CN102093970A - Low-temperature aniline-degrading strain - Google Patents
Low-temperature aniline-degrading strain Download PDFInfo
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- CN102093970A CN102093970A CN2010105783550A CN201010578355A CN102093970A CN 102093970 A CN102093970 A CN 102093970A CN 2010105783550 A CN2010105783550 A CN 2010105783550A CN 201010578355 A CN201010578355 A CN 201010578355A CN 102093970 A CN102093970 A CN 102093970A
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Abstract
The invention relates to an aniline-degrading strain, in particular relating to a low-temperature aniline-degrading strain. The low-temperature aniline-degrading strain A1 with the collection number CGMCC No. 5011) belongs to Pseudomonas and was collected in China General Microbiological Culture Collection (CGMCC) on December 1st, 2010. The low-temperature aniline-degrading strain A1 provided by the invention is used for biological enhanced treatment of aniline-containing waste water in low-temperature conditions with 100% aniline degradation rate, no secondary environmental pollution and no need of addition of other chemicals, so that the waste water treatment cost is greatly reduced, and the application prospect is wide. The low-temperature aniline-degrading strain A1 is favorable for solving the problem of aniline-containing waste water biological treatment system in low-temperature conditions, and has high value of application.
Description
Technical field
The present invention relates to a kind of microphenomenon of degradation of aniline.
Background technology
Aniline is important chemical material, mainly as medicine and thiofide, also is the raw material of making resin and coating, and phenyl amines toxic pollutant toxicity is big, difficult degradation, and ecotope is caused severe contamination.Therefore, the biological degradation of research amino benzenes compounds is significant for protection environment and raising human health.
The waste water that low temperature contains aniline belongs to one of unmanageable waste water in the industry.Domestic research to low temperature degrading aniline microorganism is rarely found, be directed to this, aniline efficient degrading bacteria under the screening cold condition is also understood its degradation characteristic, the industrial treatment that can be the low temperature amino benzene analog waste water provides the microbiobacterial agent resource, thereby the problem that solves low temperature amino benzene analog waste water biological treatment system is laid the foundation.
Summary of the invention
The invention provides a kind of low temperature microphenomenon of degradation of aniline; Purpose is the biological reinforced processing that is applied to amino benzene analog waste water, for the low temperature aniline category matter in the degrading waste water biological treatment system is offered help.
The low temperature microphenomenon of degradation of aniline, it is low temperature microphenomenon of degradation of aniline A1, belongs to Rhodopseudomonas (Psendomonas) novel species, in China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation, deposit number is CGMCC No.5011, and preservation date is on December 01st, 2010; It is the Gram-positive aerobic bacteria, and is shaft-like, and length is 3.0 μ m, and wide is 0.8 μ m; On beef-protein medium, form white colony, circular, translucent, smooth surface, it is wavy that the edge is, and is difficult for provoking.
Low temperature microphenomenon of degradation of aniline A1 catalase reacting positive among the present invention, produce the indole test positive, produce the ammonia test positive, methyl red test reaction negative, starch hydrolysis feminine gender, the fat hydrolysis positive, acetylmethyl alcohol experiment positive, nitrate reduction reaction negative, the gelatine liquefication positive, produce that the sulfuration experiment is negative, the litmus milk experiment peptonizes, the citric acid utilization positive, can utilize glucose, can not oxidation ethanol or aldose, growth temperature be 5～35 ℃, the pH value of growing is 5.5～9.5.
Low temperature microphenomenon of degradation of aniline A1 of the present invention compares through 16SrDNA order-checking and homology, and obtaining the kind the most close with it is Rhodopseudomonas (Pseudomonas), and homology reaches 99%.In conjunction with ne ar feature, growth conditions, Physiology and biochemistry qualification result, determine that low temperature microphenomenon of degradation of aniline A1 is the novel bacterial of Rhodopseudomonas.
Low temperature microphenomenon of degradation of aniline among the present invention, bacterial strain number is A1, belongs to Rhodopseudomonas (Pseudomonas) novel species, in China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation, deposit number is CGMCC No.5011, and preservation date is on December 01st, 2010.
Low temperature microphenomenon of degradation of aniline A1 of the present invention has the biological reinforced processing capacity of amino benzene analog waste water under the cold condition, the degrading aniline rate of bacterial strain can reach 100%, do not cause secondary environmental pollution, and need not to add other chemicals, reduced the processing cost of waste water significantly, had broad application prospects.Low temperature microphenomenon of degradation of aniline A1 of the present invention is helpful for the problem that solves amino benzene analog waste water biological treatment system under the cold condition, has higher using value.
Description of drawings
Fig. 1 is the form atom mechanics microscopic examination figure of low temperature microphenomenon of degradation of aniline A1 colony in the embodiment one, Fig. 2 is low temperature microphenomenon of degradation of aniline A1 atom mechanics microscopic examination figure in the embodiment one, Fig. 3 is low temperature microphenomenon of degradation of aniline A1 atom mechanics microscopic examination figure in the embodiment one, and Fig. 4 is the phylogenetic tree spectrogram that the 16SrDNA gene order of low temperature microphenomenon of degradation of aniline A1 and close bacterial strain makes up in the embodiment one.
Embodiment
Embodiment one: present embodiment low temperature microphenomenon of degradation of aniline, it is low temperature microphenomenon of degradation of aniline A1, belong to Rhodopseudomonas (Pseudomonas) novel species, in China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation, deposit number is CGMCC No.5011, and preservation date is on December 01st, 2010; It is the Gram-positive aerobic bacteria, and is shaft-like, and length is 3.0 μ m, and wide is 0.8 μ m; On beef-protein medium, form white colony, circular, translucent, smooth surface, it is wavy that the edge is, and is difficult for provoking.
Low temperature microphenomenon of degradation of aniline A1 carries out conventional Physiology and biochemistry according to " uncle Jie Shi bacteriology identification handbook " to low temperature microphenomenon of degradation of aniline A1 to identify that experimental result is in the present embodiment: the catalase reacting positive, produce the indole test positive, produce the ammonia test positive, methyl red test reaction negative, starch hydrolysis feminine gender, the fat hydrolysis positive, acetylmethyl alcohol experiment positive, nitrate reduction reaction negative, the gelatine liquefication positive, produce that the sulfuration experiment is negative, the litmus milk experiment peptonizes, citric acid utilizes the positive.
The suitableeest growth pH value 7.0 of low temperature microphenomenon of degradation of aniline A1 in the present embodiment, 15 ℃ of optimum growth temperatures.
In the present embodiment low temperature microphenomenon of degradation of aniline A1 be from chemical plant, Jilin sewage disposal plant aeration tank active sludge through primary dcreening operation, and then carry out that multiple sieve obtains;
Primary dcreening operation is according to the growing state of bacterial strain in low temperature aniline substratum, filter out the adaptive phase weak point, the bacterium that metabolism is vigorous, the primary dcreening operation concrete steps are as follows: 1. get 10mL and insert fully vibration in the sterilized Erlenmeyer flask that granulated glass sphere and 90mL sterilized water be housed through the active sludge of domestication by low temperature, to break up zoogloea, make bacterium be unicellular state and be scattered in the liquid; 2. get the 1mL bacteria suspension the sludge bacteria suspension in Erlenmeyer flask, being inoculated in aniline with coubling dilution is in the solid selection substratum of sole carbon source, is inverted in 10 ℃ of constant incubators and cultivates 3d; 3. distinguish the different typical single bacterium colony of picking, on plate, carry out mark, and streak culture in new culture dish, repeat above operation 3～4 times, obtain the pure bacterium flat board of colony characteristics unanimity; 4. the 4 ℃ of preservations of transferring in preserving substratum of the pure bacterium of gained are standby.
Multiple sieve promptly be on the basis of primary dcreening operation further under 7 ℃ of low temperature, whether having higher degrading aniline rate with bacterial strain is foundation, filters out degradation efficiency good low-temperature bacterial strain.
Wherein every liter of aniline selects substratum by the NaCl of 1g, the K of 1g
2HPO
4, 1g KH
2PO
4, 10mL salts solution, the aniline solution after the agar powder of 20g and the sterilization that adds is as required after cooling formed, the pH value is 7.0～7.2,121 ℃ of autoclaving 20min.
According to " uncle Jie Shi bacteriology identification handbook " low temperature microphenomenon of degradation of aniline A1 is carried out conventional Physiology and biochemistry and identify, adopt the CTAB method to carry out the extraction of bacteria total DNA.Gained bacterial genomes DNA is carried out pcr amplification with bacterial 16 S rDNA universal primer, the PCR product is reclaimed in rubber tapping behind the electrophoresis, entrust Dalian bio-engineering corporation to carry out the 16SrDNA order-checking, sequencing result is carried out homology relatively with listed 16SrDNA sequence among BLAST software and the Genbank, find that the most close kind of low temperature microphenomenon of degradation of aniline A1 that the present embodiment screening obtains is Rhodopseudomonas (Pseudomonas), homology reaches 99%, in conjunction with the ne ar feature, growth conditions, the Physiology and biochemistry qualification result, find that belonging to other bacterial classification with this has any different, and determines that low temperature microphenomenon of degradation of aniline A1 is the novel bacterial of Rhodopseudomonas; According to the 16SrDNA sequencing result, the Blast instrument and the related softwares such as Cluastx, PhyloDraw that utilize NCBI to provide have found homologous sequence in the GeneBank database, and have set up phylogenetic tree (as shown in Figure 4).
Low temperature microphenomenon of degradation of aniline A1 in the present embodiment, below 1500mg/L aniline concentration, when cultivating 24h, the degrading aniline rate reaches 40%; When cultivating 84h, the degrading aniline rate can reach more than 87.5%, and when cultivating 96h, the degrading aniline rate reaches 100%; Experimental results show that low temperature microphenomenon of degradation of aniline A1 has the function that subzero treatment contains aniline waste water, and need not to add other chemicals, reduced the processing cost of waste water significantly, have broad application prospects.
Embodiment two: what present embodiment and embodiment one were different is low temperature microphenomenon of degradation of aniline A1 catalase reacting positive, the product indole test positive, the product ammonia test positive, the methyl red test reaction negative, starch hydrolysis feminine gender, the fat hydrolysis positive, the experiment of acetylmethyl alcohol is positive, the nitrate reduction reaction negative, the gelatine liquefication positive, it is negative to produce the sulfuration experiment, the litmus milk experiment peptonizes, citric acid utilizes positive, can utilize glucose, can not oxidation ethanol or aldose, growth temperature is 5～35 ℃, growth pH value is 5.5～9.5.Other is identical with embodiment one.
Claims (2)
1. low temperature microphenomenon of degradation of aniline, it is characterized in that it is low temperature microphenomenon of degradation of aniline A1, belong to Rhodopseudomonas (Pseudomonas) novel species, in China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation, deposit number is CGMCC No.5011, and preservation date is on December 01st, 2010; It is the Gram-positive aerobic bacteria, and is shaft-like, and length is 3.0 μ m, and wide is 0.8 μ m; On beef-protein medium, form white colony, circular, translucent, smooth surface, it is wavy that the edge is, and is difficult for provoking.
2. a kind of low temperature microphenomenon of degradation of aniline according to claim 1, it is characterized in that low temperature microphenomenon of degradation of aniline A1 catalase reacting positive, produce the indole test positive, produce the ammonia test positive, methyl red test reaction negative, starch hydrolysis feminine gender, the fat hydrolysis positive, acetylmethyl alcohol experiment positive, nitrate reduction reaction negative, the gelatine liquefication positive, produce that the sulfuration experiment is negative, the litmus milk experiment peptonizes, the citric acid utilization positive, can utilize glucose, can not oxidation ethanol or aldose, growth temperature be 5～35 ℃, the pH value of growing is 5.5～9.5.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102649943A (en) * | 2012-03-28 | 2012-08-29 | 哈尔滨师范大学 | Bacterium for high-efficiently degrading aniline under low-temperature condition |
| CN102872955A (en) * | 2012-09-29 | 2013-01-16 | 佛山市海天(高明)调味食品有限公司 | Grinding method and cultivation method for microbial solid culture medium |
| CN103497909A (en) * | 2013-08-27 | 2014-01-08 | 中国科学院成都生物研究所 | Bacterial strain JY9 and use thereof |
| CN110373352A (en) * | 2019-07-19 | 2019-10-25 | 武汉理工大学 | Efficient microphenomenon of degradation of aniline and its application in being handled containing aniline waste water |
| CN111693687A (en) * | 2020-05-28 | 2020-09-22 | 长沙都正生物科技有限责任公司 | Preparation method of biological negative sample and biological standard substance and TMAO detection method |
| CN115322919A (en) * | 2021-08-16 | 2022-11-11 | 沈阳大学 | Low-temperature-resistant p-nitrophenol degrading strain and application thereof |
-
2010
- 2010-12-08 CN CN2010105783550A patent/CN102093970A/en active Pending
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102649943A (en) * | 2012-03-28 | 2012-08-29 | 哈尔滨师范大学 | Bacterium for high-efficiently degrading aniline under low-temperature condition |
| CN102649943B (en) * | 2012-03-28 | 2013-03-13 | 哈尔滨师范大学 | Bacterium for high-efficiently degrading aniline under low-temperature condition |
| CN102872955A (en) * | 2012-09-29 | 2013-01-16 | 佛山市海天(高明)调味食品有限公司 | Grinding method and cultivation method for microbial solid culture medium |
| CN102872955B (en) * | 2012-09-29 | 2016-01-20 | 佛山市海天(高明)调味食品有限公司 | A kind of cultural method of microbe in solid state culture medium |
| CN103497909A (en) * | 2013-08-27 | 2014-01-08 | 中国科学院成都生物研究所 | Bacterial strain JY9 and use thereof |
| CN110373352A (en) * | 2019-07-19 | 2019-10-25 | 武汉理工大学 | Efficient microphenomenon of degradation of aniline and its application in being handled containing aniline waste water |
| CN111693687A (en) * | 2020-05-28 | 2020-09-22 | 长沙都正生物科技有限责任公司 | Preparation method of biological negative sample and biological standard substance and TMAO detection method |
| CN115322919A (en) * | 2021-08-16 | 2022-11-11 | 沈阳大学 | Low-temperature-resistant p-nitrophenol degrading strain and application thereof |
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Application publication date: 20110615 |