CN114410503A - Manganese oxidizing bacteria and screening method and application thereof - Google Patents
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Abstract
The invention discloses a manganese oxidizing bacterium, a screening method and an application thereof, wherein the manganese oxidizing bacterium belongs to Morganella (Morganella Morganii) and is preserved in the China general microbiological culture Collection center (CGMCC) on 7-month and 2-month 2021 with the preservation number of CGMCC No. 22815. The manganese oxidizing bacteria obtained by screening is a strain of Morganella discovered to have manganese oxidizing capability for the first time, and the strain has the advantages of strong manganese oxidizing capability, good environmental adaptability and extremely high research value; the method for preparing the biological manganese oxide by oxidizing the manganese oxidizing bacteria has the advantages of simple operation, low cost and the like, and is favorable for the application of the biological manganese oxide in the field of arsenic detoxification.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to manganese oxidizing bacteria and a screening method and application thereof.
Background
Arsenic (As) is a global pollutant, and after excessive arsenic enters a human body, the redox capability of human cells can be damaged, the normal metabolism of the cells is influenced, tissue damage and body disorder are caused, and the serious people can directly cause toxic death.
Arsenic As in the environment3+And As5+Exist in two forms, and As3+Is far more toxic than As5+Wherein: as3+Toxicity is about As5+60 times as toxic. In addition, related studies have found that As is associated with5+In contrast, As3+The mobility is higher and the adsorption rate is lower; the reason for this is that As is present at a pH of between 0 and 9.03+Is H3AsO3The surface is not charged, so that the material is not easily adsorbed by charged substances, the mobility is higher, and As5+In the condition of pH > 2.0, it is mainly present in the form of H2AsO4-、HAsO4 2-And AsO4 3-And the material is easy to combine with substances with positive charges on the surface, so that the mobility of the material is reduced.
Therefore, highly toxic and easily migrating As3+Oxidation to As of low toxicity and low mobility5+Is a better choice for reducing the toxicity of arsenic in water.
In the prior art, As3+The oxidation techniques of (a) include an oxidizing agent oxidation method, a photocatalytic oxidation method and a biological method.
Among them, biological methods are receiving more and more attention because of their advantages of low cost and no secondary pollution. The manganese oxidizing microorganisms are the prejudices in microbial oxidation, and biological manganese oxides generated by the oxidation of the manganese oxidizing microorganisms have the advantages of low charge zero point, large surface area, high negative charge, high activity and strong oxidation adsorption capacity, are usually wrapped on the surfaces of other minerals and have strong reaction activity. As an important natural adsorption carrier, a redox main body and a catalyst for chemical reaction, the biological manganese oxide plays a decisive role in the aspects of biological effectiveness, physiological toxicity, migration and conversion in the environment, degradation of organic pollutants and the like of inorganic pollutants and plays an important role in the biogeochemical cycle of various substances. As can be effectively reduced by high-valence biological manganese oxide3+Oxidized to As5+Compared with chemical manganese oxide, the biological manganese oxide has higher oxidation speed and higher efficiency, and the manganese oxide reduces Mn formed by trivalent arsenic2+Can be continuously reacted by manganese oxidizing bacteria to form biological manganese oxide, thereby achieving the effect of continuously detoxifying arsenic.
However, the common manganese oxidizing bacteria have low efficiency of oxidizing manganese and low tolerance to arsenic, and cannot achieve good arsenic detoxification effect.
Therefore, screening of a manganese oxidizing bacterium with high oxidation efficiency on manganese and high tolerance on arsenic is a necessary measure for making up the current deficiency and improving the application of biological manganese oxide in the field of arsenic detoxification.
In view of this, the invention is particularly proposed.
Disclosure of Invention
In view of the above, the invention provides a manganese oxidizing bacterium, a screening method and an application thereof.
In order to achieve the purpose, the technical scheme of the invention is as follows:
the invention provides a manganese oxidizing bacterium, which belongs to Morganella (Morganella Morganii) and is named as Morganella (Morganella sp) MnOx-1, and is preserved in the China general microbiological culture Collection center on 7-month and 2-month 2021 with the preservation number of CGMCC No. 22815.
Specifically, the manganese oxidizing bacteria are obtained by screening and separating the sludge collected from the wetland in the ocean lake of Changsha city, Hunan province, and are currently preserved in the general microbiological center of China Committee for culture Collection of microorganisms, addresses: western road No. 1, north chen, chaoyang district, beijing, ministry of sciences, china, institute of microbiology, zip code 100101; the 16S rRNA gene sequence was identified to have 99% similarity to the Morganella Morganii 16S rRNA gene sequence.
In another aspect, the present invention provides a method for screening the manganese oxidizing bacteria, comprising:
s1, taking supernatant after the sludge sample is stood, adding the supernatant into MnSO4·H2O and HEPES buffer in LB liquid medium;
s2, after constant temperature shaking culture for 5-9d, the culture medium is checked and screened primarily by LBB indicator.
Further, in the above technical solution, the method for screening manganese-oxidizing bacteria further includes:
s3, diluting and spreading the culture solution which shows blue color to LBB indicator in step S2 to MnSO-containing culture solution4·H2Culturing in solid culture medium containing O and HEPES buffer solution at constant temperature, further checking and rescreening with LBB indicator, separating and purifying the LBB indicator showing blue color, mixing the obtained strain with glycerol at a ratio of 1: 1, and preserving at-80 deg.C.
Specifically, in the above technical solution, in step S1, the LB liquid medium includes the following components:
10g of tryptone, 5g of yeast extract powder, 5g of sodium chloride and 1000ml of distilled water, wherein the pH value is 7, and the mixture is MnSO4·H2O1 mmol/L, HEPES buffer 20 mmol/L.
Specifically, in the above technical solution, in step S2, the LBB indicator is prepared as follows:
0.04g LBB powder is weighed and dissolved in 0.25ml of 45mmol/L glacial acetic acid water solution, deionized water is added, the volume is adjusted to 100ml, and the mixture is stored in a dark place at 4 ℃.
In one embodiment of the present invention, step S3 contains MnSO4·H2The preparation method of the solid culture medium of O and HEPES buffer solution is as follows:
dissolving tryptone 10g, yeast extract 5g, sodium chloride 5g and agar 15g in 1000ml distilled water, adjusting pH to 7, sterilizing at 121 deg.C for 20min, filtering with 0.22 μm filter head, and adding MnSO 1mol/L4·H2O and 1mol/L HEPES buffer solution to make Mn2+And HEPES buffer solution with final concentration of 1mmol/L and 20mmol/L, respectively, cooling to 50 deg.C, pouring the plate on a sterile operating table, and condensing the solid plate for use.
The invention also provides a microbial inoculum containing the manganese oxidizing bacteria.
The invention also provides the application of the manganese oxidizing bacteria or the microbial inoculum in preparing biological manganese oxide for oxidizing trivalent arsenic.
On the other hand, the invention also provides the application of the manganese oxidizing bacteria or the microbial inoculum in oxidizing trivalent arsenic.
Specifically, in the application of the trivalent arsenic oxide, the manganese oxidizing bacteria oxidize Mn by2+Preparing biological manganese oxide, and oxidizing trivalent arsenic into pentavalent arsenic by the biological manganese oxide.
Compared with the prior art, the invention has the following advantages:
(1) the manganese oxidizing bacteria obtained by screening is a strain of Morganella discovered to have manganese oxidizing capability for the first time, and the strain has the advantages of strong manganese oxidizing capability, good environmental adaptability and extremely high research value;
(2) the method for preparing the biological manganese oxide by oxidizing the manganese oxidizing bacteria has the advantages of simple operation, low cost and the like, and is favorable for the application of the biological manganese oxide in the field of arsenic detoxification.
Drawings
FIG. 1 is a colony morphology of manganese oxidizing bacteria according to an embodiment of the present invention;
FIG. 2 is a phylogenetic tree of manganese oxidizing bacteria according to an embodiment of the present invention;
FIG. 3 is a graph showing the oxidation rate of manganese-oxidizing bacteria to 1mmol/L manganese in the example of the present invention;
FIG. 4 is a graph of the manganese oxidation rate of manganese oxidizing bacteria at different pH values according to an embodiment of the present invention;
FIG. 5 is a graph showing the activity of manganese oxidizing bacteria at different pH levels in an example of the present invention;
FIG. 6 is a graph of the manganese oxidation rate of manganese-oxidizing bacteria with different divalent manganese contents according to an embodiment of the present invention;
FIG. 7 is a graph showing the activity of manganese oxidizing bacteria at different divalent manganese contents according to an embodiment of the present invention;
FIG. 8 is an SEM photograph of biological oxides of manganese produced by an oxidizing bacteria of the present invention;
FIG. 9 shows a pair of manganese oxidizing bacteria 100mg/LAs in an embodiment of the present invention3+Oxidation rate chart.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail 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 examples, the means used are conventional in the art unless otherwise specified.
The terms "comprises," "comprising," or any other variation thereof, as used herein, are intended to cover a non-exclusive inclusion. For example, a composition, process, method, article, or apparatus that comprises a list of elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such composition, process, method, article, or apparatus.
In addition, the technical features involved in the embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.
Unless otherwise specified, the raw materials used in the examples of the present invention are all commercially available products.
A manganese oxidizing bacterium is preserved in China general microbiological culture Collection center on 7-month and 2-month 2021, with the preservation address: beijing, Chaoyang, West Lu No. 1, Beijing, the institute for microbiology, Chinese academy of sciences, zip code 100101, with the number CGMCC No. 22815.
In order to facilitate a further understanding of the present invention, the technical solutions of the present invention will now be described in detail with reference to the preferred embodiments.
Example 1 screening, isolation and purification of manganese oxidizing bacteria
The method comprises the following specific steps:
(1) preparing a liquid culture medium:
weighing 10g of tryptone, 5g of yeast extract powder and 5g of sodium chloride, adding into 1000ml of distilled water, adjusting the pH to 7, subpackaging 100ml of the mixture in a 250ml conical flask, sterilizing the mixture at 121 ℃ for 20min in a sterilizing pot, filtering the mixture by using a 0.22 mu m filter head, and adding 1mol/L of MnSO4 & H2O aqueous solution and 1mol/L HEPES buffer solution, and allowing Mn to exist2+And HEPES buffer at final concentrations of 1mmol/L and 20mmol/L, respectively.
(2) Preparing a solid culture medium:
weighing 10g of tryptone, 5g of yeast extract powder, 5g of sodium chloride and 15g of agar, adding into 1000ml of distilled water, adjusting the pH to 7, sterilizing at 121 ℃ for 20min in a sterilizing pot, filtering by using a 0.22 mu m filter head, and adding 1mol/L of MnSO4 & H2O aqueous solution and 1mol/L HEPES buffer solution to make Mn2+And HEPES buffer solution final concentration of 1mmol/L and 20mmol/L, cooling the culture medium to 50 deg.C, pouring the plate on a sterile operating table, and condensing the solid plate for use.
(3) Screening and separating manganese oxidizing bacteria:
collecting sludge from the ocean lake wetland of Changsha city, Hunan province, standing, taking supernatant, adding the supernatant into the liquid culture medium in the step (1) according to the inoculation amount of 1%, performing shake culture on a constant-temperature shaking table at 30 ℃ and 150rpm for 7 days, qualitatively detecting whether manganese oxide is generated by adopting an LBB indicator, and performing primary screening on the culture solution with the LBB indicator showing that the color of the culture solution is changed into blue.
The LBB indicator is prepared by the following method: 0.04g of LBB powder was weighed, dissolved in 0.25ml of glacial acetic acid aqueous solution (45mmol/L), added with deionized water and made to volume of 100ml, and stored at 4 ℃ in the dark.
(4) Purification of manganese oxidizing bacteria
Diluting the culture solution for developing the blue color of the LBB indicator in the step (3), coating and separating the diluted culture solution into the solid culture medium in the step (2), placing the solid culture medium in a 30 ℃ oven for constant temperature culture, observing the colony morphology, carrying out a color development experiment by using the LBB indicator, further carrying out streak separation and purification on the blue colony, uniformly mixing the purified strain and glycerol according to the ratio of 1: 1, and then storing the mixture in a refrigerator at-80 ℃.
Example 2 morphological characteristics and molecular characterization of manganese oxidizing bacteria
Morphological characteristics:
as shown in figure 1, according to the result of a scanning electron microscope, the strain is rod-shaped as a whole, the colony is circular, the center is opaque and milky, the surface is convex and wet, and the edge is brown regularly.
Molecular identification and phylogenetic evolutionary tree alignment:
and (3) carrying out DNA extraction on the separated and purified manganese oxidizing bacteria, finishing relevant extraction operation by adopting a bacterial DNA extraction kit produced by magenta company, and then carrying out PCR amplification, wherein the primers are bacterial 16S rDNA universal primers (1492R- 'GGTTACCTTGTTACGA CTT', 27F- 'AGAGTTTGATCMTGGCTCAG'). Sequencing of the PCR products was performed by Shanghai bioengineering, Inc. The 16S rDNA results obtained by sequencing (as shown in Seq ID No: 1) were compared with the existing 16S rDNA nucleic acid sequences for similarity by the Blast program of NCBI (national center for Biotechnology information). The phylogenetic evolutionary tree is analyzed and drawn by Mega4.0 software, and is shown in figure 2.
Sequence results analysis showed that the manganese oxidizing bacterium has the highest similarity to Morganella Morganii, was identified as belonging to the genus Morganella, and was named MnOx-1.
Example 3 determination of manganese oxidizing ability of manganese oxidizing bacteria MnOx-1
(1) A250 ml Erlenmeyer flask was selected to prepare 100m1 manganese-containing liquid medium in the same manner as in step (1) of example 1, namely: weighing 10g of tryptone, 5g of yeast extract powder and 5g of sodium chloride, adding into 1000ml of distilled water, and adjustingpH of 7, packaging 100ml into 250ml conical flask, sterilizing at 121 deg.C for 20min, filtering with 0.22 μm filter head, and adding 1moL/L MnSO 4. H2O aqueous solution and 1moL/L HEPES buffer solution, and allowing Mn to exist2+And HEPES buffer at final concentrations of 1mmol/L and 20mmol/L, respectively.
(2) Adding manganese oxidizing bacteria into the manganese-containing liquid culture medium in the step (1) according to the inoculation amount of 1%, placing each conical flask in a constant-temperature shaking table at 30 ℃ and 180rpm for shake culture, wherein 3 conical flasks are parallel to each other, and 1 blank reference is adopted; sampling every 24 hours in a 10ml centrifuge tube, centrifuging for 3min at 8000rpm of a centrifuge, sucking supernatant, and measuring the residual Mn in the culture solution by using an inductively coupled plasma emission spectrometer (ICP-OES)2+Concentration, sampling was continued for 7 days until the end of the experiment.
(3) The method for calculating the manganese oxidation rate comprises the following steps:
manganese oxidation rate ═ (initial manganese content-residual manganese content)/initial manganese content ═ 100%.
As a result, as shown in FIG. 3, it was found from the analysis of FIG. 3 that the manganese oxidizing bacteria can oxidize Mn2+High oxidation rate, Mn at 1-2 days2+Is rapidly oxidized, Mn at 2-7 days2+The oxidation gradually tends to be smooth, the maximum manganese oxidation rate reaches 88.17%, and the manganese oxidation capacity is extremely strong.
Example 4 determination of manganese Oxidation Capacity at different pH and divalent manganese content
(1) Determination of manganese Oxidation Capacity under different pH conditions
A manganese-containing liquid medium was prepared in the same manner as in the step (1) in example 3, the pH in the manganese-containing liquid medium was adjusted so that the pH gradient was 5, 6, 7, 8 and 9, 100ml of the manganese-containing liquid medium was put into a 250ml Erlenmeyer flask, the inoculum size of the inoculum was 1% and inoculated into manganese-containing liquid media of different pH, and shaking culture was carried out at 180rpm at 30 ℃ in a constant temperature, 3 parallel groups, and 1 blank control.
Sampling from different culture media according to a certain time, sampling for 7 days totally, measuring the residual amount of Mn in a centrifuge tube after filtration by an atomic absorption spectrometry, measuring an OD600 value by an ultraviolet spectrophotometer, and respectively determining the manganese oxidation rate and the activity of manganese oxidizing bacteria under different pH values.
As shown in FIG. 4, the manganese oxidizing bacteria MnOx-1 has a good manganese oxidation rate at pH 5-9, and reaches about 80% at about 72 h; the manganese oxidation rate is slightly lower when the pH value is 5, the removal effect of the manganese oxidation rate is similar when the pH value is 6, 7, 8 and 9, and the oxidation efficiency of the manganese oxide bacteria manganese oxide in a neutral alkaline environment is better.
As shown in FIG. 5, the activity of the manganese oxidizing bacteria is similar when the pH value is 5-9, which shows that the manganese oxidizing bacteria has wide adaptability and can grow well in acidic to alkaline environments.
(2) Determination of manganese Oxidation Capacity at different divalent manganese contents
A manganese-containing liquid medium was prepared in the same manner as in the step (1) in example 3, and Mn in the manganese-containing liquid medium was adjusted2+ contents of 1, 5, 10, 15 and 20mmol/L respectively, adding 100ml manganese-containing liquid culture medium into a 250ml conical flask, inoculating the bacterial liquid into liquid culture medium with different divalent manganese contents according to the inoculation amount of 1%, performing shake cultivation at constant temperature of 30 ℃ at 180rpm, and performing shake cultivation on 3 parallel groups and 1 blank control.
Sampling from different culture media according to a certain time, sampling for 7 days totally, measuring the residual amount of Mn in a centrifuge tube after filtration by using an atomic absorption spectrometry, measuring an OD600 value by using an ultraviolet spectrophotometer, and respectively determining the manganese oxidation rate and the manganese oxidizing bacteria activity under different divalent manganese contents.
As shown in FIG. 6, the manganese oxidizing bacteria are in Mn2+The manganese oxidation rates at contents of 1, 5, 10, 15, 20mmo/L are very different, depending on Mn2+The content is increased, when the content is 5mmol/L, the manganese oxidation rate reaches the maximum, and Mn is continuously added2+The content and the manganese oxidation rate are gradually reduced, and the manganese oxidation rate is the lowest at 20mmol/L and is only about 20 percent.
As shown in FIG. 7, the change of the content of divalent manganese has a large influence on the activity of manganese-oxidizing bacteria, and a certain amount of Mn is added2+Has certain promotion effect on the growth of manganese oxidizing bacteria, but when Mn is used2+When the content is excessive, the growth of manganese oxidizing bacteria is obviously inhibited, such as Mn2+When the content is 20mmol/L, the growth of the manganese oxidizing bacteria is obviously slower.
Example 5 biological OxidationFormation of manganese and p-As3+Oxidation experiment of
(1) Production of biological manganese oxide
100ml of manganese-containing liquid medium was prepared in the manner described in step (1) in example 1, namely: weighing 10g of tryptone, 5g of yeast extract powder and 5g of sodium chloride, adding into 1000ml of distilled water, adjusting the pH to 7, sterilizing at 121 ℃ for 20min in a sterilizing pot, filtering by using a 0.22 mu m filter head, and adding 1mol/L of MnSO4 & H2O aqueous solution and 1mol/L HEPES buffer solution, and allowing Mn to exist2+And HEPES buffer final concentrations of 1mmol/L and 20mmol/L respectively; adding manganese oxidizing bacteria into a manganese-containing liquid culture medium according to the inoculation amount of 1%, carrying out shake culture in a constant temperature shaking table at 30 ℃ and 180rpm for 7 days, centrifuging at 8000rpm for 3min by using a centrifuge, discarding supernatant, washing and precipitating with deionized water for 3 times, and obtaining precipitate, namely biological manganese oxide.
(2) Scanning Electron Microscopy (SEM) analysis of biological manganese oxide
The biological oxide is formed by adding manganese-oxidizing bacteria into a divalent manganese-containing liquid culture medium, a Scanning Electron Microscope (SEM) of the biological oxide is shown in figure 8, under the magnification of 10000 times, the scanning electron microscope shows that the surface of the biological manganese oxide is irregular and is in a strip shape or a smooth spar shape, and a large amount of strip-shaped manganese-oxidizing bacteria are filled around the biological manganese oxide, which shows that the biological manganese oxide is mixed with the manganese-oxidizing bacteria.
(3) Biological manganese oxide pair As3+Oxidation experiment of
100ml of manganese-containing liquid medium was prepared in the manner described in step (1) in example 1, namely: weighing 10g of tryptone, 5g of yeast extract powder and 5g of sodium chloride, adding the weighed materials into 1000m1 distilled water, adjusting the pH to 7, sterilizing the materials at 121 ℃ for 20min in a sterilizing pot, and carrying out filtration by using a 0.22 mu m filter head and adding 1mol/L of MnSO 4. H2O aqueous solution, 1mol/L HEPES buffer solution and As3+Solution and making Mn2+HEPES buffer and As3+The final concentrations of (A) are 1mmol/L, 20mmol/L and 100mg/L respectively; adding manganese oxidizing bacteria into the Mn-containing solution according to the inoculation amount of 1%2+HEPES buffer and As3+The liquid medium of (4) was subjected to shaking culture at 30 ℃ on a constant temperature shaker at 180 rpm.
Sampling every 24h, sucking 1 microliter of culture solution to dilute to a 10ml centrifuge tube, sucking 1ml of the diluted 10ml centrifuge tube, continuously diluting to a new 10ml centrifuge tube by 1000 times, and sampling for 7 days.
Inductively coupled plasma mass spectrometer (ICP-MS) is adopted to measure As in the reaction process3+/As5+The content of (b) is measured.
As can be seen in FIG. 9, the biological manganese oxide pairs As produced by the strains of the invention3+The oxidation is fast, and 50.98 percent of As can be oxidized after the initial reaction is carried out for 3 hours3+And As over time3+Gradually becomes further oxidized and after 7 days, it is oxidized to As3+Finally, the oxidation effect of about 74 percent is achieved. The manganese oxidizing bacteria produced by the strain of the invention show As3+Good oxidation effect, can oxidize As3+Oxidation to less toxic As5+Is favorable for application in the field of arsenic detoxification.
In conclusion, the novel manganese oxidizing bacteria is screened out, is the bacteria with the manganese oxidizing capability found for the first time in Morganella, is high in manganese oxidizing capability, good in environmental adaptability and high in research value; as can be produced by the biological manganese oxide produced by the bacterium3+Oxidized to As5+The method is beneficial to being applied to the field of arsenic detoxification, and provides theoretical basis for the possible application potential of biological manganese oxide.
It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (10)
1. A manganese oxidizing bacterium characterized by comprising a microorganism,
belongs to Morganella (Morganella Morganii) and is named as MnOx-1, and is preserved in the China general microbiological culture Collection center on 7-month and 2-month 2021 with the preservation number of CGMCC No. 22815.
2. The method of screening for a manganese-oxidizing bacterium according to claim 1,
the method comprises the following steps:
s1, taking supernatant after the sludge sample is stood, adding the supernatant into MnSO4·H2O and HEPES buffer in LB liquid medium;
s2, after constant temperature shaking culture for 5-9d, the culture medium is checked and screened primarily by LBB indicator.
3. The method of screening for a manganese-oxidizing bacterium according to claim 2,
further comprising:
s3, diluting and spreading the culture solution which shows blue color to LBB indicator in step S2 to MnSO-containing culture solution4·H2Culturing in solid culture medium containing O and HEPES buffer solution at constant temperature, further checking and rescreening with LBB indicator, separating and purifying the LBB indicator showing blue color, mixing the obtained strain with glycerol at a ratio of 1: 1, and preserving at-80 deg.C.
4. The method of screening for a manganese-oxidizing bacterium according to claim 2 or 3,
in step S1, the LB liquid medium comprises the following components:
10g of tryptone, 5g of yeast extract powder, 5g of sodium chloride and 1000m1 of distilled water with pH of 7, and MnSO4·H2O1 mmol/L, HEPES buffer 20 mmol/L.
5. The method of screening for a manganese-oxidizing bacterium according to claim 2 or 3,
in step S2, the LBB indicator is formulated as follows:
0.04g LBB powder is weighed and dissolved in 0.25ml of 45mmol/L glacial acetic acid water solution, deionized water is added, the volume is adjusted to 100ml, and the mixture is stored in a dark place at 4 ℃.
6. The method of screening for a manganese-oxidizing bacterium according to claim 3,
in step S3, MnSO is contained4·H2The preparation method of the solid culture medium of O and HEPES buffer solution is as follows:
dissolving tryptone 10g, yeast extract 5g, sodium chloride 5g and agar 15g in 1000ml distilled water, adjusting pH to 7, sterilizing at 121 deg.C for 20min, filtering with 0.22 μm filter head, and adding MnSO 1mol/L4·H2O and 1mol/L HEPES buffer solution to make Mn2+And HEPES buffer solution with final concentration of 1mmol/L and 20mmol/L, respectively, cooling to 50 deg.C, pouring the plate on a sterile operating table, and condensing the solid plate for use.
7. A microbial agent comprising the manganese oxidizing bacterium according to claim 1.
8. Use of the manganese oxidizing bacteria of claim 1 or the microbial inoculum of claim 7 for the preparation of biological oxides of manganese that oxidize trivalent arsenic.
9. Use of the manganese oxidizing bacterium according to claim 1 or the microbial agent according to claim 7 for oxidizing trivalent arsenic.
10. Use according to claim 9,
the manganese oxidizing bacteria oxidize Mn2+Preparing biological manganese oxide, and oxidizing trivalent arsenic into pentavalent arsenic by the biological manganese oxide.
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