CN115927044A - Achromobacter A-1 and application thereof - Google Patents
Achromobacter A-1 and application thereof Download PDFInfo
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- CN115927044A CN115927044A CN202210823996.0A CN202210823996A CN115927044A CN 115927044 A CN115927044 A CN 115927044A CN 202210823996 A CN202210823996 A CN 202210823996A CN 115927044 A CN115927044 A CN 115927044A
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W10/00—Technologies for wastewater treatment
- Y02W10/10—Biological treatment of water, waste water, or sewage
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses Achromobacter sp A-1 and application thereof. The strain is preserved in the China general microbiological culture Collection center of the Committee for culture Collection of microorganisms with the preservation number of CGMCC No.24031. The leucobacter A-1 can realize the biodegradation of tetracycline in a wide concentration range (10-80 mg/L) under the condition of a wide environment temperature (10-30 ℃), has high degradation efficiency which can reach 57% -81%, is complete in biodegradation, and can effectively reduce the potential risk of tetracycline in the environment. The strain can also realize the synchronous removal of tetracycline and ammonia nitrogen, and no accumulation of nitrogenous intermediate products. Therefore, the method has wide application range in the aspect of treating the wastewater containing tetracycline and ammonia nitrogen and has good application prospect.
Description
Technical Field
The invention belongs to the technical field of application environment microorganisms, and particularly relates to achromobacter A-1 and application thereof.
Background
Antibiotics (antibiotics) are a class of secondary metabolites produced by microorganisms or higher animals and plants during life and having anti-pathogenic or other activities, and are chemical substances capable of interfering with the development functions of other living cells. There are many types of antibiotics, of which tetracycline antibiotics are a broad spectrum synthetic antibiotic widely used in human medicine and animal husbandry. Tetracycline drugs administered to humans and animals are generally not completely absorbed, most often with fecal material either in the form of their parent or metabolites, entering the sewer network or directly discharged into the surrounding receiving environment (e.g., surface water). In addition, the loss of tetracycline in the manufacturing and equipment processes and the effluent from pharmaceutical plant processes are also important sources of influence on the concentration of tetracycline in water. Tetracyclines have been detected with high frequency in different environments. The concentration of tetracycline in a certain treatment unit of a pharmaceutical wastewater treatment plant reaches 61.0 +/-12.2 mg/L, and the total concentration of tetracycline antibiotics in water and sediments of a certain seawater farm (southeast China) is 0.2-259.1 ng/L and 3.45-74.84 ng/g respectively. The wide existence of antibiotics not only seriously affects the growth and metabolism of microorganisms in the environment, but also can induce the microorganisms to generate drug resistance, so that the appearance and the transmission of antibiotic resistant bacteria and drug resistant genes are caused, the treatment potential of the antibiotics on human and animal pathogens is reduced, and the problem of antibiotic pollution gradually becomes one of the biggest public health threats in the world. On the other hand, the ammonia nitrogen exceeding standard is the most common pollution problem in water body pollution, which causes water body eutrophication, leads to mass propagation of algae, reduces dissolved oxygen in the water body, blackens the water body, endangers the water safety of human beings and seriously influences the ecological balance of the water body. Antibiotics and ammonia nitrogen may coexist in the organic wastewater to form an antibiotic and ammonia nitrogen composite organic polluted water body. Therefore, the exploration of the efficient, economic and safe water treatment technology has important significance for degrading residual antibiotics and ammonia nitrogen in sewage and protecting human health and ecological environment.
At present, researches on the removal of tetracycline in wastewater mainly focus on physical and chemical methods, such as adsorption, membrane separation, ultraviolet irradiation, ozone oxidation and the like. In recent years, biodegradation methods have also become a research hotspot due to the characteristics of long-acting effect, low cost, low toxicity of degradation products and the like. For example, the invention patent CN111154685B discloses a Klebsiella variicola strain for degrading tetracycline and application thereof, wherein the Klebsiella variicola TC3 in the invention can degrade tetracycline in a co-metabolism mode, so that the Klebsiella variicola strain can be applied to removal of low-concentration tetracycline in soil and water. The invention patent CN104403965B discloses pseudomonas hibiscus for degrading tetracycline pollutants in water and application thereof, wherein the pseudomonas hibiscus DT1 has strong growth capacity and good tetracycline degradation capacity in an environment containing tetracycline. However, the biodegradation of tetracycline is carried out at 30-35 ℃, because the degradation of tetracycline is obviously inhibited under low temperature. In view of the wide range and complexity of antibiotic contamination, there is still a need to obtain more strains with high antibiotic-degrading ability, especially strains that degrade antibiotics under relatively scarce low temperature conditions. In addition, a single strain for synchronously degrading ammonia nitrogen and tetracycline combined pollution is also rarely reported. In the face of increasingly stringent safety detection standards and extensive public concerns about ecological environments, a new efficient and safe technology and a new method are urgently needed to be developed to synchronously solve the problems of tetracycline and ammonia nitrogen pollution, so as to relieve the environmental pressure, inhibit the development of bacterial drug resistance and guarantee the safety of the ecological environment.
Disclosure of Invention
Aiming at the defects in the prior art, the invention aims to provide a novel achromobacter A-1, solve the problem of inhibiting the biodegradation of tetracycline under the existing low-temperature condition and realize the synchronous removal of tetracycline and ammonia nitrogen in organic sewage; meanwhile, more choices are provided for the biodegradation of tetracycline in the organic sewage.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows: the invention provides an achromobacter (Achromobacter) A-1, wherein the strain is preserved in the ordinary microorganism center of China Committee for culture Collection of microorganisms (the address: no. 3 of Xilu No.1 of Beijing, the microbiological research institute of Chinese academy of sciences, the zip code: 100101) within 12 months and 3 days in 2021, and the preservation number is CGMCC No.24031. Further, the nucleotide sequence of the 16S rDNA of the achromobacter A-1 has a sequence shown as SEQ ID NO. 1.
The invention also aims to provide a degrading bacterium preparation, and the effective component of the preparation comprises the leucobacter A-1. Preferably, the living bacteria amount of the Achromobacter A-1 in the preparation is more than or equal to 1 x 10 6 CFU/mL. The preparation can be wet thallus obtained by liquid culture of Achromobacter A-1, such as culture solution, culture concentrated solution, and culture separated solution, lyophilized cell obtained by freeze drying wet thallus of Achromobacter A-1, or re-suspended solution obtained by lyophilized cell of Achromobacter A-1.
Further, the present invention provides the use of the above Achromobacter A-1 or the above degrading bacteria preparation for degrading tetracycline and/or ammonia Nitrogen (NH) 4 + -N). Preferably, the temperature of the degradation is 10 to 30 ℃.
It is another object of the present invention to provide a pharmaceutical composition containing tetracycline and/or NH 4 + -N waste water treatment method, which comprises the step of putting the Achromobacter A-1 with the preservation number of CGMCC No.24031 or a degradation bacterium preparation prepared from the Achromobacter A-1 into the waste water. Under aerobic conditions, the achromobacter A-1 takes sodium acetate and tetracycline as a mixed organic carbon source to realize the treatment of tetracycline and/or NH in the wastewater 4 + -biological removal of N.
Preferably, the above Achromobacter A-1 is used for treating a composition containing tetracycline and/or NH 4 + And in the case of-N organic wastewater, the concentration of tetracycline in the wastewater is 10-80 mg/L.
Preferably, the above Achromobacter A-1 is used for treating a composition containing tetracycline and/or NH 4 + -N organic waste water, NH in said waste water 4 + The concentration of-N is 70-130 mg/L.
Preferably, the above Achromobacter A-1 is used for treating a composition containing tetracycline and/or NH 4 + And in the case of-N organic wastewater, the temperature of the wastewater is 10-30 ℃.
Preferably, the above Achromobacter A-1 is used for treating a composition containing tetracycline and/or NH 4 + And in the case of organic wastewater of-N, the C/N of the wastewater is 9-16.
Preferably, the above Achromobacter A-1 is used for treating a composition containing tetracycline and/or NH 4 + And in the case of-N organic wastewater, the pH value of the wastewater is 6-8.
The invention has the following beneficial effects:
1. the achromobacter A-1 provided by the invention can realize the biological removal of tetracycline under the condition of a wider environment temperature (10-30 ℃). The tetracycline removal efficiency in 7d is higher and can reach 57-81%. Especially under the low temperature condition of 10 ℃, the achromobacter A-1 still keeps good growth performance, can carry out biological removal on tetracycline with wider concentration range (10-80 mg/L), and has application potential for treating tetracycline-containing wastewater under the low temperature condition. The strain has low requirements on environmental temperature conditions and wide application range, thereby having strong application value.
2. The achromobacter A-1 provided by the invention can realize the biological removal of tetracycline and the synchronous removal of ammonia nitrogen within the temperature range of 10-30 ℃, and does not have the accumulation of nitrogenous intermediate products. The invention has wide application prospect in the aspect of treating the compound organic wastewater containing tetracycline and ammonia nitrogen.
3. The leucobacter A-1 provided by the invention can be used for biodegradation of tetracycline mainly through hydroxylation, dehydroxylation, demethylation, decarbonylation, carbon ring opening and other reactions, can be subjected to ring-opening reaction, and is relatively thorough in biodegradation. Can effectively reduce the potential risk of the tetracycline in the environment, and has important significance for controlling the migration and accumulation of the tetracycline in the environment.
Drawings
FIG. 1 is a colony morphology of Achromobacter A-1.
FIG. 2 is a graph showing the growth of Achromobacter A-1 at 10 ℃ and the tetracycline elimination effect.
FIG. 3 is a graph showing the growth of Achromobacter A-1 at 20 ℃ and the tetracycline elimination effect.
FIG. 4 is a graph showing the growth of Achromobacter A-1 at 30 ℃ and the tetracycline elimination effect.
FIG. 5 is a graph showing the effect of Achromobacter A-1 on tetracycline removal at 10 ℃ at various concentrations.
FIG. 6 is a diagram showing the effect of Achromobacter A-1 in synchronously removing tetracycline and ammonia nitrogen.
FIG. 7 is a chromatogram of total ions of products obtained after biodegradation of tetracycline by Achromobacter A-1.
FIG. 8 is a schematic diagram of a possible pathway for degrading tetracycline by Achromobacter A-1.
Detailed Description
In order to make the technical problems, technical solutions and advantageous effects to be solved by the present invention more clearly apparent, the present invention is further described in detail below with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
In the description of the present invention, it should be noted that: the experimental methods in the following examples are all conventional experimental methods unless otherwise specified; the test materials used in the following examples were purchased from conventional biochemical stores unless otherwise specified; each experiment referred to in the examples below was set up in triplicate and the results averaged.
The media referred to in the following examples:
LB liquid medium (per liter): 10g of tryptone, 5g of yeast extract, 10g of NaCl and NaH 2 PO 4 ·2H 2 O and Na 2 HPO 4 ·12H 2 And O, adjusting the pH = 6-8.
Basal medium (per liter): CH (CH) 3 COONa 1.5g、NH 4 Cl 0.5g, naCl 0.5g, trace elements 1mL, naH 2 PO 4 ·2H 2 O and Na 2 HPO 4 ·12H 2 O, adjusting the pH = 6-8; adding a certain amount of tetracycline into a basic culture mediumObtaining the basic culture medium containing tetracycline.
Trace elements (per liter): feSO 4 ·7H 2 O 0.2g、CoCl 2 ·6H 2 O 0.09g、CuSO 4 ·5H 2 O 0.086g、 HBO 3 0.094g、H 8 MoN 2 O 4 ·4H 2 O 0.03g、MnCl 2 ·7H 2 O 0.075g、KI 0.022g。
LB solid medium (per liter): 10g of tryptone, 5g of yeast extract, 10g of NaCl, 15-20 g of agar powder and NaH 2 PO 4 ·2H 2 O and Na 2 HPO 4 ·12H 2 And O, adjusting the pH = 6-8.
The culture medium is sterilized before use under the following conditions: sterilizing with high pressure steam at 121 deg.C for 30min; the sampling, inoculation and dilution processes of the invention are all carried out in a sterile environment. The basic culture medium added with tetracycline is covered by aluminum foil paper, so that the influence of photodegradation tetracycline on the experiment is prevented.
Example 1: separation, purification and identification of Achromobacter A-1
1. Isolation and purification of Achromobacter A-1
The activated sludge used in the embodiment is taken from an aeration tank of a crowned rock sewage treatment plant in Chongqing, and the screening of the achromobacter A-1 adopts a plate-drawing method, wherein the liquid culture condition is that the temperature is 10-30 ℃, the rotating speed of a shaking table is 90-150 rpm, and the solid plate culture condition is that a biochemical incubator is used for aseptic culture at 20-30 ℃. The specific separation and purification are carried out according to the following steps:
1) Primary screening and enrichment of the strain: adding a certain amount of glass beads for scattering activated sludge into a triangular flask filled with 95mL of sterile water, after sterilization treatment, taking 5mL of activated sludge taken from an aeration tank of a Chongqing cockscomb stone sewage plant, inoculating the activated sludge into the triangular flask, and placing the activated sludge in a constant-temperature shaking incubator for aerobic culture for 1h to obtain a dispersed and uniformly mixed bacterial suspension. And then transferring 1mL of the bacterial suspension to an LB liquid culture medium for activation for 12h, transferring 1mL of the activated bacterial liquid to a basic culture medium added with 5mg/L tetracycline, and placing the basic culture medium in a constant-temperature shaking incubator for aerobic culture for 72h to perform first domestication primary screening of the tetracycline degrading bacteria to obtain a dispersed and uniformly mixed culture solution. Then transferring 1mL of culture solution to a fresh basal culture medium added with 10mg/L tetracycline, and placing the culture solution in a constant-temperature shaking incubator for aerobic culture for 72h to perform second acclimation and screening of the tetracycline degrading bacteria. The inoculation and domestication are sequentially repeated, and the concentration of the tetracycline in the culture medium is sequentially increased by 5mg/L. After multiple inoculations, the tetracycline concentration in the basal medium reached around 70 mg/L. Then inoculating 1mL of the domesticated and screened culture solution into an LB liquid culture medium, and placing the culture solution into a constant-temperature shaking incubator for aerobic culture for 12-24 h to obtain an enriched culture solution.
2) Re-screening and purifying the strain: the enrichment culture solution is diluted to 10 degrees in gradient -4 ～10 -7 The culture solution is diluted, and then the following steps are carried out: (1) taking diluted culture solutions with different concentration gradients to respectively inoculate and streak on an LB solid culture medium, and inversely culturing for 2-3 d in a biochemical incubator; (2) selecting a single colony growing well in the flat plate to inoculate in an LB liquid culture medium, and then placing the inoculated culture solution in a constant-temperature shaking incubator to aerobically culture for 12h to obtain an enriched culture solution; (3) and (3) transferring 1mL of the enrichment culture solution obtained in the step (2) to a fresh basal culture medium added with 50mg/L tetracycline, and placing the enrichment culture solution in a constant-temperature shaking incubator for aerobic culture for 72h. And (4) measuring the concentration of the tetracycline in the basic culture medium after the culture, selecting a culture solution with a good tetracycline treatment effect, carrying out gradient dilution on the culture solution, and repeating the steps (1) to (3) until a strain with a good and stable tetracycline removal effect is obtained. It was named A-1 and stored for a long period.
2. Identification of Achromobacter A-1
Achromobacter A-1 of this example is a gram-negative bacterium, rod-shaped. The colonies obtained by culturing on the solid medium are shown in FIG. 1, and are round, slightly raised, neat in edge, moist, and translucent. And (3) homology analysis and identification: extracting strain genome DNA by using an OMEGA bacterial DNA kit as a template, and obtaining 16S rDNA of the strain through Polymerase Chain Reaction (PCR). The 16S rDNA primers were universal primers 27F (5 'AGAGAGTTTGATCMTGGCTCAG 3') and 1492R (5 'GGTTACCTTGTTACGACTT 3'). PCR amplification procedure: performing pre-denaturation at 98 ℃ for 2min; 10s at 98 ℃, 15s at 55 ℃, 15s at 72 ℃, and 38 cycles; stretching for 5min at 72 ℃. The PCR product is subjected to agarose gel tapping recovery and then submitted to Shanghai Meiji biological medicine science and technology limited company for sequencing analysis (the second generation DNA sequencing technology is applied, the reading length of the sequencing method is limited and generally does not exceed 500 bp). <xnotran> A-1 16S rDNA SEQ ID NO.1 (TGGGGAATTTTGGACAATGGGGGAAACCCTGATCCAGCCATCCCGCGTGTGCGATG AAGGCCTTCGGGTTGTAAAGCACTTTTGGCAGGAAAGAAACGTCGTGGGTTAATACC CCGCGAAACTGACGGTACCTGCAGAATAAGCACCGGCTAACTACGTGCCAGCAGCCG CGGTAATACGTAGGGTGCAAGCGTTAATCGGAATTACTGGGCGTAAAGCGTGCGCAG GCGGTTCGGAAAGAAAGATGTGAAATCCCAGAGCTTAACTTTGGAACTGCATTTTTA ACTACCGGGCTAGAGTGTGTCAGAGGGAGGTGGAATTCCGCGTGTAGCAGTGAAATG CGTAGATATGCGGAGGAACACCGATGGCGAAGGCAGCCTCCTGGGATAACACTGACG CTCATGCACGAAAGCGTGGGGAGCAAACAGG), 429bp ( , A-1 16S rDNA ). </xnotran> And comparing the sequencing result with the sequence of the existing bacteria in GenBank, wherein the similarity of the comparison result with the achromobacter is highest. Therefore, the strain A-1 was identified as a novel strain Achromobacter (Achromobacter sp.) by combining the colony morphology, metabolic characteristics and 16S rDNA partial gene sequence analysis of the strain A-1. In 2021, 12 months and 3 days, the strain is preserved in China general microbiological culture Collection center (CGMCC) (address: no. 3 of Xilu No.1 of Beijing, chaoyang, north Chen, institute of microbiology, chinese academy of sciences, zip code: 100101), and the preservation number is CGMCC No.24031.
Example 2: growth of Achromobacter A-1 at different temperatures and tetracycline-removing properties
The strain A-1 is inoculated in an LB liquid culture medium and activated for 12h. The resulting suspension was inoculated into a minimal medium supplemented with 50mg/L tetracycline, and the COD/N (C/N) was about 9. Then placing the inoculated basal culture medium in constant temperature shaking incubators at 10 ℃, 20 ℃ and 30 ℃ respectively, and culturing for 7d in a dark place at the rotating speed of 120 rpm. Timed sampling to determine tetracycline concentration and OD 600 (the absorbance of the culture medium at a wavelength of 600nm, which indicates the cell density).
Specifically, as shown in FIG. 2, FIG. 3 and FIG. 4, the effects of Achromobacter A-1 on the treatment of tetracycline-containing wastewater at temperatures of 10 ℃, 20 ℃ and 30 ℃ are shown, respectively. As shown in FIG. 2, achromobacter A-1 was cultured at 10 ℃ for about 4 days and then OD was added thereto 600 The indicated growth amount reached a peak of 1.152; as shown in FIG. 3, after Achromobacter A-1 was continuously cultured at 20 ℃ for about 2 days, the maximum growth amount reached 1.069; as shown in FIG. 4, achromobacter A-1 reached a maximum growth of 1.134 after continuous culture at 30 ℃ for about 21 hours. As can be seen from FIGS. 2 to 4, when the initial content of tetracycline was 50mg/L, the removal efficiency of 7d tetracycline by Achromobacter A-1 was 60%, 63% and 81% at 10 ℃, 20 ℃ and 30 ℃, respectively. The results show that the achromobacter A-1 can grow well and remove tetracycline under the condition of 10-30 ℃. And as the temperature is increased, the growth rate is increased, and the tetracycline removal rate is increased.
Example 3: removal performance of achromobacter A-1 on tetracycline with different concentrations under low temperature condition of 10 DEG C
The strain A-1 is inoculated in an LB liquid culture medium and activated for 12h. The resulting bacterial liquid was then inoculated into a minimal medium, to which 10, 30, 50 and 80mg/L tetracycline, having a C/N of about 9, were additionally added, respectively. Then the inoculated basal medium is placed in a constant temperature shaking incubator at 10 ℃, and is cultured for 7d in a dark place at the rotating speed of 120 rpm. The tetracycline concentration was determined by sampling at regular intervals and the results are shown in FIG. 5.
As can be seen from FIG. 5, when the initial concentration of tetracycline in the wastewater was 10, 30, 50 and 80mg/L, and Achromobacter A-1 was used at 10 ℃, the removal efficiency of 7d tetracycline was 57%, 69%, 60% and 68%, respectively. The result shows that the achromobacter A-1 has good removal effect on tetracycline with the concentration of 10-80 mg/L under the low temperature condition of 10 ℃. The achromobacter A-1 is proved to have the application potential of treating the tetracycline-containing wastewater under the low-temperature condition.
Example 4: achromobacter A-1 capacity of synchronously removing tetracycline and ammonia nitrogen
Inoculating the strain A-1 into LB liquid culture medium for activation for 12h, then inoculating the obtained bacterial liquid into a basal culture medium added with 20mg/L tetracycline, and inoculating the bacterial liquid into the basal culture medium CThe inoculated basal medium is placed in a constant temperature shaking incubator at 30 ℃ and is cultured in the dark at the rotating speed of 120 rpm. Measurement of OD by timed sampling 600 Tetracycline concentration and ammonia nitrogen concentration, the results are shown in figure 6.
As can be seen from FIG. 6, the strain A-1 grew well as the culture time increased. NH within 72h 4 + Reduction of-N from 70.4mg/L to 34.9mg/L, NH 4 + The N removal efficiency reaches 50.4 percent, and no nitrogen-containing intermediate product is accumulated. Meanwhile, the removal rate of tetracycline reaches 74.9 percent. The results show that the strain A-1 can synchronously remove tetracycline and ammonia nitrogen, and has application potential in treating wastewater containing both tetracycline and ammonia nitrogen.
Example 5: research on approach of degrading tetracycline by Achromobacter A-1
Detecting the product of the achromobacter A-1 for degrading tetracycline by using a high-resolution liquid chromatograph-mass spectrometer (Q active Plus) of an orbitrap, wherein the result shows that the tetracycline biodegradation product obtains better response in an ESI-MS positive ion mode, a total mass spectrogram is shown in figure 7, and ESI-MS high-resolution data is analyzed and processed by using Thermo Xcalibur Qual Browser software, so that 7 possible biodegradation products A (C) are finally obtained (C) 22 H 22 N 2 O 7 ；m/z＝427)、B(C 18 H 39 NO 6 ； m/z＝365)、C(C 12 H 11 O 4 ；m/z＝218)、D(C 22 H 25 N 2 O 9 ；m/z＝461)、E(C 19 H 18 O 5 ； m/z＝326)、F(C 21 H 43 NO 9 (ii) a m/z = 453) and G (C) 20 H 20 N 2 O 9 ；m/z＝432)。
Based on the above test results, the pathway of tetracycline biodegradation by Achromobacter A-1 was deduced, as shown in FIG. 8. The first way is as follows: the parent compound, tetracycline (TEC) (C) 22 H 24 N 2 O 8 (ii) a m/z = 445) dehydroxylation to Compound A (C) 22 H 22 N 2 O 7 (ii) a m/z = 427), compound a is converted into compound B (C) by a ring-opening reaction 18 H 39 NO 6 (ii) a m/z = 365), compound B is converted by further ring opening into compound C (C) 12 H 11 O 4 (ii) a m/z = 218), on the other hand, the compound a is converted into the compound E (C) by a reaction such as demethylation, decarbonylation, or the like 19 H 18 O 5 (ii) a m/z = 326). And (2) a second way: the parent compound Tetracycline (TEC) (C) 22 H 24 N 2 O 8 (ii) a m/z = 445) hydroxylation to give compound D (C) 22 H 25 N 2 O 9 (ii) a m/z = 461), compound D is converted into compound F (C) by a ring-opening reaction 21 H 43 NO 9 (ii) a m/z = 453); demethylation of compound D to give compound G (C) 20 H 20 N 2 O 9 (ii) a m/z = 432); in another aspect, compound D is dehydroxylated to compound a, which is then converted to compound C and compound E by the pathway described as pathway one.
As can be seen, the strain A-1 can be used for the biodegradation of tetracycline mainly through hydroxylation, dehydroxylation, demethylation, decarbonylation, carbocyclic ring opening and other reactions. Wherein the compound C (C) 12 H 11 O 4 (ii) a m/z = 218) is higher in the mass spectrum, which indicates that the achromobacter A-1 can realize ring opening reaction on tetracycline, and the degradation is more thorough. The result also provides certain theoretical basis for the achromobacter A-1 to reduce the potential risk of tetracycline in the environment.
Finally, the above embodiments are only for illustrating the technical solutions of the present invention and not for limiting, although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions may be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all of them should be covered in the claims of the present invention.
Claims (10)
1. Achromobacter (Achromobacter sp.) A-1 is preserved in China general microbiological culture Collection center with the preservation number of CGMCC No.24031.
2. A degrading bacterium preparation, wherein the active ingredient of the preparation comprises Achromobacter A-1 according to claim 1.
3. The degrading bacteria preparation according to claim 2, wherein the amount of the Achromobacter A-1 in the preparation is 1X 10 or more 6 CFU/mL。
4. Use of the Achromobacter A-1 according to claim 1 or the degrading bacterial preparation according to claim 2 for degrading tetracycline and/or ammonia nitrogen.
5. Use according to claim 4, wherein the degradation temperature is between 10 and 30 ℃.
6. Containing tetracycline and/or NH 4 + The method for treating wastewater containing-N is characterized in that Achromobacter A-1 with the preservation number of CGMCC No.24031 or a degradation bacterium preparation prepared from the Achromobacter A-1 is thrown into the wastewater.
7. The method according to claim 6, wherein the concentration of tetracycline in the wastewater is 10-80 mg/L; NH in the wastewater 4 + The concentration of-N is 70-130 mg/L.
8. The method of claim 6, wherein the wastewater has a pH of 6 to 8.
9. The method of claim 6, wherein the initial C/N in the wastewater is from 9 to 16.
10. The method of claim 6, wherein the temperature of the wastewater is 10 to 30 ℃.
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