KR20200081043A - Modified microorganism including genetic modification that increases a protein gene expression and method for reducing concentration of fluorine containing compounds in sample - Google Patents

Abstract

Provided are a modified microorganism comprising genetic modification increasing expression of a protein, a composition for use in reducing a concentration of fluorinated methane in a sample containing the modified microorganism, and a method for reducing the concentration of fluorinated methane in the sample. The modified microorganism of the present invention includes the genetic modification increasing the expression of a gene encoding the protein having 85% or more of sequence identity to an amino acid sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, or 7, or a combination thereof.

Description

Modified microorganisms including genetic modification that increases a protein gene expression and method for reducing concentration of fluorine containing compounds in sample}

Modified microorganisms comprising genetic modifications that increase the expression of protein genes, compositions for use in reducing the concentration of fluorine-containing compounds in samples comprising the modified microorganisms, and methods of reducing concentrations of fluorine-containing compounds in samples It is about.

Greenhouse gas emissions that accelerate global warming are one of the most serious environmental issues, and regulations on greenhouse gas emissions are being strengthened to regulate and prevent them. Among them, fluorinated gases (F-gas) such as perfluorocarbons (PFCs), hydrofluorocarbons (HFCs), and sulfur hexafluoride (SF ₆ ) have low absolute emissions but long half-lives. It is reported that the global warming coefficient is so high that it has a more serious effect. In the semiconductor and electronic industries, which are the main sources of F-gas, the amount of F-gas generated has increased over the quota for GHG emissions, and accordingly, the cost burden for disassembling greenhouse gas and purchasing emission rights is increasing every year.

However, the existing F-gas decomposition uses a thermal decomposition or catalytic thermal oxidation process, but there are limitations such as limited removal rate, secondary harmful substance emission, and high cost. To solve this problem, the introduction of a biological F-gas decomposition process using a bio-catalyst will overcome the limitations of the existing chemical decomposition process, and more economical and environmentally friendly F-gas treatment will be possible.

However, even according to the above-described prior art, there is a need for a composition and method for reducing the concentration of a fluorine-containing compound in a sample using the microorganism, which has an activity of reducing the concentration of the fluorine-containing compound in the sample.

One aspect is a genetic modification that increases the expression of a gene encoding a protein having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, or 7, or a combination thereof. It provides a modified microorganism comprising a.

Other aspects are SEQ ID NO: 1, 2, 3, 4, 5, 6, or a protein having a sequence identity of 85% or more with the amino acid sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, Or a modified microorganism comprising a genetic modification that increases the expression of a gene encoding a protein having at least 85% sequence identity with the amino acid sequence of 7, or a combination thereof, or a lysate thereof , Provides a composition for use in reducing the concentration of a fluorine-containing compound in a sample.

Other aspects are SEQ ID NO: 1, 2, 3, 4, 5, 6, or a protein having a sequence identity of 85% or more with the amino acid sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, Or a modified microorganism comprising a genetic modification that increases the expression of a gene encoding a protein having a sequence identity of 85% or more with the amino acid sequence of 7, or a combination thereof, or a lysate thereof. A method for reducing the concentration of a fluorine-containing compound in a sample is provided, comprising contacting the compound-containing sample to reduce the concentration of the fluorine-containing compound in the sample.

As used herein, the term "increase in activity", or "increased activity" refers to a detectable increase in the activity of a cell, protein, or enzyme. "Increase in activity", or "increased activity" is a cell, protein, or enzyme that does not have a given genetic modification (eg, native or "wild-type") "Indicates the activity of a modified (eg, genetically engineered) cell, protein, or enzyme above the level of a comparable cell, protein, or enzyme of the same type, such as a cell, protein, or enzyme. "Cell activity" refers to the activity of a specific protein or enzyme in a cell. For example, the activity of the modified or engineered cell, protein, or enzyme is at least about 5% greater than the activity of the same type of unengineered cell, protein, or enzyme, eg, wild-type cell, protein, or enzyme. , About 10% or more, about 15% or more, about 20% or more, about 30% or more, about 50% or more, about 60% or more, about 70% or more, or about 100% or more. The activity of a specific protein or enzyme in a cell is about 5% or more, about 10% or more, about 15% or more, about 20% or more, about 30% of the activity of the same protein or enzyme in a parent cell, e.g., an unmanaged cell Or more, about 50% or more, about 60% or more, about 70% or more, or about 100% or more. Cells with increased activity of proteins or enzymes can be identified using any method known in the art.

Increased activity of enzymes or polypeptides can be obtained by increased expression or increased specific activity. The increase in expression may be due to introduction of a polynucleotide encoding an enzyme or polypeptide into a cell, an increase in the number of copies, or a variation in the regulatory region of the polynucleotide. The microorganism into which the gene is introduced may implicitly or not contain the gene. The gene may be operably linked to a regulatory sequence that enables its expression, such as a promoter, enhancer, poly adenylation site, or combinations thereof. Polynucleotides introduced externally or whose copy number is increased can be endogenous or exogenous. The endogenous gene refers to a gene existing on a genetic material contained in a microorganism. The exogenous gene refers to a gene that is introduced into the cell from the outside, and the introduced gene may be homologous or heterologous to the host cell to be introduced. “Heterologous” may mean foreign, not native.

The term "copy number increase" may be due to introduction or amplification of the gene, and also includes a case in which a gene that does not exist in an unmanaged cell is genetically manipulated. The introduction of the gene can be accomplished by mediating a vehicle such as a vector. The introduction may be a transient introduction in which the gene is not integrated into the genome or may be inserted into the genome. The introduction may be achieved, for example, by introducing a vector into which the polynucleotide encoding the desired polypeptide is inserted into the cell, and then the vector is cloned in the cell or the polynucleotide is integrated into the genome.

The introduction of the gene can be performed by known methods such as transformation, transfection, and electroporation. The gene may be introduced through a vehicle or may be introduced by itself. As used herein, "carrier" includes nucleic acid molecules capable of delivering other nucleic acids to which they are linked. In view of the nucleic acid sequence that mediates the introduction of a particular gene, it is contemplated herein that the carrier can be used interchangeably with vectors, nucleic acid constructs, and cassettes. Vectors include, for example, plasmid or virus-derived vectors. Plasmids include circular double-stranded DNA rings to which additional DNA can be linked. Vectors can include, for example, plasmid expression vectors, viral expression vectors, such as replication-defective retroviruses, adenoviruses, and adeno-associated viruses, or combinations thereof.

The manipulation of genes used herein can be by molecular biological methods known in the art.

The term “parent cell” refers to an original cell, eg, a cell that has not been genetically engineered of the same type with respect to the engineered microorganism. For a particular genetic modification, the "parent cell" is a cell that does not have the specific genetic modification, but may be the same for other situations. Thus, the parental cells are genetically having increased activity of a given protein (e.g., a protein having at least about 85% sequence identity with the amino acid sequence of SEQ ID NOs: 1, 2, 3, 4, 5, 6, or 7). It may be a cell used as a starting material to produce engineered microorganisms. The same comparison applies to other genetic modifications.

As used herein, the term “gene” refers to a nucleic acid fragment that expresses a specific protein, and 5′-non coding sequence and/or 3′-non coding sequence. The sequence may or may not contain a regulatory sequence.

In the present specification, “sequence identity” of a nucleic acid or a polypeptide refers to the same degree of base or amino acid residues between sequences after aligning the two sequences in the specific comparison region so as to be maximally matched. Sequence identity is a value measured by optimally aligning and comparing two sequences in a specific comparison region, and a part of the sequence in the comparison region may be added or deleted compared to a reference sequence. Percent sequence identity is, for example, comparing two optimally aligned sequences across the comparison region, determining the number of positions where the same amino acid or nucleotide appears in both sequences to obtain the number of matched positions Can be calculated by the step of dividing the number of matched positions by the total number of positions in the comparison range (i.e. range size), and multiplying the result by 100 to obtain a percentage of sequence identity. The percentage of sequence identity can be determined using known sequence comparison programs, examples of such programs include BLASTN (NCBI), BLASTP (NCBI), CLC Main Workbench (CLC bio), MegAlign ^TM (DNASTAR Inc), etc. Can.

The term "genetic modification" as used herein includes artificially altering the composition or structure of a cell's genetic material.

In the present specification,% represents w/w% unless otherwise specified.

One aspect is a genetic modification that increases the expression of a gene encoding a protein having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, or 7, or a combination thereof. It provides a modified microorganism comprising a.

Proteins having at least 85% sequence identity to the amino acid sequence of SEQ ID NOs: 1, 2, 3, 4, 5, 6, or 7 may be those whose expression in the presence of CF4 in microbial cells increases compared to the absence of CF4 have.

Genes encoding proteins having the amino acid sequence of SEQ ID NOs: 1, 2, 3, 4, 5, 6, or 7 are also referred to as Gene_01585, Gene_04413, Gene_00569, Gene_00598, Gene_04421, cspD, and yttP_1. Gene_04421 may be a transposase for the insertion sequence IS3. The cspD protein may be a cold shock-like protein CspD. yttP_1 may be a possible HTH-type transcription regulator YttP protein. The genetic modification may be to increase the activity of each protein. The genetic modification may be to increase the activity of the transposase for IS3.

The protein may be foreign or intrinsic to the modified microorganism. The protein may be selected from the group consisting of Pseudomonas, Bacillus, Pseudomonas, Azotobacter, Agrobacterium, and Escherichia. The protein may be derived from a strain selected from the group consisting of Pseudomonas saitens, Bacillus cereus, Bacillus thuringiensis, and Bacillus megaterium.

In the microorganism, the genetic modification may be to increase the expression of the gene encoding the protein. The genetic modification may be to increase the number of copies of the gene encoding the protein. The genetic modification may be a modification of the regulatory region sequence of the gene encoding the protein. The regulatory region can be a promoter, a terminator, or an operator. The gene may have a sequence identity of at least 85% with the nucleotide sequence of SEQ ID NO: 8, 9, 10, 11, 12, 13, or 14, respectively. The genetic modification may be that a gene encoding the protein is introduced, for example, introduced through a vehicle such as a vector. The gene encoding the protein may exist within or outside the chromosome. The introduced gene encoding the protein may be plural, for example, 2 or more, 5 or more, 10 or more, 10 or more, 50 or more, 100 or more, or 1,000 or more.

The microorganism may be of Escherichia genus, Bacillus genus, Xanthobacter genus, Pseudomonas genus, Azotobacter genus, or Agrobacterium genus. The microorganism may be Pseudomonas saitens, E. coli, or X. autotrophicus.

The modified microorganism may reduce the concentration of the fluorine-containing compound in the sample. The reduction may be achieved by introducing the hydroxyl group into carbon by the protein acting on the C-F or C-H bond of the fluorine-containing compound, or by accumulating the fluorine-containing compound in the cells of microorganisms. In addition, the reduction may include cutting the C-F bond of the fluorine-containing compound, converting the fluorine-containing compound to another substance, or accumulating the fluorine-containing compound in cells. The sample may be in a liquid or gaseous state. The sample may be factory wastewater or waste gas. Any of the above samples may be included as long as it contains the fluorine-containing compound.

The fluorine-containing compound may be a compound represented by the following Chemical Formulas 1 to 3.

<Formula 1> <Formula 2>

C(R ₁ )(R ₂ )(R ₃ )(R ₄ ) (R ₅ )(R ₆ )(R ₇ )C-[C(R ₁₁ )(R ₁₂ )] _n -C(R ₈ )( R ₉ )(R ₁₀ )

<Formula 3>

S(R ₁₃ )(R ₁₄ )(R ₁₅ )(R ₁₆ ) (R ₁₇ )(R ₁₈ )

In the above equations,

n is an integer from 0 to 10, and when n is 2 to 10, R11 among the plurality of R11 may have each other or be different, and R12 among the plurality of R12 may have each other or different,

R ₁ , R ₂ , R ₃ and R ₄ are independently of each other F, Cl, Br, I, or H, and at least one of R ₁ , R ₂ , R ₃ and R ₄ is F,

R ₅ , R ₆ , R ₇ , R ₈ , R ₉ , R ₁₀ , R ₁₁ and R ₁₂ are independently F, Cl, Br, I, or H, and R ₅ , R ₆ , R ₇ , R ₈ , R ₉ , R ₁₀ , one or more of R ₁₁ and R ₁₂ is F,

R ₁₃ , R ₁₄ , R ₁₅ , R ₁₆ , R ₁₇ and R ₁₈ are each independently F, Cl, Br, I, or H, and R ₁₃ , R ₁₄ , R ₁₅ , R ₁₆ , R ₁₇ and R ₁₈ One or more of them are F.

The fluorine-containing compound may be a fluorinated methane compound represented by CHnF4-n (n is an integer of 0 to 3). The fluorine-containing compound may include, for example, one or more selected from the group consisting of CH ₃ F, CH ₂ F ₂ , CHF ₃ , CF ₄ and SF ₆ .

Other aspects are SEQ ID NO: 1, 2, 3, 4, 5, 6, or a protein having a sequence identity of 85% or more with the amino acid sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, Or a modified microorganism comprising a genetic modification that increases the expression of a gene encoding a protein having at least 85% sequence identity with the amino acid sequence of 7, or a combination thereof, or a lysate thereof , As a composition for use in reducing the concentration of a fluorine-containing compound in a sample, the fluorine-containing compound provides a composition that is a compound represented by the following Chemical Formulas 1 to 3.

In the composition, the protein, the modified microorganism, the sample, and the fluorine-containing compound are as described above.

In the composition, the lysate breaks down the cell membrane and/or cell wall of the modified microorganism to indicate a state in which the cell contents are exposed to the outside. The destruction can be accomplished by physical, chemical, or enzymatic methods. The physical method can be achieved by applying a shear stress to the cells. The chemical method can be performed by contacting a cell membrane or a material such as a surfactant that can destroy a wall. Enzymatic methods involve contacting the cell with an enzyme that has the activity to cleave components of the cell membrane or wall. The enzyme may be a lipase, protease or glycolytic enzyme.

In the above composition, the term "reduction" is to reduce the concentration of the fluorine-containing compound in the sample, which includes completely removing it. The sample may be gas or liquid. The sample may be one that does not contain the microorganism. The composition may further include a substance that increases the solubility of the fluorine-containing compound in the medium or culture.

The composition may be to reduce the concentration of the fluorine-containing compound in the sample by contacting the sample. The contact may be made in a liquid or solid phase. The contact may be made, for example, by contacting the culture with a culture of microorganisms cultured in the medium. The culture may be performed under conditions for propagating microorganisms. The contact may be performed in a closed container. The contacting may include culturing or incubating the modified microorganism while contacting a sample containing a fluorine-containing compound. The contact includes culturing in a condition in which the transformed microorganism grows in a closed container.

Other aspects are SEQ ID NO: 1, 2, 3, 4, 5, 6, or a protein having a sequence identity of 85% or more with the amino acid sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, Or a modified microorganism comprising a genetic modification that increases the expression of a gene encoding a protein having a sequence identity of 85% or more with the amino acid sequence of 7, or a combination thereof, or a lysate thereof. A method for reducing the concentration of a fluorine-containing compound in a sample is provided, comprising contacting the compound-containing sample to reduce the concentration of the fluorine-containing compound in the sample.

In the above method, the sample containing the protein, the modified microorganism and the fluorine-containing compound is as described above.

In the above method, the contact may be made in a liquid or solid phase. The contact may be made, for example, by contacting the culture with a culture of microorganisms cultured in the medium. The culture can be performed under the condition that the microorganisms proliferate. The contact may be performed in a closed container. The contact may be made when the growth stage of the microorganism is an exponential phase or a stationary phase. The culture may be performed under aerobic or anaerobic conditions. The contact may be performed in a condition in which the microorganism transformed in the sealed container is viable. The viable condition may be a condition in which the modified microorganism is in a proliferative condition or a resting sate.

In the above method, the sample may be in a liquid or gaseous state. The sample can be factory wastewater or waste. The sample includes not only passive contact with the culture of the microorganism, but also active contact. The sample may be, for example, sparging in the culture medium of the microorganism. That is, the sample may be blown through a medium or a culture medium. The sparging may be blown from the bottom of the medium or culture medium to the top. The sparging may be injecting while dropping the sample.

In the above method, the contacting can be carried out batchwise or continuously. The contacting may include, for example, contacting the contacted sample obtained in the reducing step with a modified microorganism comprising a genetic modification that increases the activity of the fresh protein. The contact with the fresh microorganism may be made two or more times, for example, 2, 3, 5, or 10 times or more. The contact can be continued or repeated until a desired reduced concentration of the fluorine-containing compound in the sample is achieved.

Another aspect provides a method of producing a microorganism, comprising introducing a genetic modification to increase the activity of the protein in the microorganism. The genetic modification may be an increase in the number of copies of the gene encoding the protein or a modification of the regulatory region sequence of the gene encoding the protein. The method may be a method for producing a microorganism, comprising introducing a gene encoding the protein into the microorganism. The step of introducing a gene encoding the protein may be to introduce a vehicle containing the gene into the microorganism. In the above method, the genetic modification may include amplifying the gene, manipulating the regulatory sequence of the gene, or manipulating the sequence of the gene itself. The manipulation may be nucleotide insertion, substitution, conversion or addition.

Another aspect provides an isolated polypeptide having 85% sequence identity to the amino acid sequence of SEQ ID NOs: 1, 2, 3, 4, 5, 6, or 7.

Another aspect provides an isolated polynucleotide having at least 85% sequence identity to the nucleotide sequence of SEQ ID NOs: 8, 9, 10, 11, 12, 13, or 14.

Another aspect provides a vector comprising a polynucleotide encoding an isolated polypeptide having 85% sequence identity with the amino acid sequence of SEQ ID NOs: 1, 2, 3, 4, 5, 6, or 7. The vector can be a plasmid, or a virus-derived vector.

Another aspect provides a modified host cell comprising a vector comprising a polynucleotide encoding an isolated polypeptide having 85% sequence identity with the amino acid sequence of SEQ ID NOs: 1, 2, 3, 4, 5, 6, or 7 do. The host cell may be of Pseudomonas genus, Escherichia genus, Bacillus genus, or Xanthobacter genus. The host cell may be heterologous to Pseudomonas saitens.

Another aspect is culturing the host cell to produce a protein; And isolating the protein from the culture.

Modified microorganisms according to one aspect can be used to remove fluorine-containing compounds from a sample.

Compositions according to other aspects can be used to reduce the concentration of a fluorine-containing compound in a sample.

According to another aspect, a method of reducing the concentration of a fluorine-containing compound in a sample can effectively reduce the concentration of a fluorine-containing compound in a sample.

1 shows a vector map of pBBR122_selected gene.

Hereinafter, the present invention will be described in more detail through examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.

Example One: Pseudomonas saitens Modified microorganism expressing a derived protein gene and CF in a sample thereby ₄ remove

One. Pseudomonas from saitens Identification of inducible expression of protein genes

The cultured: (KCTC Korea Collection for Type Culture ) with Pseudomonas saitens entrusted to the accession number of KCTC 13107BP in conditions of CF ₄ present and CF ₄ present in September 2016 a dated international deposit under the Budapest Treaty agency Korea Bio Resource Center The gene by which expression is induced was confirmed. This strain is also called Pseudomonas saitens SF1 strain.

Inoculate P. saitens SF1 cells into 40 ml of LB medium in OD600 0.1 in a high-temperature sterilized and vertically arranged glass Dimroth screw tube reflux cooler (reactor length 350 mm, exterior diameter 35 mm, internal volume 200 mL) Then, it was incubated for 91 hours while being connected to a 30°C thermostat and maintained at 30°C. The cultivation was performed in a state where air was sealed by adding CF ₄ gas to 1,333 ppm in the space at the top of the medium of the incubator and closing the lid. After adding CF ₄ gas, the liquid phase was circulated. After the specified time, that is, after 91 hours, the CF ₄ gas content in the space above the medium was confirmed by GC-MS. The control group did not supply CF ₄ gas inside the space at the top of the medium. As a result, the concentration of CF _{4 in} the space at the top of the medium was reduced by 30.7%.

Next, as an experimental group, mRNA expressed from cultured P.saitens SF1 and a control experiment that would not inject CF ₄ gas were separated from the mRNA and subjected to transcriptomic analysis.

As a result, a gene in which the expression level of mRNA was increased by about 2 times was selected compared to when CF ₄ was injected. As a result, in the presence of CF ₄ , 7 genes whose expression is more than doubled, namely, Gene_01585, Gene_04413, Gene_00569, Gene_00598, Gene_04421, cspD, and yttP_1 were confirmed. The Gene_01585, Gene_04413, Gene_00569, Gene_00598, Gene_04421, cspD, and yttP_1 proteins have amino acid sequences of SEQ ID NOs: 1, 2, 3, 4, 5, 6, and 7, respectively. Genes encoding the Gene_01585, Gene_04413, Gene_00569, Gene_00598, Gene_04421, cspD, and yttP_1 proteins have nucleotide sequences of SEQ ID NOs: 8, 9, 10, 11, 12, 13, and 14, respectively.

2. Pseudomonas from saitens Expression of 7 genes Increased CF among microorganisms and samples using the same ₄ Removal of

(1) Production of microorganisms each containing 7 genes

After culturing P.saitens SF1 in LB medium at 30°C at 230 rpm overnight, the genomic DNA was separated by a method using a total DNA extraction kit (Invitrogen Biotechnology), and this genomic DNA was used as a template. And SEQ ID NOs: 15 and 16; SEQ ID NOs: 17 and 18; SEQ ID NOs: 19 and 20; SEQ ID NOs: 21 and 22; SEQ ID NOs: 23 and 24; SEQ ID NOs: 25 and 26; And PCR was performed using the nucleotide sequences of SEQ ID NOs: 27 and 28 as a primer set to amplify the genes encoding 7 genes, namely Gene_01585, Gene_04413, Gene_00569, Gene_00598, Gene_04421, cspD, and yttP_1 proteins.

The amplified gene was linked to the pBBR122 vector (MoBiTec GmbH, Germany) fragments amplified by PCR using SEQ ID NOs: 6 and 7, respectively, through an InFusion Cloning Kit (Clontech Laboratories, Inc.). The pBBR122 vector was developed by Dr. Camille of the Pasteur Institute in France and is commercially available. The pBBR122 vector has a broad bacterial host range.

The base vector pBBR122 is constructed to be resistant to chloramphenicol and kanamycin antibiotics, and pBBR122_selected where expression is induced by the original promoter by introducing 7 genes together with the original promoter at the chloramphenicol locus. gene.

1 shows a vector map of pBBR122_selected gene.

This vector is introduced through electroporation into a fluorine-containing hydrocarbon, that is, a CF ₄ spontaneous microorganism, Pseudomonas saitens (SF1) strain, and cultured in LB plate medium containing 50 μg/mL of kanamicin to enhance kanamicin resistance. Visible strains were selected. Modified Pseudomonas thus prepared The saitens (SF1) strains were named SF1_pBBR122 and pBBR122_selected genes, respectively. SF1_pBBR122 is a control strain into which an empty PBBR122 vector is introduced.

(2) CF using modified microorganisms ₄ remove

The three strains obtained in section (1) were subjected to primary evaluation in a 50 mL flask. The strain was inoculated in a 50 mL flask in 10 mL in the form of LB medium (OD ₆₀₀ 3.0) and 220 ppm CF ₄ was injected, followed by shaking culture at 30°C and 230 rpm. After about 3 days, the reduced CF ₄ amount was confirmed by GC-MS (Gas Chromatography Mass-Spectrum) to calculate the removal rate. The removal rate was calculated from Equation 1 below and the results are shown in Table 1.

<Equation 1>

CF ₄ removal rate = [(Initial CF ₄ content-CF ₄ content after reaction) / Initial CF ₄ content] × 100

Strain CF4 removal rate (%) 0h 72h SF1_pBBR122 0 0 SF1_pBBR122_01585 0 4.06 SF1_pBBR122_04413 0 4.36 SF1_pBBR122_00569 0 4.59 SF1_pBBR122_00598 0 3.95 SF1_pBBR122_04421 0 9.59 SF1_pBBR122_cspD 0 3.44 SF1_pBBR122_yttP_1 0 7.96

As shown in Table 1, when performing the flask experiment, the experimental group significantly decreased the concentration of CF ₄ by 3.4 to 9.6 (w/v)% higher than the control group.

SEQUENCE LISTING <110> Samsung Electronics Co., Ltd. <120> Modified microorganism including genetic modification that increases a protein gene expression and method for reducing concentration of fluorine containing compounds in sample <130> PN124109KR <160> 28 <170> PatentIn version 3.5 <210> 1 <211> 206 <212> PRT <213> Pseudomonas saitens SF1 <400> 1 Met Asn Leu Glu Gln Ser Leu Ala Ile Leu His Glu His Phe Asp Lys 1 5 10 15 Val Phe Arg Asp Ala Thr Leu Ala Ser Ser Ile Ser His Ala Glu Val 20 25 30 Arg Arg Ile Ile Cys Leu Ile Ile Asp Gln Gln Asn Ser Ala Pro Ala 35 40 45 Glu Asp Arg Leu Leu Ser His Tyr Tyr Gln Phe Ile Phe Ala Thr Glu 50 55 60 Thr Leu His Ala Ser Tyr Arg Ile Tyr Glu Leu Thr Asp Met Gly Ser 65 70 75 80 Trp Ile Gly Trp Ala Leu Ser Glu Asp Thr Arg Asp Ser His Ser Lys 85 90 95 Asn Leu His Leu Gly Arg Ile Phe Asp Phe Tyr Ser Ser Ala Val Val 100 105 110 Arg Thr Trp Pro Gly Phe Val Pro Val Met Val Lys Phe Phe Ser Ala 115 120 125 Phe Tyr Tyr Tyr Ala Arg Glu Arg Thr Ala Met Asn Asn Ile Ala Arg 130 135 140 Glu Leu Trp Pro Phe Ala Ala Ser Thr Phe Thr Asp Thr Met Asn Leu 145 150 155 160 Pro Ala Glu Tyr Ile Tyr Ile Asp Glu Ala Asn Leu Gly Ser Gln Met 165 170 175 Ala Cys Trp Ala Ala Lys Glu Ala Pro Asp Leu Ala Lys Thr Phe Val 180 185 190 Pro Tyr Leu Glu Ser Val Ala Gly Lys Asn Thr Leu Pro Asp 195 200 205 <210> 2 <211> 57 <212> PRT <213> Pseudomonas saitens SF1 <400> 2 Met Val Val Arg Leu Ala Val Leu Ala Met Leu Ser Arg Pro Asp Val 1 5 10 15 Ser Asp Leu Ala Val Val Ser Ile Gly Ile Gly Leu Ala Ser Tyr Val 20 25 30 Ala Val Met Ser Asp Leu Gly Phe Lys Lys Leu Asp Glu Thr Leu Asn 35 40 45 Asp Leu Val Arg Val Arg Val Val Arg 50 55 <210> 3 <211> 88 <212> PRT <213> Pseudomonas saitens SF1 <400> 3 Met Asn Thr Phe Ala Val Ala Leu Met Leu Ala Ala Ser Leu Gly Leu 1 5 10 15 Ala Ala Cys Asp Lys Lys Ser Glu Asp Lys Ala Gln Asp Ala Ala Gln 20 25 30 His Ser Glu Gln Ala Gln Glu Lys Met Ser Glu Ala Gln Asp Lys Val 35 40 45 Asn Asp Ala Ala Lys Glu Asn Ala Glu Ala Ala Lys Ala Gln Ala Glu 50 55 60 Ser Asn Lys Ala Ala Ala Glu Glu Ala Gly Gln Thr Ala Pro Val Ala 65 70 75 80 Pro Ala Ala Thr Pro Glu Ser Lys 85 <210> 4 <211> 60 <212> PRT <213> Pseudomonas saitens SF1 <400> 4 Met Phe Glu Thr Thr Ala Phe His Ser Met Met Ile Pro Ile Ile Ser 1 5 10 15 Gly Met Leu Leu Leu Thr Ile Gly Phe Asn Phe Arg Glu Arg Ser Ser 20 25 30 Gly Ile Gly Val Leu Met Ile Trp Ala Gly Met Leu Leu Ile Phe Gly 35 40 45 Thr Ile Val Phe Lys Ile Leu Ala Lys Leu Asn Glu 50 55 60 <210> 5 <211> 290 <212> PRT <213> Pseudomonas saitens SF1 <400> 5 Met Tyr Ala Phe Ile Lys Gln Arg Ala Gly Asp Tyr Ser Ile Arg Arg 1 5 10 15 Leu Cys Leu Thr Leu Lys Val His Pro Ser Gly Tyr Tyr Ala Trp Leu 20 25 30 Ser Glu Pro Gln Ser Ala Arg Ala Lys Asp Asp Gln Arg Leu Leu Gly 35 40 45 Leu Ile Lys His Ser Trp Leu Glu Ser Gly Gly Val Tyr Gly Tyr Arg 50 55 60 Lys Ile His Asp Asp Leu Arg Glu Val Gly Glu Asp Cys Gly Arg Asn 65 70 75 80 Arg Val Ala Arg Leu Met Arg Leu Glu Gly Leu Arg Ser Gln Thr Gly 85 90 95 Tyr Arg Arg Arg Pro Gly Lys Tyr Gly Gly Lys Pro Ala Val Ala Ser 100 105 110 Pro Asn Leu Leu Lys Arg Gln Phe Asp Val Ala Glu Pro Asn Lys Val 115 120 125 Trp Val Thr Asp Ile Thr Tyr Ile Arg Thr Tyr Glu Gly Trp Leu Tyr 130 135 140 Leu Ala Val Val Leu Asp Leu Phe Ser Arg Gln Phe Val Gly Trp Ser 145 150 155 160 Met Lys Ser Gln Met Thr Ser Asp Leu Ala Ile Asp Ala Leu Leu Met 165 170 175 Ala Val Trp Arg Arg Lys Pro Lys Gln Glu Val Met Val His Ser Asp 180 185 190 Gln Gly Ser Gln Tyr Ser Ser Ser Asp Trp Arg Ser Phe Leu Lys Ala 195 200 205 Asn Asn Leu Val Ala Ser Met Ser Arg Arg Gly Asn Cys His Asp Asn 210 215 220 Ala Val Ala Glu Ser Phe Phe Gln Leu Leu Lys Arg Glu Arg Ile Lys 225 230 235 240 Arg Lys Ile Tyr Asn Thr Arg Gln Asp Ala Arg Ser Asp Val Phe Asp 245 250 255 Tyr Ile Glu Met Phe Tyr Asn Ala Lys Arg Arg His Gly Phe Asn Asn 260 265 270 Arg Leu Ser Pro Val Glu Phe Glu Lys Arg Tyr Ala Met Ser Leu Gln 275 280 285 Gly Val 290 <210> 6 <211> 89 <212> PRT <213> Pseudomonas saitens SF1 <400> 6 Met Val Lys Trp Phe Asn Asn Ala Lys Gly Phe Gly Phe Ile Asn Thr 1 5 10 15 Gln Ala Lys Glu Gly Ile Asp Glu His Gly Asn Pro Ile Asp Phe Phe 20 25 30 Ala His Phe Ser Ala Ile Gln Met Asp Gly Tyr Lys Thr Leu Lys Ala 35 40 45 Gly Gln Pro Val Ser Phe Glu Ile Ile Gln Gly Pro Lys Gly Leu His 50 55 60 Ala Val Ala Ile Thr Asn Ala Gln Val Pro Ala Ser Ala Gln Ala Pro 65 70 75 80 Ala Gln Glu Val Thr Ser Leu Ser Val 85 <210> 7 <211> 234 <212> PRT <213> Pseudomonas saitens SF1 <400> 7 Met Gln Ser Glu Thr Val Glu Arg Ile Leu Asp Ala Ala Glu Gln Leu 1 5 10 15 Phe Ala Glu Lys Gly Phe Ala Glu Thr Ser Leu Arg Leu Ile Thr Ser 20 25 30 Lys Ala Gly Val Asn Leu Ala Ala Val Asn Tyr His Phe Gly Ser Lys 35 40 45 Lys Ala Leu Ile Gln Ala Val Phe Ser Arg Phe Leu Gly Pro Phe Cys 50 55 60 Ala Asn Leu Asp Arg Glu Leu Glu Arg Arg Gln Gly Lys Pro Glu Asn 65 70 75 80 Arg Pro Thr Leu Glu Glu Leu Leu Glu Met Leu Val Glu Gln Ala Leu 85 90 95 Val Ile Gln Pro Arg Ser Gly Asn Asp Leu Ser Ile Phe Met Arg Leu 100 105 110 Leu Gly Leu Ala Phe Ser Gln Ser Gln Gly His Leu Arg Arg Tyr Leu 115 120 125 Glu Asp Met Tyr Gly Lys Val Phe Arg Arg Tyr Met Leu Leu Val Asn 130 135 140 Glu Ala Ala Pro Gln Ile Pro Pro Ile Glu Leu Phe Trp Arg Val His 145 150 155 160 Phe Met Leu Gly Ala Ala Ala Phe Ser Met Ser Gly Ile Lys Ala Leu 165 170 175 Arg Ala Ile Ala Glu Thr Asp Phe Gly Val Asn Thr Ser Ile Glu Gln 180 185 190 Val Met Arg Leu Met Val Pro Phe Leu Ala Ala Gly Met Arg Ala Glu 195 200 205 Ser Gly Val Thr Asp Pro Ala Met Ala Ser Ala Gln Leu Arg Pro Arg 210 215 220 Ser Lys Thr Ala Pro Val Ser Ala Lys Val 225 230 <210> 8 <211> 921 <212> DNA <213> Pseudomonas saitens SF1 <400> 8 tcctgccgat tgcaggtagg attgacagcg gccaaaacat ccacagccag tttaggcgct 60 cgacattgac agtgcccata gagaggagca gtccgactga ggccaacgga tacataaatt 120 gatcaatgca gactagaagg aagcaatgta ttgaacctcg aacaaagcct agcgattctg 180 catgagcact ttgacaaagt tttccgtgac gccactttag cttcatccat ctcgcatgca 240 gaagtccgac gcataatctg ccttataatt gatcagcaaa atagtgcccc tgccgaagac 300 agactattgt cccactatta ccagttcatt tttgctaccg aaacacttca cgcgtcatac 360 cgtatatatg aactaactga tatggggagc tggattggat gggcgttatc agaagacacc 420 agagacagtc actcaaaaaa tttacatctg gggagaatct ttgacttcta ttcatcagcg 480 gtggtaagaa cctggccagg gtttgtgccg gtgatggtga aatttttcag cgccttctac 540 tactacgctc gcgagcgaac agccatgaac aatatagcac gcgagctatg gcccttcgcc 600 gcatcgactt ttacagacac gatgaatctg cctgcggaat acatttacat tgacgaagca 660 aatcttggaa gtcaaatggc atgctgggca gcaaaagaag cgcctgactt ggccaagact 720 ttcgttccat acctagagtc tgtcgctggc aaaaatactc taccagatta agtccgtgcc 780 gttcattatt acaccttgtc tactacagca agtgcgttct caagcaaagc atcgcatgag 840 tgggcagcaa aggcactcgc cgacttcaat gaaaacctcg ctgagccgca taagctgcaa 900 ctgctagcca ctatctatgc a 921 <210> 9 <211> 474 <212> DNA <213> Pseudomonas saitens SF1 <400> 9 cggcgggatc tcgaccaagc cgctgacagc ccctactgat aggcccaagg cgctggccca 60 tgcctccacc agacggttga acatattgtc cctgccgggc tcaaacttac ccagcgcgtt 120 gaaggcatcg tagttcaaca ggatagccag atggtggtgc ggcttgcagt gctggccatg 180 ctctcgcgcc cagacgtatc ggaccttgct gtcgtgagta tagggattgg attggcgagc 240 tatgttgcgg ttatgtcgga tttaggcttt aaaaaactcg atgaaacgct caatgacctg 300 gttcgtgtac gtgtagtcag gtagctgaac tcccacaggc atccggtata ccgcgcacag 360 gattgcatcg gcgcagcggt caatggtgag tgtcatggca gcaccggcgc agcgcaaccc 420 caagcggcct gctatggccg gatgctcaac ggccaatgca ctgggccaca gttc 474 <210> 10 <211> 570 <212> DNA <213> Pseudomonas saitens SF1 <400> 10 gcaaatttca aggcctgatg gagaagctca agtttttacc agtagtttca gcccatgctg 60 ctttacttat atatgtgttc gagcatggaa cttaagcgat cagcccatgc tcaaacgccc 120 cgtagatcta ctcacaagga gaaacacccc atgcgtaata cttttgctgt tgccttgatg 180 ctcgctgctt cgcttggcct ggcggcctgc gataaaaaat ccgaagacaa agcccaggat 240 gctgctcagc actctgagca agctcaagaa aaaatgtctg aagcacaaga caaagtgaac 300 gacgctgcca aagaaaacgc cgaagctgca aaagcacaag ccgagtccaa taaagccgct 360 gctgaagaag ccggccagac tgccccggtt gcacctgcgg caactccaga aagcaagtaa 420 caccactttc tgcgtaataa aaccaagccg ccaagtgcgg gctttgttgt atctgaacgt 480 taatggcgac tgatggcatg tataaataca cggccattaa aaaagccctg catccattaa 540 gatgcagggc ttttttattg actcggcaac 570 <210> 11 <211> 483 <212> DNA <213> Pseudomonas saitens SF1 <400> 11 gaaaaagcag ggtcgctgcg caacccgtcg cagcctgcgg cagcggctac aaggttctgc 60 gtctggctga ttcatttttg atacacatca ccacacatct gcgcaaaccg acctagactt 120 ggggcattca tttgttcagg tccccttacc atgttcgaaa ccactgcatt ccactccatg 180 atgatcccca tcatcagcgg catgttgctg ctgaccattg gcttcaactt tcgcgaacgc 240 agtagcggca ttggcgtgct gatgatctgg gccggcatgc tgctgatttt cggcaccatc 300 gtgttcaaga tcctcgccaa gctcaacgaa tagacccgac gctgttccgc ccaggggccg 360 cgatcattta tttatgtgac aagaaatgaa tgatcactgg ccctagcgta cactcggcca 420 attcaccccc tcgaggttgg tcgcccagtg ttttcccgtc ttttcgcctt gccctgttac 480 ctg 483 <210> 12 <211> 1176 <212> DNA <213> Pseudomonas saitens SF1 <400> 12 gataaagcgc tacagcaaac ctcaagaaga acggcagcag gacgatgatc agcacgctga 60 actacgtcgt ctgagagccg agctaaagcg cgtcactgaa gaacgagaca tcttaaaaaa 120 ggccgccgcg tactttgcca aggagtgcgg gtgaagtacg cctttatcaa gcagcgagcc 180 ggcgactatt ccattcgacg gctttgcctg acgctgaaag tccatcccag tggctattac 240 gcttggttgt ctgagccgca atctgcacgc gccaaagacg accaacgatt gctgggtttg 300 atcaagcatt cgtggctgga gagcggcggc gtttatggct atcgcaaaat ccatgacgac 360 ctgcgcgagg tcggtgagga ttgtggtcgg aatcgtgtgg caaggctgat gcgtcttgaa 420 ggtctgcgct ctcagacagg gtatcgacgc cgccctggca agtacggcgg taagccagcg 480 gtcgcctctc ccaatttgct gaagcgccag ttcgatgtcg ctgaaccaaa caaagtttgg 540 gtcaccgaca tcacctatat tcgcacctat gaaggctggc tgtatttggc ggtggtgctg 600 gatctgtttt ctcgtcagtt tgttggctgg tcaatgaagt cgcagatgac cagtgatttg 660 gccattgatg cgctattgat ggcggtttgg aggcgaaaac cgaaacagga ggtgatggtt 720 cactcagacc agggcagcca gtacagcagc tccgattggc gcagcttttt gaaggcgaac 780 aatttggttg ccagcatgag ccgtcgaggt aactgtcatg acaacgccgt agccgagagc 840 tttttccagc ttctgaagcg ggaacggatt aagcgaaaaa tctacaacac acggcaagat 900 gctcgcagcg atgtgttcga ttacatcgag atgttctaca acgcaaaacg ccgccatggt 960 ttcaacaatc ggctgtcacc ggtagagttt gaaaagcgtt acgcaatgag cttgcaaggt 1020 gtctagagaa tccggggcga ttcacagctc taacccgacg acctacacat cttggacttt 1080 ttcaaggtgg tgagaagccc actcgtcagc ggcgaatgtg gatttttcgc tgagttaacg 1140 tactgggaaa agtccccgta tcgcctctaa atgcat 1176 <210> 13 <211> 579 <212> DNA <213> Pseudomonas saitens SF1 <400> 13 accttgacta tcggtaaaac ggtgttacaa ccatatagta cccacagtgg gaaaaatggt 60 ccgcacagtc aaccatattt acgggtttct gtgcgagttg actggataat actccagtga 120 tggagtggtt tgcagcagag ggatatgagt atgcaaggca aggtcaagtg gttcaacaac 180 gccaaaggct tcggctttat taatacgcag gccaaggaag gcatcgatga acacggaaac 240 ccgatcgact tttttgcgca tttttcagca atccagatgg acggttacaa gacgctcaag 300 gcaggccagc ctgtgagctt cgaaataatc cagggcccca agggcctgca tgcggttgcc 360 atcaccaacg ctcaagttcc agccagtgct caagcgcctg cacaagaggt gaccagcctc 420 tcggtctgac ccgccgcaga agaaaccggc cgattcaatc acgcgaatcg gccggttttt 480 ttatgcccgc gccctgcacg gggaaaatgc agattgccga aataccgcgt aaacactggg 540 cgaatcagca aaaatcaggg cattttgccg cagggactt 579 <210> 14 <211> 1008 <212> DNA <213> Pseudomonas saitens SF1 <400> 14 cccgcacttt atcaggtggg gcctggctgg ccgaggctgt ggttaaatgc gaaaccgccc 60 ggtcgcaaaa cgaatacgtc atcttgactt ttcacaagct gaaacgtatg tttcaaacaa 120 gtgtttaaat gtttgtcagg cggagtagtc atggcccagt cggaaaccgt tgaacgtatc 180 cttgatgctg ccgagcagct gttcgcggaa aaagggtttg ctgaaacctc attgcggctg 240 attaccagca aggccggggt caatctggcg gcggtcaatt accattttgg ttcaaagaaa 300 gcgctgatcc aggcggtgtt ctcacgcttc ctggggcctt tctgtgccaa cctggaccgt 360 gagctggagc gccgtcaggg caagccggag aatcgtccga cgcttgagga gttgctggag 420 atgctggtcg agcaggcgct ggtgatccag ccacgcagcg gcaacgacct gtcgatcttc 480 atgcgtttgc tggggctggc cttcagccag agccagggcc acttgcgccg ctaccttgaa 540 gacatgtacg gcaaggtgtt ccgtcgctac atgttgctgg tcaacgaagc agccccgcag 600 atcccgccca tcgagctgtt ctggcgcgtg catttcatgc tcggcgccgc ggccttcagc 660 atgtccggga tcaaggcgtt gcgtgccatt gccgagaccg attttggcgt caacacctcg 720 atcgagcagg tgatgcgcct gatggtgccg ttcctcgcgg ccggcatgcg tgccgagagc 780 ggtgtcacgg atccggcaat ggccagtgcc cagcttcgcc cgcgcagtaa aaccgcaccg 840 gtcagcgcca aggtttgacc tgctgcgggt gggcgcggca gctttgttcc gctaagctag 900 ctgccatgcc atgcccagcc ctgtcttgtt cccgtctgtt gattcgtcgc tgcctctctg 960 gagtgcggcg cgcaggcccg tgcccgcgca tccggaccgg gcaacaag 1008 <210> 15 <211> 37 <212> DNA <213> Artificial <220> <223> primer <400> 15 actgccttaa aaaaatcctg ccgattgcag gtaggat 37 <210> 16 <211> 36 <212> DNA <213> Artificial <220> <223> primer <400> 16 gctaaggaag ctaaatgcat agatagtggc tagcag 36 <210> 17 <211> 38 <212> DNA <213> Artificial <220> <223> primer <400> 17 actgccttaa aaaaacggcg ggatctcgac caagccgc 38 <210> 18 <211> 39 <212> DNA <213> Artificial <220> <223> primer <400> 18 gctaaggaag ctaaagaact gtggcccagt gcattggcc 39 <210> 19 <211> 37 <212> DNA <213> Artificial <220> <223> primer <400> 19 actgccttaa aaaaagcaaa tttcaaggcc tgatgga 37 <210> 20 <211> 37 <212> DNA <213> Artificial <220> <223> primer <400> 20 gctaaggaag ctaaagttgc cgagtcaata aaaaagc 37 <210> 21 <211> 36 <212> DNA <213> Artificial <220> <223> primer <400> 21 actgccttaa aaaaagaaaa agcagggtcg ctgcgc 36 <210> 22 <211> 37 <212> DNA <213> Artificial <220> <223> primer <400> 22 gctaaggaag ctaaacaggt aacagggcaa ggcgaaa 37 <210> 23 <211> 38 <212> DNA <213> Artificial <220> <223> primer <400> 23 actgccttaa aaaaagataa agcgctacag caaacctc 38 <210> 24 <211> 38 <212> DNA <213> Artificial <220> <223> primer <400> 24 gctaaggaag ctaaaatgca tttagaggcg atacgggg 38 <210> 25 <211> 38 <212> DNA <213> Artificial <220> <223> primer <400> 25 actgccttaa aaaaaacctt gactatcggt aaaacggt 38 <210> 26 <211> 38 <212> DNA <213> Artificial <220> <223> primer <400> 26 gctaaggaag ctaaaaagtc cctgcggcaa aatgccct 38 <210> 27 <211> 38 <212> DNA <213> Artificial <220> <223> primer <400> 27 actgccttaa aaaaacccgc actttatcag gtggggcc 38 <210> 28 <211> 37 <212> DNA <213> Artificial <220> <223> primer <400> 28 gctaaggaag ctaaacttgt tgcccggtcc ggatgcg 37

Claims (20)

Genetic modification to increase the expression of a gene encoding a protein having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, or 7, or a combination thereof Transformed microorganism. The microorganism according to claim 1, wherein the genetic modification is a genetic modification that increases the number of copies of the gene. The microorganism according to claim 1, wherein the protein is derived from Bacillus, Pseudomonas, Azotobacter, Agrobacterium, or Escherichia. The method according to claim 1, wherein the gene has a sequence identity of at least 85% with the nucleotide sequence of SEQ ID NO: 8, 9, 10, 11, 12, 13, or 14. The microorganism according to claim 1, which belongs to the genus Pseudomonas, the genus Escherichia, the genus Bacillus, or the genus Xanthobacterium. Protein having at least 85% sequence identity to the amino acid sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, or 7, or a combination thereof, or SEQ ID NO: 1, 2, 3, 4, 5, 6, or 7 A fluorine-containing compound in a sample comprising a modified microorganism comprising genetic modification that increases the expression of a protein having a sequence identity of 85% or more with a sequence identity of 85% or more, or a lysate thereof As a composition for use in reducing the concentration of, the fluorine-containing compound is a composition represented by any one of the following formulas 1 to 3:
<Formula 1><Formula2>
C(R ₁ )(R ₂ )(R ₃ )(R ₄ ) (R ₅ )(R ₆ )(R ₇ )C-[C(R ₁₁ )(R ₁₂ )] _n -C(R ₈ )( R ₉ )(R ₁₀ )
<Formula 3>
S(R ₁₃ )(R ₁₄ )(R ₁₅ )(R ₁₆ ) (R ₁₇ )(R ₁₈ )
In the above equations,
n is an integer from 0 to 10, and when n is 2 to 10, R11 among the plurality of R11 may have each other or be different, and R12 among the plurality of R12 may have each other or different,
R ₁ , R ₂ , R ₃ and R ₄ are independently of each other F, Cl, Br, I, or H, and at least one of R ₁ , R ₂ , R ₃ and R ₄ is F,
R ₅ , R ₆ , R ₇ , R ₈ , R ₉ , R ₁₀ , R ₁₁ and R ₁₂ are independently F, Cl, Br, I, or H, and R ₅ , R ₆ , R ₇ , R ₈ , R ₉ , R ₁₀ , one or more of R ₁₁ and R ₁₂ is F,
R ₁₃ , R ₁₄ , R ₁₅ , R ₁₆ , R ₁₇ and R ₁₈ are each independently F, Cl, Br, I, or H, and R ₁₃ , R ₁₄ , R ₁₅ , R ₁₆ , R ₁₇ and R ₁₈ One or more of them are F. The composition of claim 6, wherein the genetic modification is a genetic modification that increases the number of copies of the gene. The composition according to claim 6, wherein the protein is derived from the genus Bacillus, Pseudomonas, Azotobacter, Agrobacterium, or Escherichia. The method according to claim 6, wherein the gene has a sequence identity of at least 85% with the nucleotide sequence of SEQ ID NO: 8, 9, 10, 11, 12, 13, or 14. The method according to claim 6, wherein the microorganism is Pseudomonas genus, Escherichia genus, Bacillus genus, or composition that belongs to the genus Xanthobacterium. The composition according to claim 6, wherein the sample is in a liquid or gaseous state. Protein having a sequence identity of 85% or more with the amino acid sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, or 7, or a combination thereof, of SEQ ID NO: 1, 2, 3, 4, 5, 6, or 7 A sample containing a fluorine-containing compound containing a modified microorganism comprising genetic modification that increases the expression of a gene encoding a protein having a sequence identity of 85% or more with an amino acid sequence, or a combination thereof, or a lysate thereof. A method of reducing the concentration of a fluorine-containing compound in a sample, comprising contacting with to decrease the concentration of a fluorine-containing compound in a sample,
The fluorine-containing compound is a method represented by any one of the following formulas 1 to 3:
<Formula 1><Formula2>
C(R ₁ )(R ₂ )(R ₃ )(R ₄ ) (R ₅ )(R ₆ )(R ₇ )C-[C(R ₁₁ )(R ₁₂ )] _n -C(R ₈ )( R ₉ )(R ₁₀ )
<Formula 3>
S(R ₁₃ )(R ₁₄ )(R ₁₅ )(R ₁₆ ) (R ₁₇ )(R ₁₈ )
In the above equations,
n is an integer from 0 to 10, and when n is 2 to 10, R11 among the plurality of R11 may have each other or be different, and R12 among the plurality of R12 may have each other or different,
R ₁ , R ₂ , R ₃ and R ₄ are independently of each other F, Cl, Br, I, or H, and at least one of R ₁ , R ₂ , R ₃ and R ₄ is F,
R ₅ , R ₆ , R ₇ , R ₈ , R ₉ , R ₁₀ , R ₁₁ and R ₁₂ are independently F, Cl, Br, I, or H, and R ₅ , R ₆ , R ₇ , R ₈ , R ₉ , R ₁₀ , one or more of R ₁₁ and R ₁₂ is F,
R ₁₃ , R ₁₄ , R ₁₅ , R ₁₆ , R ₁₇ and R ₁₈ are each independently F, Cl, Br, I, or H, and R ₁₃ , R ₁₄ , R ₁₅ , R ₁₆ , R ₁₇ and R ₁₈ One or more of them are F. The method of claim 12, wherein the genetic modification is a genetic modification that increases the number of copies of the gene. The method according to claim 12, wherein the protein is derived from the genus Bacillus, Pseudomonas, Azotobacter, Agrobacterium, or Escherichia. The method according to claim 12, wherein the gene has a sequence identity of at least 85% with the nucleotide sequence of SEQ ID NO: 8, 9, 10, 11, 12, 13, or 14. The method according to claim 12, wherein the microorganism belongs to the genus Pseudomonas, Escherichia, Bacillus, or Xanthobacter. The method of claim 12, wherein the sample is in liquid or gaseous state. The method of claim 12, wherein the contacting is performed in an air-tight container. The method of claim 12, wherein the contacting comprises culturing the modified microorganism while contacting the sample containing the fluorine-containing compound. The method of claim 12, wherein the contacting comprises culturing in a condition in which the transformed microorganism grows in an air-tight container.

Images