Plant-dwelling bacillus panlonei, composition containing same, preparation and application

Abstract

The present invention provides a plant-dwelling Bacillus pannonianus, a composition comprising the same, preparation and application, the plant-dwelling Bacillus pannonianus, named MnO‑3, belongs to the genus Pannonianus (Pannonibacter), has been deposited in the General Microbiological Center (CGMCC) of the China Microbiological Culture Collection Administration Committee, with a deposit date of April 27, 2023, and a deposit number of CGMCC No.27230. Compared with the prior art, the plant-dwelling Bacillus pannonianus obtained by the present invention has a high degradation efficiency for organic decabromodiphenyl ether, high tolerance to decabromodiphenyl ether concentration, and high environmental adaptability; and its manganese oxidation ability is strong, and has a high research value. The preparation method of its composition is simple, easy to operate, low cost, green and pollution-free. And the application of the composition to degrade decabromodiphenyl ether is highly efficient, which is conducive to large-scale application in the field of degrading organic matter.

Description

Technical Field

[0001] The present invention relates to the technical field of environmental microorganisms, and in particular to a plant-dwelling Bacillus panlongus, a composition containing the same, and preparation and application thereof. Background Art

[0002] Polybrominated diphenyl ethers (PBDES) are widely used in industries such as petrochemicals and electronic devices due to their excellent flame retardancy, stable physical and chemical properties, and low price and availability. PBDES has stable chemical properties, strong lipophilicity, is insoluble in water, and has endocrine disrupting effects and neurotoxicity to organisms exposed to such pollutants. There are many compounds belonging to the PBDES class, but decabromodiphenyl ether (BDE-209) is the main one in commercial production. BDE-209 has become the most widely used brominated flame retardant in the world. According to incomplete statistics, my country uses an average of about 20-40 million kilograms of commercial BDE-209 each year, mainly concentrated in electronic waste dismantling sites and surrounding areas.

[0003] Due to its huge amount and long history of use, PBDES is widely distributed in the atmosphere, water, sediment or soil through global activities. Organisms enter the food chain through contact, ingestion, etc., which ultimately endangers human health. PBDES has low acute toxicity, but after enrichment in the food chain, it eventually produces toxic effects on organisms, mainly concentrated in thyroid toxicity, neurotoxicity, reproductive developmental toxicity, etc. PBDES is an endocrine disruptor that affects the balance of thyroid hormones and interferes with the normal operation of the immune system. PBDES pollution in electronic waste dismantling sites has seriously endangered human survival and healthy development and urgently needs to be repaired and treated.

[0004] At present, the treatment and removal technologies for PBDES mainly include biodegradation, photodegradation, zero-valent iron reduction degradation and combined treatment of multiple methods, all of which have the problem of low degradation efficiency. Among them, biodegradation has the advantages of being green and pollution-free, but so far, the single microorganism has a low tolerance to BDE-209 concentration and the efficiency of degrading organic matter is generally low. Summary of the invention

[0005] The main purpose of the present invention is to provide a plant-dwelling Bacillus panloniae, a composition comprising the same, and a preparation and application thereof, aiming to solve the problems of poor oxidation ability of plant-dwelling Bacillus panloniae to manganese and low degradation efficiency of decabromodiphenyl ether in the prior art.

[0006] To achieve the above-mentioned purpose, the present invention provides a plant-dwelling Pannonibacter, the plant-dwelling Pannonibacter, named MnO-3, belongs to the genus Pannonibacter (Pannonibacter), has been deposited in the General Microbiology Center (CGMCC) of the China Microorganism Culture Collection Administration, the preservation date is April 27, 2023, and the preservation number is CGMCC No.27230.

[0007] The present invention also provides an application of the above-mentioned plant-dwelling Lac. panloniae to oxidize manganese.

[0008] The present invention also provides a manganese oxide / plant-dwelling Bacillus panulosa composition, comprising an amorphous manganese oxide and the plant-dwelling Bacillus panulosa as described above; wherein the amorphous manganese oxide and the plant-dwelling Bacillus panulosa are aggregated into a block.

[0009] The present invention also provides a method for preparing the composition as described above, comprising:

[0010] The plant-dwelling Panlongian alkaline lake bacillus as described above is inoculated at an inoculation rate of 1 to 10% per 100 mL of manganese oxidizing bacteria ion culture medium, and shaken culture is carried out at 25 to 35° C. for 5 to 10 days to obtain the manganese oxide / plant-dwelling Panlongian alkaline lake bacillus composition; wherein the manganese oxidizing bacteria ion culture medium includes 0.1 to 4 mmol / LMn 2+ .

[0011] Furthermore, the manganese oxidizing bacteria ion culture medium specifically includes 1g / L yeast extract, 1g / L acid hydrolyzed casein, 1g / L glucose, 0.222g / L CaCl2, 0.396g / L MgSO4, 0.0006g / L FeCl3, 1mL / L trace element solution and 0.1-4mmol / L MnSO4·H2O.

[0012] Furthermore, the trace element solution includes 10 mg / L CuSO4·5H2O, 44 mg / L ZnSO4·7H2O, 20 mg / L CoCl2·6H2O, and 13 mg / L Na2MoO4·2H2O.

[0013] The present invention also provides a use of the plant-dwelling Lac. panloniae in treating organic wastewater; the organic wastewater includes decabromodiphenyl ether.

[0014] The present invention also provides an application of the above-mentioned composition in treating organic wastewater; the organic wastewater includes decabromodiphenyl ether.

[0015] The beneficial effects achieved by the present invention are:

[0016] The plant-dwelling Lac. panloniae strain screened by the present invention has high degradation efficiency for organic decabromodiphenyl ether, high tolerance for decabromodiphenyl ether concentration, high environmental adaptability, and strong manganese oxidation ability, and has high research value.

[0017] The preparation method of the manganese oxide / plant-dwelling Bacillus panloniae composition provided by the present invention is simple and easy to operate, low in cost, green and pollution-free. The composition has high efficiency in degrading decabromodiphenyl ether, which is conducive to large-scale application in the field of organic matter degradation. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the structures shown in these drawings without paying creative work.

[0019] Figure 1 These are SEM images of the manganese oxide / plant-dwelling Lac. panloniae composition in Example 2 of the present invention at different magnifications; wherein (a) is a SEM image magnified 6000 times and (b) is a SEM image magnified 13000 times;

[0020] Figure 2 TEM images of the manganese oxide / plant-dwelling Lac. panlongenica composition in Example 2 of the present invention; wherein (a) is a TEM image of the composition (scale bar, 2 μm), and (b) is a local enlarged TEM image of (a) (scale bar, 500 nm);

[0021] Figure 3 This is the phylogenetic analysis tree of the plant-dwelling Lacobacillus panloniae in Example 1 of the present invention;

[0022] Figure 4 is the XRD diagram of the manganese oxide / plant-dwelling Lac. panloniae composition in Example 2 of the present invention;

[0023] Figure 5 This is a HRTEM image of the manganese oxide / plant-dwelling Lac. panloniae composition in Example 2 of the present invention;

[0024] Figure 6 This is the XPS graph of the manganese oxide / plant-dwelling Lac. panloniae composition in Example 2 of the present invention;

[0025] Figure 7The growth curve and pH change diagram of the plant-dwelling Bacillus panloniae at different pH values ​​in Example 3 of the present invention; wherein (a) is its growth curve diagram; (b) is its pH change diagram;

[0026] Figure 8 This is a graph of the residual Mn(II) concentration of the plant-dwelling Lac. panloniae at different pH values ​​in Example 3 of the present invention;

[0027] Fig. 9 The growth curve and residual Mn(II) concentration diagram of the plant-dwelling Bacillus panlongensis at different inoculation doses in Example 3 of the present invention; wherein (a) is its growth curve diagram; (b) is its residual Mn(II) concentration diagram;

[0028] Fig.10 The plant-dwelling Lac. panlongenica in Example 3 of the present invention has different Mn 2+ The growth curve and residual Mn(II) concentration diagram under the concentration; (a) is its growth curve diagram; (b) is its residual Mn(II) concentration diagram;

[0029] Fig.11 The comparison charts are of the degradation rate of 5 mg / L decabromodiphenyl ether and the removal rate of Mn(II) by the manganese oxide / plant-dwelling Bacillus panloniae composition (MnO-3+Mn) experimental group, the plant-dwelling Bacillus panloniae (MnO-3) experimental group, and the blank control (CK) group in Example 4 of the present invention; wherein (a) is a degradation rate comparison chart; and (b) is a removal rate comparison chart.

[0030] The realization of the purpose, functional features and advantages of the present invention will be further explained in conjunction with the implementation methods and with reference to the accompanying drawings. DETAILED DESCRIPTION

[0031] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0032] It should be noted that, in the absence of conflict, the following embodiments and features in the embodiments can be combined with each other. It should also be understood that the terms used in the embodiments of the present invention are intended to describe specific embodiments rather than to limit the scope of protection of the present invention.

[0033] Unless otherwise defined, all technical and scientific terms used in the present invention are consistent with the prior art mastery of the technicians in this technical field and the records of the present invention. Any methods, equipment and materials of the prior art that are similar or equivalent to the methods, equipment and materials in the embodiments of the present invention can also be used to implement the present invention. It should be known by those skilled in the art that as an explanation of the present application document, without affecting the actual understanding of the technical scheme of the present application, "TEM" can be expressed as a transmission electron microscope, "XRD" can be expressed as an X-ray diffractometer, "HRTEM" can be expressed as a high-resolution transmission electron microscope, "XPS" can be expressed as an X-ray photoelectron spectrometer, "Intensity / (au)" can be expressed as intensity, "2θ / degree" can be expressed as a diffraction angle, "Intensity(au)" can be expressed as intensity, "Bindingenergy(ev)" can be expressed as binding energy, "satellite" can be expressed as satellite peak, "OD 600 " can be expressed as the optical density value measured at a wavelength of 600nm.

[0034] When the embodiments give numerical ranges, it should be understood that unless otherwise specified in the present invention, both endpoints of each numerical range and any value between the two endpoints can be selected. The test methods in the following embodiments without specifying specific conditions are usually carried out under conventional conditions or under conditions recommended by the manufacturers. The materials or reagents required in the following embodiments are commercially available unless otherwise specified.

[0035] In order to solve the problems that the existing plant-dwelling Pannonibacterium has poor oxidation ability of manganese and low degradation efficiency of decabromodiphenyl ether, the present invention provides a plant-dwelling Pannonibacterium, the plant-dwelling Pannonibacterium, named MnO-3, belongs to the genus Pannonibacter (Pannonibacter), has been deposited in the General Microbiology Center (CGMCC) of China Microorganism Culture Collection Administration, with a preservation date of April 27, 2023, and a preservation number of CGMCC No. 27230.

[0036] Specifically, soil samples from the Xiangtan manganese mine in Changsha City, Hunan Province can be collected. The supernatant of the soil sample after standing is taken and added to the LB liquid culture medium containing MnSO4·H2O. After 7 days of constant temperature shaking culture, the culture is spread on a solid plate containing 3mmol / L MnSO4·H2O, and the colonies grown on the plate are used for preliminary screening. Select well-growing colonies and inoculate them into the LB liquid culture medium of MnSO4·H2O to continue culturing to purify the strains. Samples are taken at the beginning and after 7 days of culture, and the Mn content is determined by inductively coupled plasma optical emission spectroscopy (ICP-OES). 2+Removal rate. The culture fluid of the strain with high removal rate was further tested and screened by LBB indicator, and the strain that showed blue to LBB indicator was separated and purified. After that, the culture fluid of the strain that can change color was further diluted and applied to the solid culture medium containing 3mmol / L MnSO4·H2O. After constant temperature culture, the obtained strain was mixed with 50% glycerol at a ratio of 1:1 and stored at -80°C. It is specifically preserved in the General Microbiology Center of China Microbiological Culture Collection Administration, address: No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, Postal Code 100101; after identification, its 16SrRNA gene sequence has the highest similarity with Pannonibacter phragmitetus 16S rRNA gene sequence.

[0037] The plant-dwelling Lac. panloniae strain screened by the present invention has high degradation efficiency for organic decabromodiphenyl ether, high tolerance for decabromodiphenyl ether concentration, high environmental adaptability, and strong manganese oxidation ability, and has high research value.

[0038] The present invention also provides an application of the above-mentioned plant-dwelling Bacillus panloniae to oxidize manganese. Specifically, the selected plant-dwelling Bacillus panloniae can be inoculated with an inoculum amount of 1 to 10% on 100 mL of manganese oxidizing bacteria ion culture medium with a pH value of 5 to 8, and the residual Mn in the supernatant can be measured by an inductively coupled plasma optical emission spectrometer (ICP-OES). 2+ The removal rate at different sampling times was calculated and the removal effect of Mn(Ⅱ) was good. 2+ The concentration can be 0.1~4mmol / L.

[0039] The present invention also provides a manganese oxide / plant-dwelling Bacillus panulosa composition, comprising an amorphous manganese oxide and the above-mentioned plant-dwelling Bacillus panulosa; wherein the amorphous manganese oxide and the plant-dwelling Bacillus panulosa are aggregated into a block.

[0040] The present invention also provides a method for preparing the above composition, comprising:

[0041] The above plant-dwelling Panlonga alkaline lake bacillus is inoculated with 1-10% inoculum per 100 mL of manganese oxidizing bacteria ion culture medium, and shaken culture is carried out at 25-35° C. for 5-10 days to obtain a manganese oxide / plant-dwelling Panlonga alkaline lake bacillus composition; wherein the manganese oxidizing bacteria ion culture medium includes 0.1-4 mmol / L Mn 2+ .

[0042] Specifically, when the inoculation amount is <1%, the microorganisms grow slowly and the rate of biogenic manganese oxide generation is slow; when the inoculation amount is >10%, the dissolved oxygen is insufficient, more metabolic waste is introduced, and the oxidation rate is affected. 2+ When the concentration is 0.1~4mmol / L, more manganese oxides can be generated; during the preparation process, it was found that when Mn 2+ When the concentration is >4mmol / L, it will inhibit the growth of the plant-dwelling Bacillus panlongenae and prevent the formation of manganese oxides.

[0043] Furthermore, the manganese oxidizing bacteria ion culture medium specifically includes 1g / L yeast extract, 1g / L acid hydrolyzed casein, 1g / L glucose, 0.222g / L CaCl2, 0.396g / L MgSO4, 0.0006g / L FeCl3, 1mL / L trace element solution and 0.1-4mmol / L MnSO4·H2O. The manganese oxidizing bacteria ion culture medium can just meet the generation conditions of manganese oxides and is suitable for the growth of the plant-dwelling Bacillus panloniae.

[0044] The preparation method of the manganese oxide / plant-dwelling Lac. panlonicum composition provided by the invention is simple, easy to operate, low in cost, and green and pollution-free.

[0045] Further, the trace element solution includes 10 mg / L CuSO4·5H2O, 44 mg / L ZnSO4·7H2O, 20 mg / LCoCl2·6H2O, and 13 mg / L Na2MoO4·2H2O.

[0046] The present invention also provides an application of the above plant-dwelling Bacillus panloniae in treating organic wastewater; the organic wastewater includes decabromodiphenyl ether. The plant-dwelling Bacillus panloniae screened by the present invention has high degradation efficiency for organic decabromodiphenyl ether, high tolerance to decabromodiphenyl ether concentration, and high environmental adaptability.

[0047] The present invention also provides an application of the above composition in treating organic wastewater; the organic wastewater includes decabromodiphenyl ether. Compared with the concentration of decabromodiphenyl ether degraded by bacteria in other studies, the degradation rate of the composition for 5 mg / L decabromodiphenyl ether reaches more than 50%, indicating that the composition has good environmental adaptability and great potential for removing and degrading other organic pollutants, and can be widely used in the field of organic pollution.

[0048] For further understanding of the present invention, examples are given below:

[0049] Example 1

[0050] Screening, Isolation and Purification of Plant-dwelling Lacobacillus panlonei

[0051] 1. Preparation of LB liquid medium:

[0052] Weigh 5 g of yeast extract, 10 g of tryptone, and 10 g of sodium chloride, add 1000 ml of distilled water, adjust the pH to 7, and dispense 100 ml into 250 ml conical flasks. Sterilize in an autoclave at 121 °C for 20 min, filter with a 0.22 μm sterile filter, and add 3 ml of 100 mmol / L MnSO4·H2O mother solution to make Mn 2+ The final concentration is 3mmol / L.

[0053] 2. Preparation of solid culture medium:

[0054] Dissolve 5 g of yeast extract, 10 g of tryptone, 10 g of sodium chloride and 15 g of agar in 1000 ml of distilled water, adjust the pH to 7, sterilize in an autoclave for 20 min, filter with a 0.22 μm sterile filter, add 3 ml of 100 mmol / L MnSO4·H2O stock solution, and make Mn 2+ The final concentration is 3 mmol / L. After cooling to 50°C, pour the plate on a UV sterilized operating table and set aside after the solid plate condenses.

[0055] 3. Screening and isolation of plant-dwelling Lacobacillus panloniae:

[0056] Soil was collected from the Xiangtan manganese mine in Hunan Province. After standing, the supernatant sludge liquid was taken and added to the LB liquid culture medium in step 1 at a 1% inoculation amount. After shaking and culturing on a constant temperature shaker at 30°C and 150rpm for 7 days, it was spread and cultured on a solid plate containing 3mmol / LMnSO4·H2O, and the colonies grown on the plate were used for preliminary screening. After constant temperature culture, well-growing colonies were selected and inoculated into LB liquid culture medium containing MnSO4·H2O to continue culturing to purify the strain. Samples were taken at the beginning and after 7 days of culture, and Mn was determined using an inductively coupled plasma optical emission spectrometer (ICP-OES). 2+ Removal rate. The culture fluid of the strain with high removal rate is further tested and screened by LBB indicator, and the strains that show blue color to LBB indicator are separated.

[0057] The preparation method of the LBB indicator is as follows: weigh 0.04 g of LBB powder, dissolve it in 0.25 ml of glacial acetic acid aqueous solution (45 mmol / L), add deionized water and make up to 100 ml, and store at 4° C. away from light.

[0058] 4. Purification of the plant-dwelling Bacillus panloniae

[0059] The culture solution that turns blue to the LBB indicator in step 3 is diluted and spread on the solid culture medium separated in step 2, and cultured in a 30°C constant temperature incubator for purification. The purified strain is mixed with 50% glycerol at a ratio of 1:1 and stored in a -80°C refrigerator.

[0060] Example 2

[0061] Preparation, characterization and molecular identification of manganese oxide / plant-dwelling Lac. panlonicola composites

[0062] 1. Preparation of manganese oxide / plant-dwelling Lac. panlonicum composition

[0063] (1) Preparation of manganese oxidizing bacteria ion culture medium:

[0064] Referring to the adapted medium for manganese oxidizing bacteria isolated from the freshwater system, Leptothrix medium, a certain degree of modification was made on this basis, namely: weigh 1g / L yeast extract, 1g / L acid hydrolyzed casein, 1g / L glucose, 0.222g / L CaCl2, 0.396g / L MgSO4, 0.0006g / L FeCl3, 1mL / L trace element solution (10mg / LCuSO4·5H2O, 44mg / LZnSO4·7H2O, 20mg / L CoCl2·6H2O, 13mg / L Na2MoO4·2H2O) and 3mmol / L MnSO4·H2O, add them to 1000mL deionized water, and sterilize them in an autoclave at 121℃ for 20min. Among them, MnSO4·H2O is filtered with a sterile filter and added to 100mL culture medium.

[0065] (2) Production of manganese oxide / plant-dwelling Lac. panlonicum composites

[0066] A 250 mL volumetric flask was selected, and 100 mL of the manganese oxide mineralization medium prepared in step (1) was added. The plant-dwelling Panlongia alkaline lake bacillus obtained in Example 1 was added to the manganese oxidizing bacteria ion medium at an inoculum rate of 1%, and the culture was placed in a constant temperature shaker at 30° C. and 150 rpm for 7 days, and then centrifuged at 8000 rpm for 3 minutes, the supernatant was discarded, and the mixture was washed with deionized water 3 times. The resulting precipitate was the manganese oxide / plant-dwelling Panlongia alkaline lake bacillus composition.

[0067] 2. Characterization analysis

[0068] The SEM image of the manganese oxide / plant-dwelling Lac. panloniae composition obtained in step 1 is as follows: Figure 1 As shown, Figure 1 (a) is a SEM image magnified 6000 times. Figure 1(b) is a SEM image magnified 13,000 times. The TEM image of the manganese oxide / plant-dwelling Bacillus panlongus alkaline lake composition is as follows Figure 2 As shown, Figure 2 (a) is a TEM image of the composition (scale bar, 2 μm). Figure 2 (b) Figure 2 (a) Locally enlarged TEM image (scale bar, 500 nm).

[0069] according to Figure 1 and Figure 2 It can be observed that the plant-dwelling Panlongian alkaline lake Bacillus strain is rod-shaped as a whole, with a length of about 700nm and a diameter of about 312nm. In the manganese oxide / plant-dwelling Panlongian alkaline lake Bacillus composition, the plant-dwelling Panlongian alkaline lake Bacillus and the manganese oxide mineral particles are aggregated together to form a larger block. And the surface of the manganese oxide / plant-dwelling Panlongian alkaline lake Bacillus composition is uneven, which can provide more attachment sites.

[0070] The manganese oxide / plant-dwelling Lac. panlongensis composition was subjected to XRD and XPS analysis, and its XRD graph, HRTEM graph and XPS graph were as follows: Figure 4 , Figure 5 , Figure 6 As shown. Figure 4 and Figure 5 It can be seen that the manganese oxide in the manganese oxide / plant-dwelling Lac. panlongensis composition is in an amorphous state, with no diffraction peaks in XRD and chaotic and disordered diffraction stripes. Moreover, the amorphous biological manganese oxide has a stronger reaction activity than the chemical manganese oxide with obvious crystal form. Figure 6 XPS analysis shows that the content of high-valent manganese (III, IV) in the manganese oxide is 54.53%, which is 45.46% higher than that of Mn (II), and has a strong oxidizing ability.

[0071] 3. Molecular identification and phylogenetic tree comparison:

[0072] DNA extraction and sequencing were performed on the plant-dwelling Bacillus panloniae isolated in Example 1, and the entire process was completed by Shenzhen Micro-Technology Group Co., Ltd. The 16S rRNA results obtained by sequencing were compared with the existing 16S rRNA nucleic acid sequences using the Blast program of NCBI (National Center for Biotechnology Information). The phylogenetic tree was analyzed and drawn using Mega11.0 software, and the phylogenetic tree is shown in Figure 3 .

[0073] The sequence results showed that the manganese oxidizing bacteria had the highest similarity with Pannonibacter phragmitetus, and it was identified as belonging to the genus Pannonibacter and named MnO-3.

[0074] Example 3

[0075] Physiological characteristics of plant-dwelling Lactobacillum panlone

[0076] 1. Manganese oxidation capacity of the plant-dwelling Bacillus panloniae (MnO-3) at different pH values

[0077] Select a 250 ml conical flask and prepare 100 ml of manganese oxidizing bacteria ion culture medium according to step (1) in Example 2, and adjust the pH to 5, 6, 7, and 8 respectively. Add MnO-3 to the manganese oxidizing bacteria ion culture medium at a 5% inoculum amount, and shake and culture in a constant temperature shaker at 30°C and 150 rpm; take samples at regular intervals, centrifuge at 8000 rpm for 3 minutes, and filter with a 0.22 μm filter. The residual Mn in the supernatant was determined by inductively coupled plasma optical emission spectrometry (ICP-OES). 2+ The concentration of microorganisms was measured by ultraviolet spectrophotometer at a wavelength of 600 nm.

[0078] The growth curves of MnO-3 and pH changes at pH 5, 6, 7, and 8 are shown in Figure 7 As shown; among them, Figure 7 (a) is its growth curve; Figure 7 (b) is the pH change diagram. Figure 7 It can be seen that different pH values ​​have almost no effect on the growth activity of MnO-3. After the pH value decreased at 12h, the final pH value reached about 8.7 as time went by.

[0079] The residual Mn(II) concentration at different pH values ​​is shown in Figure 2. Figure 8 Analysis Figure 8 The results show that under the condition of pH = 7, the removal effect of Mn(Ⅱ) is the best, reaching a removal rate of 80%.

[0080] 2. Manganese oxidation capacity of MnO-3 at different inoculation doses

[0081] Select a 250ml conical flask, prepare 100ml of manganese oxidizing bacteria ion culture medium according to step (1) in Example 2, adjust the pH value to 7, add the plant-dwelling Lac. panlongensis screened in Example 1 to the manganese oxidizing bacteria ion culture medium at a bacterial inoculation amount of 1%, 2%, 5%, and 10%, respectively, and shake and culture in a constant temperature shaker at 30°C and 150rpm; take samples at regular intervals, centrifuge at 8000rpm for 3min, and filter with a 0.22μm filter. The residual Mn in the supernatant was determined by inductively coupled plasma optical emission spectrometry (ICP-OES). 2+ The concentration of microorganisms was measured by ultraviolet spectrophotometer at a wavelength of 600 nm.

[0082] The growth curves and residual Mn(Ⅱ) concentrations at different inoculation doses are shown in Figure 2. Fig. 9 As shown; among them, Fig. 9 (a) is its growth curve; Fig. 9 (b) is the residual Mn(II) concentration diagram. Fig. 9 It can be seen that MnO-3 reached an OD600 value of 1 in about 10 hours at different inoculation doses, and different inoculation doses had almost no effect on the growth activity of the microorganism (Bacillus panlongensis). However, different inoculation doses had a significant effect on the removal rate of Mn(Ⅱ), and the Mn(Ⅱ) removal effect was the best at 10% inoculation dose, reaching 84.4%.

[0083] 3. MnO-3 in different Mn 2+ Mn oxidation capacity at concentration

[0084] Select a 250 ml conical flask and follow the method of step (1) in Example 2, except that Mn 2+ The final concentration of Mn was 100 ml of manganese oxidizing bacteria ion medium. 2+ The concentrations were set to 0.1, 0.3, 0.5, 1, and 2 mmol / L, respectively. The pH value was adjusted to 7, and the plant-dwelling Lac. panlongensis screened in Example 1 was added at a 10% inoculum to each of the different Mn 2+ The conical flasks were placed in a constant temperature shaker at 30°C and 150 rpm for shaking culture in a medium containing manganese oxidizing bacteria with a concentration of 1.5 Mn. Samples were taken at regular intervals, centrifuged at 8000 rpm for 3 min, and filtered with a 0.22 μm filter. The remaining Mn in the supernatant was determined by inductively coupled plasma optical emission spectrometry (ICP-OES). 2+ The concentration of microorganisms was measured by ultraviolet spectrophotometer at a wavelength of 600 nm.

[0085] Different Mn 2+ The growth curve and residual Mn(Ⅱ) concentration graph under the concentration are shown in Fig.10 As shown; among them, Fig.10 (a) is its growth curve; Fig.10 (b) is the residual Mn(II) concentration diagram. Fig.10 It can be seen that different Mn 2+ The concentration had little effect on the growth activity of MnO-3, but the growth activity was the highest at 2 mM Mn concentration, indicating that MnO-3 can tolerate higher Mn concentrations. 2+ The removal effect of Mn(Ⅱ) in the presence of 2-nitrogen was the best, reaching a removal rate of 94%.

[0086] Example 4

[0087] Experimental study on the oxidative degradation of organic matter decabromodiphenyl ether by plant-dwelling Bacillus panloniae and manganese oxide / plant-dwelling Bacillus panloniae combination

[0088] Select a 250 ml conical flask and refer to step (1) in Example 2 to change the Mn 2+ The final concentration was 2mmol / L, and 100ml of manganese oxidizing bacteria ion culture medium was prepared. After sterilization, the pH was adjusted to 7, and the plant-dwelling Bacillus panloniae selected in Example 1 was inoculated at a 10% inoculum amount; wherein the final concentration of decabromodiphenyl ether was 5mg / L. The culture was shaken in a constant temperature shaker at 30°C and 150rpm, as the manganese oxide / plant-dwelling Bacillus panloniae composition (MnO-3+Mn) experimental group.

[0089] Select a 250 ml conical flask and refer to the preparation method of the manganese oxidizing bacteria ion culture medium in step (1) of Example 2, without adding Mn 2+ , prepare 100 ml of plant-dwelling Bacillus panloniae experimental culture medium. After sterilization, adjust the pH to 7, and inoculate the plant-dwelling Bacillus panloniae screened in Example 1 at a 10% inoculum amount; wherein the final concentration of decabromodiphenyl ether is 5 mg / L. Shake and culture in a constant temperature shaker at 30°C and 150 rpm, as the plant-dwelling Bacillus panloniae (MnO-3) experimental group.

[0090] Select a 250 ml conical flask and refer to step (1) in Example 2 to change the Mn 2+ The final concentration was 2mmol / L. 100ml of manganese oxidizing bacteria ion culture medium was prepared. After sterilization, the pH was adjusted to 7, and the plant-dwelling Bacillus panlongensis screened in Example 1 was not inoculated; wherein the final concentration of decabromodiphenyl ether was 5mg / L. The medium was shaken and cultured in a constant temperature shaker at 30°C and 150rpm, as a blank control (CK) group.

[0091] The three groups of conical flasks were sampled at regular intervals, and the samples were pretreated after sampling to meet the conditions for the determination of decabromodiphenyl ether. That is, the samples were centrifuged at 8000rpm for 3min, and the supernatant was filtered through a 0.22μm filter head. The sample and the extractant were extracted for 10min in a volume ratio of 1:1 using n-hexane and dichloromethane in a volume ratio of 1:1, and the extraction was repeated three times, and the organic phases were combined. In ultra-high performance liquid chromatography (Acquity UPLC H-class), the mobile phase was methanol: water = 95:5, the flow rate was 0.3mL / min, the injection volume was 10μL, the detection wavelength was 233nm, and the retention time was 5min.

[0092] The degradation rate of 5 mg / L decabromodiphenyl ether and the removal rate of Mn(Ⅱ) in each group are as follows Fig.11 shown; Fig.11 (a) is a comparison chart of its degradation rates; Fig.11(b) is a comparison chart of the removal rates. Fig.11 It can be seen that after 7 days of culture, the degradation effect of the MnO-3+Mn experimental group was the best, and the CK group had no degradation effect. 2+ The degradation effect in the presence of 57% was achieved, and the removal rate of Mn(Ⅱ) was only 55%, indicating that the presence of decabromodiphenyl ether inhibited the manganese oxidation effect of MnO-3. However, the degradation effect of the MnO-3+Mn experimental group was higher than that of the CK group and the MnO-3 experimental group, indicating that the manganese oxide in the manganese oxide / plant-dwelling Bacillus panlongus alkaline lake composition played a key role in the degradation of decabromodiphenyl ether.

[0093] In summary, the above technical solutions of the present invention are only preferred embodiments of the present invention, and the patent scope of the present invention is not limited thereto. All equivalent structural changes made by using the contents of the present invention specification and drawings under the technical concept of the present invention, or directly / indirectly applied in other related technical fields are included in the patent protection scope of the present invention.

Claims

1. A plant-dwelling Bacillus panloniae, characterized in that: The plant-dwelling Lac. panloniae ( Pannonibacter phragmitetus ), named MnO-3, belongs to the genus Panlongea alkalinebacillin ( Pannonibacter ), has been deposited in the General Microbiology Center of China Microorganism Culture Collection (CGMCC), the deposit date is April 27, 2023, and the deposit number is CGMCC No.27230.

2. Use of the plant-dwelling Lacobacillus panlongus as claimed in claim 1 for oxidation of manganese.

3. A manganese oxide / plant-dwelling Lac. panlonicum composition, characterized in that: It comprises amorphous manganese oxide and the plant-dwelling Bacillus panulosa as claimed in claim 1; wherein the amorphous manganese oxide and the plant-dwelling Bacillus panulosa are aggregated into a block.

4. A method for preparing the composition according to claim 3, characterized in that: include: The plant-dwelling Bacillus panloniae as claimed in claim 1 is inoculated at an inoculation rate of 1-10% per 100 mL of manganese oxidizing bacteria ion culture medium, and shake-cultured at 25-35° C. for 5-10 days to obtain the manganese oxide / plant-dwelling Bacillus panloniae composition; Wherein, the manganese oxidizing bacteria ion culture medium includes 0.1~4mmol / L Mn 2+ .

5. The preparation method according to claim 4, characterized in that: The manganese oxidizing bacteria ion culture medium specifically comprises 1 g / L yeast extract, 1 g / L acid hydrolyzed casein, 1 g / L glucose, 0.222 g / L CaCl2, 0.396 g / L MgSO4, 0.0006 g / L FeCl3, 1 mL / L trace element solution and 0.1-4 mmol / L MnSO4·H2O.

6. The preparation method according to claim 5, characterized in that: The trace element solution includes 10 mg / L CuSO4·5H2O, 44 mg / L ZnSO4·7H2O, 20 mg / L CoCl2·6H2O, and 13 mg / L Na2MoO4·2H2O.

7. A use of the plant-dwelling Lac. panloniae as claimed in claim 1 in treating organic wastewater; The organic wastewater includes decabromodiphenyl ether.

8. Use of the composition as claimed in claim 3 in treating organic wastewater; The organic wastewater includes decabromodiphenyl ether.

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