CN108467842B - Bacterial strain and application thereof - Google Patents
Bacterial strain and application thereof Download PDFInfo
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Abstract
The invention relates to the technical field of microorganisms, in particular to a strain and application thereof. The strain is used for treating the pollutant chlorate in the water body, and after the microbial treatment, the pollutant chlorate in the water body is completely converted into chloride ions, so that the strain has strong bioremediation capability. The strain has wide tolerance pH range during chlorate respiration, can tolerate the pH range of 4.5-8.0, keeps the specific growth rate at a higher level under the condition of pH of 4.5-8.0, grows rapidly, and can reduce chlorate by 100% under the condition of pH of 5.0-8.0. The strain can resist NaClO during chlorate respiration3High concentration, highest tolerance to NaClO3The concentration is 132 mM; within 120h, 100%, 88%, 47%, 48% degradation rate can be achieved for chlorate with initial concentration of 9mM, 19mM, 24mM, 48mM respectively.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to a strain and application thereof.
Background
Chlorate anion (ClO3-), or "chlorate" for short, is a strong oxidizing agent with toxicity, can stably exist in water environment, has the characteristics of high solubility, mobility and the like, and is not easy to be adsorbed or biologically aggregated by soil and sediment particles. Chlorate has been produced industrially for many years and is used in agricultural quantities as a herbicide, defoliant, soil fungicide and as a regulator of the production period in the production of out-of-season longan; in industry, it is used to make perchlorates, chlorites, as oxidizing and mordant agents for aniline dyeing, as bleaching agents in chlorine dioxide production; it is also a by-product of the disinfection of water using chlorine dioxide.
There are currently no established discharge standards for chlorate, but according to previous studies chlorate has been found to have a significant toxic effect on some organisms. It is highly toxic to many microorganisms and algae, and higher doses of chlorate in drinking water may be one of the causes of anemia.
At present, the method for removing the environmental pollutants such as chlorate mainly comprises a physical chemical method and a biological method, wherein the physical chemical method comprises adsorption, ion exchange, membrane technology, chemical reduction and the like. The general physical and chemical methods are expensive and require secondary treatment. The method for removing chlorate by microorganisms has low energy consumption and high efficiency, does not need secondary treatment, has good feasibility and receives more and more attention. An important part of bioremediation methods, which need to be investigated, is microorganisms that can perform chlorate reduction. The bacteria mainly used for chlorate reduction at present include chlorate-reducing bacteria and perchlorate-reducing bacteria, most of which belong to the class of α, β, γ, proteobacteria in the phylum proteobacteria. The variety of chlorate-reducing bacteria is an important factor influencing the chlorate-reducing effect, so that research on screening and culturing of the chlorate-reducing bacteria is necessarily enhanced to obtain different high-efficiency dominant bacterial strains so as to solve the problem that large-scale chlorate-polluted water bodies can be treated. Currently, although many bacteria have been screened for chlorate-reducing activity, such as: bacteria of the genera Deslorospirillum, Magnetospirillum, Desloromonas, Deslorobacter, Propionivibrio, Azospira, Deslorosoma, Pseudomonas, Desloromarinus. However, the tolerance range of the bacteria reported to date to pH during chlorate respiration is not broad enough, and the tolerance of most of the pure cultured bacteria to chlorate at higher concentrations is unclear.
Disclosure of Invention
In view of the above, the present invention provides a strain and an application thereof. Aiming at the defects that the tolerance range of the prior strain to pH is not wide enough and the tolerance condition to chlorate with higher concentration is unclear when the prior strain is used for chlorate respiration, the invention provides a novel strain which can tolerate chlorate with wider pH and higher concentration and can reduce chlorate, and provides a method for eliminating chlorate which is an environmental pollutant by using the strain.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a strain, which has any one of the nucleotide sequences shown as follows:
i, has a nucleotide sequence shown as SEQ ID NO. 1;
II, a nucleotide sequence obtained by modifying, substituting, deleting or adding one or more bases in the nucleotide sequence shown in SEQ ID NO. 1;
III, a sequence with at least 80 percent of homology with the nucleotide sequence shown in SEQ ID NO. 1;
IV, the complement of the sequence shown in I, II or III.
In some embodiments of the invention, the strain preservation number provided by the invention is CCTCC NO: m2018048.
The invention also provides application of the strain in reduction of chlorate.
The invention also provides the application of the strain in sewage treatment.
In some embodiments of the invention, the wastewater contains chlorate.
The invention also provides a method for reducing chlorate and/or treating sewage containing chlorate, which is to activate and expand the strain provided by the invention and then inoculate the strain in a culture medium or sewage containing chlorate.
In some embodiments of the invention, the strain has a minimum tolerance value to pH of 4.5.
In some embodiments of the invention, the pH of the culture medium or wastewater is 4.5 to 8.0.
In some embodiments of the invention, the strain is resistant to chloride at a concentration of 132 mM.
In some embodiments of the invention, the method is specifically: the single bacterial colony of the bacterial strain provided by the invention is inoculated in 3mL LB to be cultured for 12h, then is transferred to 100mL LB to be cultured for 12h, the collected bacteria are added into an inorganic salt culture medium taking sodium chlorate as an electron acceptor to be cultured for 240h, and the bacteria growth condition, the chlorate concentration change and the chloride ion concentration change in the solution are measured by sampling at regular time.
In some embodiments of the invention, the LB medium composition is (g/L): tryptone 10; yeast extract 5; NaCl 10.
In some embodiments of the invention, the mineral salts medium is (g/L): NH (NH)4Cl0.25；NaH2PO4·2H2O 0.78；KCl0.1；NaHCO32.5；NaClO31.064; glucose 1.802.
In some embodiments of the invention, the initial OD of the strain is transferred into an inorganic salt medium600About 0.1.
In some embodiments of the invention, OD is determined using a spectrophotometer600And (4) drawing a bacterial growth curve, and measuring the chlorate concentration change and the chloride ion concentration change in the solution by using an ion chromatography instrument. The strain can grow under the condition of pH4.5-8.0, and sodium chlorate is used as an electron acceptor. Under the condition of pH 4.5-8.0, the specific growth rate of bacteria is kept at a high level, and the bacteria grow rapidly. The bacteria can only reduce 74% of chlorate under the condition of pH4.5, and can reduce 100% of chlorate under the condition of pH 5.0-8.0.
In some embodiments of the invention, the method is specifically: inoculating the single bacterial colony of the bacterial strain provided by the invention into 3mL LB to be cultured for 12h, then transferring the bacterial strain into 100mL LB to be cultured for 12h, collecting bacteria, adding the bacteria into inorganic salt culture media with different pH values and taking sodium chlorate as an electron acceptor, sampling at regular time, and measuring the growth curve and chlorate reduction curve of the bacteria.
In some embodiments of the invention, the buffer system in the inorganic salt medium is 100mM phosphate.
In some embodiments of the invention, the strain is most resistant to NaClO3The concentration was 132 mM. Within 120h, 100%, 88%, 47%, 48% degradation rate can be achieved for chlorate with initial concentration of 9mM, 19mM, 24mM, 48mM respectively.
In some embodiments of the invention, the method is specifically: inoculating the single colony of the separated strain into 3mL LB for culturing for 12h, then transferring into 100mL LB for culturing for 12h, collecting bacteria, adding into inorganic salt culture medium with different concentrations of sodium chlorate, and sampling at regular time to determine the growth condition of the bacteria in the solution.
The invention also provides a preparation for reducing chlorate and/or treating chlorate-containing sewage, which comprises the strain provided by the invention and acceptable auxiliary materials.
The invention screens and obtains the human ochrobactrum strain XM-1 capable of reducing chlorate from microbial flora, wherein the strain is the first bacterium found to have the ability of reducing chlorate in the human ochrobactrum. The bacteria can completely reduce the chlorate of the pollutant in the water body into chloride ions under proper conditions.
The invention discloses a microbial strain, which is XM-1 and is classified and named as human ochrobactrum anthropi (Ochrobactrum anthropi), wherein the preservation number of the microbial strain in China center for type culture collection is CCTCC NO: m2018048.
The pH tolerance range of the screened human ochrobactrum anthropi strain XM-1 during chlorate respiration is wide and is 4.5-8.0. Compared with the previously screened chlorate reducing bacteria, the chlorate reducing bacteria has obviously stronger ability of tolerating acidic conditions, and the specific growth rate of the bacteria is kept at a higher level and grows rapidly under the condition of pH 4.5-8.0, while the specific growth rate of the bacteria is obviously reduced under the condition that the pH of the previously separated chlorate reducing bacteria is lower than 7; the bacteria only degrade 74% of chlorate under the condition of pH4.5, and can degrade 100% of chlorate under the condition of pH 5.0-8.0. Therefore, the strain has application prospect in reducing chlorate of practical water body with wide pH value, especially water body in an acid range.
The screened human ochrobactrum strain XM-1 resists NaClO when carrying out chlorate respiration3High concentration, highest tolerance to NaClO3At a concentration of 132mM, about 48% degradation of 132mM chlorate was achieved in 120 h. The strain has application prospect in reducing high-concentration chlorate water.
Biological preservation Instructions
Biological material: human ochrobactrum XM-1; and (3) classification and naming: human Ochrobactrum anthropi XM-1 (Ochrobactrum anthropi XM-1); the culture is preserved in China center for type culture Collection in 2018, month 01 and day 22, and the addresses of the preservation centers are as follows: wuhan university in Wuhan, China; the preservation number is CCTCC NO: m2018048.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below.
FIG. 1 shows a phylogenetic tree of a strain according to the invention;
FIG. 2 shows chlorate degradation curve, chloride ion generation curve and bacteria growth curve in inorganic salt medium of the strain of the invention;
FIG. 3 shows growth curves of the strains according to the invention under different pH conditions;
FIG. 4 shows the maximum specific growth rates of the strains according to the invention under different pH conditions;
FIG. 5 shows chlorate degradation curves of strains according to the invention under different pH conditions;
FIG. 6 shows growth curves of strains according to the invention in different concentrations of chlorate;
figure 7 shows the chlorate degradation curves of the strains according to the invention in different concentrations of chlorate.
Detailed Description
The invention discloses a strain and application thereof, and can be realized by appropriately improving process parameters by referring to the content in the text by a person skilled in the art. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the methods and applications of this invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications in the methods and applications described herein, as well as other suitable variations and combinations, may be made to implement and use the techniques of this invention without departing from the spirit and scope of the invention.
Aiming at the defects that the tolerance range of the prior strain on pH is not wide enough when chlorate respiration is carried out, and the tolerance condition on chlorate with higher concentration is unclear, the novel strain which can tolerate chlorate with wider pH and higher concentration and can reduce chlorate is provided, and the method for eliminating the chlorate which is an environmental pollutant is provided by using the strain.
The initial inoculum used by the invention is derived from activated sludge of an oxidation ditch process of a sewage treatment plant of a hope pond of the Anhui province fertilizer city, and is used within one week after sampling.
The invention adopts a liquid selective medium enrichment method to enrich the required strains. Inoculating 5g sludge into anaerobic serum bottle, adding 50mL inorganic salt culture medium into serum bottle, adding NaClO3(10mM) as electron acceptor, CH3COONa (10mM) as an electron donor. The inorganic salt culture medium comprises the following components in percentage by weight (g/L): NH (NH)4Cl 0.25；NaH2PO4·2H2O 0.78；KCl0.1；NaHCO32.5; 10mL of trace elements and 10mL of vitamins. The trace elements consist of (g/L): 1.5 parts of nitrilotriacetic acid; MgSO (MgSO)43.0；MnSO4·H2O0.5；NaCl 1.0；FeSO4·7H2O 0.1；CaCl2·2H2O 0.1；CoCl2·6H2O 0.1；ZnCl 0.13；CuSO40.01；AlK(SO4)2·12H2O 0.01；H3BO20.01；Na2MoO40.025；NiCl2·6H2O 0.024；Na2WO4·2H2O0.025. The vitamin composition is (mg/L): biotin 2; 2 parts of folic acid; vitamin B25; vitamin B15; 5 parts of nicotinic acid; vitamin B55; 120.1 of vitamin B; p-aminobenzoic acid 5. The pH of the medium was about 7.2, and no additional pH adjustment step was required. With N2/CO2Gas (N)2:CO2When the ratio is 8: 2) the medium in the serum bottle was aerated for 20min to remove oxygen from the bottle to maintain an anaerobic environment in the bottle, and then sterilized (121 ℃, 20 min). Inoculating sludge into the serum bottle for the first time, and culturing in a shaking table (30 ℃, 200rpm) for 10 days; transferring for the second time, taking 10% of the mixture in the serum bottle transferred for the last time, transferring into a fresh culture medium, and culturing in a shaking table (30 ℃, 200 rpm); then the continuous switching is carried out according to the same method,and after the transfer is carried out for more than or equal to three times, the solution in the serum bottle is in a turbid state after overnight culture, and the enrichment step is stopped.
The present invention uses a solid selection medium to isolate the desired strain. Taking the culture medium in the serum bottle transferred last time in the enrichment step, and coating the culture medium on an inorganic salt solid culture medium which is NaClO3(10mM) as electron acceptor, CH3COONa (10mM) as electron donor, 1.5% Agar was added. The single colonies with different forms growing on the plate are respectively picked out, and then the pure bacteria are obtained through continuous purification steps.
In the invention, the chlorate reducing capability of the pure bacteria is obtained by verification and separation, and the target bacterial strain is obtained. A single colony of the obtained bacterium was selected and placed in 3mL of LB medium, the composition of which was (g/L): tryptone 10; yeast extract 5; NaCl10, placed in a shaker (30 ℃, 200rpm) for 12 h. Quantitative removal (1mL, OD)6002.472) was transferred into 100mL of LB medium and cultured on a shaker (30 ℃ C., 200rpm) for 12 hours. Centrifuging the cultured bacteria in 100mL LB (6000g, 5min), removing supernatant, and collecting thallus; washing with inorganic salt culture medium once, centrifuging (6000g, 5min), removing supernatant, and collecting thallus; resuspend bacteria using mineral salts media and inject into serum flasks containing 50mL of mineral salts media (control of initial OD of bacteria)600About 0.1), NaClO3(10mM) as an electron acceptor and glucose (10mM) as an electron donor were incubated in a shaker (30 ℃ C., 200rpm) and sampled periodically. The target strain can grow in the culture medium, and the chlorate concentration in the culture medium is reduced along with time, so that the target strain is judged to be obtained.
The bacterial classification was determined by determining the 16S rDNA sequence of the isolated strain. Extracting the strain DNA, amplifying the 16S rDNA gene of the strain by using primers 27F and 1492R, connecting the product with a T vector, and confirming the sequence of the fragment by sequencing. The sequence was aligned with the relevant data in GenBank, and the homology of the strain with human ochrobactrum anthropi (Ochrobactrum anthropi) reached 99%. The strain was identified as Ochrobactrum anthropi (Ochrobactrum anthropi) and named as XM-1.
The microorganism is preserved in China center for type culture Collection with the preservation number of CCTCC NO: m2018048.
The invention changes the pH of the inorganic salt culture medium and determines the growth condition and chlorate reduction condition of the obtained bacteria under different pH conditions. NaClO in inorganic salt culture medium3(10mM) as an electron acceptor and glucose (10mM) as an electron donor, the pH of the medium was adjusted to 4.0, 4.5, 5.0, 6.0, 6.5, 7.0, 7.5, 8.0, and 9.0 by changing the buffer salt of bicarbonate-dihydrogen phosphate to 100mM phosphate buffer salt in the original medium, respectively. The result shows that the bacterium can tolerate the pH range of 4.5-8.0, and the specific growth rate of the bacterium is kept at a higher level and grows rapidly under the condition of the pH of 4.5-8.0; the bacteria can only reduce 74% of chlorate under the condition of pH4.5, and can reduce 100% of chlorate under the condition of pH 5.0-8.0.
The invention changes electron donor NaClO3Determining the concentration of the resulting bacteria at different concentrations of NaClO3Growth under the conditions of (1) and reduction of chlorate. The results show that the bacterium can tolerate NaClO to the highest degree3The concentration is 132 mM; within 120h, 100%, 88%, 47%, 48% degradation rate can be achieved for chlorate with initial concentration of 9mM, 19mM, 24mM, 48mM respectively.
The invention has the beneficial effects that:
the invention relates to a method for screening a human ochrobactrum strain XM-1 capable of reducing chlorate from a microorganism group, wherein the microorganism is the first bacterium found to have the ability of reducing chlorate in the human ochrobactrum. The bacteria can completely reduce the chlorate of the pollutant in the water body into chloride ions under proper conditions.
Secondly, the pH tolerance range of the screened human ochrobactrum anthropi strain XM-1 is wide when chlorate respiration is carried out, and the tolerance pH range is 4.5-8.0. Compared with the previously screened chlorate reducing bacteria, the chlorate reducing bacteria has obviously stronger acid condition tolerance capability, the specific growth rate of the bacteria is kept at a higher level and grows rapidly under the condition of pH 4.5-8.0, and the specific growth rate of the bacteria is obviously reduced under the condition that the pH value of the previously separated chlorate reducing bacteria is lower than 7; the bacteria only degrade 74% of chlorate under the condition of pH4.5, and can degrade 100% of chlorate under the condition of pH 5.0-8.0. Therefore, the strain has application prospect in reducing chlorate of practical water body with wide pH value, especially water body in an acid range.
Thirdly, the human ochrobactrum anthropi strain XM-1 obtained by screening of the invention is resistant to NaClO in chlorate respiration3High concentration, highest tolerance to NaClO3At a concentration of 132mM, about 48% degradation of 132mM chlorate was achieved in 120 h. The strain has application prospect in reducing high-concentration chlorate water.
The strain and the raw materials and reagents used in the whole test process of the application of the strain provided by the invention can be purchased from the market.
The invention is further illustrated by the following examples:
example 1: screening of the microorganism strains of the present invention
Liquid selective medium enrichment method: the initial inoculum uses activated sludge of oxidation ditch process of Wangquan pond sewage treatment plant of Anhui province, and 5g of sludge is inoculated into an anaerobic serum bottle, 50mL of inorganic salt culture medium is added into the serum bottle, and NaClO is used as the culture medium3(10mM) as electron acceptor, CH3COONa (10mM) as an electron donor. The pH of the medium was about 7.2, and no additional pH adjustment step was required. With N2/CO2Gas (N)2:CO2When the ratio is 8: 2) the medium in the serum bottle was aerated for 20min to remove oxygen from the bottle to maintain an anaerobic environment in the bottle, and then sterilized (121 ℃, 20 min). Inoculating sludge into the serum bottle for the first time, and culturing in a shaking table (30 ℃, 200rpm) for 10 days; transferring for the second time, taking 10% of the mixture in the serum bottle transferred for the last time, transferring into a fresh culture medium, and culturing in a shaking table (30 ℃, 200 rpm); and then, carrying out continuous transfer by the same method, and after the transfer is carried out for more than or equal to three times, carrying out overnight culture to ensure that the solution in the serum bottle is in a turbid state, which indicates that the target strain possibly is enriched.
Solid selective medium separation method: taking the culture medium in the serum bottle transferred last time in the enrichment step, coating the culture medium on an inorganic salt solid culture mediumAdding NaClO to culture medium, adding inorganic salt solid culture medium3(10mM) as electron acceptor, CH3COONa (10mM) as electron donor, 1.5% Agar was added. The single colonies growing on the plate with different forms are picked out respectively, and then the obtained bacteria are purified by continuously streaking on a solid culture medium. By the method, the microbial strain XM-1 is screened out.
The inorganic salt culture medium comprises the following components in percentage by weight (g/L): NH (NH)4Cl 0.25；NaH2PO4·2H2O 0.78；KCl0.1；NaHCO32.5; 10mL of trace elements and 10mL of vitamins. The trace elements consist of (g/L): 1.5 parts of nitrilotriacetic acid; MgSO (MgSO)43.0；MnSO4·H2O 0.5；NaCl 1.0；FeSO4·7H2O 0.1；CaCl2·2H2O 0.1；C℃l2·6H2O 0.1；ZnCl0.13；CuSO40.01；AlK(SO4)2·12H2O 0.01；H3BO20.01；Na2MoO40.025；NiCl2·6H2O0.024；Na2WO4·2H2O0.025. The vitamin composition is (mg/L): biotin 2; 2 parts of folic acid; vitamin B25; vitamin B15; 5 parts of nicotinic acid; vitamin B55; 120.1 of vitamin B; p-aminobenzoic acid 5.
Example 2: identification of the microorganism strains according to the invention
Extracting the strain DNA according to the steps of a genome DNA extraction kit of the live Ezup columnar bacteria, and performing the following steps by using a primer: 27F: 5'-AGAGTTTGATCCTGGCTCAG-3' and 1492R: 5'-GGTTACCTTGTTACGACTT-3' amplifying the 16S rDNA gene of the strain, connecting the product with T carrier, and sequencing to confirm that the actual length of the fragment is 1443 bp. The sequence is compared with related data in GenBank, and in the result of the highest comparison matching degree, the homology of the 16S rDNA sequence part measured by the strain and the human ochrobactrum anthropi reaches 99 percent, so that the strain is confirmed to be the human ochrobactrum anthropi and named as XM-1.
Based on the measured 16S rDNA gene sequence of the isolate (shown as SEQ ID No. 1) and the 16SrDNA gene sequences of other bacteria with higher matching degree with the alignment result in GenBank and some (high) chlorate-reducing bacteria reported in other literatures, a phylogenetic tree of the isolate was made according to the adjacency method by using MEGA6 software, as shown in FIG. 1. As can be seen from FIG. 1, most of the (per) chlorate-reducing bacteria known so far belong to Proteobacteria, and are mainly distributed in the beta and alpha Proteobacteria, and the strain of bacteria isolated by the present invention belongs to the genus Ochrobactrum which is below the alpha Proteobacteria, and no other bacteria in the genus Ochrobactrum have been reported to have the ability to reduce chlorate.
Example 3: method for reducing chlorate by using bacterial strain XM-1
The strain was selected and single-cloned into 3mL of LB medium and cultured on a shaker (30 ℃ C., 200rpm) for 12 hours. Quantitative removal (1mL, OD)6002.472) was transferred into 100mL of LB medium and cultured on a shaker (30 ℃ C., 200rpm) for 12 hours. Centrifuging the cultured bacteria in 100mL LB (6000g, 5min), removing supernatant, and collecting thallus; washing with inorganic salt culture medium once, centrifuging (6000g, 5min), removing supernatant, and collecting thallus; resuspend bacteria using mineral salts media and inject into serum flasks containing 50mL of mineral salts media (control of initial OD of bacteria)600About 0.1), NaClO3As an electron acceptor, glucose as an electron donor was co-cultured in a shaker (30 ℃ C., 200rpm) for 240 hours, and samples were taken periodically.
Determination of OD of sample Using Spectrophotometer600The growth curve was plotted, and the chlorate concentration in the medium was measured by an ion chromatography apparatus, and the results are shown in fig. 2 and table 1. Within 120h of culture, the strain can utilize glucose and NaClO3Respiration is carried out to realize the growth of bacteria, and the strain can convert all sodium chlorate in the culture medium into chloride ions.
TABLE 1
Example 4: growth of strain XM-1 under different pH conditions and reduction of chlorate
The isolated pure culture strains were selected and monocloned into 3mL of LB medium and cultured for 12 hours on a shaker (30 ℃, 200 rpm). A certain amount of the bacteria were transferred to 100mL of LB medium and cultured in a shaker (30 ℃ C., 200rpm) for 12 hours. Centrifuging the cultured bacteria in 100mL LB (6000g, 5min), removing supernatant, and collecting thallus; washing with inorganic salt culture medium once, centrifuging (6000g, 5min), removing supernatant, and collecting thallus; the bacteria were resuspended in mineral salt medium and injected into 50mL of mineral salt medium with pH 4.0, 4.5, 5.0, 6.0, 6.5, 7.0, 7.5, 8.0, 9.0 (to control initial OD of bacteria)600About 0.1). The inorganic salt culture medium is NaClO3(10mM) as an electron acceptor and glucose (10mM) as an electron donor, the buffer salt of bicarbonate-dihydrogen phosphate in the original medium was replaced with 100mM phosphate buffer salt, and the pH was adjusted to 4.0, 4.5, 5.0, 6.0, 6.5, 7.0, 7.5, 8.0, and 9.0, respectively.
The samples were taken periodically and the growth curve and chlorate reduction curve of the bacteria were determined. As shown in FIG. 3 and tables 2 to 3, the bacterium can grow under the condition of pH4.5 to 8.0. As shown in FIG. 4 and Table 4, the specific growth rate of the bacteria was maintained at a high level and the growth was rapid under the conditions of pH4.5 to 8.0. As shown in FIG. 5 and tables 5 to 6, the bacteria can reduce chlorate by 100% under the condition of pH 5.0 to 8.0 except that the bacteria can reduce chlorate by 74% under the condition of pH 4.5.
TABLE 2
TABLE 3
TABLE 4
TABLE 5
TABLE 6
Example 5: growth of strain XM-1 in chlorate environment with different concentrations and reduction of chlorate
The isolated pure culture strains were selected and monocloned into 3mL of LB medium and cultured for 12 hours on a shaker (30 ℃, 200 rpm). A certain amount of the bacteria were transferred to 100mL of LB medium and cultured in a shaker (30 ℃ C., 200rpm) for 12 hours. Centrifuging the cultured bacteria in 100mL LB (6000g, 5min), removing supernatant, and collecting thallus; washing with inorganic salt culture medium once, centrifuging (6000g, 5min), removing supernatant, and collecting thallus; the bacteria were resuspended in mineral salt medium and separately injected into mineral salt medium containing 50mL of different chlorate concentration (control of initial OD of bacteria)600About 0.1). The inorganic salt medium used glucose (10mM) as an electron donor.
The samples were taken periodically and the growth curve and chlorate reduction curve of the bacteria were determined. As shown in FIG. 6 and Table 7, the bacterium was found to be the most resistant to NaClO3The concentration was 132 mM. As shown in FIG. 7 and Table 8, the initial concentrations of chlorate of 9mM, 19mM, 24mM and 48mM can be 100%, 88%, 47% and 48% respectively within 120 h.
TABLE 7
TABLE 8
Comparative example
The culture conditions were the same as in example 4.
The maximum growth rate of the bacteria at different pH values is shown in table 9:
TABLE 9
Under the condition of the same pH value, the maximum growth rate of the strain of the comparative example is very different from that of the strain provided by the invention (P < 0.01), which shows that the tolerance of the strain provided by the invention to the pH value is very much better than that of the strain of the comparative example.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Sequence listing
<110> university of science and technology in China
<120> bacterial strain and use thereof
<130>MP1800943
<160>1
<170>SIPOSequenceListing 1.0
<210>1
<211>1443
<212>DNA
<213> human Ochrobactrum anthropic
<400>1
gagtttgatc ctggctcaga acgaacgctg gcggcaggct taacacatgc aagtcgagcg 60
ccccgcaagg ggagcggcag acgggtgagt aacgcgtggg aacgtacctt ttgctacgga 120
ataactcagg gaaacttgtg ctaataccgt atgtgccctt cgggggaaag atttatcggc 180
aaaggatcgg cccgcgttgg attagctagt tggtgaggta aaggctcacc aaggcgacga 240
tccatagctg gtctgagagg atgatcagcc acactgggac tgagacacgg cccagactcc 300
tacgggaggc agcagtgggg aatattggac aatgggcgca agcctgatcc agccatgccg 360
cgtgagtgat gaaggcccta gggttgtaaa gctctttcac cggtgaagat aatgacggta 420
accggagaag aagccccggc taacttcgtg ccagcagccg cggtaatacg aagggggcta 480
gcgttgttcg gatttactgg gcgtaaagcg cacgtaggcg gacttttaag tcaggggtga 540
aatcccgggg ctcaaccccg gaactgcctt tgatactgga agtcttgagt atggtagagg 600
tgagtggaat tccgagtgta gaggtgaaat tcgtagatat tcggaggaac accagtggcg 660
aaggcggctc actggaccat tactgacgct gaggtgcgaa agcgtgggga gcaaacagga 720
ttagataccc tggtagtcca cgccgtaaac gatgaatgtt agccgttggg gagtttactc 780
ttcggtggcg cagctaacgc attaaacatt ccgcctgggg agtacggtcg caagattaaa 840
actcaaagga attgacgggg gcccgcacaa gcggtggagc atgtggttta attcgaagca 900
acgcgcagaa ccttaccagc ccttgacata ccggtcgcgg acacagagat gtgtctttca 960
gttcggctgg accggataca ggtgctgcat ggctgtcgtc agctcgtgtc gtgagatgtt 1020
gggttaagtc ccgcaacgag cgcaaccctc gcccttagtt gccagcattt agttgggcac 1080
tctaagggga ctgccggtga taagccgaga ggaaggtggg gatgacgtca agtcctcatg 1140
gcccttacgg gctgggctac acacgtgcta caatggtggt gacagtgggc agcgagcacg 1200
cgagtgtgag ctaatctcca aaagccatct cagttcggat tgcactctgc aactcgagtg 1260
catgaagttg gaatcgctag taatcgcgga tcagcatgcc gcggtgaata cgttcccggg 1320
ccttgtacac accgcccgtc acaccatggg agttggtttt acccgaaggc gctgtgctaa 1380
ccgcaaggag gcaggcgacc acggtagggt cagcgactgg ggtgaagtcg taacaaggta 1440
acc 1443
Claims (7)
1. The strain is characterized in that the preservation number is CCTCC NO: m2018048.
2. Use of a strain according to claim 1 for the reduction of chlorate.
3. The use of the strain of claim 1 for treating wastewater.
4. Use according to claim 3, wherein the effluent contains chlorate.
5. A method for reducing chlorate and/or treating chlorate-containing wastewater, comprising activating and expanding the strain of claim 1 and inoculating the strain in a chlorate-containing medium or wastewater.
6. The method according to claim 5, wherein the pH of the culture medium or the wastewater is 4.5 to 8.0.
7. A preparation for reducing chlorate and/or treating chlorate-containing wastewater comprising the strain of claim 1 and an acceptable adjuvant.
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| CN101676224A (en) * | 2008-09-18 | 2010-03-24 | 中国科学院福建物质结构研究所 | Method for treating industrial chromium-containing waste water by bacteria |
| CN105670955A (en) * | 2015-10-14 | 2016-06-15 | 四川农业大学 | Ochrobacterum sp. MGJ11, and method of immobilizing heavy metal cadmium in soil therewith |
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