CN117185499B - Synechocystis PCC 6803 capable of tolerating sulfonamide antibiotics and application thereof - Google Patents
Synechocystis PCC 6803 capable of tolerating sulfonamide antibiotics and application thereof Download PDFInfo
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- 229940123317 Sulfonamide antibiotic Drugs 0.000 title claims abstract description 45
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- 229960005404 sulfamethoxazole Drugs 0.000 claims description 28
- JLKIGFTWXXRPMT-UHFFFAOYSA-N sulphamethoxazole Chemical compound O1C(C)=CC(NS(=O)(=O)C=2C=CC(N)=CC=2)=N1 JLKIGFTWXXRPMT-UHFFFAOYSA-N 0.000 claims description 28
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- 150000003456 sulfonamides Chemical class 0.000 claims 1
- 241000192581 Synechocystis sp. Species 0.000 abstract description 3
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- FABPRXSRWADJSP-MEDUHNTESA-N moxifloxacin Chemical compound COC1=C(N2C[C@H]3NCCC[C@H]3C2)C(F)=CC(C(C(C(O)=O)=C2)=O)=C1N2C1CC1 FABPRXSRWADJSP-MEDUHNTESA-N 0.000 description 5
- 229960003702 moxifloxacin Drugs 0.000 description 5
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- IWVCMVBTMGNXQD-PXOLEDIWSA-N oxytetracycline Chemical compound C1=CC=C2[C@](O)(C)[C@H]3[C@H](O)[C@H]4[C@H](N(C)C)C(O)=C(C(N)=O)C(=O)[C@@]4(O)C(O)=C3C(=O)C2=C1O IWVCMVBTMGNXQD-PXOLEDIWSA-N 0.000 description 5
- 235000019366 oxytetracycline Nutrition 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- FDDDEECHVMSUSB-UHFFFAOYSA-N sulfanilamide Chemical compound NC1=CC=C(S(N)(=O)=O)C=C1 FDDDEECHVMSUSB-UHFFFAOYSA-N 0.000 description 5
- 229940124530 sulfonamide Drugs 0.000 description 5
- IWVCMVBTMGNXQD-UHFFFAOYSA-N terramycin dehydrate Natural products C1=CC=C2C(O)(C)C3C(O)C4C(N(C)C)C(O)=C(C(N)=O)C(=O)C4(O)C(O)=C3C(=O)C2=C1O IWVCMVBTMGNXQD-UHFFFAOYSA-N 0.000 description 5
- 230000003698 anagen phase Effects 0.000 description 4
- 238000005286 illumination Methods 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 230000009036 growth inhibition Effects 0.000 description 3
- 239000002028 Biomass Substances 0.000 description 2
- ULGZDMOVFRHVEP-RWJQBGPGSA-N Erythromycin Chemical compound O([C@@H]1[C@@H](C)C(=O)O[C@@H]([C@@]([C@H](O)[C@@H](C)C(=O)[C@H](C)C[C@@](C)(O)[C@H](O[C@H]2[C@@H]([C@H](C[C@@H](C)O2)N(C)C)O)[C@H]1C)(C)O)CC)[C@H]1C[C@@](C)(OC)[C@@H](O)[C@H](C)O1 ULGZDMOVFRHVEP-RWJQBGPGSA-N 0.000 description 2
- 229960004475 chlortetracycline Drugs 0.000 description 2
- CYDMQBQPVICBEU-XRNKAMNCSA-N chlortetracycline Chemical compound C1=CC(Cl)=C2[C@](O)(C)[C@H]3C[C@H]4[C@H](N(C)C)C(O)=C(C(N)=O)C(=O)[C@@]4(O)C(O)=C3C(=O)C2=C1O CYDMQBQPVICBEU-XRNKAMNCSA-N 0.000 description 2
- 235000019365 chlortetracycline Nutrition 0.000 description 2
- 230000008878 coupling Effects 0.000 description 2
- 238000010168 coupling process Methods 0.000 description 2
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- 239000003344 environmental pollutant Substances 0.000 description 2
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 241000195628 Chlorophyta Species 0.000 description 1
- 241000192700 Cyanobacteria Species 0.000 description 1
- 241000192710 Microcystis aeruginosa Species 0.000 description 1
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- 239000003120 macrolide antibiotic agent Substances 0.000 description 1
- ZYZCGGRZINLQBL-GWRQVWKTSA-N microcystin-LR Chemical compound C([C@H](OC)[C@@H](C)\C=C(/C)\C=C\[C@H]1[C@@H](C(=O)N[C@H](CCC(=O)N(C)C(=C)C(=O)N[C@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H]([C@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N1)C(O)=O)C(O)=O)C)C1=CC=CC=C1 ZYZCGGRZINLQBL-GWRQVWKTSA-N 0.000 description 1
- DIDLWIPCWUSYPF-UHFFFAOYSA-N microcystin-LR Natural products COC(Cc1ccccc1)C(C)C=C(/C)C=CC2NC(=O)C(NC(CCCNC(=N)N)C(=O)O)NC(=O)C(C)C(NC(=O)C(NC(CC(C)C)C(=O)O)NC(=O)C(C)NC(=O)C(=C)N(C)C(=O)CCC(NC(=O)C2C)C(=O)O)C(=O)O DIDLWIPCWUSYPF-UHFFFAOYSA-N 0.000 description 1
- 238000001000 micrograph Methods 0.000 description 1
- 239000008239 natural water Substances 0.000 description 1
- 231100000719 pollutant Toxicity 0.000 description 1
- LISFMEBWQUVKPJ-UHFFFAOYSA-N quinolin-2-ol Chemical compound C1=CC=C2NC(=O)C=CC2=C1 LISFMEBWQUVKPJ-UHFFFAOYSA-N 0.000 description 1
- 230000008261 resistance mechanism Effects 0.000 description 1
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Abstract
The invention relates to the technical field of microalgae, in particular to a sulfonamide antibiotic-resistant Synechocystis sp PCC 6803 and application thereof. The invention discovers that the synechocystis PCC 6803 has good tolerance to high-concentration sulfonamide antibiotics, has semi-lethal concentration of more than 1mg/L, can be used as a tolerant strain for stress on the sulfonamide antibiotics, and has important value for biological treatment of the sulfonamide antibiotics.
Description
Technical Field
The invention relates to the technical field of microalgae, in particular to a sulfonamide antibiotic-resistant Synechocystis sp PCC 6803 and application thereof.
Background
Antibiotics are widely used in many fields such as medicine, aquaculture, animal husbandry, and crop production. The traditional sewage treatment process has limited removal efficiency of antibiotics, and a large amount of antibiotics are discharged into natural water. At present, the existence of various antibiotics is detected in rivers, lakes and groundwater worldwide, which poses a great threat to water environment and human health. Wherein the sulfonamide antibiotics are the main residual antibiotics in the water environment, and the residual concentration of the sulfonamide antibiotics reported in surface water and sewage treatment plants can reach as high as 10 mug/L.
In recent years, a coupled system of bacteria and algae has been found to have great potential in sewage treatment, especially for some difficult-to-treat micro-pollutants. Compared with the traditional activated sludge process, the algae-bacteria coupling treatment process has stronger impact load resistance and better pollutant removal efficiency, so that the algae-bacteria coupling treatment process has remarkable advantages in the process of removing antibiotics. The toxic stress of antibiotics on organisms is one of the important factors limiting the efficiency of biological treatment, so screening out bacteria and algae with high antibiotic tolerance is the key to optimizing the efficiency of the process.
In general, bacteria resistant to antibiotics have been screened in the prior art, for example, published chinese patents CN107201328A, CN115322934A, CN116333902a, but no algae resistant to sulfonamide antibiotics have been available.
Disclosure of Invention
In order to solve the problems, the invention provides a sulfanilamide antibiotics-resistant synechocystis PCC 6803 and application thereof in treating wastewater containing sulfanilamide antibiotics. The invention discovers the tolerance function (semi-lethal concentration is more than 1 mg/L) of the synechocystis PCC 6803 to high-concentration sulfonamide antibiotics, and has important value for biological treatment of the sulfonamide antibiotics.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides application of synechocystis PCC 6803 as a strain of algae resistant to sulfanilamide antibiotics.
The invention provides an application of synechocystis PCC 6803 in treatment of wastewater containing sulfonamide antibiotics.
Preferably, the wastewater comprises production wastewater and/or domestic wastewater containing sulfonamide antibiotics.
Preferably, the sulfonamide antibiotic comprises sulfamethoxazole and/or sulfamethoxazole.
Preferably, the concentration of the sulfonamide antibiotics in the wastewater is less than or equal to 1mg/L.
The beneficial effects are that:
the invention provides application of synechocystis PCC 6803 as a strain of algae resistant to sulfanilamide antibiotics. The invention discovers that the synechocystis PCC 6803 has good tolerance to high-concentration sulfonamide antibiotics, has semi-lethal concentration of more than 1mg/L, can be used as a strain with tolerance to the stress of the sulfonamide antibiotics, and has important value for the biological treatment of the sulfonamide antibiotics.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are required to be used in the embodiments will be briefly described below.
FIG. 1 is a schematic diagram of a process for finding that Synechocystis PCC 6803 is resistant to high concentration of sulfonamide antibiotics according to the present invention;
FIG. 2 is an electron microscope image of Synechocystis PCC 6803;
FIG. 3 shows the results of the resistance of Synechocystis PCC 6803 to sulfonamide antibiotics.
Detailed Description
The invention provides application of synechocystis PCC 6803 as a strain of algae resistant to sulfanilamide antibiotics.
The invention provides an application of synechocystis PCC 6803 in treatment of wastewater containing sulfonamide antibiotics. The synechocystis PCC 6803 is preferably purchased from a fresh water algae seed library of China academy of sciences, and the algae seed number is FACHB-898.
In the present invention, the wastewater preferably includes wastewater from production and/or domestic sewage containing sulfonamide antibiotics; the sulfonamide antibiotic preferably comprises sulfamethoxazole and/or sulfamethoxazole; the concentration of the sulfonamide antibiotics in the wastewater is preferably less than or equal to 1mg/L.
Embodiments of the invention relate to an embodiment of the invention wherein the Synechocystis PCC 6803 is in an exponential growth phase at 10 4 Initial biomass of cells/L is exposed to four eight antibiotics in the concentration range of 0-100 mug/L, and the resistance of Synechocystis PCC 6803 to different antibiotics is measuredThe selected antibiotics were Ciprofloxacin (CIP) or Moxifloxacin (MOX) representing quinolone antibiotics, azithromycin (AZM) and Clarithromycin (CTM) representing macrolide antibiotics, aureomycin (CTC) and Oxytetracycline (OTC) representing tetracycline antibiotics, and Sulfamethoxazole (SMZ) and Sulfamethoxazole (SMX) representing sulfonamide antibiotics, respectively, and the results showed that the growth inhibition rate of the sulfonamide antibiotics (SMX and SMZ) on synechocystis PCC 6803 was the lowest; and then the tolerance threshold value of the synechocystis PCC 6803 to the sulfonamide antibiotics is measured, and the result shows that the semi-lethal concentration of the sulfonamide antibiotics (SMX and SMZ) to the synechocystis PCC 6803 is higher than 1mg/L. It can be seen that the synechocystis PCC 6803 has good tolerance to high-concentration sulfonamide antibiotics, can be used as an algae strain for tolerating the stress of the sulfonamide antibiotics, and has important value for biological treatment of the sulfonamide antibiotics (such as treatment of wastewater containing the sulfonamide antibiotics).
For further explanation of the present invention, a sulfonamide antibiotic resistant synechocystis PCC 6803 and its use according to the present invention will be described in detail below with reference to the accompanying drawings and examples, but they should not be construed as limiting the scope of the present invention.
Example 1
(1) Synechocystis sp PCC 6803 was purchased from a fresh water algae seed stock of China academy of sciences and subjected to stable culture, which is:
culturing with BG-11 medium, and placing in a light incubator at 28+ -1deg.C, 50μmol.m -2 ·s -1 The culture was carried out for 4 days with a light cycle of 14h light/10 h dark and a rotation speed of 120rpm, and the synechocystis PCC 6803 cultured on the 4 th day was in the exponential growth phase, and was used as an algal species for the antibiotic resistance study.
(2) Sections of synechocystis PCC 6803 cells were prepared and their morphology and internal structure were characterized by transmission electron microscopy (fig. 2).
(3) The synechocystis PCC 6803 in exponential growth phase was selected and its resistance to eight four major classes of antibiotics, ciprofloxacin (CIP), moxifloxacin (MOX), azithromycin (AZM), clarithromycin (CTM), aureomycin (CTC), oxytetracycline (OTC), sulfamethoxazole (SMZ) and Sulfamethoxazole (SMX) was determined.
90ml of BG11 medium was placed in a 300ml Erlenmeyer flask, and the medium was sterilized by autoclaving. The synechocystis PCC 6803 in the exponential growth phase was diluted with BG11 medium after sterilization. Diluted synechocystis PCC 6803 (initial biomass 1×10) 4 cells/l，OD 680 Approximately 0.1) as algal species. 10ml of algae seeds are added into each 90ml of sterilized culture medium to obtain 100ml of algae liquid.
Standards of representative antibiotics CIP, MOX, AZM, CTM, CTC, OTC, SMX and SMZ were purchased (purity>98%) of various antibiotic mother solutions with concentration of 500mg/L are prepared by methanol, the mother solutions are diluted, antibiotic working solutions with different concentrations are prepared, 100ml of algae solution is added, and the final exposure concentration is 0, 0.01, 0.1, 1, 10 and 100 mug/L. Three biological replicates per group. The antibiotic treatment groups with different concentrations and different types are all placed in an illumination incubator at the temperature of 28+/-1 ℃ and 50 mu mol.m -2 ·s -1 Is cultured for 4 days under the conditions of illumination intensity of 14h illumination/10 h dark illumination period and rotation speed of 120 rpm.
On day 4 of culture, 3ml of algae solution was taken for each group and placed in a cuvette. Determination of OD for each sample with UV Spectrophotometer 680 Absorbance at antibiotic concentration of 0 group and OD of control group 680 Values obtained for each sample were normalized to yield four major classes of growth inhibition dose response curves for eight antibiotics versus synechocystis PCC 6803, the results are shown in tables 1 and 2.
TABLE 1 survival of Synechocystis PCC 6803 at different concentrations of antibiotic exposure (%)
TABLE 2 survival of Synechocystis PCC 6803 at different concentrations of antibiotic exposure (%)
From tables 1 and 2, it can be seen that the growth inhibition rate of the sulfonamide antibiotics (SMX and SMZ) on synechocystis PCC 6803 was the lowest.
Example 2
The upper limit threshold of the concentration of the sulfonamide antibiotic is set to be 1mg/L, and the experimental procedure is the same as that of example 1. The threshold of tolerance of synechocystis PCC 6803 was determined and the results are shown in table 2, table 3 and fig. 3.
TABLE 3 tolerance of different algae to sulfamethoxazole
As can be seen from Table 2, table 3 and FIG. 3, the semi-lethal concentrations of the sulfonamide antibiotics (SMX and SMZ) on the Synechocystis PCC 6803 were higher than 1mg/L. Review of the literature [ Zhang et al (2023) & gt The distinct resistance mechanisms of cyanobacteria and green algae to sulfamethoxazole and its implications for environmental risk assembly Science of The Total Environment, 854, 158723 ] and [ Zhang et al (2020) & gt Effects of erythromycin and sulfamethoxazole on Microcystis aeruginosa: cytotoxic endpoints, production and release of microcystin-LR. Journal of Hazardous Materials, 399, 123021 ] shows that the resistance of synechocystis PCC 6803 to sulfonamide antibiotics is greater than the resistance of other algae.
Example 3
The 4-day removal rate of Synechocystis PCC 6803 was determined for 0.5mg/L and 1mg/L of sulfonamide antibiotic (SMX and SMZ) solutions, and the experimental procedure was as in example 1. The results are shown in Table 4.
TABLE 4 removal rate of sulfonamide antibiotics by Synechocystis PCC 6803
As shown in Table 4, the synechocystis PCC 6803 has good removal effect on the sulfonamide antibiotics, and can be used for treating sewage containing the sulfonamide antibiotics.
Although the foregoing embodiments have been described in some, but not all, embodiments of the invention, it should be understood that other embodiments may be devised in accordance with the present embodiments without departing from the spirit and scope of the invention.
Claims (2)
1. Sea algaeSynechocystissp.) the use of PCC 6803 in the treatment of wastewater containing sulphonamides; the sulfonamide antibiotic is sulfamethoxazole; the concentration of the sulfonamide antibiotics in the wastewater is less than or equal to 1mg/L.
2. Use according to claim 1, characterized in that the waste water comprises production waste water and/or domestic sewage containing sulfonamide antibiotics.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005074986A2 (en) * | 2004-02-10 | 2005-08-18 | Genobiotix Aps | Bioactive species capable of interfering with a microbial toxin-antitoxin complex and methods for evaluation and use of said bioactive species |
| CN102497883A (en) * | 2009-06-03 | 2012-06-13 | 阿尔吉法玛公司 | Alginate oligomers for use in overcoming multidrug resistance in bacteria |
| CN103304029A (en) * | 2012-03-15 | 2013-09-18 | 中国科学院城市环境研究所 | Artificial soil infiltration system for treating rural distributed sewage |
| CN113862127A (en) * | 2021-09-28 | 2021-12-31 | 山东大学 | Device and method for culturing oil-producing blue algae by using sewage containing low-concentration antibiotics |
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| US10773978B2 (en) * | 2016-08-19 | 2020-09-15 | Noblegen, Inc. | Methods and uses of dissolved organic material fractions for binding metal ions |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005074986A2 (en) * | 2004-02-10 | 2005-08-18 | Genobiotix Aps | Bioactive species capable of interfering with a microbial toxin-antitoxin complex and methods for evaluation and use of said bioactive species |
| CN102497883A (en) * | 2009-06-03 | 2012-06-13 | 阿尔吉法玛公司 | Alginate oligomers for use in overcoming multidrug resistance in bacteria |
| CN103304029A (en) * | 2012-03-15 | 2013-09-18 | 中国科学院城市环境研究所 | Artificial soil infiltration system for treating rural distributed sewage |
| CN113862127A (en) * | 2021-09-28 | 2021-12-31 | 山东大学 | Device and method for culturing oil-producing blue algae by using sewage containing low-concentration antibiotics |
Non-Patent Citations (2)
| Title |
|---|
| Efficient degradation of sulfadiazine by photosynthetic cyanobacteria Synechocystis sp. PCC 6803 coupling with recombinant laccase strategy;Ping Wu等;Chemical Engineering Journal;第474卷;1145974 * |
| The transcriptomic analysis revealed sulfamethoxazole stress at environmentally relevant concentration on the mechanisms of toxicity of cyanobacteria Synechococcus sp;Yibo Zhang等;The transcriptomic analysis revealed sulfamethoxazole stress at environmentally relevant concentration on the mechanisms of toxicity of cyanobacteria Synechococcus sp;第2022卷(第10期);107637 * |
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