Antibacterial proteins having potent lysolytic activity against staphylococcus including staphylococcus aureus

Abstract

The present invention relates to an antibacterial protein S2200 having an antibacterial activity specific to Staphylococcus aureus, including Staphylococcus aureus, and a method for preparing the antibacterial protein S2200, the antibacterial protein S2200 having an antibacterial activity specific to Staphylococcus aureus, the antibacterial protein S2200 having an antibacterial activity specific to Staphylococcus aureus, and the antibacterial protein S2200 having an antibacterial activity specific to Staphylococcus aureus. More particularly, the present invention relates to: an antibacterial protein S2200 specific to staphylococcus aureus including staphylococcus aureus, which is characterized by having an ability to specifically lyse the staphylococcus aureus including staphylococcus aureus and having an amino acid sequence represented by SEQ ID NO: 1; and a pair comprising the antibacterial protein S2200 as an active ingredient, said antibacterial protein S2200 being specific to staphylococcus aureus including staphylococcus aureus. The present invention relates to a pharmaceutical composition for treating infections induced by staphylococcus, including Staphylococcus aureus, of an antibacterial protein S2200 specific to staphylococcus, including Staphylococcus aureus.

Description

Technical Field

[0001] The present invention relates to an antimicrobial protein S2200 having bacteriolytic activity against staphylococci including Staphylococcus aureus, and more particularly to an antimicrobial protein S2200 specific for staphylococci characterized by having the ability to specifically lyse staphylococci that may cause infectious diseases and having an amino acid sequence represented by sequence number 1, and a pharmaceutical composition for treating infectious diseases induced by staphylococci comprising the antimicrobial protein S2200 specific for staphylococci as an active ingredient. Background Art

[0002] Staphylococcus aureus is a facultatively anaerobic, Gram-positive coccus that was first discovered by German scientist Robert Koch in 1878 in specimens of purulent inflammation and named Staphylococcus by Alexander Ogston of Scotland in 1881. It is the most common pathogen of Staphylococcus infections.

[0003] Staphylococcus aureus is one of the pathogens that cause human infections in hospitals and communities, and is the cause of fatal diseases such as pneumonia, endocarditis, and sepsis. In particular, infectious diseases mediated by methicillin-resistant Staphylococcus aureus (MRSA) (accounting for more than 50% of Staphylococcus aureus infections and with a mortality rate of approximately 15%) pose a serious threat to human health.

[0004] Antibiotics are widely used in the treatment of Staphylococcus aureus infections as described above, but recently, with the increasing resistance of Staphylococcus aureus to antibiotics, the therapeutic effect of antibiotics has been significantly reduced. Therefore, it is necessary to develop new antibacterial substances that are also very effective in treating infections caused by Staphylococcus aureus that are resistant to existing antibiotics. In particular, it is urgently necessary to develop pharmaceutical preparations with a new mechanism of action that can quickly exert antibacterial power. Summary of the invention

[0005] Therefore, as a solution to the problems caused by the use of existing antibiotics against harmful pathogenic bacteria, namely, Staphylococci including Staphylococcus aureus, the present inventors intend to provide an antibacterial protein that can specifically lyse Staphylococci including Staphylococcus aureus, and to provide a pharmaceutical composition comprising the antibacterial protein as an effective ingredient and which can be used to treat Staphylococcal infections, and further to provide a method for effectively treating Staphylococcal infections using the pharmaceutical composition.

[0006] Therefore, an object of the present invention is to provide an antimicrobial protein S2200 having an amino acid sequence represented by SEQ ID NO: 1 and having antimicrobial activity capable of specifically lysing Staphylococci including Staphylococcus aureus.

[0007] Another object of the present invention is to provide a method for effectively producing the antimicrobial protein S2200 having antimicrobial activity that can specifically lyse staphylococci including Staphylococcus aureus and comprising an amino acid sequence represented by sequence number 1. In this regard, Escherichia coli Endo-22 is provided as a production strain of the antimicrobial protein S2200. The production strain, i.e., Escherichia coli Endo-22, was deposited in the Bioresource Center of the Korea Institute of Biotechnology on July 12, 2022 (deposit number KCTC15031BP).

[0008] Another object of the present invention is to provide a pharmaceutical composition for the purpose of treating staphylococcal infection, comprising as an active ingredient the antimicrobial protein S2200 that can specifically lyse staphylococci including Staphylococcus aureus.

[0009] In addition, another object of the present invention is to provide a method for treating staphylococcal infection including Staphylococcus aureus using a pharmaceutical composition comprising the antimicrobial protein S2200 as an active ingredient.

[0010] In order to achieve the above-mentioned objectives, the inventors of the present invention utilized the information of various antimicrobial proteins known to have antimicrobial activity against various bacterial species to maximize their own knowledge and professional capabilities and manufactured various antimicrobial protein candidates into recombinant protein forms, and screened out antimicrobial proteins with excellent lytic activity by detecting their lytic activity against staphylococci including Staphylococcus aureus, and further developed a method for effectively manufacturing the antimicrobial proteins, and finally developed a pharmaceutical composition containing the antimicrobial proteins as active ingredients that can be used for the purpose of treating staphylococcal infections including Staphylococcus aureus, thereby completing the present invention.

[0011] Therefore, in one aspect of the present invention, the present invention provides an amino acid sequence of an antimicrobial protein S2200 that can specifically lyse Staphylococci including Staphylococcus aureus. Specifically, it is an amino acid sequence corresponding to SEQ ID NO: 1. The antimicrobial protein S2200 that can specifically lyse Staphylococci including Staphylococcus aureus is composed of 345 amino acid residues and has a molecular weight of approximately 38.0 kDa.

[0012] It is obvious that the amino acid sequence of sequence number 1 can be partially deformed by relevant practitioners using known techniques. The deformation includes partial substitution of the amino acid sequence, partial addition of the amino acid sequence, and partial deletion of the amino acid sequence. However, it is preferable to apply the amino acid sequence of sequence number 1 disclosed in the present invention.

[0013] In addition, the present invention provides a strain, namely Escherichia coli Endo-22, which can be used for producing the antimicrobial protein S2200 having the amino acid sequence of SEQ ID NO. 1. The strain, namely Escherichia coli Endo-22, is a strain that can produce S2200 by transforming Escherichia coli with a plasmid having the nucleic acid sequence (5,178 bp) of SEQ ID NO. 2 produced by the present inventors.

[0014] In another aspect of the present invention, the present invention provides a pharmaceutical composition that can be effectively used to treat staphylococcal infections including Staphylococcus aureus, comprising as an active ingredient an antimicrobial protein S2200 characterized by the amino acid sequence of SEQ ID NO: 1 and having specific lytic ability against staphylococci including Staphylococcus aureus.

[0015] The antimicrobial protein S2200 according to the present invention, which is characterized by the amino acid sequence of sequence number 1 and has a specific lytic ability against staphylococci including Staphylococcus aureus, contained in the pharmaceutical composition of the present invention, can specifically lyse staphylococci including Staphylococcus aureus as described above, and therefore has the effect of treating a variety of diseases induced by staphylococci including Staphylococcus aureus. Therefore, the pharmaceutical composition of the present invention can be used to treat diseases induced by staphylococci including Staphylococcus aureus. In addition, the pharmaceutical composition of the present invention can be implemented as an antibiotic, disinfectant, bactericide, and therapeutic agent, and is used to treat a variety of diseases induced by staphylococci including Staphylococcus aureus.

[0016] Therefore, in another aspect of the present invention, the present invention provides a method for treating various diseases induced by Staphylococci by administering an antimicrobial protein S2200 characterized by having an amino acid sequence represented by sequence number 1 and specific to Staphylococci including Staphylococcus aureus to a subject infected with Staphylococci including Staphylococcus aureus.

[0017] In the present specification, "diseases induced by Staphylococcus aureus" refers to diseases induced by Staphylococcus aureus infection, including skin diseases, bacteremia, sepsis, endocarditis, ulcers, etc., but is not limited thereto.

[0018] In the present specification, Staphylococci including Staphylococcus aureus may be sensitive to existing antibiotics or may be resistant to existing antibiotics. In other words, whether or not the existing antibiotics are resistant has no effect.

[0019] The term "treatment" or "treatment" as used in the present specification refers to all actions that can inhibit infection induced by Staphylococcus including Staphylococcus aureus and alleviate the pathological state of diseases induced by Staphylococcus including Staphylococcus aureus.

[0020] In order to improve the efficiency for the above-mentioned application purposes, antibacterial substances that can provide antibacterial activity against other bacterial species may be added to the pharmaceutical composition of the present invention.

[0021] The pharmaceutically acceptable carriers included in the pharmaceutical composition of the present invention are substances commonly used in formulation, including lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinyl pyrrolidone, cellulose, water, syrup, methylcellulose, methylparaben, propylparaben, talc, magnesium stearate and mineral oil, but are not limited thereto. In addition to the above ingredients, the pharmaceutical composition of the present invention may additionally include lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents and preservatives.

[0022] The pharmaceutical composition of the present invention can be administered orally or parenterally. In the case of parenterally, it can be administered by intravenous administration, intraperitoneal administration, intramuscular administration, subcutaneous administration or topical administration. In addition, it can also be used by coating or spraying on the diseased area, but is not limited to this.

[0023] The pharmaceutical composition of the present invention can be formulated using pharmaceutically acceptable carriers and / or excipients in a method that can be easily implemented by a person having general knowledge of the technical field to which the present invention belongs, so as to be manufactured into a unit dosage form or a form to be loaded into a multi-dose container. In this case, the dosage form can be a solution, suspension or emulsion in an oily or aqueous medium, or a paste, powder, granule, tablet or capsule, and can also contain a dispersant or stabilizer.

[0024] In addition, the appropriate coating, spraying or administration amount of the pharmaceutical composition can vary depending on factors such as formulation method, administration method, age, body weight, sex, disease symptoms, diet, administration time, administration route, excretion rate and reaction sensitivity. Generally speaking, an experienced doctor or veterinarian can easily determine and prescribe an effective dosage according to the desired therapeutic effect.

[0025] The pharmaceutical composition of the present invention can be implemented as an antibiotic, disinfectant, bactericide, and therapeutic agent.

[0026] According to the present invention, the method for treating infection of Staphylococcus aureus including Staphylococcus aureus using a pharmaceutical composition containing the antimicrobial protein S2200 having the amino acid sequence represented by sequence number 1 as an active ingredient can effectively act on Staphylococcus aureus that is resistant to existing antibiotics or antimicrobial substances. In addition, the antimicrobial protein S2200 specific to Staphylococcus aureus including Staphylococcus aureus of the present invention does not affect other normal symbiotic bacteria except Staphylococcus aureus, thereby minimizing the side effects caused by the use of the pharmaceutical composition containing the antimicrobial protein S2200 as an active ingredient. For existing antibiotics, there is a problem of inducing multiple side effects because of adverse effects on probiotics. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 This is an electrophoresis photograph illustrating the results of producing the antimicrobial protein S2200, which is characterized by the amino acid sequence represented by sequence number 1 and is specific to Staphylococci including Staphylococcus aureus, in the form of a recombinant protein. Lane M is a protein size marker, lane 1 is a cell lysis solution, lane 2 is a chromatography eluate during the initial purification, and lane 3 is a chromatography eluate during the secondary purification. DETAILED DESCRIPTION

[0028] Next, the present invention will be described in more detail with reference to examples. However, these examples are merely illustrative of the present invention, and the scope of the present invention is not limited by these examples.

[0029] Example 1: Preparation of antimicrobial protein S2200 specific to Staphylococci including Staphylococcus aureus

[0030] In the following, the production of antimicrobial protein S2200 specific to Staphylococcus aureus including Staphylococcus aureus will be described. In this example, a strain produced by transforming Escherichia coli with a plasmid having a nucleic acid sequence of SEQ ID NO: 2 produced by the present inventors, namely Escherichia coli Endo-22, was used as a production strain.

[0031] After adding 20 μl of Escherichia coli Endo-22 to 20 ml of LB medium (Tryptone 10 g / L, yeast extract 5 g / L and NaCl 10 g / L) containing 50 μg / ml kanamycin for inoculation, shaking culture was performed overnight at 37°C. The next day, the culture solution that had been cultured overnight was added to 1 L of LB medium containing 50 μg / ml kanamycin at a volume ratio of 1 / 100. Culture was performed at a temperature of 37°C with a stirring speed of 200 rpm. When the cell concentration reached 0.3 to 0.35 based on the absorbance at 600 nm, the culture temperature was changed to 19°C. Then, after about 30 minutes, when the cell concentration reached 0.5 based on the absorbance at 600 nm, L-arabinose was added to a final concentration of 0.2% to induce the expression of the antimicrobial protein S2200 having the amino acid sequence represented by sequence number 1, and the culture was continued for an additional 16 hours.

[0032] After the culture was completed, the cell culture solution was taken and the cell precipitate was recovered by centrifugation at 6,000 rpm at 4°C for 10 minutes, and the recovered cell precipitate was suspended in 40 ml of a buffer solution (50 mM Na2HPO4, 10 mM ethylenediaminetetraacetic acid (EDTA) and 1 mM dithiothreitol (DTT), pH 7.5). The cell suspension prepared as described above was disrupted by ultrasonic disruption, which was repeated for 15 minutes under the condition of applying ultrasonic waves for 3 seconds and then stopping for 3 seconds, and was performed in an ice bath.

[0033] After the cells were pulverized, the supernatant obtained by centrifuging the cell lysate at 13,000 rpm at 4° C. for 20 minutes was purified by general cation-exchange chromatography and hydrophobic interaction chromatography purification processes.

[0034] Next, the purification process will be briefly described. In this example, 5 ml of HiTrap was used as a cation exchange resin. TMSP FF (GE Healthcare) and 5 ml of Toyopearl PPG-600M (Tosoh Bioscience) were used as a hydrophobic interaction resin. As the primary purification process, the cation exchange resin chromatography operation was performed after the chromatography column was equilibrated in advance with buffer A (Buffer A, 25 mM Na2HPO4, 10 mM EDTA, pH 7.5). After the sample was dripped onto the chromatography column, 1.5 column volumes (CV) of buffer A, 30 column volumes (CV) of buffer B (buffer B, 25 mM Na2HPO4, 10 mM EDTA, 50 mM NaCl and 0.5% Triton X-100, pH 7.5), and buffer C in buffer A (buffer C, 25 mM Na2HPO4, 10 mM EDTA and 0.7 M NaCl, pH 7.5) were used at a flow rate of 5 ml / min. 7.5) was washed using 5 column volumes (CV) at 31%. After washing, the primary purification process, i.e., cation exchange resin chromatography, was performed at a flow rate of 5 ml / min with a concentration gradient from 31% to 100% from buffer A to buffer C. Next, the secondary purification process, i.e., hydrophobic interaction chromatography, was performed using the obtained primary purification eluate. The hydrophobic interaction chromatography was performed after the chromatography column was equilibrated with buffer D (buffer D, 25 mM Na2HPO4 and 1.5 M NaCl, pH 7.5) in advance, and 4 M NaCl was added to the sample obtained in the primary purification process until it had the same conductivity as buffer D and added dropwise to the chromatography column, and then washed with 10 column volumes (CV) of buffer E (10 mM L-histadine and 1.5 M NaCl, pH 7.5) at a flow rate of 5 ml / min. After the washing was completed, the chromatography was performed at a flow rate of 5 ml / min under the conditions of a concentration gradient from 0% to 100% from buffer E to buffer F (buffer F, 10 mM L-histadine and 1.0 M urea, pH 7.5). In the process, the desired antimicrobial protein S2200 having the amino acid sequence represented by sequence number 1 was eluted. Figure 1The figure shows the electrophoresis analysis results of the antimicrobial protein S2200 purified by the chromatography purification process.

[0035] The fractions containing the antimicrobial protein S2200 at a high concentration were collected from the purified fractions and dialyzed against a buffer solution (10 mL of L-histadine, pH 6.0) to perform medium exchange, thereby obtaining an antimicrobial protein S2200 solution with a purity of more than 90%.

[0036] The "S2200 pharmaceutical composition" was produced by buffer exchange of the S2200 solution concentrated as described above using a buffer (buffer, 10 mL of L-histadine, 5% (w / v) sorbitol and 0.1% (w / v) poloxamer 188, pH 6.0).

[0037] Example 2: Antimicrobial activity detection of antimicrobial protein S2200

[0038] The antibacterial activity of the antibacterial protein S2200 specific to Staphylococcus aureus produced according to Example 1 was tested. The information of the target strain used in the antibacterial activity test is shown in Table 1.

[0039] Table 1

[0040] Antimicrobial activity test strains

[0041]

[0042] In the antibacterial activity test, turbidity reduction assay and minimum inhibitory concentration (MIC) assay were used. Turbidity reduction assay was implemented as described below. The test strain was suspended in physiological saline so that its absorbance at 600nm reached about 0.7, and then 0.1ml of antimicrobial protein S2200 solution (final concentration: 2.5μM) was added to 0.9ml of the suspension, and then the absorbance was measured at 600nm for 30 minutes. As a negative control group, a buffer solution not containing antimicrobial protein S2200 (10mM L-histadine, 5% (w / v) sorbitol and 0.1% (w / v) poloxamer 188, pH 6.0) was used instead of the antimicrobial protein S2200 solution. The minimum inhibitory concentration (MIC) was performed and derived according to the general minimum inhibitory concentration (MIC) test method specified in the Clinical and Laboratory Standards Institute (CLSI) guidelines.

[0043] The results showed that the antimicrobial protein S2200 exhibited bacteriolytic activity only against Staphylococcus aureus and other staphylococci, but not against other test subject strains. The results are shown in Table 2.

[0044] Table 2

[0045] Antimicrobial activity test results

[0046]

[0047]

[0048] The above results confirm that the antimicrobial activity of the antimicrobial protein of the present invention has very significant specificity for Staphylococci including Staphylococcus aureus.

[0049] The above results confirm that the antimicrobial protein S2200 of the present invention, which is specific to staphylococci including Staphylococcus aureus, can lyse staphylococci including Staphylococcus aureus and eventually kill them. The above characteristics indicate that the pharmaceutical composition containing the antimicrobial protein S2200, which is specific to staphylococci including Staphylococcus aureus, can be used for the purpose of killing staphylococci when infected by staphylococci including Staphylococcus aureus, and can also be used for the purpose of treating staphylococcal infection in the same manner as general antibiotics.

[0050] Example 3: Properties of the antimicrobial protein S2200 specific to Staphylococci including Staphylococcus aureus investigation

[0051] The immunogenicity of antimicrobial protein S2200 was tested. The immunogenicity test was performed in two steps, the first step was the preparation of peripheral blood mononuclear cells (PBMC) labeled with carboxyfluorescein diacetate succinimidyl ester (CFSE), and the second step was T-cell proliferation assay and fluorescence-activated cell sorting (FACS) analysis. The preparation of peripheral blood mononuclear cells (PBMC) labeled with carboxyfluorescein diacetate succinimidyl ester (CFSE) was performed as shown in Table 3 below.

[0052] Table 3

[0053] Preparation of carboxyfluorescein succinimidyl ester (CFSE) labeled peripheral blood mononuclear cells (PBMC)

[0054]

[0055]

[0056] T-cell proliferation assay and fluorescence activated cell sorting (FACS) analysis were performed as shown in Table 4 below.

[0057] Table 4

[0058] T-cell proliferation assay and fluorescence activated cell sorting (FACS) analysis

[0059]

[0060]

[0061] The results of immunogenicity test are shown in Table 5.

[0062] Table 5

[0063] Immunogenicity test results

[0064]

[0065] The above results confirm that the immunogenicity of the antimicrobial protein S2200 is very low. The clinical impact of immune responses in patients varies greatly, ranging from extremely mild to causing considerable harm. In addition, considering its significant impact on pharmacokinetics, pharmacodynamics, safety or efficacy, the low immunogenicity of S2200 can be considered a relatively large advantage when used as a pharmaceutical.

[0066] Example 4: Antimicrobial protein S2200 specific to Staphylococcus aureus including Staphylococcus aureus Testing the therapeutic effect of Staphylococcus aureus infection

[0067] The therapeutic effect of the antimicrobial protein S2200, which is specific to Staphylococci, including Staphylococcus aureus, on Staphylococcus aureus infection was tested by an infection animal model experiment. Specifically, female ICR mice weighing 23 g ± 20% (5 weeks after birth) were used. A total of 20 mice were divided into 2 groups (10 mice in each group) and intravenously injected with Staphylococcus aureus MW2 (1×10 8CFU / mouse). For the control group animals, buffer (buffer, 10mM L-histadine, 5% (w / v) sorbitol and 0.1% (w / v) poloxamer 188, pH 6.0) was intravenously injected at 30 minutes, 12 hours and 24 hours after bacterial infection. For the treatment group animals, the pharmaceutical composition prepared in Example 1 (dose: 25mg / kg) was intravenously injected at 30 minutes, 12 hours and 24 hours after bacterial infection. The dead individuals were tested and the clinical symptoms were observed every day. The bacterial killing ability of the pharmaceutical composition of the present invention in the blood was tested by the general colony counting method using blood collected at 5 days after bacterial infection. The observation results of clinical symptoms showed that the animals in the treatment group behaved normally throughout the test period, but the animals in the control group began to show various clinical symptoms such as erythema of lid margin and decreased activity from 2 days after bacterial infection. The results of the detection of dead individuals showed that the survival rate of the animals in the treatment group injected with the pharmaceutical composition of the present invention was significantly improved compared with that of the control group (Table 6).

[0068] Table 6

[0069] Therapeutic effect on Staphylococcus aureus infection

[0070]

[0071] In addition, the number of bacteria in the blood was significantly reduced when the pharmaceutical composition of the present invention was injected. The number of blood bacteria in the blood collected from the control group animals was an average of 1×10 6 CFU / ml or more, but no bacteria were observed in the blood collected from animals in the treatment group.

[0072] The above results confirm that the pharmaceutical composition prepared according to the present invention has a therapeutic effect on Staphylococcus aureus infection. Therefore, it can be concluded that the pharmaceutical composition of the present invention can be effectively used to treat Staphylococcus aureus infection.

[0073] Example 5: Antimicrobial protein S2200 specific to Staphylococcus aureus Testing the effectiveness of treatment for coccal infections

[0074] The therapeutic effect of the antimicrobial protein S2200, which is specific to Staphylococci, including Staphylococcus aureus, on Staphylococcal infection was tested by an infection animal model experiment. Specifically, female ICR mice weighing 22g±20% (5 weeks after birth) were used. A total of 20 mice were divided into 2 groups (10 mice in each group) and then intravenously injected with Staphylococcus hemolyticus CCARM 3733 (1×10 8 CFU / mouse) were used to infect the animals in the control group. For the animals in the control group, buffer (buffer, 10mM L-histadine, 5% (w / v) sorbitol and 0.1% (w / v) poloxamer 188, pH 6.0) was injected intravenously at 30 minutes, 12 hours and 24 hours after the bacterial infection. For the animals in the treatment group, the pharmaceutical composition prepared in Example 1 (dose: 25mg / kg) was injected intravenously at 30 minutes, 12 hours and 24 hours after the bacterial infection. The dead individuals were tested and the clinical symptoms were observed every day. The bacterial killing ability of the pharmaceutical composition of the present invention in the blood was tested by the general colony counting method using blood collected at 5 days after the bacterial infection. The clinical symptom observation results confirmed that the animals in the treatment group behaved normally throughout the test period, but the animals in the control group began to show various clinical symptoms such as eyelid redness and reduced activity from 2 days after the bacterial infection. According to the results of the dead individual detection, it can be confirmed that the injection of the pharmaceutical composition of the present invention can significantly improve the survival rate of animals (Table 7).

[0075] Table 7

[0076] Effects of treatment on staphylococcal infection

[0077]

[0078] In addition, the number of bacteria in the blood was significantly reduced when the pharmaceutical composition of the present invention was injected. The number of blood bacteria in the blood collected from the control group animals was an average of 1×10 5 CFU / ml or more, but no bacteria were observed in the blood collected from animals in the treatment group.

[0079] The above results confirm that the pharmaceutical composition prepared according to the present invention also has a therapeutic effect on staphylococcal infection. Therefore, it can be concluded that the pharmaceutical composition of the present invention can be effectively used to treat staphylococcal infection.

[0080] In the above content, specific parts of the present invention are described in detail, but people with general knowledge of the industry should understand that these specific descriptions are only preferred implementation examples, and the scope of the present invention is not limited thereby. Therefore, the substantive scope of the present invention should be defined by the attached claims and their equivalents.

[0081] Name of depository institutionKCTC

[0082] Accession number: KCTC15031BP

[0083] Collection date: 20220712

[0084] Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure International Form

[0085] Certificate of acceptance for the original deposited item is issued in accordance with Rule 7.1

[0086] Depositor Name: Inton Biotechnology

[0087] Address: 137, Sagimokgok-ro, Jungwon-gu, Seongnam-si, Gyeonggi-do, South Korea

[0088]

Claims

1. An antimicrobial protein S2200 having antimicrobial activity specific to Staphylococci including Staphylococcus aureus and having an amino acid sequence represented by SEQ ID NO:

1.

2. The antimicrobial protein S2200 according to claim 1, characterized in that: The antimicrobial protein S2200 consists of 345 amino acids and has a molecular weight of 38.0 kDa.

3. A pharmaceutical composition for treating staphylococcal infection including Staphylococcus aureus, comprising the antimicrobial protein S2200 according to claim 1 as an active ingredient.

4. The pharmaceutical composition for treating staphylococcal infection including Staphylococcus aureus according to claim 3, characterized in that: The pharmaceutical composition is used as an antibiotic, disinfectant, bactericide or therapeutic agent.

5. A method for producing the antimicrobial protein S2200 according to claim 1 using the production strain Escherichia coli Endo-22 (Deposit No. KCTC 15031BP) produced by transforming Escherichia coli with the plasmid represented by the nucleic acid sequence of SEQ ID NO: 2.

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