of 10% (volumn concentration), stirring velocity is 220 rev/mins, and sterile air feeding amount is 1: 0.8 (volume ratio).
4, produce jar (producing 5 tons of a tankages) used medium component identical with fermention medium (4.5 tons of charging capacitys), the production jar 1.1kg/cm after feeding intake
2Pressure under, 121 ℃ of high pressure moist heat sterilizations, below the sterilization postcooling to 35 ℃, logical sterile air keeps sterile state standby; The seed liquor that arrives logarithmic phase is produced jar by the inoculum size access of 10% (volumn concentration), a postvaccinal production jar temperature is controlled at 30-35 ℃, the air flow of sterile air is 1 in the culturing process of production jar: (0.6-1.2), stirring velocity is 180-240 rev/min (as 240 rev/mins), and the whole process flow incubation time is 48-60 hour; After the fermentation ends thalline quantity reach 1,000,000,000/more than the ml.
5, fermentation is finished the back nutrient solution and is gone out jar and directly be distributed into liquid dosage form with plastic barrel or packing bottle or adopt peat to adsorb and be distributed into the solid fungicide formulation with packing bag.
Bacterial strain SG-1 provided by the invention has stronger resistance and adaptability to chromium and other hazardous and noxious substances, can in waste water, survive for a long time and work, has wider degraded spectrum, compounds such as aniline degradation, phenol, ortho-methyl phenol, m-methyl phenol, p-methyl phenol, para-chlorophenol, naphthalene, 1-naphthols simultaneously, degradation rate reaches more than 75%, and can reduce chromium (VI), reach the effect of dechromisation (VI).Microbial inoculum of the present invention has low, easy to use, the wider degraded spectrum of production use cost, advantage that removal effect is good.Bacterial strain provided by the invention and microbial inoculum can be used for the organic toxic composition in the degrading waste water and remove chromium (VI), be applicable to the chromate waste water that process hides and dyeing process are produced, and produce aniline in the waste water in other petrochemical process, phenol, ortho-methyl phenol, m-methyl phenol, p-methyl phenol, para-chlorophenol, naphthalene, the biological treatment of organic pollutants such as 1-naphthols and chromium (VI) can guarantee the aniline in the waste water, phenol, ortho-methyl phenol, m-methyl phenol, p-methyl phenol, para-chlorophenol, naphthalene, organism such as 1-naphthols and chromium (VI) content meet wastewater discharge standard.The present invention has successfully solved and has contained the not high problem of toxic substance removal effect in the chromium treatment of Organic Wastewater process; reduced the workload in production and the use; reduced production and use cost, for preserving the ecological environment, the protection people health has great importance.
Embodiment
Following embodiment is convenient to understand better the present invention, but does not limit the present invention.Experimental technique among the following embodiment if no special instructions, is ordinary method.Used test materials among the following embodiment if no special instructions, is to buy from routine biochemistry reagent shop and obtains.Below experiment all is provided with three repetitions, results averaged." % " in following examples if no special instructions, is the quality percentage composition.
Degradation rate (clearance)=(amount/amount of compound when reaction is initial of compound when the 1-reaction finishes) * 100%.
Each organic method for measurement of concentration in the salt culture medium of basis: get 0.5ml liquid, add (the fully vibration of isopyknic methyl alcohol, make compound dissolution), be equipped with the high performance liquid chromatography of UV-detector to detect with using behind the filtering with microporous membrane of 0.22 μ m; Testing conditions is as follows: chromatographic column, C18 reversed-phase column (4.6mm * 150mm, 5 μ m); Moving phase, methyl alcohol: water=5: 2, volume ratio; Flow velocity is 0.7ml/min; Applied sample amount is 20 μ l; The detection of each compound (quantitatively) wavelength: phenol, 271nm; Cresols (m-methyl phenol, ortho-methyl phenol, p-methyl phenol), 276nm; Para-chlorophenol, 281nm; The 1-naphthols, 280nm; Aniline, 230nm; Naphthalene, 218nm.Concentration quantitatively adopts external standard method.
The method for measurement of concentration of phenol and cresols (m-methyl phenol, ortho-methyl phenol, p-methyl phenol) in the coking chemical waste water (or leather-making waste water): get 0.5ml liquid, add isopyknic methyl alcohol (fully vibration), be equipped with the high performance liquid chromatography of UV-detector to detect with using behind the filtering with microporous membrane of 0.22 μ m; Testing conditions is as follows; Chromatographic column, C18 reversed-phase column (4.6mm * 150mm, 5 μ m); Moving phase, methyl alcohol: water, the original volume ratio is 2: 5, at the uniform velocity is adjusted into 5: 2 in 15min, adjusts to initial ratio after keeping 2min; Flow velocity is 0.7ml/min; Applied sample amount is 20 μ l; The detection of each compound (quantitatively) wavelength is as follows: phenol, 271nm; Cresols, 276nm.Concentration quantitatively adopts external standard method.
Diphenyl carbazide spectrophotometry (GB 7467-87) is adopted in chromium in the leather-making waste water (VI) concentration determination.
The acquisition of embodiment 1, bacterial strain SG-1 and evaluation
One, the acquisition of bacterial strain SG-1
In August, 2008 from Shaoguan, Guangdong get coking chemical waste water active sludge 2g join the basic salt culture medium that contains phenol cultivate 7 days after, the dilution back is coated with containing on the basic salt culture medium flat board of phenol, cultivates after 7 days, picking list bacterium colony is rule and is preserved behind the purifying.
Two, the evaluation of bacterial strain SG-1
Morphological specificity: thalline is shaft-like.
Physiological and biochemical property: main biological characteristics is G
-, the Gram-negative bacterium; It is positive that nitrate reduction and glucose fermentation produce acid; Catalase reaction, gelatin hydrolysis, nitrous acid reduction reaction, clark and Lubsreaction, oxydase, V.P. react, indole reaction is negative; Utilize citric acid, can not utilize tartrate and propionic salt, can not hydrolyzed starch
Molecular Identification result: the partial sequence of 16S rDNA is seen the sequence 1 of sequence table.
Three, the preservation of bacterial strain SG-1
By above qualification result, confirm that bacterial strain SG-1 for coming from the new bacterium of Rhodopseudomonas (Pseudomonas), with its called after SG-1, is preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center, preserving number is CGMCC No.3556.
Embodiment 2, bacterial strain in basic salt liquid substratum to the degradation rate of different compounds
Basis salt liquid substratum: with 1.0g NaCl, 1.0g NH
4NO
3, 1.5g K
2HPO
4, 0.5g KH
2PO
4With 0.2g MgSO
47H
2The O water is settled to 1L.
The single bacterium colony of picking SG-1 is in 3ml LB liquid nutrient medium, and 30 ℃, 165 rev/mins shaking culture 24h obtain fresh bacterium liquid.In 100ml solution A (or solution B or solution C or solution D or solution E or solution F or solution G or Solution H), insert the fresh bacterium liquid of 5ml, 30 ℃ of shaking tables are cultivated (170 rev/mins), the degradation rate of (72 hours) sampling and measuring compound after 3 days.The results are shown in Table 1.
The preparation method of solution A: phenol and chromium (VI) are added in the basic salt liquid substratum, and the final concentration of phenol is 100mg/L, and the final concentration of chromium (VI) is 2mg/L.
The preparation method of solution B: ortho-methyl phenol and chromium are added in the basic salt liquid substratum, and the final concentration of ortho-methyl phenol is 100mg/L, and the final concentration of chromium (VI) is 2mg/L.
The preparation method of solution C: m-methyl phenol and chromium (VI) are added in the basic salt liquid substratum, and the final concentration of m-methyl phenol is 100mg/L, and the final concentration of chromium (VI) is 2mg/L.
The preparation method of solution D: p-methyl phenol and chromium (VI) are added in the basic salt liquid substratum, and the final concentration of p-methyl phenol is 100mg/L, and the final concentration of chromium (VI) is 2mg/L.
The preparation method of solution E: para-chlorophenol and chromium (VI) are added in the basic salt liquid substratum, and the final concentration of para-chlorophenol is 100mg/L, and the final concentration of chromium (VI) is 2mg/L.
The preparation method of solution F: naphthalene and chromium (VI) are added in the basic salt liquid substratum, and the final concentration of naphthalene is 100mg/L, and the final concentration of chromium (VI) is 2mg/L.
The preparation method of solution G: 1-naphthols and chromium (VI) are added in the basic salt liquid substratum, and the final concentration of 1-naphthols is 100mg/L, and the final concentration of chromium (VI) is 2mg/L.
The preparation method of Solution H: aniline and chromium (VI) are added in the basic salt liquid substratum, and the final concentration of aniline is 100mg/L, and the final concentration of chromium (VI) is 2mg/L.
Table 1 bacterial strain SG-1 in basic salt liquid substratum to the degradation rate (3 days) of different compounds
| Substrate | Phenol | Ortho-methyl phenol | M-methyl phenol | P-methyl phenol |
| Degradation rate (%) | 100 | 100 | 100 | ＞90 |
| Substrate | Para-chlorophenol | Naphthalene | The 1-naphthols | Aniline |
| Degradation rate (%) | ＞90 | ＞75 | ＞80 | ＞95 |
Embodiment 3, bacterial strain Pyrogentisinic Acid's in leather-making waste water degradation rate and to chromium (VI) clearance
The single bacterium colony of picking SG-1 is in 3ml LB liquid nutrient medium, and 30 ℃, 165 rev/mins shaking culture 24 hours obtain fresh bacterium liquid.Measure the 100ml leather-making waste water, detect the wherein concentration (starting point concentration) of chromium (VI).The about 2mg/L of starting point concentration of chromium (VI).Add phenol in leather-making waste water, making its final concentration is 100mg/L; Insert the fresh bacterium liquid of 4ml, mixing places 30 ℃, and 165 rev/mins shaking table is cultivated, the degradation rate of (72 hours) sampling and measuring compound after 3 days.The results are shown in Table 2.
Table 2 bacterial strain SG-1 Pyrogentisinic Acid's in leather-making waste water degradation rate and to chromium (VI) clearance (3 days)
| Substrate | Phenol | Cr(VI) |
| Degradation rate/clearance (%) | ＞99 | ＞50 |
Embodiment 4, bacterial strain in coking chemical waste water to the degradation rate of different compounds
The single bacterium colony of picking SG-1 is in 3ml LB liquid nutrient medium, and 30 ℃, 165 rev/mins shaking culture 24 hours obtain fresh bacterium liquid.Measure the 100ml coking chemical waste water, detect the wherein concentration (starting point concentration) of phenol and cresols.The starting point concentration of phenol is 400mg/L, and the starting point concentration of cresols is 100mg/L.Insert the fresh bacterium liquid of 2ml in coking chemical waste water, mixing places 30 ℃, and 165 rev/mins shaking table is cultivated, the degradation rate of (72 hours) sampling and measuring compound after 3 days.The results are shown in Table 3.
Table 3 bacterial strain SG-1 is degradation of phenol and cresols (3 days) in coking chemical waste water
| Substrate | Phenol | Cresols |
| Degradation rate (%) | ＞99 | ＞99 |
Sequence table
＜110〉Peking University
＜120〉a pseudomonas bacterial strain and an application thereof
<130>CCGNARY102082
<160>1
<210>1
<211>1500
<212>DNA
＜213〉pseudomonas (Pseudomonas sp.)
<400>1
tagagtttga?tcctggctca?gattgaacgc?tggcggcagg?cctaacacat?gcaagtcgag 60
cggatgagag?gagcttgctc?cttgatttag?cggcggacgg?gtgagtaatg?cctaggaatc 120
tgcctggtag?tgggggataa?cgtccggaaa?cgggcgctaa?taccgcatac?gtcctacggg 180
agaaagcagg?ggaccttcgg?gccttgcgct?atcagatgag?cctaggtcgg?attagctagt 240
tggtgaggta?atggctcacc?aaggcgacga?tccgtaactg?gtctgagagg?atgatcagtc 300
acactggaac?tgagacacgg?tccagactcc?tacgggaggc?agcagtgggg?aatattggac 360
aatgggcgaa?agcctgatcc?agccatgccg?cgtgtgtgaa?gaaggttttc?ggattgtaaa 420
gcactttaag?ttgggaggaa?gggcagtaag?ttaatacctt?gctgttttga?cgttaccgac 480
agaataagca?ccggctaact?tcgtgccagc?agccgcggta?atacgaaggg?tgcaagcgtt 540
aatcggaatt?actgggcgta?aagcgcgcgt?aggtggttcg?ttaagttgga?tgtgaaagcc 600
ccgggctcaa?cctgggaact?gcatccaaaa?ctggcgagct?agagtacggt?agagggtggt 660
ggaatttcct?gtgtagcggt?gaaatgcgta?gatataggaa?ggaacaccag?tggcggaggc 720
gaccacctgg?actgatactg?acactgaggt?gcgaaagcgt?ggggagcaaa?caggattaga 780
taccctggta?gtccacgctg?taaacgatgt?caactagccg?ttgggttcct?tgagaactta 840
gtggcgcagc?taacgcatta?agttgaccgc?ctggggagta?cggccgcaag?gttaaaactc 900
aaatgaattg?acgggggccc?gcacaagcgg?tggagcatgt?ggtttaattc?gaagcaacgc 960
gaagaacctt?acctggcctt?gacatgctga?gaactttcca?gagatggatt?ggtgccttcg 1020
ggaactcaga?cacaggtgct?gcatggctgt?cgtcagctcg?tgtcgtgaga?tgttgggtta 1080
agtcccgtaa?cgagcgcaac?ccttgtcctt?agttaccagc?acgttatggt?gggcactcta 1140
aggagactgc?cggtgacaaa?ccggaggaag?gtggggatga?cgtcaagtca?tcatggccct 1200
tacggccagg?gctacacacg?tgctacaatg?gtcggtacaa?agggttgcca?agccgcgagg 1260
tggagctaat?cccataaaac?cgatcgtagt?ccggatcgca?gtctgcaact?cgactgcgtg 1320
aagtcggaat?cgctagtaat?cgtgaatcag?aatgtcacgg?tgaatacgtt?cccgggcctt 1380
gtacacaccg?cccgtcacac?catgggagtg?ggttgctcca?gaagtagcta?gtctaacctt 1440
cggggggacg?gttaccacgg?agtgattcat?gactggggtg?aagtcgtaac?aaggtaacca 1500
Claims (10)
1. pseudomonas (Pseudomonas sp.) SG-1 CGMCC No.3556.
2. microbial inoculum, its activeconstituents is pseudomonas (Pseudomonas sp.) SG-1 CGMCC No.3556.
3. described bacterial strain of claim 1 and/or the described microbial inoculum of claim 2 application in the degraded toxic substance; Described toxic substance is at least a in phenol, ortho-methyl phenol, m-methyl phenol, p-methyl phenol, para-chlorophenol, naphthalene, 1-naphthols and the aniline.
4. described bacterial strain of claim 1 and/or the described microbial inoculum of claim 2 application in removing chromium (VI).
5. the method for the toxic substance of degrading is with described bacterial strain of claim 1 and/or the described microbial inoculum degraded of claim 2 toxic substance; Described toxic substance is at least a in phenol, ortho-methyl phenol, m-methyl phenol, p-methyl phenol, para-chlorophenol, naphthalene, 1-naphthols and the aniline.
6. described bacterial strain of claim 1 and/or the application of the described microbial inoculum of claim 2 in wastewater treatment.
7. application as claimed in claim 6 is characterized in that: described waste water is chromate waste water.
8. application as claimed in claim 6 is characterized in that: described waste water is coking chemical waste water or leather-making waste water.
9. a method of handling waste water is that described bacterial strain of claim 1 and/or the described microbial inoculum of claim 2 are added in the described waste water.
10. method as claimed in claim 9 is characterized in that: described waste water is coking chemical waste water or leather-making waste water.
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| CN2010101084561A CN101935622B (en) | 2010-02-05 | 2010-02-05 | Pseudomonas strain and application thereof |
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|---|---|---|---|
| CN2010101084561A CN101935622B (en) | 2010-02-05 | 2010-02-05 | Pseudomonas strain and application thereof |
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| CN101935622B CN101935622B (en) | 2012-01-11 |
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| CN102409009A (en) * | 2011-09-19 | 2012-04-11 | 北京大学 | Pseudomonas and application thereof in carbazole degradation |
| CN102476894A (en) * | 2011-10-20 | 2012-05-30 | 常州亚环环保科技有限公司 | Method for processing chromium-containing wastewater by using biological agent |
| CN103243057A (en) * | 2013-05-27 | 2013-08-14 | 江苏南资环保科技有限公司 | Pseudomonas SYA-1 for degrading triethylamine and application of pseudomonas SYA-1 |
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| CN106754512A (en) * | 2016-12-22 | 2017-05-31 | 天津凯英科技发展股份有限公司 | One plant of orthoresol degradation bacteria and its application |
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| CN102409009B (en) * | 2011-09-19 | 2012-12-19 | 北京大学 | Pseudomonas and application thereof in carbazole degradation |
| CN102409009A (en) * | 2011-09-19 | 2012-04-11 | 北京大学 | Pseudomonas and application thereof in carbazole degradation |
| CN102476894A (en) * | 2011-10-20 | 2012-05-30 | 常州亚环环保科技有限公司 | Method for processing chromium-containing wastewater by using biological agent |
| CN103243057A (en) * | 2013-05-27 | 2013-08-14 | 江苏南资环保科技有限公司 | Pseudomonas SYA-1 for degrading triethylamine and application of pseudomonas SYA-1 |
| CN103243057B (en) * | 2013-05-27 | 2015-03-25 | 江苏南资环保科技有限公司 | Pseudomonas SYA-1 for degrading triethylamine and application of pseudomonas SYA-1 |
| CN106244502B (en) * | 2016-09-27 | 2019-06-18 | 湖北大学 | The pseudomonad of one plant of efficient dephosphorization and degradation lecithin |
| CN106244502A (en) * | 2016-09-27 | 2016-12-21 | 湖北大学 | One strain efficient dephosphorization and the pseudomonas of degraded lecithin |
| CN106754512A (en) * | 2016-12-22 | 2017-05-31 | 天津凯英科技发展股份有限公司 | One plant of orthoresol degradation bacteria and its application |
| CN111662851A (en) * | 2020-07-14 | 2020-09-15 | 南京工业大学 | Bacterial strain for oxidizing aromatic compounds and application thereof |
| CN111662851B (en) * | 2020-07-14 | 2022-09-09 | 南京工业大学 | Bacterial strain for oxidizing aromatic compounds and application thereof |
| CN112920967A (en) * | 2020-12-31 | 2021-06-08 | 中南大学 | Microbial agent for reducing chroma of aniline-containing printing and dyeing wastewater, preparation method, method for reducing chroma of aniline-containing printing and dyeing wastewater and application |
| CN118028185A (en) * | 2024-04-12 | 2024-05-14 | 广州大学 | Salt-tolerant spring pseudomonas strain D4, microbial agent and application thereof |
| CN118028185B (en) * | 2024-04-12 | 2024-06-07 | 广州大学 | Salt-tolerant spring pseudomonas strain D4, microbial agent and application thereof |
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| CN101935622B (en) | 2012-01-11 |
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