CN113862178B - Korean bacillus LQB126, immobilized microbial agent and application thereof - Google Patents
Korean bacillus LQB126, immobilized microbial agent and application thereof Download PDFInfo
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- CN113862178B CN113862178B CN202111068096.1A CN202111068096A CN113862178B CN 113862178 B CN113862178 B CN 113862178B CN 202111068096 A CN202111068096 A CN 202111068096A CN 113862178 B CN113862178 B CN 113862178B
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C1/00—Reclamation of contaminated soil
- B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/10—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a carbohydrate
- C12N11/12—Cellulose or derivatives thereof
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/14—Enzymes or microbial cells immobilised on or in an inorganic carrier
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/20—Heavy metals or heavy metal compounds
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/20—Heavy metals or heavy metal compounds
- C02F2101/22—Chromium or chromium compounds, e.g. chromates
Abstract
The invention particularly discloses a Korean bacillus LQB126, an immobilized microbial agent and application thereof. In the invention, a Korean bacillus LQB126 is firstly screened and obtained, and the strain is preserved in China Center for Type Culture Collection (CCTCC) in the year 6 and 4 of 2021, and the preservation number is CCTCC NO: m2021667, deposit address: university of martial arts, taxonomic naming: the bacillus koraiensis (Bacillus koreensis) has a certain adsorption effect on heavy metals As, cd, cr and Pb, and in addition, the strain is added into an immobilized carrier for immobilization to obtain an immobilized microbial agent, compared with free bacillus koraiensis LQB126, the immobilized bacillus koraiensis LQB126 has a great improvement on the adsorption performance on the heavy metals, so that the strain and the immobilized microbial agent have good application prospects in the treatment of heavy metal pollution environments.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to a Korean bacillus LQB126, an immobilized microbial agent and application thereof.
Background
The heavy metal pollution of the soil exploded out due to the excessive deposition of heavy metal in the soil caused by human activities is particularly serious, and the heavy metal sources in the soil are various, including mining, metal smelting, chemical production, sewage irrigation and the like. Compared with water pollution and air pollution, the soil pollution is not easy to be perceived by human senses, people often need to pay attention to the soil pollution through animal and plant receptor expression and special detection, and the soil pollution has concealment and hysteresis, and meanwhile, heavy metals are difficult to naturally degrade in the soil and are almost irreversible once the heavy metal pollution occurs in the soil. Therefore, it is important to effectively prevent, control, repair and treat the heavy metal pollution of the soil. The existing soil heavy metal pollution restoration technology comprises the following steps: 1) Thermal desorption technique: the technology has high removal rate and can recycle heavy metals, but has high energy consumption, large investment, high operation cost and soil structure damage; 2) Electric repair technology: the technology has high efficiency, convenient operation and less secondary pollution, but is only suitable for repairing the soil in a small-area polluted area, and has long repairing period and high cost; 3) Soil leaching technology: the technology has stable repairing effect, short period and simple operation, but has higher repairing cost and is easy to cause leaching and precipitation of soil nutrient elements; 4) Curing treatment technology: the technology has the advantages of wide application range, low cost and simple operation, but has high requirements on the cured material and limited treatment depth; 5) Chemical oxidation technology: the technology has high efficiency and high speed, is insensitive to the type and concentration of pollutants, but has high chemical consumption, and can produce toxic action on soil to influence the growth and development of other plants; 6) Phytoremediation technology: the technology has low repair cost, easy operation and capability of maintaining the physicochemical properties of the original soil, but has long repair period and too high heavy metal concentration can influence the repair effect.
The repairing technology has certain defects when being applied. In recent years, microbial remediation treatment technology has become a research hotspot, and microorganisms living in heavy metal pollution environments for a long time form certain resistance and resistance to heavy metals, so that the microorganisms can well adapt to the toxic action of heavy metals on the growth of cells. By means of the adsorption and accumulation of the microorganism to the heavy metal, the method can forcefully repair the heavy metal pollution and has the advantages of low treatment cost, small influence on environment, high efficiency and the like. Therefore, adsorption of heavy metals in soil by means of microbial methods is an important topic of research on heavy metal pollution remediation technologies.
Korean bacillus (Bacillus koreensis) has been reported in the literature (Lim, J. -M.Bacillus koreensis sp.nov.a spore-forming bacterium, isolated from the rhizosphere of willow roots in Korea [ J ]. International Journal of Systematic & Evolutionary Microbiology,2006,56 (1): 59-63.), which belongs to the genus Bacillus and is an aerobic, spore-forming, mesophilic bacterium isolated from the rhizosphere, but recently has been reported for the function of Korean bacillus.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides a Korean bacillus LQB126, an immobilized microbial agent and application thereof. The bacterial colony of the strain is round light yellow opaque, flat protruding, smooth in surface and dry, is a gram-positive bacterium, has an optimal growth temperature of 25 ℃, has an optimal growth pH of 9.0, and has a certain adsorption effect on heavy metals As, cd, cr and Pb, so that the strain has a good application prospect in treating heavy metal polluted environments.
An object of the present invention is to provide a korean bacillus LQB126.
The strain of Korean bacillus LQB126 has been preserved in China Center for Type Culture Collection (CCTCC) at 2021, 6 and 4 days, and the preservation number is CCTCC NO: m2021667, deposit address: university of martial arts, taxonomic naming: korean Bacillus (Bacillus koreensis).
Korean bacillus (Bacillus koreensis) LQB126 is cultured on LB agar medium at 37deg.C for 24 hr, the colony is round light yellow, opaque, flat, protruding, smooth and dry, and is a gram positive bacterium with optimal growth temperature of 25deg.C and optimal growth pH of 9.0; the amplified 16s rDNA consists of 1420bp bases, and the homology alignment analysis shows that the sequence similarity of the strain LQB126 and Bacillus koreensis strain B11 (Accession: MN 880411.1) is 100.00%, the sequence similarity of the strain LQB126 and Bacillus koreensis strain WAB2227 (Accession: MH 169307.1) is 99.79%, and the sequence similarity of the strain LQB126 and Bacillus flexus strain T-17 (Accession: MN 330431.1) is 96.34%. The analysis of the evolutionary tree shows that the strains LQB126 and Bacillus koreensis are gathered on the same branch, and the sequence of the 16S rDNA of the Korean bacillus LQB1262 is shown as SEQ ID NO:1, based on the cell morphology, the 16S rDNA sequencing comparison analysis, the strain LQB126 was found to belong to Korean bacillus (Bacillus koreensis).
Further, the invention provides an application of the Korean bacillus LQB126 in treating heavy metal polluted environments.
Further, the heavy metal is one or more of As, cd, cr and Pb.
Further, the polluted environment is one of a water sample or a soil sample.
Another object of the present invention is to provide an immobilized microbial agent.
The immobilized microbial agent includes the above-mentioned Korean bacillus LQB126 and an immobilization carrier.
Further, the immobilization carrier is one of sepiolite, diatomite, peat soil, corncob and chestnut shells.
Furthermore, the invention provides application of the immobilized microbial agent in treating heavy metal pollution environments.
Further, the heavy metal is one or more of As, cd, cr and Pb.
Further, the polluted environment is one of a water sample or a soil sample.
Compared with the prior art, the invention has the following advantages:
according to the invention, the Korean bacillus LQB126 is obtained by screening, the bacterial colony of the strain is round light yellow opaque, flat protruding, smooth in surface and dry, is a gram positive bacterium, has an optimal growth temperature of 25 ℃, has an optimal growth pH of 9.0, has a certain adsorption effect on heavy metals As, cd, cr and Pb, and has a good application prospect in treating heavy metal pollution environment compared with free Korean bacillus LQB126 after being added into an immobilized carrier for immobilization, and the immobilized Korean bacillus LQB126 has a great improvement on the adsorption performance on heavy metals.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for the description of the embodiments will be briefly described below, and it is apparent that the drawings in the following description are only some embodiments of the present invention, and other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a colony morphology map (Panel A) and gram stain map (Panel B) of strain LQB 126;
FIG. 2 is a graph of the result of the phylogenetic tree of strain LQB 126;
FIG. 3 is a graph showing the effect of temperature on the growth of strain LQB 126;
FIG. 4 is a graph showing the effect of pH on the growth of strain LQB 126;
FIG. 5 is an agarose gel electrophoresis of a 16S rDNA fragment of strain LQB 126;
FIG. 6 is a graph showing the result of the change in adsorption of heavy metals in solution by strain LQB 126;
FIG. 7 is a graph showing the change of the adsorption of immobilized LQB126 microbial inoculum to heavy metals in solution;
FIG. 8 shows the adsorption effect of sepiolite immobilized LQB126 bacteria on heavy metals in soil; wherein, the graph (A) is the adsorption result of Cr polluted soil; the graph (B) shows the adsorption result of Cd-polluted soil; FIG. (C) shows the result of adsorption of Pb-contaminated soil; FIG. D shows the result of adsorption onto As-contaminated soil.
Detailed Description
The technical scheme of the present invention will be clearly and completely described in connection with specific embodiments. It will be apparent that the described embodiments are only some, but not all, embodiments of the invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to fall within the scope of the invention.
The formula of the culture medium and the components of the reagent involved in the embodiment of the invention are as follows:
LB liquid medium: 3g of beef extract, 10g of peptone and 5g of NaCl, distilled water is fixed to 1L, the pH is regulated to 7.0-7.2, and the beef extract is autoclaved for 30min at 121 ℃.
LB agar medium: 2% agar was added on the basis of LB liquid medium.
Other conventional reagents and equipment, unless otherwise specified, are commercially available.
Example 1 screening and identification of Bacillus koraiensis LQB126
Screening: taking a surface layer sediment sample of 0-50 cm by a sediment sampler (the sediment sample is from the bottom sediment of a Row bridge street in Dayum city of Hubei province), and collecting 5-10 g of samples into a sterile tube at multiple points in the sampling process; taking out 5g of sediment sample from the aseptic tube, adding the sediment sample into 45ml of aseptic water, oscillating for 2 hours, standing for 10 minutes, and taking supernatant to prepare suspension. Taking proper amount of suspension (50-100 mu L) and adopting dilute solutionThe release coating method is respectively and uniformly coated on the Cd-containing film 2+ The concentration is 50 mg.L -1 、Cr 6+ The concentration is 50 mg.L -1 、Pb 2+ The concentration is 50 mg.L -1 And As 3+ The concentration is 50 mg.L -1 On an LB solid culture medium plate, carrying out enrichment culture to obtain 117 bacteria;
single colony of the 117 bacteria was continuously selected and inoculated with Cd-containing bacteria 2+ The concentration is 200 mg.L -1 、Cr 6+ The concentration is 200 mg.L -1 、Pb 2+ The concentration is 200 mg.L -1 And As 3+ The concentration is 200 mg.L -1 On an LB solid culture medium plate, carrying out enrichment culture to obtain 60 bacteria;
single colony of the above 60 bacteria was continuously selected and inoculated with Cd-containing bacteria 2+ The concentration is 400 mg.L -1 、Cr 6+ The concentration is 400 mg.L -1 、Pb 2+ The concentration is 400 mg.L -1 And As 3+ The concentration is 300 mg.L -1 On an LB solid culture medium plate, carrying out enrichment culture to obtain 32 bacteria;
single colony of the 32 bacteria is continuously selected to be inoculated on the strain containing Cd 2+ The concentration is 1000 mg.L -1 、Cr 6+ The concentration is 1000 mg.L -1 、Pb 2+ The concentration is 1000 mg.L -1 And As 3+ The concentration is 500 mg.L -1 On a LB solid medium plate, a bacterium was obtained by enrichment culture, and the strain was designated as LQB126.
Morphology observation of strains: the purified LQB126 strain was inoculated on LB liquid medium for overnight activation, spread on LB agar medium, cultured at 37℃for 24 hours, and the surface morphology of the colony was observed.
Molecular biology identification: the 16S rRNA gene of LQB126 strain was sequenced as follows: using the genomic DNA of LQB126 strain as template, selecting general primer 27F and 1492R to amplify 16S rDNA sequence, and analyzing the amplified 16S rDNA sequence by agarose gel electrophoresis to obtain the target fragment with about 1.5kb, the result is shown in figure 5; the sequences are purified and inserted into a pMD18-T vector (Takara), then transformed into E.coli DH5 alpha, positive clones are obtained and sent to Shanghai Meiji biological medicine science and technology Co., ltd for sequencing, homologous comparison analysis is carried out on the sequences obtained by sequencing and bacterial database sequences in on-line software NCBI (https:// www.ncbi.nlm.nih.gov /), cluster analysis and systematic evolution tree construction are carried out by using MEGA6.5 software through an N-J method, sampling is repeated 1000 times, the calculated topological structural stability of the evolution tree is checked through a Bootstrap (Bootstrap) method, and the nodes of the development tree only display values of which the Bootstrap value is larger than 50%.
The primer sequences are as follows:
27F：5'-AGAGTTTGATCCTGGCTCAG-3'(SEQ ID NO.2)；
1492R：5'-TACGGCTACCTTGTTACGACTT-3'(SEQ ID NO.3)
specific identification information of the LQB126 strain is as follows:
morphological characteristics: the bacterial colony of the strain is round light yellow, opaque, flat protruding, smooth in surface and dry, is observed to be in a short rod shape under a microscope, is purple in gram staining and is a gram positive bacterium after being cultured for 24 hours at 37 ℃, and the specific result is shown in figure 1.
16s rDNA sequencing: the amplified 16s rDNA consisted of 1420bp, and the sequence similarity of strain LQB126 to Bacillus koreensis strain B11 (Accession: MN 880411.1) was found to be 100.00%, to Bacillus koreensis strain WAB2227 (Accession: MH 169307.1) to be 99.79%, and to Bacillus flexus strain T-17 (Accession: MN 330431.1) to be 96.34%. The treelet analysis showed that strain LQB126 and Bacillus koreensis aggregated in the same branch, see FIG. 2. The 16S rDNA sequence of the Korean bacillus LQB126 is shown in SEQ ID NO: 1.
Based on the cell morphology, the 16S rDNA sequencing comparison analysis, the strain LQB126 was attributed to Korean bacillus (Bacillus koreensis).
Example 2 experiment of influence of different conditions on growth of Korean bacillus LQB126
2.1 experiment of influence of different temperatures on growth of Korean Bacillus LQB126
Single colonies of LQB126 strain were individually picked from LB solid plates and inoculated into 250ml flasks containing 100ml of LB liquid medium, withoutShake flask culture at the same temperature (25 ℃, 30 ℃, 35 ℃ and 40 ℃) and 150r/min, sampling at intervals, and measuring the growth (OD) of the strain 600 )。
Growth of strain (OD) 600 ) As a result, as can be seen from FIG. 3, strains at different temperatures grew rapidly within 0 to 10 hours, and after the stationary phase, the growth amount of the strain was highest at 25℃and thus the optimum growth temperature of Bacillus koraiensis LQB126 was 25 ℃.
2.2 experiment of the effect of different pH on the growth of Korean Bacillus LQB126
Single colonies of LQB126 strain were picked up from LB solid plates respectively and inoculated into 250ml flasks containing 100ml of LB liquid medium, the pH values of the LB liquid medium were adjusted to 3.0, 5.0, 7.0, 9.0 and 11.0 respectively, shake flask culture was carried out at 37℃under 150r/min, samples were taken at intervals, and the growth (OD) of the strain was measured 600 )。
Growth of strain at different pH values (OD 600 ) As a result, as can be seen from FIG. 4, the strain grew rapidly in the range of 0 to 10 hours under different pH values, and the strain grew most in the condition of 9.0 pH value after the stationary phase, so that the optimum growth pH value of Korean bacillus LQB126 was 9.0.
EXAMPLE 3 adsorption experiment of Bacillus koraiensis LQB126 on heavy metals in solution
Picking single colony of LQB126 strain from LB solid plate, inoculating into 250ml flask containing 100ml LB liquid medium, shake flask culturing at 37deg.C under 150r/min to logarithmic phase, inoculating the bacterial liquid grown to logarithmic phase into 100mg/L single heavy metal solution (Cd) according to 5% inoculum size 2+ 、Cr 6+ 、Pb 2+ 、As 3+ ) In the mixed heavy metal solution with the neutralization concentration of 25mg/L, a culture medium without bacterial liquid is used as a blank control, shake flask culture is continued for 3d, then the concentration of heavy metal ions in the solution is measured, the concentration of the heavy metal ions is measured by adopting an atomic absorption spectrometry, and the calculation formula of the heavy metal adsorption rate is as follows:
η＝(C 0 -C e )/C 0 ×100％
c in the formula 0 The concentration of heavy metal in a blank control sample (the unit of soil is mg/kg, the unit of solution is mg/L) C e The concentration of heavy metal in the experimental sample (the unit of soil is mg/kg, and the unit of solution is mg/L).
The adsorption result of the Korean bacillus LQB126 on the heavy metal in the solution is shown in FIG. 6, and As can be seen from the graph, in the single heavy metal solution with the concentration of 100mg/L, the adsorption rate of the LQB126 on Cd and Pb is higher and is 26.64% and 28.28% respectively, but the adsorption effect on Cr and As is less than 2%; in the mixed heavy metal solution with the concentration of 25mg/L, the adsorption rate of the LQB126 strain on Cr, cd and Pb is more than 25%, the adsorption rates of the LQB126 strain on As are respectively 32.11%, 29.54% and 35.33%, and the adsorption rate of the LQB126 strain on As is 12.18%, so that the result shows that the LQB126 strain has a certain adsorption effect on high-concentration heavy metals.
Example 4 adsorption experiment of immobilized Korean bacillus LQB126 microbial agent on heavy metals in solution
4.1 pretreatment of immobilized Carrier
Placing sepiolite in a muffle furnace, heating and activating at 400 ℃ for 48h, sieving with a 40-mesh sieve, and preserving for later use; soaking diatomite in 5% hydrochloric acid for 2h, washing with distilled water to neutrality, soaking the diatomite in 5% sodium hydroxide for 2h, washing with distilled water to neutrality, oven drying, and sieving with 40 mesh sieve; soaking peat soil in 0.01 mol/L NaOH solution with a solid-to-liquid ratio of 1:5, placing in a constant temperature shaking table, shaking for 12-24 h, then adjusting pH to 6.5-7.5, centrifuging, taking precipitate, drying at 60 ℃, grinding, sieving with a 40-mesh sieve, and preserving for later use; sieving corncob and chestnut shell with 40 mesh sieve; placing the above 5 carriers into 250mL conical flask (100 mL carrier is placed in each flask), sealing with gauze and kraft paper, sterilizing with high pressure steam at 121deg.C for 30min, and storing in sterile room temperature environment.
4.2 immobilization of Bacillus koraiensis LQB126 microbial inoculum
Single colonies of LQB126 strain are respectively picked from LB solid plates and inoculated into 5 250ml flasks containing 100ml LB liquid medium, shake flask culture is carried out for 48h under the condition of 30 ℃ and 120r/min, 10ml of the immobilized carrier is respectively added into the 5 flasks under the aseptic condition, the flasks are placed on a shaking table under the condition of 120r/min, and after fixation for 48h, the flasks are placed in a refrigerator at 4 ℃ for preservation.
4.3 adsorption of the immobilized Korean bacillus LQB126 microbial inoculum to heavy metals in the solution
The immobilized Korean bacillus LQB126 microbial inoculum was inoculated into a single heavy metal solution (Cd) having a concentration of 100mg/L at an inoculum size of 5% 2+ 、Cr 6+ 、Pb 2+ 、As 3+ ) And (3) continuously shaking and culturing for 3d in a mixed heavy metal solution with the neutralization concentration of 25mg/L, and then measuring the concentration of heavy metal ions in the solution, wherein the measurement and calculation of the concentration of the heavy metal ions are the same as those in the embodiment 3.
The adsorption result of the immobilized bacillus koraiensis LQB126 microbial agent on heavy metals in the solution is shown in fig. 7, and it can be seen from the graph that in a single heavy metal solution with the concentration of 100mg/L, the adsorption rate of the sepiolite immobilized microbial agent on heavy metals Cr is 99.61%, which is superior to that of other carrier immobilized microbial agents, the adsorption rate of heavy metals Cd is 29.76%, and the adsorption rate of heavy metals Pb (16.34%) is slightly higher than that of other carrier immobilized microbial agents; the adsorption rates of the peat soil and the chestnut shell immobilized bacterial agent on Cd are 70.85 percent and 44.16 percent respectively, which are slightly higher than those of the sepiolite immobilized bacterial agent; the adsorption rate of the 5 carrier immobilized microbial agents to the heavy metal As is lower than 5 percent.
In mixed heavy metal solution with the concentration of 25mg/L, the adsorption rate of the sepiolite immobilized microbial agent to heavy metals Cr, pb and As is superior to that of other carrier immobilized microbial agents, and the adsorption rate of the sepiolite immobilized microbial agent to heavy metals Cd is only 4.53 percent, namely 44.55 percent, 65.08 percent and 60.53 percent; the adsorption rate of the peat soil immobilized microbial agent to heavy metals Cr and Pb reaches 12.21% and 42.02%, but is lower than that of sepiolite immobilized microbial agent.
The above results indicate that the immobilized microorganism microbial inoculum is obtained after the strain is added into the immobilized carrier for immobilization, and compared with free Korean bacillus LQB126, the immobilized Korean bacillus LQB126 has greatly improved adsorption performance on heavy metals.
Example 5 adsorption experiment of sepiolite-immobilized Bacillus koraiensis LQB126 microbial inoculum on heavy metals in soil
The immobilized Korean bacillus LQB126 microbial inoculum was inoculated into a single heavy metal soil (Cd) having a concentration of 100mg/L at an inoculum size of 5% 2+ 、Cr 6+ 、Pb 2+ 、As 3+ ) In the above, only the immobilized carrier and Korean bacillus LQB126 were inoculated as a control, and the culture was performed in shake flask at 30℃and 120r/min, and samples were taken at 0, 5, 10, 20 and 30d to perform the measurement of heavy metals in soil, and the soil was dissolved first, and the supernatant was collected by centrifugation, and the concentration of heavy metals in the supernatant was measured and calculated as in example 3.
The adsorption effect of sepiolite immobilized Korean bacillus LQB126 bacteria on heavy metals in soil is shown in fig. 8, and as can be seen from fig. 8 (A), the immobilized bacterial strain bacteria and single carrier are stable in about 10 days, and the adsorption rate is 49.19% and 48.48%; the adsorption effect of the single strain is slow in adsorption rate increase after 20 days, and the adsorption rate of the sepiolite immobilized bacterial agent to Cr is 60.50% when the sepiolite immobilized bacterial agent is adsorbed for 30 days; as can be seen from fig. 8 (B), the adsorption rate of the immobilized strain microbial agent reached a relatively stable level after 5 days, and was 35.96%; the adsorption rate of the single carrier reached a peak (47.19%) at day 10, after which the adsorption rate was slightly decreased; the adsorption rate of the single strain reached a peak (46.61%) at day 20, and was reduced to 24.50% at day 30; as can be seen from fig. 8 (C), the adsorption rate of the immobilized bacterial strain agent slowly increased to a peak value over 0 to 20 days, followed by a slow decrease in adsorption rate, the adsorption rate peak value being 19.67%; the adsorption rate of the single carrier reaches 20.71% on the 5 th day, slowly falls to 15.39%, and the maximum adsorption rate on the 30 th day is 27.77%; the adsorption rate of the single strain reaches a peak value on the 10 th day, and the highest adsorption rate is 25.60%; as can be seen from FIG. 8 (D), the adsorption rate of the immobilized strain microbial inoculum rapidly increased within 5-10 days, followed by a slow increase, and at day 30 the adsorption rate was 77.67%; the adsorption rate of the single carrier is stable after 20 days, and the peak value of the adsorption rate is 39.02%; the adsorption rate of the single strain rapidly rises in 5-10 days and then gradually stabilizes, and the peak value of the adsorption rate is 59.83%.
The results show that the bacillus koraiensis LQB126 and sepiolite immobilized bacillus koraiensis LQB126 microbial agents have certain adsorption capacity to heavy metal ions with high concentration in soil, so that the strain and the immobilized microbial agents have good application prospects in the treatment of heavy metal pollution environments.
The above examples are only specific embodiments of the present invention for illustrating the technical solution of the present invention, but not for limiting the scope of the present invention, and although the present invention has been described in detail with reference to the foregoing examples, it will be understood by those skilled in the art that the present invention is not limited thereto: any person skilled in the art may modify or easily conceive of the technical solution described in the foregoing embodiments, or perform equivalent substitution of some of the technical features, while remaining within the technical scope of the present disclosure; such modifications, changes or substitutions do not depart from the spirit and scope of the technical solutions of the embodiments of the present invention, and are intended to be included in the scope of the present invention.
Sequence listing
<110> Wang Shanxian
Hubei province ecological environment science institute (province ecological environment engineering evaluation center)
<120> a Korean bacillus LQB126, immobilized microbial agent and application thereof
<160> 3
<170> SIPOSequenceListing 1.0
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<211> 1420
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<213> Korean bacillus (Bacillus koreensis)
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gcattaagtt gggcactcta aggtgactgc cggtgacaaa ccggaggaag gtggggatga 1140
cgtcaaatca tcatgcccct tatgacctgg gctacacacg tgctacaatg gatgatacaa 1200
agggttgcga agccgcgagg tgaagctaat ctcataaaat cattctcagt tcggattgta 1260
ggctgcaact cgcctacatg aagctggaat cgctagtaat cgcggatcag catgccgcgg 1320
tgaatacgtt cccgggcctt gtacacaccg cccgtcacac cacgagagtt tgtaacaccc 1380
gaagtcggtg gggtaaccgt aaggagccag ccgcctaagg 1420
<210> 2
<211> 20
<212> DNA
<213> Artificial sequence (Artificial Sequence)
<400> 2
agagtttgat cctggctcag 20
<210> 3
<211> 22
<212> DNA
<213> Artificial sequence (Artificial Sequence)
<400> 3
tacggctacc ttgttacgac tt 22
Claims (9)
1. A bacillus korea LQB126, characterized by the taxonomic designation bacillus korea (Bacillus koreensis) having a accession number cctccc NO: m2021667.
2. Use of korean bacillus LQB126 according to claim 1 for the treatment of heavy metal contaminated environments.
3. The use of korean bacillus LQB126 for the treatment of heavy metal contaminated environments according to claim 2, wherein the heavy metal is one or more of As, cd, cr and Pb.
4. The use of korean bacillus LQB126 for the treatment of heavy metal contaminated environments according to claim 2, wherein the contaminated environments are one of water samples or soil samples.
5. An immobilized microbial agent comprising the korean bacillus LQB126 of claim 1 and an immobilization carrier.
6. The immobilized microbial inoculant of claim 5, wherein the immobilization carrier is one of sepiolite, diatomaceous earth, peat soil, corn cob and chestnut shells.
7. The use of the immobilized microbial agent of claim 5 or 6 in the treatment of heavy metal polluted environments.
8. The use of korean bacillus LQB126 for the treatment of heavy metal contaminated environments according to claim 7, wherein the heavy metal is one or more of As, cd, cr and Pb.
9. The use of korean bacillus LQB126 for the treatment of heavy metal contaminated environments according to claim 7, wherein the contaminated environments are one of water samples or soil samples.
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