CN110835617B

CN110835617B - Burkholderia cepacia and application thereof

Info

Publication number: CN110835617B
Application number: CN201911181380.2A
Authority: CN
Inventors: 李佳; 陈振兴; 杜冬云; 叶恒朋; 李学成
Original assignee: South Central University for Nationalities
Current assignee: South Central Minzu University
Priority date: 2019-11-22
Filing date: 2019-11-27
Publication date: 2021-04-13
Anticipated expiration: 2039-11-27
Also published as: CN110835617A

Abstract

The invention belongs to the technical field of biology, and particularly relates to Burkholderia cepacia and application thereof. The invention obtains the best process parameters by screening and inducing bacteria, utilizing the bred high-efficiency activated silicon bacteria and optimizing silicon leaching conditions, preliminarily verifies the selectivity of the silicon leaching effect of the bacteria by adopting different characterization means, and finally breeds a strain with good silicon activation effect: the burkholderia cepacia is preserved in the China center for type culture Collection, the preservation date is 2019, 9 and 17 days, and the preservation number is CCTCC NO: m2019732, which was classified and named Burkholderia cepacia (Burkholderia contaminans T).

Description

Burkholderia cepacia and application thereof

Technical Field

The invention belongs to the technical field of biology, and particularly relates to Burkholderia cepacia and application thereof.

Background

Electrolytic manganese slag (EMR, abbreviated as "manganese slag") is a solid waste generated in the production process of Electrolytic manganese metal. According to statistics, each 1 ton of electrolytic MnO is produced₂The production of 7-9t of manganese slag is accompanied. The accumulation of a large amount of manganese slag easily causes serious environmental pollution, thereby limiting the benign development of the electrolytic manganese industry. Meanwhile, the electrolytic manganese slag contains a plurality of beneficial elements, and is recycled by a reasonable means, so that the electrolytic manganese slag has economic benefits and can be used for realizingThe reduction treatment of the existing electrolytic manganese slag is the key of sustainable development of the electrolytic manganese industry.

Silicon is used as a fourth major nutrient element required by plant growth, and has great recovery value. The electrolytic manganese slag is a solid waste with high silicon content, but most of the waste is SiO which cannot be directly absorbed by plants₂The state of (A) exists, and the silicon that can be absorbed by plants is only monosilicic acid (H)₄SiO₄) The activated silicon is also called as activated silicon (effective silicon), so that the problem of serious shortage of effective silicon resources can be solved by researching the activation technology of silicon in the electrolytic manganese slag, and the method is also an effective way for realizing reduction treatment of the electrolytic manganese slag from the source.

At present, a chemical method is mainly adopted for resource utilization of silicon in electrolytic manganese slag to activate the silicon in the electrolytic manganese slag, but the method has the defects of high energy consumption, high cost and the like, and the use of chemical reagents can cause secondary pollution to the environment. Therefore, with the increasing maturity of the microbial leaching method, the application of the microbial leaching to the activation of the silicon in the electrolytic manganese slag becomes an effective way for solving the economic silicon release of the electrolytic manganese slag.

Disclosure of Invention

In order to obtain an effective way for solving the problem of economic silicon release of the electrolytic manganese slag, the invention takes the electrolytic manganese slag as an ore leaching object, bacteria with a silicon activation effect, namely the burkholderia cepacia, are screened and separated, and the burkholderia cepacia can be effectively applied to the activation of the silicon in the electrolytic manganese slag through enrichment culture, optimized culture, strain domestication and ore leaching evaluation.

In order to achieve the technical purpose, the invention adopts the following technical scheme:

the burkholderia cepacia is preserved in the China center for type culture Collection, the preservation date is 2019, 9 and 17 days, and the preservation number is CCTCC NO: m2019732, which is classified and named Burkholderia cepacia contaminans T.

The preparation method of the burkholderia cepacia of the invention comprises the following operations:

placing the soil sample around the electrolytic manganese slag heap leaching slag into an Alexander Raov liquid culture medium for culturing for 1-2d, standing for a period of time, sucking supernatant on a sterile operating platform, and then separating and screening silicon activated bacteria from the supernatant;

the separation and screening of the silicon activated bacteria are carried out according to the following steps:

(1) preparing the supernatant into bacterial liquids with different dilutions;

(2) coating the bacterial liquid obtained in the step (1) on an Alexander macro-loff liquid culture medium, carrying out inverted culture in a 30 ℃ biochemical incubator for 2-3d to observe colony morphology, preliminarily screening bacteria with characteristics similar to those of Bacillus mucilaginosus, and selecting strains with larger and mellow colonies;

(3) carrying out streak culture on the strain for 2-3 times, and carrying out microscopic examination until pure bacteria are obtained;

(4) culturing the pure strain obtained in the step (3) in an Alexander Raov liquid culture medium to logarithmic phase, centrifuging to prepare a bacterial suspension, and controlling the bacterial number to be 10⁸Per mL; the bacterial suspension is diluted to 10 degrees in a gradient way^-7Then placing the mixture in a culture dish with the diameter of 60mm, respectively coating the mixture on an LB culture medium, then placing the mixture in a constant temperature incubator at the temperature of 30 ℃, selecting a plate with the colony number of 100-200 after the colony grows fully, and determining the dilution concentration of the plate as the optimal dilution concentration; and (3) placing the flat plate with the optimal dilution concentration under an ultraviolet lamp at 30W for 30cm for irradiating for 120s, immediately taking out the flat plate after irradiation is finished, and storing in a refrigerator at 4 ℃ to obtain the burkholderia cepacia.

Further, the dilution degree in the step (1) is 10^-1、10^-2、10^-3、10^-4Or 10^-5Preferably 10^-3。

Further, the colony selection mode in the step (2) is as follows: bacillus mucilaginosus (B.M) is used as one of silicate bacteria, has good acid production capacity, thereby improving the mineral leaching and silicon releasing effect, preliminarily screens out bacteria with similar characteristics to the Bacillus mucilaginosus through the comparison of the screened bacteria and the Bacillus mucilaginosus in colony morphology and physiological and biochemical characteristics, reduces the bacteria screening range, shows a hemispherical shape by referring to the characteristics that the bacterial colony surface of the bacterial strain B.M on a flat plate is smooth, the edge is neat, transparent, colorless, glossy, moist and elastic, and is selected by the characteristic of wire drawing when being picked up by an inoculating needle.

The invention also provides application of the burkholderia cepacia in activation of silicon in electrolytic manganese residues.

Further, the specific steps of the application are as follows:

adding an alexander liquid culture medium into the electrolytic manganese slag, adjusting the pH value to 7.0-7.5, sterilizing, inoculating the screened Burkholderia cepacia, and reacting for a period of time to complete the silicon leaching process of the electrolytic manganese slag.

Further, the inoculation amount of the burkholderia cepacia in the application is 5% -15%, and preferably 10%.

Compared with the prior art, the invention has the advantages and beneficial effects that:

the method obtains the optimal process parameters by screening and inducing bacteria, utilizing the selected high-efficiency activated silicon bacteria and optimizing the silicon leaching conditions, adopts different characterization means to preliminarily verify the selectivity of the silicon leaching effect of the bacteria, proves that the burkholderia cepacia finally bred by the method has good silicon activation effect, and provides theoretical basis for resource utilization of electrolytic manganese slag.

Drawings

FIG. 1 is SiO in example 2₂A standard curve.

FIG. 2 shows SiO in the leachate of the silicon-activated bacteria of example 2₂The tendency of the content to change with time.

FIG. 3 is the final SiO in the leachate of silicon-activated bacteria of example 2₂And (4) content.

FIG. 4 is a graph showing the pH of the leachate of the silicon-activated bacteria in example 2 as a function of time.

FIG. 5 is the final pH of the silicon-activated bacterial leachate of example 2.

FIG. 6 is a colony morphology and a microscopic morphology of the strain Y-03 of example 3 on a solid medium.

FIG. 7 is a graph showing the growth of strain Y-03 in Alexander liquid medium in example 3.

FIG. 8 shows the lethality of the strains in example 3 under different UV irradiation times.

FIG. 9 shows the silicon activation effect of five strains after UV mutagenesis in example 3.

FIG. 10 shows the lethality of the strains in example 3 at different NTG mass concentrations.

FIG. 11 shows the effect of silicon activation of the strains in example 3 at different NTG mass concentrations.

FIG. 12 is an XRD pattern of the electrolytic manganese residue before and after leaching with Burkholderia cepacia in example 4.

FIG. 13 shows the effect of different bacteria on the activation of silicon in the electrolytic manganese slag in example 4.

Detailed Description

In the following examples, the electrolytic manganese slag is prepared from manganese ore of Retown under Daxin county of Chongzuo city of Zhuang nationality in Guangxi province₂The main components and contents of the waste generated after the decomposition of manganese slag are shown in Table 1.

TABLE 1 main chemical composition of electrolytic manganese slag

And (3) placing the electrolytic manganese slag in an electric heating air blast drying oven, drying for 6h at 105 ℃, taking the dried sample, and grinding to uniformly distribute sample particles, thereby obtaining the electrolytic manganese slag to be used.

Example 1 separation and screening of Burkholderia cepacia was carried out according to the following steps:

the media required during experiment a was prepared as follows:

liquid medium of alexander macroroff: 5.0g of sucrose, 5.0g of dipotassium phosphate dodecahydrate, 0.5g of magnesium sulfate heptahydrate, 0.1g of calcium carbonate, 0.005g of ferric chloride hexahydrate, 1000mL of distilled water and 20g of agar, and sterilizing the mixture in a sterilizing pot at 115 ℃ for 30min for later use.

LB culture medium: 10.0g of tryptone, 5.0g of yeast extract, 10.0g of sodium chloride, 1000mL of distilled water and 20g of agar, and sterilizing in a sterilization pot at 121 ℃ for 20min for later use.

B separation and screening of silicon-activated bacteria

And grinding the collected soil sample around the electrolytic manganese slag heap leaching slag warehouse to ensure that soil particles are fine and uniformly distributed. 5g of ground soil sample was put into 5 250mL dry Erlenmeyer flasks, 100mL Alexander Raov liquid medium was added to each Erlenmeyer flask, and after shaking for 1 day in a constant temperature shaker at a rotation speed of 200r/min and a temperature of 30 ℃, the Erlenmeyer flasks were taken out, left to stand for a while, and then the supernatant was separately aspirated on a sterile operating table, and then the silicon activated bacteria were separated and screened from the supernatant.

The separation and screening of the silicon activated bacteria are specifically carried out according to the following steps:

(1) respectively sucking 100 μ L of supernatant from 5 conical flasks into 1.5mL centrifuge tube, adding 900 μ L of sterile water, and mixing to obtain dilution 10^-1The bacterial liquid of (4).

(2) Then the same analogy is repeated to prepare 10^-2、10^-3、10^-4、10^-5And (4) diluted bacteria liquid.

(3) Placing the prepared Alexander macroroff culture medium into a culture dish, uniformly distributing the culture medium on the bottom of the whole culture dish, condensing to obtain a flat culture medium, pouring a flat plate on a sterile table, cooling and solidifying the flat plate, and numbering the culture dish.

(4) 0.1mL of each of the above-prepared bacterial solutions with different dilutions was pipetted and plated on a petri dish.

(5) The plate is inversely cultured in a biochemical incubator at 30 ℃ for 2-3d to observe colony morphology, and strains with larger and mellow colonies are selected.

(6) The strain is streaked and cultured for 2-3 times, and microscopic examination is carried out until pure bacteria are obtained.

The colony selection mode in the step (5) is as follows: the bacillus mucilaginosus (B.M) is one of silicate bacteria, has good acid production capacity, thereby improving the mineral leaching and silicon releasing effect, preliminarily screens the bacteria with similar characteristics to the bacillus mucilaginosus through the comparison of the bacterial colony morphology and the physiological and biochemical characteristics of the screened bacteria and the bacillus mucilaginosus, and reduces the bacteria screening range. The colony of reference strain B.M on the plate was smooth, clean, transparent, colorless, glossy, moist, elastic, and semi-spherical, with stringiness when picked up with an inoculating needle.

Finally, 1 strain of silicon activated bacteria is obtained in each conical flask, and the serial numbers are respectively as follows: y-01, Y-02, Y-03, Y-04, Y-05.

Example 2 acid production test by silicon-activated bacteria and measurement of silicon activation effect

Conditions for fixed bacterial culture: the temperature was 30 ℃, the rotation speed was 200rpm, the pH was 7.5, and the inoculum size was 10%.

Acid production experiment by silicon activated bacteria:

adding 5.0g of electrolytic manganese slag into a dry 250mL conical flask, adding 90mL of Alexander Raov liquid culture medium prepared in example 1, adjusting pH to 7.5 with 0.05mol/L NaOH solution, sealing the bottle mouth with newspaper to avoid contamination, placing into a sterilization pot, sterilizing at 115 deg.C for 30min, cooling, inoculating 10mL of bacterial liquid in logarithmic phase on a sterile operating platform, placing into a constant temperature oscillator with 200rpm and 30 deg.C, culturing for 14d, and measuring pH value of the leachate with pH meter every 2 d.

Determination of the silicon activation effect of bacteria:

SiO₂standard curve determination of standard solutions: taking 0.0mL, 1.0mL, 2.0mL, 3.0mL, 4.0mL, 6.0mL of 1000. mu.g/mL SiO₂Respectively putting the standard solution into 6 50mL volumetric flasks, and measuring SiO with different concentrations by adopting a silicon-molybdenum blue spectrophotometry₂Absorbance of the standard solution at 680nm, recording data and drawing SiO₂Standard curve: y ═ 0.1306X +0.011 (Y: absorbance, X: SiO)₂ mg·L^-1)，SiO₂The standard curve is shown in figure 1.

Adding 5.0g of electrolytic manganese slag into a dry 250mL conical flask, adding 100mL of Alexander Raov liquid culture medium prepared in example 1, adjusting pH to 7.5, sealing the flask mouth with newspaper to avoid contamination, sterilizing in a sterilization pot at 115 deg.C for 30min, cooling, inoculating 10mL of bacterial liquid in logarithmic phase on a sterile operating platform, culturing in a constant temperature oscillator at 200rpm and 30 deg.C for 14d, measuring absorbance of the leachate once every 2d by silicon-molybdenum blue spectrophotometry, and measuring the absorbance of the leachate according to the SiO₂Calculating SiO in the leaching solution by using a standard curve₂The content, 3 samples per strain of bacteria, was averaged.

The above experiments were carried out using the strains of silicon-activated bacteria Y-01, Y-02, Y-03, Y-04, and Y-05 obtained in example 1, respectively.

In the 14d leaching system, the SiO content of the leaching solution is measured every 2d₂The content and the silicon activation effect of each strain at the early stage are continuously improved, and the silicon activation effect at the later stage tends to be stable, as shown in figure 2.

At the end of leaching 14d, the final silicon activation effect of each strain is shown in fig. 3, with SiO in the leachate₂The strains with higher content are Y-01, Y-02 and Y-03, and the leaching solution is SiO₂The strains with lower content are Y-04 and Y-05. Wherein SiO in the leaching solution of the strain Y-03₂The highest content is 130.30 mg.L^-1Secondly, strains Y-02 and Y-01, SiO in the leaching solution₂The content is 128.17 mg.L respectively^-1、126.11mg·L^-1. SiO in leaching solution of strain Y-04₂The content is less, 117.84 mg.L^-1SiO in leachate of strain Y-05₂The lowest content is 111.64 mg.L^-1。

In the leaching process of 14d, the pH value of the leaching solution is shown in fig. 4, and the result shows that a certain amount of acid is generated in the leaching process of the bacterial strain, and the bacterial strain generates more acid in the early stage of leaching along with the increase of the leaching time; in the later stage of leaching, the acid production capacity of the bacterial strain tends to be stable.

In the leaching system of 14d, the pH of leachate of each strain is reduced, and the final pH of leachate of each strain at the end of leaching is shown in FIG. 5. The strains Y-02 and Y-03 produce more acid, and the final pH values are 5.28 and 5.25 respectively, wherein the pH value of the strain Y-03 is reduced most, which indicates that the acid-producing capability of the strain Y-03 is strongest. The strains Y-01, Y-04 and Y-05 have relatively less acid production, and the final pH values are 5.38, 5.51 and 5.64 respectively, wherein the pH value of Y-05 is reduced the least, which indicates that the acid production capability is the weakest.

In a 14d leaching system, the bacterial strain with better acid production capability has better silicon activation effect; the acid-producing ability of the strain Y-03 with the best silicon activation effect is not the best from the individual point of view, so that the silicon activation effect of the strain is related to the acid-producing ability of the strain, but the silicon activation effect of the strain is not determined by the single acid-producing ability.

Example 3 mutagenesis experiment of silicon-activated bacteria

The strain Y-03 having the best silicon activation effect was used as the starting strain for Ultraviolet (UV) and Nitrosoguanidine (NTG) mutagenesis experiments to culture a mutant strain having better silicon activation ability, and the form of the strain Y-03 on the Alexander Raov solid medium prepared in example 1 is shown in FIG. 6.

Culturing the strain Y-03 with optimal silicon activation effect in Alexander Raov liquid culture medium prepared in example 1 (fixed bacteria culture conditions: temperature 30 ℃, rotation speed 200rpm, pH 7.5, inoculum size 10%), measuring OD600 value every 4h until the OD600 value tends to be stable, drawing a growth curve of bacteria, taking the bacterial suspension in logarithmic phase, centrifuging for 10min in a 8000rpm high-speed centrifuge, removing supernatant, washing with sterile water to obtain bacterial suspension, and controlling the bacterial number to 10⁸And (4) obtaining bacterial suspension required by mutagenesis experiments. The growth curve of strain Y-03 in Alexander liquid medium is shown in FIG. 7.

Ultraviolet (UV) mutagenesis:

the bacterial suspension concentration gradient required by the mutagenesis experiment is diluted to 10^-7Then placing the culture medium in a culture dish with the diameter of 60mm, coating the culture medium on LB solid medium, placing the culture medium in a constant temperature incubator at the temperature of 30 ℃, selecting a plate with the colony number of 100-200 after the colony grows fully, and determining the dilution concentration of the plate as the optimal dilution concentration. Selecting an ultraviolet lamp with power of 30W, placing the flat plate with the optimal dilution concentration under the lamp for 30s, 60s, 120s and 180s respectively, taking out the flat plate immediately after the irradiation is finished, and storing in a refrigerator at 4 ℃ for 12 h. Meanwhile, a bacterial plate control group which is not subjected to ultraviolet mutagenesis is set up, the experimental group and the control group are placed in a 30 ℃ constant temperature incubator to be cultured for 24 hours, colonies of the two groups of plates are counted, and the mutagenesis lethality rate is calculated according to the following formula:

the lethality is shown in FIG. 8. The positive mutant strain generally resulted in an irradiation time of 90% lethality, and the strain lethality rates were 46.02%, 65.33%, 89.77%, 94.98%, 98.12% at ultraviolet irradiation times of 0s, 30s, 60s, 120s, and 180s, respectively, so 120s was selected as the optimum ultraviolet irradiation time for UV mutagenesis.

The positive mutant strain after UV mutagenesis (120s) was subjected to scale-up culture, five strains with round colonies and larger diameter than the original strain were selected after coating, and the silicon activation experiment by leaching was performed according to the method of example 2, and the silicon activation effect is shown in FIG. 9. Among the five selected variant strains, the leaching solution SiO of the strains Y-03-U1, Y-03-U2, Y-03-U3 and Y-03-U4₂The content is respectively 116.76 mg.L^-1、105.17mg·L^-1、121.43mg·L^-1、126.21mg·L^-1SiO of only one strain Y-03-U5₂The leaching amount is higher than that of the original strain, and SiO in the leaching solution is₂The mass concentration is 149.11 mg.L^-1. The silicon activation effect of the strain after mutagenesis is 130.30 mg.L^-1Lifting to 149.11 mg.L^-1. The results show that the silicon activation effect of the strains screened by UV mutagenesis is better than that of the original strains.

Nitrosoguanidine (NTG) mutagenesis:

into 5 250mL Erlenmeyer flasks were added 90mL of the Alexandriv liquid medium prepared in example 1 and a nitrosoguanidine reagent in a predetermined amount such that the nitrosoguanidine concentration was 0, 300, 500, 600, and 700 mg.L^-1Then adding 10mL of bacterial suspension required by a mutagenesis experiment into a conical flask, culturing for 40min in a constant-temperature reciprocating oscillator at 30 ℃ and 200rpm, taking the bacterial suspension, centrifuging for 10min at 8000rpm of a high-speed centrifuge, washing out thalli by using sterile water, repeating the process for three times, and terminating mutagenesis. The mutagenic bacterium liquid is coated on the Alexander macro-loff solid culture medium prepared in example 1 and then placed in a constant temperature incubator at 30 ℃, and after 48 hours, plate colony counting is carried out, the lethality is calculated, and the optimal dose of NTG mutagenesis is determined. The lethality is shown in FIG. 10. Positive mutants typically result in mutagenesis concentrations of 90% lethality at 0, 300, 500, 600, 700 mg.L^-1The lethality of the strain is 0%, 36.84%, 78.72%, 89.96% respectively at the mass concentration of nitrosoguanidine(s)% and 98.64%, 600 mg. L was selected^-1The concentration of Nitrosoguanidine (NTG) was used as the optimum concentration for NTG mutagenesis.

Determination of the silicon activation effect of the mutagenized strain: subjecting to NTG mutagenesis (the mass concentration of nitrosoguanidine is 600 mg. multidot.L)^-1) And (3) performing amplification culture on the subsequent positive mutant strain, and selecting 5 bacteria which have mellow colonies and larger diameters than the original strain: the silicon activation effect was measured in the same manner as in example 2 using Y-03-N1, Y-03-N2, Y-03-N3, Y-03-N4 and Y-03-N5 as experimental groups and Y-03-N4 as control group, and the silicon activation effect is shown in FIG. 11. Among the five selected mutant strains, the SiO in the leachate of the strains Y-03-N1, Y-03-N2, Y-03-N4 and Y-03-N5₂The content is 128.26 mg.L respectively^-1、116.23mg·L^-1、96.34mg·L^-1、95.51mg·L^-1Compared with the silicon activation effect of the original strain, the silicon activation effects of the four strains are not as good as those of the original strain. SiO of only one strain₂The leaching amount is higher than that of the original strain, the serial number is Y-03-N3, and SiO in the leaching solution is₂The mass concentration is 145.71 mg.L^-1. The silicon activation effect of the strain after mutagenesis is 130.30 mg.L^-1Lifting to 145.71 mg.L^-1. The result shows that the strain screened by NTG mutagenesis has better silicon activation effect than the original strain.

Exploration of genetic stability of mutagenized strain

Ultraviolet mutagenesis and NTG mutagenesis were performed on the starting strain Y-03 to generate 2 bacteria with stronger silicon activation effect, Y-03-U5 and Y-03-N3, respectively, and 2 bacteria and the starting strain were continuously cultured for 7 generations in the Alexander macro-loff liquid medium prepared in example 1, and SiO in the measuring medium of 1 st, 4 th and 7 th generations of bacteria was selected₂The genetic stability of the silicon activation ability of the mutagenized strain was determined, the silicon activation effect was measured according to the method of example 2, and the SiO content of the leachate was measured at the end of leaching the electrolytic manganese residue 14d₂Mass concentration, results are shown in table 2.

TABLE 2 genetic stability of the mutagenized strains (values in mg. multidot.L in the table)^-1)

It can be known from the table that the silicon activation performance of the strain Y-03-N3 begins to degrade after the seventh generation, the other strain still has stable silicon activation effect after continuous subculture for 7 generations, and the result of ultraviolet mutagenesis and NTG mutagenesis is combined, so that the strain Y-03-U5 has the strongest silicon leaching capability and stable genetic performance, and the mutagenized strain Y-03-U5 is determined to be the leaching strain of the later experiment.

The identification of bacteria is completed by China center for type culture Collection. Reference is made to "6.5" in NY/T1736-2009. PCR amplification is carried out on a specific gene segment of the strain Y-03-U5 (primer: 27F: AGAGAGTTTGATCCTGGCTCCAG; 1492R: ACGGCTACC TTGTTACGACTT), an amplification product is recovered and sequenced, the sequence is compared with the sequence registered in a GenBank database in a homology way, and the homology of the 16S rDNA sequence of the strain and the Burkholderia cepacia reaches 100 percent. The strain should be Burkholderia cepacia (Burkholderia continans). The Burkholderia cepacia is preserved in the China center for type culture Collection, the preservation date is 2019, 9 and 17 months, and the preservation number is CCTCC NO: m2019732, which was classified and named Burkholderia cepacia (Burkholderia contineans T).

16S rDNA sequence of the strain:

AACGGCAGCACGGGTGCTTGCACCTGGTGGCGAGTGGCGAACGGGTGAGTAATACATCGGAACATGTCCTGTAGTGGGGGATAGCCCGGCGAAAGCCGGATTAATACCGCATACGATCTACGGATGAAAGCGGGGGACCTTCGGGCCTCGCGCTATAGGGTTGGCCGATGGCTGATTAGCTAGTTGGTGGGGTAAAGGCCTACCAAGGCGACGATCAGTAGCTGGTCTGAGAGGACGACCAGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATTTTGGACAATGGGCGAAAGCCTGATCCAGCAATGCCGCGTGTGTGAAGAAGGCCTTCGGGTTGTAAAGCACTTTTGTCCGGAAAGAAATCCTTGGCTCTAATACAGTCGGGGGATGACGGTACCGGAAGAATAAGCACCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGTGCGAGCGTTAATCGGAATTACTGGGCGTAAAGCGTGCGCAGGCGGTTTGCTAAGACCGATGTGAAATCCCCGGGCTCAACCTGGGAACTGCATTGGTGACTGGCAGGCTAGAGTATGGCAGAGGGGGGTAGAATTCCACGTGTAGCAGTGAAATGCGTAGAGATGTGGAGGAATACCGATGGCGAAGGCAGCCCCCTGGGCCAATACTGACGCTCATGCACGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCCTAAACGATGTCAACTAGTTGTTGGGGATTCATTTCCTTAGTAACGTAGCTAACGCGTGAAGTTGACCGCCTGGGGAGTACGGTCGCAAGATTAAAACTCAAAGGAATTGACGGGGACCCGCACAAGCGGTGGATGATGTGGATTAATTCGATGCAACGCGAAAAACCTTACCTACCCTTGACATGGTCGGAATCCCGCTGAGAGGTGGGAGTGCTCGAAAGAGAACCGGCGCACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGTCCTTAGTTGCTACGCAAGAGCACTCTAAGGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAGTCCTCATGGCCCTTATGGGTAGGGCTTCACACGTCATACAATGGTCGGAACAGAGGGTTGCCAACCCGCGAGGGGGAGCTAATCCCAGAAAACCGATCGTAGTCCGGATTGCACTCTGCAACTCGAGTGCATGAAGCTGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGTCTTGTACACACCGCCCGTCACACCATGGGAGTGGGTTTTACCAGAAGTG

example 4 Leaching of silicon in electrolytic manganese sludge by Burkholderia cepacia Y-03-U5 Strain

The leaching effect of the Burkholderia cepacia Y-03-U5 strain on silicon in the electrolytic manganese residue was measured according to the method for measuring the activation effect of bacterial silicon in example 2.

The residue morphology of the Burkholderia cepacia Y-03-U5 strain after ore leaching for 14d was characterized by SEM and compared with the original manganese residue morphology, and the graph is omitted.

SEM pictures of electrolytic manganese residues before and after leaching of the Burkholderia cepacia Y-03-U5 strain can be seen: most of original manganese slag particles are in a regular and smooth columnar structure, the edges and corners are clear, the crystal structure is complete, and clear outlines and smooth surfaces can be seen; however, the manganese slag leached by Burkholderia cepacia undergoes obvious change, the smooth surface and the columnar structure of the particles are seriously damaged and corroded, the surface becomes uneven, the outline becomes a little fuzzy, and the electrolytic manganese slag particles are corroded under the action of bacteria acid production and self metabolism, so that the particle structure and the surface form are changed.

After the Burkholderia cepacia strain Y-03-U5 was subjected to UV mutagenesis and subjected to mineral leaching for 14d, the mineral composition of the leaching residue was characterized by XRD and compared with that of the original manganese residue, and the result is shown in FIG. 12.

FIG. 12 XRD patterns of the electrolytic manganese slag before and after bacterial leaching show that the original manganese slag without bacterial action contains quartz, gypsum and brushite as main mineral components. After the electrolytic manganese slag is leached by the Burkholderia cepacia and the strain Y-03-U5, the XRD pattern shows that the characteristic peak of quartz is obviously reduced, which indicates that the quartz is seriously corroded after the Burkholderia cepacia Y-03-U5 is leached. Analysis shows that after the electrolytic manganese slag is leached by the Burkholderia cepacia, the electrolytic manganese slag corrodes the crystal structures of various minerals due to the metabolism and acid production of the Burkholderia cepacia. The quartz is seriously corroded, and the other minerals are less corroded, so that the Burkholderia cepacia has selective corrosion action on the minerals, and the weathering decomposition action of the Burkholderia cepacia on the surfaces of the minerals can be reflected to a certain extent.

By using the method of the silicon activation effect test experiment in example 2, the effect of burkholderia cepacia in leaching the silicon in the electrolytic manganese residue was compared with the purchased silicate bacteria SB (purchased from cangzhou wang Producer technology research institute) and bacillus mucilaginosus BM (purchased from China agricultural microbial culture Collection), and whether the burkholderia cepacia Y-03-U5 has an advantage in the activation of the silicon in the electrolytic manganese residue was investigated, and the leaching comparison result is shown in fig. 13.

As can be seen from FIG. 13, the silicon leaching effects of silicate bacteria, Bacillus mucilaginosus and Burkholderia cepacia were 93.64, 115.86 and 149.11 mg.L, respectively^-1The silicon leaching effect of the burkholderia cepacia on the electrolytic manganese residue is obviously higher than that of two purchased bacteria.

Sequence listing

<110> university of the south China nationality

<120> Burkholderia cepacia and application thereof

<141> 2019-11-22

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<170> SIPOSequenceListing 1.0

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<213> Burkholderia cepacia (Burkholderia contaminans)

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cggatgaaag cgggggacct tcgggcctcg cgctataggg ttggccgatg gctgattagc 180

tagttggtgg ggtaaaggcc taccaaggcg acgatcagta gctggtctga gaggacgacc 240

agccacactg ggactgagac acggcccaga ctcctacggg aggcagcagt ggggaatttt 300

ggacaatggg cgaaagcctg atccagcaat gccgcgtgtg tgaagaaggc cttcgggttg 360

taaagcactt ttgtccggaa agaaatcctt ggctctaata cagtcggggg atgacggtac 420

cggaagaata agcaccggct aactacgtgc cagcagccgc ggtaatacgt agggtgcgag 480

cgttaatcgg aattactggg cgtaaagcgt gcgcaggcgg tttgctaaga ccgatgtgaa 540

atccccgggc tcaacctggg aactgcattg gtgactggca ggctagagta tggcagaggg 600

gggtagaatt ccacgtgtag cagtgaaatg cgtagagatg tggaggaata ccgatggcga 660

aggcagcccc ctgggccaat actgacgctc atgcacgaaa gcgtggggag caaacaggat 720

tagataccct ggtagtccac gccctaaacg atgtcaacta gttgttgggg attcatttcc 780

ttagtaacgt agctaacgcg tgaagttgac cgcctgggga gtacggtcgc aagattaaaa 840

ctcaaaggaa ttgacgggga cccgcacaag cggtggatga tgtggattaa ttcgatgcaa 900

cgcgaaaaac cttacctacc cttgacatgg tcggaatccc gctgagaggt gggagtgctc 960

gaaagagaac cggcgcacag gtgctgcatg gctgtcgtca gctcgtgtcg tgagatgttg 1020

ggttaagtcc cgcaacgagc gcaacccttg tccttagttg ctacgcaaga gcactctaag 1080

gagactgccg gtgacaaacc ggaggaaggt ggggatgacg tcaagtcctc atggccctta 1140

tgggtagggc ttcacacgtc atacaatggt cggaacagag ggttgccaac ccgcgagggg 1200

gagctaatcc cagaaaaccg atcgtagtcc ggattgcact ctgcaactcg agtgcatgaa 1260

gctggaatcg ctagtaatcg cggatcagca tgccgcggtg aatacgttcc cgggtcttgt 1320

acacaccgcc cgtcacacca tgggagtggg ttttaccaga agtg 1364

Claims

1. The burkholderia cepacia is preserved in the China center for type culture Collection, the preservation date is 2019, 9 and 17 days, and the preservation number is CCTCC NO: m2019732, its classification name is Burkholderia cepacia (B.)Burkholderia contaminans T）。

2. Use of the burkholderia cepacia according to claim 1 in desiliconization of electrolytic manganese residues.

3. The application according to claim 2, characterized in that the specific steps of the application are as follows:

adding an alexander waffle liquid culture medium into the electrolytic manganese slag, then adjusting the pH =7.0-7.5 of the system, sterilizing, inoculating Burkholderia cepacia, and reacting for a period of time to complete the desiliconization process of the electrolytic manganese slag.

4. The use according to claim 3, wherein the amount of Burkholderia cepacia is 5% to 15% by weight.