Transformed plant expressing starry flounder l-amino-acid oxidase

Abstract

The purpose of the present invention is to provide a means for obtaining a large amount of recombinant psLAAO with better ease and economic efficiency as compared to artificial synthesis using yeast. This is achieved by, for example, a transformed plant in which a soluble starry flounder (Platicithys stellatus) L-amino-acid oxidase (psLAAO) gene and a viral RNA silencing suppressor gene are introduced as foreign genes, and which expresses the introduced genes.

Description

Transgenic plants expressing L-amino acid oxidase of the swamp flounder 【0001】 The present invention relates to a transgenic plant expressing recombinant Platichthys stellatus L-amino acid oxidase (psLAAO). 【0002】 L-amino acid oxidases (LAAOs) are found in fish mucus and other environments, and catalyze the deamination of L-amino acids. LAAOs bind to bacterial surfaces, and hydrogen peroxide produced via flavin adenine dinucleotide (FAD) creates pores in the bacterial membrane at the binding site. This allows bacterial contents to leak out, resulting in cell membrane damage and antibacterial activity against bacteria. Furthermore, LAAOs are highly conserved in various organisms, from microorganisms to mammals, and have been reported to possess a wide range of effects, including antitumor effects, apoptosis induction, antiviral activity, antibacterial activity, antiparasitic activity, and cell cycle arrest. Interestingly, LAAOs have been reported to bind to bacteria (Gram-positive and Gram-negative bacteria) and exhibit a biological activity mechanism mediated by reactive oxygen species (ROS). 【0003】 The starry flounder (Platicithys stellatus) inhabits brackish water areas and lakes, and its body surface is covered with a large amount of mucus. The mucus on the surface of the starry flounder contains the starry flounder L-amino acid oxidase (psLAAO) protein. psLAAO exhibits antibacterial activity against pyogenic bacteria such as Staphylococcus epidermidis, Staphylococcus aureus, Streptococcus pyogenes, Vibrio parahaemolyticus, and methicillin-resistant Staphylococcus aureus (MRSA), a causative agent of hospital-acquired infections (Non-patent Literature 1). 【0004】The inventors have identified the full-length gene sequence of psLAAO and have succeeded in artificially synthesizing psLAAO by inserting the psLAAO gene into the yeast genome and expressing it (Patent Document 1 and Non-Patent Document 2). 【0005】 International Publication No. 2024 / 075809 brochure 【0006】 Kasai K., et al. : Novel L-amino acid oxidase with antibacterial activity against methicili-resistant Staphylococcus aureus isolated from epidermal mucus of the flounder Platichthys stellatus. FEBS J. 277, 453-465, 2010.Kasai K., et al. : Recombinant production and evaluation of an antibacterial L-amino acid oxidase derived from flounder Platichthys stellatus. Appl Microbiol Biotechnol (2015) 99:6693-6703 【0007】 In artificial synthesis methods using bacteria such as E. coli, almost all of the resulting recombinant psLAAO becomes inclusion bodies, requiring refolding of the synthetic protein, thus complicating the synthesis process. Furthermore, artificial synthesis methods using animal cells are impractical from a productivity standpoint due to the high cost of serum mixed into the culture medium. 【0008】 In contrast, recombinant psLAAO, artificially synthesized using yeast, is characterized by its activity despite being a secreted form, and its amino acid sequence being almost identical to that of the natural protein. Thus, yeast-derived recombinant psLAAO exhibits strong antibacterial activity against various pathogenic microorganisms, similar to the natural psLAAO found in the mucus of the flounder. 【0009】However, when attempting to obtain recombinant psLAAO in large quantities through yeast fermentation, it is necessary to subject the yeast to high-density or large-scale culture treatment, and then separate and purify the recombinant psLAAO from the culture medium. This requires large-scale and multiple types of equipment, and when combined with the costs associated with cultivation and purification, it is generally not economically viable. 【0010】 Therefore, the problem that the present invention aims to solve is to provide a means for obtaining large quantities of recombinant psLAAO in a simpler and more economical way compared to artificial synthesis using yeast. 【0011】 In an effort to solve the above problems, the inventors of the present invention considered producing recombinant psLAAO using plants. Therefore, they selected rice as the plant and attempted to infect rice callus (undifferentiated cell mass) induced from rice seeds (rice) with Agrobacterium containing a binary vector with the psLAAO gene, and then collect genetically modified seeds after selecting drug-resistant callus and redifferentiation. 【0012】 Using the method described above, recombinant psLAAO was insoluble in rice seeds expressing the psLAAO gene and could hardly be obtained without denaturation treatment. Therefore, we attempted to induce callus from rice seeds and obtain soluble psLAAO from the callus. As a result, we succeeded in obtaining recombinant psLAAO with antibacterial activity from genetically modified primary (0th generation) callus. 【0013】However, soluble psLAAO was hardly obtained from callus obtained by further inducing callus from rice seeds (first generation) obtained by redifferentiating and cultivating the zero-generation callus. Therefore, the inventors continued to experiment and designed a soluble psLAAO gene by adding a signal peptide to psLAAO that is anchored to the endoplasmic reticulum, and further introduced a viral RNA silencing suppressor gene along with this gene into rice callus. Surprisingly, as a result, they succeeded in obtaining recombinant psLAAO with high expression and solubilization of antibacterial activity not only from the zero-generation callus but also from the first-generation callus. 【0014】 By redifferentiating and cultivating the first generation callus, it is possible to obtain large quantities of rice seeds. Then, by simply inducing callus from the large quantity of rice seeds obtained, soluble psLAAO can be produced in large quantities. As a result, compared to artificial synthesis using yeast, artificial synthesis using plants such as rice is simpler and more economical, allowing for the production of large quantities of soluble psLAAO. 【0015】 Based on the knowledge and successful examples obtained in this manner, the inventors have succeeded in creating transformed plants expressing soluble psLAAO, etc., as a solution to the problems of the present invention. The present invention is completed based on the knowledge and successful examples obtained for the first time by the inventors. 【0016】In other words, according to each aspect of the present invention, the following embodiments are provided: [1] A transformed plant in which a soluble Platicitys stellatus L-amino acid oxidase (psLAAO) gene and a viral RNA silencing repressor gene have been introduced as foreign genes and which express the introduced genes. [2] The transformed plant according to item [1], wherein the soluble psLAAO is psLAAO having an endoplasmic reticulum anchoring signal peptide or psLAAO from which a secretion signal peptide sequence has been removed. [3] The transformed plant according to item [1], wherein the viral RNA silencing inhibitor is at least one protein selected from the group consisting of Cucumber mosaic virus 2b protein, Tombus virus p19 protein, Turnip crinkle virus p38 protein, and Flockhouse virus B2 protein. [4] The transformed plant according to item [1], wherein the transformed plant is transformed rice, transformed strawberry, or transformed soybean. [5] Seeds or callus of a transformed plant according to any one of items [1] to [4]. [6] A method for producing soluble psLAAO, comprising the step of obtaining soluble psLAAO from seeds or callus of a transformed plant according to item [5]. [7] A DNA fragment comprising, or in combination with, a first DNA fragment containing a soluble psLAAO gene and a second DNA fragment containing a viral RNA silencing suppressor gene. [8] The DNA fragment according to item [7], wherein the soluble psLAAO is psLAAO having an endoplasmic reticulum anchoring signal peptide or psLAAO from which a secretion signal peptide sequence has been removed.[9] The DNA fragment described in item [7], wherein the viral RNA silencing inhibitor is at least one protein selected from the group consisting of Cucumber mosaic virus 2b protein, Tombus virus p19 protein, Turnip crinkle virus p38 protein, and Flockhouse virus B2 protein.

[10] A method for producing a transformed plant that expresses soluble psLAAO, comprising the step of introducing a DNA fragment described in any one of items [7] to [9] as an exogenous gene into a plant.

[11] An antimicrobial composition containing soluble psLAAO obtained from the seeds or callus of a transformed plant described in item [5] as an active ingredient.

[12] The antimicrobial composition according to item

[11] , which is used for antimicrobial action against at least one selected from the group consisting of Staphylococcus epidermidis, Staphylococcus aureus, Vibrio parahaemolyticus, methicillin-resistant Staphylococcus aureus, Pseudomonas aeruginosa, Streptococcus pyogenes, Propionibacterium acnes, Acinetobacter, Trichophyton, Yersinia cyanocarcinoma, Escherichia coli, and Enterococcus.

[13] The antimicrobial composition according to item

[11] , which is used as a topical preparation applied to the skin or eyes.

[14] A method for preventing or treating an infectious disease, comprising administering a soluble psLAAO obtained from a transformed plant according to any one of items [1] to [4] to a target organism.

[15] An antimicrobial composition having antimicrobial activity, comprising soluble Platicitys stellatus L-amino acid oxidase (psLAAO) as an active ingredient, wherein the antimicrobial activity is antimicrobial activity against at least one selected from the group consisting of Staphylococcus epidermidis, Staphylococcus aureus, Vibrio parahaemolyticus, methicillin-resistant Staphylococcus aureus, Pseudomonas aeruginosa, Streptococcus pyogenes, Propionibacterium acnes, Acinetobacter, Trichophyton, Yersinia cyanobacteria, Escherichia coli, and Enterococcus.

[16] The antimicrobial composition according to item

[15] , wherein the antimicrobial activity is antimicrobial activity against at least one selected from the group consisting of Staphylococcus aureus, Pseudomonas aeruginosa, Streptococcus pyogenes, Propionibacterium acnes, Acinetobacter, Trichophyton, Yersinia cyanocarcinoma, Escherichia coli, and Enterococcus.

[17] The antimicrobial composition according to item

[15] , wherein the soluble psLAAO is psLAAO having an endoplasmic reticulum anchoring signal peptide or psLAAO from which the secretion signal peptide sequence has been removed. 【0017】 According to the present invention, large quantities of soluble psLAAO can be produced simply and economically. That is, transgenic rice expressing soluble psLAAO can be cultivated on a large scale. Furthermore, soluble psLAAO can be produced simply and in large quantities by inducing callus from the obtained rice seeds and crushing the resulting callus in a liquid. According to the present invention, by using plant material, large quantities of soluble psLAAO can be produced without using a huge industrial plant, and furthermore, without overexploiting resources or the effects of climate change. 【0018】 According to the present invention, seed callus can be cultured in a petri dish within a growth chamber, and soluble psLAAO equivalent to 1 liter of yeast culture solution can be produced from a single 10 cm petri dish. This reduces the economic burden of soluble psLAAO production to about 1 / 100th compared to artificial synthesis using yeast, and also enables space-saving in manufacturing equipment. Furthermore, by selecting high-expression seeds and crossbreeding the selected high-expression next-generation seeds, a further increase in soluble psLAAO yield can be expected. 【0019】 This invention is applicable to rice, including major varieties such as Koshihikari, ultra-high-yielding rice such as feed rice, large-embryo rice, and dwarf rice, when rice is used as the plant. Rice seeds can be stored for a long period of time, and it is possible to maintain psLAAO-expressing rice seed lines through complete hydroponics. 【0020】Figure 1 is a vector map showing the gene composition of p35SHPTAg7-GW. Figure 2 is a diagram showing the gene composition of the DNA construct pHPT / UbiP / LAAO / KDEL. Figure 3 is a diagram showing the gene composition of (A) pHPT / UbiP / LAAO / KDEL / 2b and (B) pHPT / UbiP / LAAO / KDEL / p19. Figure 4 is a photograph of rice seeds, callus, and panicles at each step of the process of producing transformed rice callus using Agrobacterium, as described in the examples below. Figure 5 is a photograph of agar plates showing the results of the halo test of (A) recombinant psLAAO fraction and (B) recombinant psLAAO purified solution derived from transformed callus obtained using pHPT / UbiP / LAAO / KDEL, as described in the examples below. Figure 6 is a gel photograph showing the results of (A) SDS-PAGE, (B) Western blot using anti-psLAAO1 antibody, and (C) Western blot using anti-His antibody of recombinant psLAAO purified solution derived from transformed callus obtained using pHPT / UbiP / LAAO / KDEL, as described in the examples below. Figure 7 is a photograph of an agar plate showing the results of a halo test using recombinant psLAAO purified solution derived from transformed callus obtained using pHPT / UbiP / LAAO / KDEL, as described in the examples below, with (A) 20 mM Tris-HCl (pH 7.0), (B) catalase solution, or (C) anti-psLAAO1 antibody solution. Figure 8 is a photograph of agar plates showing the results of a halo test using recombinant psLAAO fractions obtained from (A) 0th generation callus and (B) 1st generation callus, as described in the examples below. Figure 9 is a photograph of agar plates showing the results of a halo test for recombinant psLAAO purified solutions derived from eight clones of transformed callus obtained using (A) pHPT / UbiP / LAAO / KDEL / 2b and one clone of transformed callus obtained using (B) pHPT / UbiP / LAAO / KDEL / p19, as described in the examples below.Figure 10A is a photograph of an agar plate showing the results of a halo test using recombinant psLAAO purified solution derived from first-generation callus obtained using pHPT / UbiP / LAAO / KDEL / 2b, as described in the examples below, and a gel photograph showing the results of a Western blot using anti-psLAAO1 antibody. Figure 10B is a photograph of an agar plate showing the results of a halo test using recombinant psLAAO purified solution derived from second-generation callus obtained using pHPT / UbiP / LAAO / KDEL / 2b, as described in the examples below. Figure 11 is a photograph of an agar plate showing the results of a halo test using recombinant psLAAO purified solution derived from zero-generation and first-generation callus obtained using pHPT / UbiP / LAAO / KDEL / p19, as described in the examples below. Figure 12 shows the estimated three-dimensional structure of psLAAO1 as described in the examples below. Figure 13 shows the estimated three-dimensional structures of psLAAO1(1-522), psLAAO1(29-522), and psLAAO1(35-505) as described in the examples below. 【0021】 The details of the present invention will be described below, but the present invention can take various forms insofar as it achieves its objective. 【0022】 In this specification, unless otherwise specified, each term is used in the sense commonly used by those skilled in the art in the fields of biology, biochemistry, biotechnology, etc., and should not be interpreted as having an unreasonably restrictive meaning. Furthermore, since the assumptions and theories made herein are based on the inventors' prior knowledge and experience, the present invention is not limited solely to such assumptions and theories. 【0023】"Includes" means that elements other than those explicitly included can be added (synonymous with "at least include"), but it also includes "consists of" and "essentially consists of." That is, "includes" can mean including the explicitly included elements and any one or more of those elements, consisting of the explicitly included elements, or essentially consisting of the explicitly included elements. Examples of elements include components, processes, conditions, parameters, and other limitations. "Has" is synonymous with "includes." The terms "and / or" mean any one of the multiple related items listed, any combination of two or more, or all of them. The "~" in a numerical range means a range that includes the numbers before and after it, and also includes the range excluding one of the limit values ​​in which they are included. For example, "0% to 100%" could be 0% or more, 100% or less, or 0% or more and 100% or less. "Approximately" means a quantity within ±10% of the quantity that follows the term. For example, "approximately 100" means 100 ± 10%, i.e., 90 to 110. "Greater than" and "less than" mean the lower and upper limits, respectively, without including the preceding number. For example, "greater than 1" means a number greater than 1, and "less than 100" means a number less than 100. The number of digits in an integer value matches the number of significant figures. For example, 1 has 1 significant figure, and 10 has 2 significant figures. Also, the number of digits after the decimal point in a decimal value matches the number of significant figures. For example, 0.1 has 1 significant figure, and 0.10 has 2 significant figures. 【0024】"Foreign gene" refers to a gene that is not naturally occurring on the chromosome (genomic) DNA of the organism being introduced, and is also called a heterogene. In this specification, genome and chromosome are synonymous. "Gene expression" means that a protein (gene-encoded protein) having the amino acid sequence encoded by the nucleotide sequence of a gene is produced through transcription, translation, etc., to possess its original structure and activity. "Wild-type" refers to an organism that exists naturally and has not been artificially genetically modified. "Transformed" refers to an organism that has been artificially genetically modified. "Wild-type gene" refers to a gene that is naturally present on the genomic DNA of a wild-type organism. "Wild-type protein" refers to a protein encoded by a wild-type gene. "Antibacterial activity" and "antibacterial" mean functioning to kill, inhibit the growth of, or slow the growth rate of target microorganisms. 【0025】 [Summary of the Invention] One aspect of the present invention is a transgenic plant expressing soluble Platicitys stellatus L-amino acid oxidase (psLAAO). In one embodiment of the present invention, the transgenic plant has the soluble psLAAO gene and a viral RNA silencing repressor gene introduced as foreign genes and expresses the introduced genes. Another aspect of the present invention is the seeds and callus of a transgenic plant expressing soluble psLAAO. 【0026】 Another aspect of the present invention is a method for producing soluble psLAAO. One embodiment of the present invention includes the step of obtaining soluble psLAAO from the seeds or callus of a transgenic plant expressing soluble psLAAO. 【0027】Another aspect of the present invention is a DNA fragment for producing a transformed plant expressing soluble psLAAO. A DNA fragment according to one aspect of the present invention comprises a first DNA fragment containing a soluble psLAAO gene and a second DNA fragment containing a viral RNA silencing suppressor gene. A DNA fragment according to another aspect of the present invention is a combination of a first DNA fragment containing a soluble psLAAO gene and a second DNA fragment containing a viral RNA silencing suppressor gene. 【0028】 Another aspect of the present invention is a method for producing a transformed plant that expresses soluble psLAAO. One embodiment of the present invention includes the step of obtaining a transformed plant that expresses soluble psLAAO by introducing a DNA fragment for producing a transformed plant that expresses soluble psLAAO as an exogenous gene into a plant. 【0029】 Another aspect of the present invention is an antimicrobial composition comprising soluble psLAAO as an active ingredient. A composition according to one embodiment of the present invention comprises soluble psLAAO obtained from the seeds or callus of a transgenic plant expressing soluble psLAAO as an active ingredient. 【0030】 [Soluble psLAAO] Three types of L-amino acid oxidase (psLAAO) have been found in the starry flounder (Platicithys stellatus). In starry flounder, psLAAO1 (SEQ ID NO: 1) is localized in the gills, psLAAO2 (SEQ ID NO: 2) is localized in the epidermis, and psLAAO3 (SEQ ID NO: 3) is localized in the liver. The psLAAO can be either psLAAO1 or psLAAO3, which have been confirmed to have antibacterial properties, or a combination thereof. 【0031】Both psLAAO1 and psLAAO3 possess antibacterial activity while sharing 80% sequence identity with each other in their amino acid sequences. Therefore, psLAAO may be a protein having an amino acid sequence that has 70% or more, preferably 75% or more, more preferably 80% or more, and even more preferably 83% or more, 85% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more sequence identity with the amino acid sequence of psLAAO1 or psLAAO3. However, psLAAO preferably has the cysteine ​​residues at positions 37 and 200 that are present in wild-type psLAAO1, and more preferably it closely conserves the structure of wild-type psLAAO1 (excluding the N-terminal signal peptide sequence). Furthermore, psLAAO1(29-522) and psLAAO1(35-505), which have less than 100% sequence identity with the wild-type psLAAO, are not wild-type psLAAO but mutant (non-natural) psLAAO. 【0032】 There are no particular limitations on the method for determining sequence identity of amino acids, but for example, it can be determined by using a program that aligns the amino acid sequences of two proteins using commonly known methods and calculates the degree of sequence agreement between the two. 【0033】As a program for calculating the coincidence rate in two amino acid sequences, for example, the algorithm of Karlin and Altschul (Proc. Natl. Acad. Sci. USA 87: 2264-2268, 1990; Proc. Natl. Acad. Sci. USA 90: 5873-5877, 1993) is known, and the BLAST program using this algorithm has been developed by Altschul et al. (J. Mol. Biol. 215: 403-410, 1990). Furthermore, Gapped BLAST, a program for determining sequence identity with higher sensitivity than BLAST, is also known (Nucleic Acids Res. 25: 3389-3402, 1997). Those skilled in the art can use the above programs to search for sequences showing high sequence identity with a given sequence from a database. These are available, for example, on the website on the Internet of the US National Center for Biotechnology Information (http: / / blast.ncbi.nlm.nih.gov / Blast.cgi). 【0034】 Examples of amino acid sequences having 70% or more sequence identity with the amino acid sequence of psLAAO1 or psLAAO3 include, for example, amino acid sequences having one or more deletions, substitutions, additions, etc. of amino acids in the amino acid sequence of psLAAO1 or psLAAO3. The range of "several" is determined by the sequence identity of the amino acid sequence. For example, if 100 amino acids in the amino acid sequence are taken as one unit, it is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21 to 30 per unit. Also, "deletion of amino acids" means the deletion or disappearance of amino acid residues in the sequence, "substitution of amino acids" means that the amino acid residues in the sequence are replaced by other amino acid residues, and "addition of amino acids" means that new amino acid residues are added so as to be inserted into the sequence. 【0035】Specific embodiments of "amino acid deletion, substitution, addition" include a mode in which an amino acid is replaced with another chemically similar amino acid. For example, when a hydrophobic amino acid is replaced with another hydrophobic amino acid, or when a polar amino acid is replaced with another polar amino acid having the same charge. Such chemically similar amino acids are known in the art for each amino acid. Specific examples include non-polar (hydrophobic) amino acids such as alanine, valine, isoleucine, leucine, proline, tryptophan, phenylalanine, methionine, etc. Polar (neutral) amino acids include glycine, serine, threonine, tyrosine, glutamine, asparagine, cysteine, etc. Basic amino acids with a positive charge include arginine, histidine, lysine, etc. Acidic amino acids with a negative charge include aspartic acid, glutamic acid, etc. 【0036】 Both psLAAO1 and psLAAO3 have antibacterial activity against Staphylococcus epidermidis, Staphylococcus aureus, Methicillin-resistant Staphylococcus aureus (MRSA) and / or Vibrio parahaemolyticus. Soluble psLAAO may have antibacterial activity against Staphylococcus epidermidis, Staphylococcus aureus, MRSA or Vibrio parahaemolyticus, and preferably has antibacterial activity against two, three or all four of these. 【0037】Specific examples of psLAAO include proteins having an amino acid sequence with 70% or more sequence identity with psLAAO1 or psLAAO3, and possessing antibacterial activity against Staphylococcus epidermidis, Staphylococcus aureus, MRSA, or Vibrio parahaemolyticus. Preferably, these proteins have an amino acid sequence with 80% or more, 85% or more, 90% or more, or 95% or more sequence identity with psLAAO1 or psLAAO3, and possess antibacterial activity against any two, three, or four of Staphylococcus epidermidis, Staphylococcus aureus, MRSA, and Vibrio parahaemolyticus. More preferably, they are psLAAO1 or psLAAO3. The psLAAO in the above specific examples preferably has cysteine ​​residues at positions 37 and 200, which are present in wild-type psLAAO1, and more preferably, the structure of wild-type psLAAO1 (excluding the N-terminal signal peptide sequence) is well preserved. 【0038】 If psLAAO is recombinant psLAAO obtained using genetic engineering technology, it is thought that it is not solubilized by binding to cysteine-rich proteins such as prolamin in the organism in which it is used, for example, in plants. Therefore, when attempting to express psLAAO in plants, it is preferable to modify the recombinant psLAAO to make it solubilized. However, if recombinant psLAAO is expressed in a solubilized state in plants, solubilization modification is not necessary, and it can be made soluble psLAAO on its own. 【0039】Methods for solubilizing recombinant psLAAO include, for example, adding peptides, proteins, etc., that do not bind to prolamin to the C-terminus and / or N-terminus of recombinant psLAAO, which function to prevent recombinant psLAAO from binding to prolamin. Examples of such peptides include endoplasmic reticulum anchoring signal peptides and intracellular translocation signal peptides, but endoplasmic reticulum anchoring signal peptides are preferred because their amino acid length is relatively short and they have little effect on the antibacterial activity of psLAAO. A specific example of an endoplasmic reticulum anchoring signal peptide is the KDEL sequence (SEQ ID NO: 4). The endoplasmic reticulum anchoring signal peptide may be directly added to the C-terminus of psLAAO, or it may be added with one or more amino acids acting as spacers. The type and number of endoplasmic reticulum anchoring signal peptides added may be one of one type, two or more of one type, or two or more of two types. However, wild-type psLAAO is transported to the Golgi apparatus via the endoplasmic reticulum and secreted extracellularly due to the presence of a secretory signal peptide sequence. If a KDEL sequence is added to the C-terminus of wild-type psLAAO, it will accumulate instead of being secreted because it binds to the inner membrane of the endoplasmic reticulum. However, since the secretory signal peptide sequence and the KDEL sequence do not directly affect psLAAO expression, they can be omitted. If the secretory signal peptide sequence is removed from psLAAO and the KDEL sequence is not added, psLAAO can be recovered by lysing the transformed plant. 【0040】 psLAAO preferably has a tag peptide to facilitate separation and purification. Examples of such tag peptides include His tags. A specific example of a His tag is a 6His tag consisting of six histidine molecules linked together. When adding a sequence such as a 6His tag, a sequence for protease cleavage may be inserted between the added sequence and the psLAAO sequence. Examples of such sequences include TEV protease recognition sequences. In the case of TEV protease, the 6His tag-TEV protease recognition sequence can be inserted at the N-terminus of position 29 (S) or position 35 (S) of psLAAO so that the C-terminus of the cleavage site is Gly / Ser. 【0041】 Specific examples of soluble psLAAO include soluble psLAAO having an endoplasmic reticulum anchoring signal peptide at the C-terminus, preferably soluble psLAAO having a tag peptide at the C-terminus and / or N-terminus, more preferably soluble psLAAO having both an endoplasmic reticulum anchoring signal peptide and a tag peptide at the C-terminus, and even more preferably soluble psLAAO having a KDEL sequence and a 6His tag at the C-terminus. The soluble psLAAO having the amino acid sequence described in SEQ ID NO: 5 is obtained by adding a KDEL sequence and a 6His tag to the C-terminus of psLAAO1. Another specific example of soluble psLAAO is soluble psLAAO from which the secretion signal peptide sequence has been removed. 【0042】 [Viral RNA silencing inhibitors] Viral RNA silencing inhibitors function to suppress RNA silencing. 【0043】 In RNA silencing, double-stranded RNA (dsRNA) is processed by the Dicer protein to form small RNA double strands of approximately 20 to 25 nucleotides. These small RNA double strands are then incorporated into the Argonaut (AGO) protein. One strand (guide strand) remains with the AGO, while the other strand (passenger strand) is eliminated. In this way, the RNA-induced silencing complex (RISC) is formed. The RISC finds a single-stranded mRNA complementary to the guide strand, cleaves and degrades it, or inhibits its translation. 【0044】 Viral RNA silencing inhibitors function to suppress RNA silencing by participating in the RNA silencing pathway and blocking mRNA degradation or translation inhibition by RISCs. 【0045】For example, the cucumber mosaic virus (Cucumber mosaic virus) 2b protein and the turnip crinkle virus (Turnip crinkle virus) p38 protein interact with AGO and inhibit RISC formation. The Flockhouse virus (Flockhouse virus) B2 protein interacts with double-stranded RNA and prevents cleavage by the Dicer protein. The Tombus virus (Tombus virus) p19 protein binds to small RNA molecules and inhibits RISC formation. Thus, viral RNA silencing inhibitors only need to function by blocking any part of the RNA silencing pathway, thereby preventing RNA silencing. 【0046】 Preferred examples of viral RNA silencing inhibitors include cucumber mosaic virus 2b protein (SEQ ID NO: 6), tombas virus p19 protein (SEQ ID NO: 7), cabbage crinkle virus p38 protein (SEQ ID NO: 8), and flockhouse virus B2 protein (SEQ ID NO: 9), which may be used individually or in combination of two or more. These proteins only need to function to suppress RNA silencing, and may, for example, have amino acid sequences that have 70% or more, preferably 80% or more, more preferably 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more sequence identity with the amino acid sequences described in SEQ ID NOs: 6 to 9. 【0047】[Soluble psLAAO gene and viral RNA silencing suppressor gene] The soluble psLAAO gene has a nucleotide sequence that codes for the amino acid sequence of the soluble psLAAO protein. The viral RNA silencing suppressor gene has a nucleotide sequence that codes for the amino acid sequence of the viral RNA silencing suppressor. These nucleotide sequences in the soluble psLAAO gene and the viral RNA silencing suppressor gene may be coding sequences (CDS) or open reading frames (ORF), but ORF is preferred. For example, the mRNA of the psLAAO1 gene is registered in the DDBJ database as accession number AB495360 (current version is AB495360.1) (http: / / getentry.ddbj.nig.ac.jp / getentry / na / AB495360?filettype=html). The mRNA for the psLAAO2 gene is registered in the DDBJ database with accession number AB495361 (current version AB495361.1) (https: / / www.ncbi.nlm.nih.gov / nuccore / AB495361.1). The mRNA for the psLAAO3 gene is registered in the DDBJ database with accession number AB495362 (current version AB495362.1) (https: / / www.ncbi.nlm.nih.gov / nuccore / AB495362.1). 【0048】 Soluble psLAAO genes and viral RNA silencing repressor genes may consist of nucleotide sequences that code for each protein, or they may also include transcriptional regulatory sequences such as promoters and terminators, and non-coding sequences such as introns. A gene is not merely sequence information, but a DNA molecule (fragment) made up of linked nucleotides. 【0049】The soluble psLAAO gene and the viral RNA silencing suppressor gene may be nucleotide sequences with codon modifications, taking advantage of the fact that there are several types of codons corresponding to a single amino acid. Examples of codon-modified nucleotide sequences include nucleotide sequences that encode the same amino acid sequence as the nucleotide sequence encoded by the wild-type gene, but which are different from the wild-type gene, specifically nucleotide sequences with optimized codons, secondary structure, GC content, etc. Preferably, the nucleotide sequences have been codon-modified to facilitate expression in the host plant. 【0050】 The soluble psLAAO gene and the viral RNA silencing repressor gene are introduced into plants as exogenous genes. Therefore, the soluble psLAAO gene and the viral RNA silencing repressor gene are genes that do not exist in the genomic DNA of the host plant. 【0051】 A non-specific example of a soluble psLAAO gene is the soluble psLAAO gene (SEQ ID NO: 10), which encodes a soluble psLAAO having a KDEL sequence and a 6His tag at the C-terminus of psLAAO1, modified with codons to facilitate expression in rice. A non-specific example of a viral RNA silencing repressor gene is the 2b gene, which has been modified with codons to facilitate expression in rice. ｏｐｔ Gene (SEQ ID NO: 11) and p19 ｏｐｔ It is a gene (SEQ ID NO: 12). 【0052】 The soluble psLAAO gene and the viral RNA silencing repressor gene are introduced to be expressed in plants. Gene expression may be autonomously regulated by the transcriptional regulatory sequences of the genes, or it may be regulated using the transcriptional regulatory mechanisms of the host plant, but it is preferably autonomously regulated by the transcriptional regulatory sequences of the genes. 【0053】For the soluble psLAAO gene and the viral RNA silencing repressor gene to be expressed autonomously in plants, it is preferable that they include a promoter, ORFs for each protein, and terminators. For example, it is preferable to use a promoter suitable for the host and the psLAAO gene, to use a psLAAO gene modified to a codon suitable for the host, and to appropriately determine the terminator in accordance with the host and the psLAAO gene. 【0054】 Promoter and terminator can be appropriately selected to suit the host plant. For example, when the host is rice, possible promoters include the cauliflower mosaic virus (CaMV) 35S promoter (35S pro), the maize ubiquitin promoter (ZmUbi pro), and the NOS promoter (Nos P); and possible terminators include the Agrobacterium gene 7 terminator (Ag7 T), the rice glutelin B1 gene terminator (GluB1 T), and the NOS terminator (Nos T). The nucleotide sequences of these promoters and terminators are already known; for example, 35S pro, ZmUbi pro, Ag7 T, and GluB1 T are described in Sequence IDs 13-16, respectively. 【0055】For example, if the host is a strawberry, the promoters would be 35S pro, SVBV (Jiang L et al., Functional analysis of a viral promoter from a strawberry vein banding virus isolate from China. 2022 31;19(1):60; Sitakanta Pattanaik et al., Plant Science 167 (2004) 427-438)), FvEF1α (Chonprakun Thagun et al., Scientific Reports (2025) 15:20392), FaExp2 (C.F. Nardi et al., Postharvest Biology and Technology 114 (2016) 17-28); Nos as a terminator T, OCS (Jun-Li et al., Mol Biol Rep (2010) 37:2157-2162), CaMV 35S (Stephanie C. et al., Current Plant Biology 24 (2020) 100179), HSP (Shingo Nagaya et al., Plant Cell Examples include Physiol. 51(2):328-332 (2010). Transgenic strawberries can express psLAAO in their leaves and fruits. 【0056】 For example, if the host is soybean, the promoters would be 35S pro, P34 AB013289 (1-4646) (Sung Cheol Koo et al., Plant Biotechnol Rep (2013) 7:331-338), Gmubi (Carlos M. et al., Plant Cell Rep (2009) 28:837-849; Carola M. et al., Plant Cell Rep (2015) 34:111-120), GmActin (Zhifen Zhang & John J. Finer, Plant (2015) 51:9-18); the terminator would be Nos Examples include T, OCS, and CaMV 35S. Transgenic soybeans can express psLAAO in the leaves and hypocotyl. 【0057】 The soluble psLAAO gene and the viral RNA silencing suppressor gene may be DNA fragments containing each gene individually, or a single DNA fragment containing both genes together. That is, the soluble psLAAO gene and the viral RNA silencing suppressor gene may be a DNA fragment containing the soluble psLAAO gene, a DNA fragment containing the viral RNA silencing suppressor gene, or a DNA fragment containing both the soluble psLAAO gene and the viral RNA silencing suppressor gene. In each DNA fragment, the copy number of the soluble psLAAO gene and the viral RNA silencing suppressor gene may be any number, such as one copy or two or more copies. The DNA fragment may also be a DNA construct such as a plasmid or vector. 【0058】 DNA fragments containing a soluble psLAAO gene and / or a viral RNA silencing repressor gene preferably contain a selection marker gene to facilitate the selection of plants into which these genes have been introduced, i.e., transgenic plants. Examples of selection marker genes include drug resistance genes such as the hygromycin B-phosphotransferase gene (HPT; SEQ ID NO: 17), neomycin phosphotransferase II (NPTII), and phosphinotricin acetyltransferase (PAT), which inactivate hygromycin, but other selection marker genes such as nutrient requirement marker genes may also be used. 【0059】 Examples of DNA fragments containing the soluble psLAAO gene include, as shown in Figure 2, a DNA fragment containing the soluble psLAAO gene (2 copies) and a selection marker gene, which include a nucleotide sequence encoding psLAAO1 having a KDEL sequence and 6His at the C-terminus. 【0060】Examples of DNA fragments containing the soluble psLAAO gene and the viral RNA silencing suppressor gene include, as shown in Figure 3, a DNA fragment containing the soluble psLAAO gene (two or more copies), the 2b protein gene and the selection marker gene, which includes a nucleotide sequence encoding psLAAO1 having a KDEL sequence and 6His at the C-terminus; and a DNA fragment containing the soluble psLAAO gene (two copies), the p19 protein gene and the selection marker gene. 【0061】 [Transformed plant] A transformed plant according to one aspect of the present invention has a soluble psLAAO gene and a viral RNA silencing repressor gene introduced as foreign genes, and expresses the introduced genes. 【0062】 Soluble psLAAO can be obtained from part or all of a transgenic plant. For example, if the host of the transgenic plant is a seed plant, the seeds of the transgenic plant can be subjected to callus induction treatment, and soluble psLAAO can be obtained by separating and purifying the soluble psLAAO from the resulting callus. In this case, the host of the transgenic plant may be angiosperms or gymnosperms, but angiosperms are preferred considering the storability of the seeds, and model plants such as rice, soybeans, tobacco, and strawberries, which have a relatively long cultivation history, are more preferred. Rice is even more preferred because it is easy to cultivate and has a large number of seeds per panicle. 【0063】 [Method for producing transformed plants] Transformed plants are produced by introducing a soluble psLAAO gene and a viral RNA silencing suppressor gene as foreign genes into a host plant, thereby transforming the plant to express these genes. 【0064】The introduction of soluble psLAAO genes and viral RNA silencing suppressor genes into plants can be performed using methods already employed for gene transfer into plants, such as the Agrobacterium method, electroporation, particle gun method, viral vector methods, and genome editing. Any method is acceptable for transient expression of the introduced gene, but the Agrobacterium method is preferred for introduction into chromosomes within the plant nucleus due to its high introduction efficiency. 【0065】 The Agrobacterium method uses microorganisms of the genus Agrobacterium. A binary vector is obtained by cloning a Ti plasmid in Agrobacterium microorganisms so that a DNA excision containing the soluble psLAAO gene and / or a viral RNA silencing repressor gene is inserted between the left border repeat sequence (LB) and the right border repeat sequence (RB). The cloned binary vector is introduced into Agrobacterium microorganisms by known gene transfer methods into microbial cells, such as electroporation, to obtain transformed Agrobacterium microorganisms. Next, the transformed Agrobacterium microorganisms are infected with plants, causing the sequence surrounded by LB and RB (T-DNA) to be excised in the Agrobacterium microorganisms, and this T-DNA is transported into the nucleus of the plant cells. The T-DNA is then randomly inserted into the genomic DNA in the nucleus. 【0066】 When using the Agrobacterium method, the DNA fragment containing the soluble psLAAO gene and / or viral RNA silencing repressor gene is preferably a T-DNA having LB and RB at both ends. 【0067】 The Agrobacterium microorganisms used in the Agrobacterium method can be any Agrobacterium microorganism capable of introducing its own T-DNA into plants. Examples include Agrobacterium tumefaciens and Agrobacterium rhizogenes, with Agrobacterium tumefaciens being preferred. 【0068】 Vectors, culture media, and procedures used to transform Agrobacterium microorganisms can be found in the examples described later. For example, methods for transforming Agrobacterium microorganisms by introducing a binary vector include electroporation, protoplastation, and calcium ion methods, but electroporation is preferred. Other molecular biological, biotechnological, and biochemical methods can be found, for example, in Molecular Cloning: A Laboratory Manual, 2nd Ed., Cold Spring Harbor Laboratory, Cold Spring Harbor, NY. You can refer to the methods described in literature such as Current Protocols in Molecular Biology, Supplement 1–38, John Wiley & Sons, 1987–1997. 【0069】 Infection of plants with transformed Agrobacterium microorganisms obtained by introducing a DNA fragment containing the soluble psLAAO gene and / or the viral RNA silencing inhibitor gene as an exogenous gene can be carried out by bringing the transformed Agrobacterium microorganisms into contact with the plants so that the DNA fragment containing the soluble psLAAO gene and / or the viral RNA silencing inhibitor gene is introduced into the plants. For example, as described in the examples below, an Agrobacterium-infected callus can be obtained by adding a suspension containing transformed Agrobacterium to plant callus, then stirring the callus with the added bacterial suspension and letting it stand, then removing the excess bacterial suspension from the callus, and then culturing the callus on a suitable solid medium. Plant callus can be obtained by subjecting plant seeds to a callus induction treatment. For example, as described in the examples below, sterilized seeds can be placed on a solid medium containing plant hormones, and the resulting dedifferentiated embryo and proliferated cells can be used to obtain callus. 【0070】For the selection of transformed Agrobacterium microorganisms and Agrobacterium-infected plants, a selective medium containing a compound corresponding to a selection marker gene incorporated into a DNA fragment or binary vector containing a soluble psLAAO gene and / or a viral RNA silencing suppressor gene may be used. 【0071】 Agrobacterium-infected callus selected as transformed cells can be cultured in a redifferentiation medium containing plant hormones such as naphthaleneacetic acid and kinetin to regenerate into a plant. The resulting redifferentiated cells can be acclimated and cultivated in a hormone-free medium or soil to obtain germinated plants. The transformed plant can be callus, redifferentiated cells, or germinated plants, as long as it has been transformed by introducing the soluble psLAAO gene and the viral RNA silencing suppressor gene. 【0072】 The transformation of a transformed plant to express soluble psLAAO can be confirmed by obtaining soluble psLAAO from the transformed plant, or by confirming the presence or expression of the soluble psLAAO gene in the genomic DNA of the transformed plant. 【0073】 [Method for producing soluble psLAAO] A method for producing soluble psLAAO according to one aspect of the present invention includes the step of obtaining soluble psLAAO from the seeds or callus of a transgenic plant that expresses soluble psLAAO. 【0074】 For a method to obtain soluble psLAAO from the seeds or callus of a transformed plant expressing soluble psLAAO, refer to the method described in the examples below. For example, callus can be induced from the seeds of a transformed plant, the protein in the obtained callus can be recovered to obtain a crude protein product, and then soluble psLAAO can be separated and eluted from the crude protein product using chromatography with a hydrophobic column, thereby obtaining soluble psLAAO as a fraction containing a large amount of soluble psLAAO. 【0075】The presence of soluble psLAAO can be confirmed by evaluating its antimicrobial activity according to the type of psLAAO used. For example, since psLAAO1 and psLAAO3 have antimicrobial activity against MRSA, the presence of soluble psLAAO can be confirmed by performing a halo test targeting MRSA using the obtained fraction and confirming the formation of an inhibition zone. The halo test can be performed by referring to the method described in the examples below. 【0076】 The yield of soluble psLAAO obtained by the production method according to one embodiment of the present invention varies depending on the efficiency of introducing the soluble psLAAO gene and the viral RNA silencing suppressor gene into the transformed plant, and is not particularly limited, but for example, it is preferably 10 μg or more, more preferably 30 μg or more, even more preferably 50 μg or more, and still more preferably 70 μg or more, 80 μg or more, 90 μg or more, or 100 μg or more, per 1 g (wet weight) of callus of the transformed plant. 【0077】 [Antibacterial Composition] An antibacterial composition according to one aspect of the present invention contains soluble psLAAO obtained from the seeds or callus of a transgenic plant expressing soluble psLAAO as an active ingredient. The active ingredient psLAAO exerts its antibacterial effect by deaminating L-amino acids to produce hydrogen peroxide. 【0078】 Depending on the antibacterial activity of soluble psLAAO, the antimicrobial composition can be used for antimicrobial purposes against various microorganisms, including Gram-positive microorganisms such as MRSA and Gram-negative microorganisms, specifically for sterilization, disinfection, or infection prevention. 【0079】 The antimicrobial composition can be applied to any microorganism in which wild-type psLAAO exhibits antimicrobial activity, such as pathogenic bacteria that cause skin inflammation, specifically Staphylococcus epidermidis, Staphylococcus aureus, Vibrio parahaemolyticus, MRSA, Pseudomonas aeruginosa, Streptococcus pyogenes, Propionibacterium acnes, Acinetobacter (e.g., Acinetobacter baumannii), Trichophyton, Yersinia infection bacteria, Escherichia coli, Enterococcus (e.g., Enterococcus), but is not limited to these. 【0080】For example, Patent Document 1 and Non-Patent Document 2 describe the minimum inhibitory concentration (MIC) of recombinant psLAAO1 expressed in yeast against various microorganisms: Staphylococcus epidermidis (0.078 μg / mL), Staphylococcus aureus (0.63 μg / mL), MRSA (0.31 μg / mL), and Vibrio parahaemolyticus (0.16 μg / mL). Furthermore, the present inventors have found that the MIC against Yersinia parahaemolyticus is 1.25 μg / mL. Therefore, it is preferable that the antimicrobial composition be used for antimicrobial action against at least one, two, three, four, or five species selected from the group consisting of Staphylococcus epidermidis, Staphylococcus aureus, Vibrio parahaemolyticus, methicillin-resistant Staphylococcus aureus, and Yersinia parahaemolyticus. However, psLAAO has relatively mild antimicrobial activity against Escherichia coli and Pseudomonas aeruginosa. 【0081】 In the antimicrobial composition, the content of the active ingredient (soluble psLAAO) at the time of use (application) is preferably 0.03 μg / ml to 50.0 μg / ml, more preferably 0.3 μg / ml to 10.0 μg / ml, and even more preferably 0.5 μg / ml to 10.0 μg / ml, 0.7 μg / ml to 10.0 μg / ml, or 1.0 μg / ml to 10.0 μg / ml, from the viewpoint of exhibiting a more sufficient antimicrobial effect. 【0082】 The antimicrobial composition may be used in any application where an antimicrobial effect is exhibited. Preferably, the application is intended to exert an antimicrobial effect on target organisms such as mammals (humans, dogs, cats, cattle, horses, sheep, birds such as chickens, and fish), and more preferably, it is intended to exert an antimicrobial effect on humans. Furthermore, the antimicrobial composition may be applied not to a part of the target organism's body, but to materials that may come into contact with the body. 【0083】 The antimicrobial composition may be used as an alternative or in combination with antibiotics against multidrug-resistant bacteria. 【0084】 The antibacterial composition may be provided in the form of a liquid (such as a suspension or emulsion), an ointment, or eye drops, for example. 【0085】The dosage and administration of the antimicrobial composition can be appropriately selected depending on the type and form of the formulation, the symptoms of the target organism, age, etc. 【0086】 Preparations containing antimicrobial compositions can be manufactured by conventional methods using one or more additives commonly used in pharmaceuticals, cosmetics, and other formulations, such as excipients, fillers, binders, wetting agents, disintegrants, lubricants, surfactants, dispersants, buffers, preservatives, solubilizers, antiseptics, flavoring and odor-masking agents, analgesics, stabilizers, and isotonic agents. The antimicrobial composition may also contain other active ingredients different from psLAAO. 【0087】 Soluble psLAAO obtained from the seeds or callus of transformed plants is less likely to contain sensitizing substances such as endotoxins compared to methods using microorganisms such as yeast. Furthermore, even if it does contain such substances, soluble psLAAO with reduced sensitizing properties can be obtained by applying the technology described in Patent Document 1. Therefore, by using soluble psLAAO with low sensitizing properties as the active ingredient, the sensitizing properties of the antimicrobial composition can be reduced. Due to its low sensitizing properties, such an antimicrobial composition can be used as a topical agent applied to the skin or eyes, and preferably as a topical agent applied to human skin or eyes. 【0088】 Antimicrobial compositions, which are topical skin preparations, can be used, for example, by applying them to the skin. Examples of topical skin preparations include medical topical skin preparations and cosmetics. The dosage form of a topical skin preparation can be any form that can be applied to the skin, and examples include topical preparations such as gels, creams, ointments, liquids, and lotions, and adhesive preparations such as poultices, tapes, and patches. 【0089】 Antimicrobial compositions used as ophthalmic topical agents can be used by local administration to a target organism, preferably to a specific area of ​​the eye such as the anterior side of the eye, the underside of the upper eyelid, or the upper side of the lower eyelid, or by dropping them into the eye. The dosage form of the ophthalmic topical agent can be any dosage form that can be applied to the eye, and examples include aqueous eye drops, aqueous suspension eye drops, viscous eye drops, non-aqueous eye drops, non-aqueous suspension eye drops, emulsion eye drops, eye ointments, eye creams, Tenon's injections, and fundus injections. 【0090】 If sensitizing properties are observed in soluble psLAAO, it is preferable to subject the soluble psLAAO to a sensitization reduction treatment. For example, in accordance with the description in Patent Document 1, any chromatographic method such as affinity chromatography, cation exchange chromatography, anion exchange chromatography, mixed-mode chromatography, HPLC, or electrophoresis is used, preferably affinity chromatography, and more preferably affinity chromatography using a polylysine-containing material such as a column packed with poly-L-lysine resin is used. 【0091】 The reduction in sensitization (confirmation of purity) can be confirmed by comparing the expression levels of the cell surface markers CD54 and / or CD86 when soluble psLAAO (e.g., supernatant and cell extract) before the reduction process is added to a human monocytic leukemia cell line (THP-1) with the expression levels of the cell surface markers CD54 and / or CD86 when soluble psLAAO after the reduction process is added to a human monocytic leukemia cell line (THP-1), and confirming that the expression levels after the reduction process are lower. Alternatively, the reduction in endotoxin can be confirmed by a standard endotoxin measurement method, such as the Limulus test. 【0092】 As for the amount of sensitization reduction, for example, it is preferable that the content of the sensitizing substance in the system after the reduction process is 1 / 2 or less, more preferably 1 / 4 or less, even more preferably 1 / 10 or less, and even more preferably substantially absent, compared to the content of the sensitizing substance in the system before the reduction process. 【0093】 As for the degree of sensitization reduction, for example, it is preferable that the system containing soluble psLAAO after the reduction step is represented by at least one selected from the group consisting of (1) and (2) below: (1) Effective concentration (EC) of cell surface marker CD54 expression in human monocytic leukemia cell line (THP-1) ２００(2) The effective concentration (EC) of the cell surface marker CD86 expression in human monocytic leukemia cell line (THP-1) is 0.35 μg / mL or more, more preferably 1.2 μg / mL or more, and even more preferably 3.0 μg / mL or more. １５０ The concentration is 4.1 μg / mL or higher, more preferably 6.0 μg / mL or higher, and even more preferably 8.0 μg / mL or higher. 【0094】 Here, the effective concentration (EC) ２００ ) refers to the concentration (μg / mL) of soluble psLAAO that results in a 200% increase in CD54 expression compared to a control in the absence of soluble psLAAO. Similarly, the effective concentration (EC) １５０ This refers to the concentration of soluble psLAAO (μg / mL) at which CD86 expression is increased by 150% compared to a control in the absence of soluble psLAAO. 【0095】 Effective concentration (EC) related to human sensitization ２００ ) and effective concentration (EC １５０ This can be measured and evaluated in accordance with OECD Guideline 442E "In vitro skin sensitization: Human cell line activation test (h-CLAT)" for the testing of chemical substances (http: / / www.nihs.go.jp / hse / chem-info / oecd / tgj / tg442ej.pdf). For details, refer to the method described in Patent Document 1. 【0096】 The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these examples, and can take various forms as long as it can solve the problems of the present invention. 【0097】 [1. Preparation of Binary Vectors pHPT / UbiP / LAAO / KDEL] pKS221MCS and p35SHPTAg7-GW, as described in the literature by Wakasa et al. (Wakasa et. al., Plant Biotechnology Journal (2006) 4: pp. 499-510), were used as basic vectors. Figure 1 shows the vector map representing the gene structure of p35SHPTAg7-GW. 【0098】Two sets of DNA fragments (UbiP / LAAO / KDEL) containing, in this order, the maize ubiquitin promoter (ZmUbi pro), the flathead flounder (Platicithes stellatus) L-amino acid oxidase (psLAAO; codon-optimized for rice), a 6×His-tag (6His) consisting of six histidine residues, an endoplasmic reticulum retention signal (KDEL), and the rice glutelin B1 gene terminator (GluB1 T) were inserted into the multi-cloning site (MCS) of pKS221MCS. ２ By inserting them, a DNA construct pUbiP / LAAO / KDEL was obtained. 【0099】 By homologous recombination of the obtained pUbiP / LAAO / KDEL and p35SHPTAg7-GW, the above DNA fragment (UbiP / LAAO / KDEL) was inserted between the CaMV 35S promoter (35S pro) and the right T-DNA border repeat sequence (RB) in p35SHPTAg7-GW. ２ A DNA construct pHPT / UbiP / LAAO / KDEL containing the above was prepared. The gene composition of DNA construct pHPT / UbiP / LAAO / KDEL from the left T-DNA border repeat sequence to the right T-DNA border repeat sequence is shown in Figure 2. In Figure 2, Ag7 T indicates the Agrobacterium gene 7 terminator, and HPT indicates the hygromycin phosphotransferase gene. 【0100】 [2. Preparation of binary vectors pHPT / UbiP / LAAO / KDEL / 2b and pHPT / UbiP / LAAO / KDEL / p19] A DNA cassette (UbiP / LAAO / KDEL), ２ and a DNA fragment (35S pro / 2b / Nos T) containing, in this order, the 35S pro, the Cucumber mosaic virus 2b protein (2b), and the NOS terminator (Nos T) were inserted into the multi-cloning site (MCS) of pKS221MCS to obtain a DNA construct pUbiP / LAAO / KDEL / 2b. 【0101】By homologous recombination of the obtained pUbiP / LAAO / KDEL / 2b and p35SHPTAg7-GW, the above DNA fragment (UbiP / LAAO / KDEL) is formed between 35S pro and RB in p35SHPTAg7-GW. ２ DNA construct pHPT / UbiP / LAAO / KDEL / 2b containing (35S pro / 2b / Nos T) was prepared. Similarly, DNA construct pHPT / UbiP / LAAO / KDEL / p19 was prepared by using p19 protein of tombus viruses (p19) instead of 2b. 【0102】 Figure 3 shows the gene structure from the left T-DNA boundary repeat to the right T-DNA boundary repeat for two types of binary vectors, (A) pHPT / UbiP / LAAO / KDEL / 2b and (B) pHPT / UbiP / LAAO / KDEL / p19. 【0103】 [3. Method for producing transformed rice callus using Agrobacterium] Figure 4 shows photographs of rice seeds, callus, and panicles at each step in the process of producing transformed rice callus. Tables 1A to 1C show the composition of the culture medium used in this example. 【0104】 【0105】 【0106】 【0107】 (3-1) Preparation of competent cells: Agrobacterium tumefaciens strain EHA105 was pre-cultured in LB-Mg agar medium at 30°C for 2 days. Then, a single colony from the agar medium was transferred to 1 mL of LB-Mg liquid medium and cultured at 30°C for 24 hours. 【0108】 Next, transfer 0.5 mL of culture medium to 50 mL of YEB-Mg liquid medium, and OD ６００The culture was incubated at 30°C for 2 hours until the concentration reached 0.6 to 1.2. The culture solution was then subjected to centrifugation at 6,000 rpm, 4°C, for 5 minutes to collect the bacterial cells. 【0109】 The collected bacterial cells were suspended in 10 mL of ice-cold 10% glycerol and then washed by centrifugation. This process was repeated twice. 【0110】 The washed bacterial cells were suspended in 250 μL of 10% glycerol, dispensed into 40 μL portions, and stored at -80°C. 【0111】 (3-2) 1-2 μL of binary vector was added to 40 μL of electroporation-competent cell and placed in an ice-cold cuvette (0.2 cm). The cuvette containing the competent cell and binary vector was then subjected to electroporation using an electric pulse ("E. coli pulser", Bio-rad; 2.5 kV) (one pulse). 【0112】 Next, 1 mL of YEB-Mg liquid medium was added to the contents of the cuvette to suspend it, and then it was incubated at 28°C for 1 hour. 【0113】 100 μL of culture medium was spread onto YEB-Mg agar medium containing spectinomycin at a final concentration of 100 μg / mL, and then incubated at 28°C for 2 days. Transformed Agrobacterium were obtained in this manner. 【0114】 (3-3) Rice seeds (paddy; 14 seeds per petri dish; Figure 4 (A)) for callus induction from seeds, sterile water, 70% ethanol, N6D agar medium, and a rice hulling machine were prepared. 【0115】 The hulls were removed from the seeds using a rice hulling machine. After hulling, discolored, deformed, and moldy seeds were discarded, and normal seeds were selected. 【0116】 The seeds were sterilized by adding 70% ethanol to the centrifuge tube containing the seeds and letting it stand for 30 seconds. Then, after removing the ethanol, the seeds were treated with hypochlorite by adding 2:1 diluted sodium hypochlorite to the centrifuge tube and shaking it for 30 minutes. 【0117】 Next, after removing the sodium hypochlorite, sterile water was added to the centrifuge tube to rinse the seeds. This was repeated five times. 【0118】 The seeds were placed on N6D agar medium in a petri dish with the embryo facing upwards (Figure 4(B)). The petri dish was then closed and sealed with surgical tape (3M). 【0119】 Sealed petri dishes were cultured under the conditions of 16 hours in light / 8 hours in darkness, 32°C, for 10 days, and then transferred to a fresh culture medium every 10 days for callus induction treatment (total of 30 days). Callus was obtained from the seeds through this callus induction treatment (Figure 4(C)). 【0120】 (3-4) The endosperm and bud portions of the pre-cultured callus seeds, fine callus, white callus, hard callus, and discolored callus were removed, and good quality callus was selected and transferred to new N6D solid medium. The petri dish was closed and sealed with surgical tape. 【0121】 (3-5) Pre-culture of transformed Agrobacterium AB agar medium was prepared. Spectinomycin-containing AB agar medium and spectinomycin-containing YEB-Mg liquid medium were also prepared by adding spectinomycin to a final concentration of 100 μg / mL. 【0122】 Transformed Agrobacterium cultured in YEB-Mg liquid medium containing spectinomycin was spread onto AB agar medium containing spectinomycin and cultured at 28°C in the dark for 3 days. 【0123】 Transformed Agrobacterium were cultured in YEB-Mg liquid medium containing spectinomycin, and the resulting culture medium was added to 70% sterile glycerol and stored at -80°C. 【0124】 (3-6) Infection treatment and co-culture of transformed Agrobacterium AAM medium (30 mL / 1 construct), acetosyringone (stock solution 2 mg / ml: acetosyringone dissolved in 100% DMSO), sterile Kimwipes and 2N6AS agar medium were prepared. 【0125】Approximately half of the transformed Agrobacterium, cultured on AB agar for 3 days, was taken using a small spatula moistened with AAM medium (without acetosyringone). This was then added to 30 mL of acetosyringone-containing AAM medium in a 50 mL centrifuge tube. The centrifuge tube was then stirred with a vortex mixer to suspend the transformed Agrobacterium in the acetosyringone-containing AAM medium, thereby obtaining an Agrobacterium suspension. The acetosyringone-containing AAM medium was prepared by adding 300 μL of acetosyringone stock solution to 30 mL of AAM medium and then adding acetosyringone to achieve a final concentration of 20 μg / mL. 【0126】 Agrobacterium suspension was poured over pre-cultured callus placed in a stainless steel mesh (Figure 4(D)). The callus was gently mixed by shaking the mesh, and then the mesh containing the callus was left to stand for 3 minutes to treat it with Agrobacterium infection. 【0127】 After standing, the mesh was placed on a sterile Kimwipe to completely remove any excess bacterial suspension. The callus in the mesh was transferred to 2N6AS agar (containing 20 μg / mL acetosyringone) and cultured at 24°C in the dark for 2 days to obtain Agrobacterium-treated callus (Figure 4(E)). 【0128】 (3-7) Selection of transformed callus N6D medium (containing antibiotics: meropenem 50 μg / mL, hygromycin B 50 μg / mL; hereafter referred to as antibiotic-containing N6D medium) was prepared. 【0129】 Agrobacterium-treated callus was transferred to antibiotic-containing N6D medium without washing. Approximately 14 callus cells were placed per petri dish. 【0130】 Agrobacterium-treated callus was cultured under the conditions of 16 hours in light / 8 hours in darkness, 32°C, for one week, and then transferred to a fresh culture medium every week for a total of 3-4 weeks. After culture, callus that continued to grow was selected to obtain transformed callus (Figure 4(F)). 【0131】[4. Redifferentiation of Transgenic Rice Callus and Recovery of Rice Seeds] Selected transgenic callus was subcultured in antibiotic-containing N6D medium, and then redifferentiated using a redifferentiation medium containing plant hormones (naphthaleneacetic acid (NAA), kinetin) (Figure 4(G)). 【0132】 The redifferentiated plants were transplanted into agripots containing hormone-free MS medium (Figure 4(H)) and cultured in a growth chamber at 32°C for 6 days under conditions of 16 hours of light / 8 hours of darkness. When sprouts and roots emerged, they were transplanted into soil and acclimatized in a tabletop greenhouse (with LEDs) at 24°C, 60% humidity, for 6 days under conditions of 12 hours of light / 12 hours of darkness. They were then cultivated for approximately 20-30 days in an LED-irradiated growth chamber or indoor greenhouse (Figure 4(I)). Seeds were collected from heading rice plants and stored (Figure 4(J)). 【0133】 [5. Purification of recombinant psLAAO derived from transformed rice callus] Transformed callus was ground in liquid nitrogen using a mortar and pestle. The ground callus was then purified using 50 mM NaH ２ PO ４ The callus solution was dissolved in 20 mL of a buffer solution (pH 7.0) containing 1 M ammonium sulfate. The callus solution was subjected to centrifugation to obtain a crude protein solution as the supernatant. 【0134】 Crude protein solution, hydrophobic column ("Capto ＴＭ The solution is passed through "High sub" (manufactured by Cytiva), followed by 50 mM NaH ２ PO ４ Elution was performed with an aqueous solution (pH 7.0). At this time, a Ni-NTA column ("Ni-NYA Agarose"; manufactured by Fujifilm Wako Pure Chemical Industries; resin: 5 mL slurry) was linked to a hydrophobic column. 【0135】 The Ni-NTA column was washed with Ni-NTA wash buffer, and then eluted with Ni-NTA elution buffer (fractionated solution: 1 mL / tube; total 10 tubes). 【0136】[6. Evaluation Method for Purified Recombinant psLAAO] (6-1) Evaluation of Antimicrobial Activity of Recombinant psLAAO Fraction and Recombinant psLAAO Purified Solution The antimicrobial activity of the fraction was evaluated by a halo test using methicillin-resistant Staphylococcus aureus (MRSA). In the antimicrobial test, MRSA 1 × 10⁶ was placed on TSA medium in a petri dish. ６ CFU was sprinkled, then holes were made in the culture medium, and 20 μL of the fractionated solution was added to each hole. The petri dish was then covered and incubated at 37°C for 16 hours, and the presence or absence of antibacterial activity was evaluated by visually confirming the presence of a halo. 【0137】 Furthermore, the recombinant psLAAO fraction, which showed antibacterial activity, was collected and subjected to dialysis and concentration according to conventional methods to obtain a recombinant psLAAO purified solution. The antibacterial activity of the recombinant psLAAO purified solution was evaluated by performing a halo test. 【0138】 (6-2) Separation and Purification Evaluation of Recombinant psLAAO The recombinant psLAAO purified solution was subjected to SDS-PAGE treatment to separate recombinant psLAAO. The SDS-PAGE after treatment was subjected to Western blotting using anti-psLAAO1 antibody and anti-His antibody. 【0139】 (6-3) Evaluation of the mechanism of action of purified recombinant psLAAO In order to confirm that the mechanism of action of the antibacterial activity of recombinant psLAAO is based on the generation of hydrogen peroxide, a halo test was performed using recombinant psLAAO purified solution + 20 mM Tris-HCl (pH 7.0), recombinant psLAAO purified solution + catalase solution, and recombinant psLAAO purified solution + anti-psLAAO1 antibody solution. 【0140】 [7. Evaluation Results of Transformed Callus Using pHPT / UbiP / LAAO / KDEL] Figures 5 to 7 show the results of evaluating the antibacterial activity, separation and purification, and mechanism of action of each recombinant psLAAO solution derived from transformed callus obtained using pHPT / UbiP / LAAO / KDEL. 【0141】As shown in Figure 5(A), fractions No. 3 to 7 of the recombinant psLAAO fractions showed antibacterial activity against MRSA. Furthermore, as shown in Figure 5(B), the recombinant psLAAO purified solution obtained by collecting, dialyzing, and concentrating the fractions that showed antibacterial activity also showed antibacterial activity against MRSA. 【0142】 As shown in Figure 6, the presence of recombinant psLAAO in the recombinant psLAAO purified solution was confirmed, and it was also confirmed that recombinant psLAAO could be separated and purified. 【0143】 As shown in Figure 7, the antibacterial activity of recombinant psLAAO was inhibited by catalase and neutralized by anti-psLAAO1 antibody. These results indicate that the antibacterial activity of the recombinant psLAAO purified solution is due to the psLAAO it contains, and that psLAAO exerts its antibacterial activity by generating hydrogen peroxide. 【0144】 Of the transformed calluses obtained using pHPT / UbiP / LAAO / KDEL, 44 clones were selected that highly expressed recombinant psLAAO. Of these, clone 34, which had the highest expression level of recombinant psLAAO, yielded 223 μg of recombinant psLAAO from 1.756 g of callus (44.7 μg / mL). Therefore, 127 μg of recombinant psLAAO could be recovered per 1 g of transformed callus. 【0145】 Clone 34 callus (0th generation callus) was redifferentiated and cultivated to produce ears of grain, and seeds were collected. The obtained seeds were used to induce callus formation to obtain 1st generation callus. Figure 8 shows the results of halo tests conducted on recombinant psLAAO fractions obtained from the 0th generation and 1st generation callus. 【0146】As shown in Figure 8, the recombinant psLAAO fraction obtained from the 0th generation callus showed antibacterial activity, while the recombinant psLAAO fraction obtained from the 1st generation callus showed almost no antibacterial activity. From this, it was found that it is difficult to produce recombinant psLAAO from seeds obtained by redifferentiation of transformed callus obtained using pHPT / UbiP / LAAO / KDEL. 【0147】 [8. Evaluation results of transformed callus using pHPT / UbiP / LAAO / KDEL / 2b and pHPT / UbiP / LAAO / KDEL / p19] Figure 9 shows the results of evaluating the antibacterial activity of recombinant psLAAO purified solution derived from eight clones of transformed callus obtained using pHPT / UbiP / LAAO / KDEL / 2b and one clone of transformed callus obtained using pHPT / UbiP / LAAO / KDEL / p19. 【0148】 As shown in Figure 9, antibacterial activity was confirmed in recombinant psLAAO purified solutions derived from all clones. 【0149】 Of the clones of transformed callus obtained using pHPT / UbiP / LAAO / KDEL / 2b, clone No. 05 was redifferentiated and cultivated to produce ears, and seeds were collected. The obtained seeds were used to induce callus formation, and first-generation callus clones (20 clones) were obtained. Figure 10A shows the results of a halo test (using a disk instead of a cup) and Western blotting using anti-psLAAO1 antibody on the recombinant psLAAO purified solution obtained from this first-generation callus. Furthermore, Figure 10B shows the results of a halo test performed using recombinant psLAAO purified solution obtained from second-generation callus corresponding to clones No. 3, No. 5, and No. 10 of the first-generation callus clones. 【0150】 As shown in Figures 10A and 10B, it was found that recombinant psLAAO purified solutions derived from any clone possessed antibacterial activity and contained recombinant psLAAO, although there were differences in the strength of activity and expression. 【0151】On the other hand, the antimicrobial activity of recombinant psLAAO purified solution derived from five clones (No. 34, 40, 43, 47, and 62) of transformed callus obtained using pHPT / UbiP / LAAO / KDEL / p19 was evaluated. Furthermore, clone No. 40 was redifferentiated and cultivated to produce ears, and seeds were collected. Callus induction was performed on the obtained seeds to obtain first-generation callus clones (6 clones). The antimicrobial activity of recombinant psLAAO purified solution obtained from this first-generation callus was evaluated. The evaluation results of the antimicrobial activity of (A) the 0th generation clones and (B) the 1st generation clones are shown in Figure 11. 【0152】 As shown in Figure 11, both the 0th generation clones and the 1st generation clones exhibited antibacterial activity in the recombinant psLAAO purified solution. 【0153】 These results show that recombinant psLAAO with antibacterial activity can be produced by using transformed callus with pHPT / UbiP / LAAO / KDEL / 2b and pHPT / UbiP / LAAO / KDEL / p19, and furthermore, recombinant psLAAO with antibacterial activity can be produced from the next generation callus of the transformed callus. In addition, since the seeds can be stored, recombinant psLAAO can be easily produced when needed. 【0154】 [9. Structural Evaluation of psLAAO] Wild-type psLAAO1-3 consist of the amino acid sequences described in SEQ ID NOs: 1-3, respectively. The three-dimensional structure of psLAAO1 was estimated using ColabFold v1.5.5:AlphaFold2 using MMseqs2. The estimated three-dimensional structure of psLAAO1 is shown in Figure 12. 【0155】 As shown in Figure 12, psLAAO has a signal peptide sequence at the N-terminus and an α-helix structure sequence at the C-terminus. For example, psLAAO1 has a signal peptide sequence from position 1 (M) to position 27 (A) and an α-helix structure sequence from position 506 (S) to position 522 (F). 【0156】Furthermore, the cysteine ​​at position 37 (C37) and position 200 (C200) of the amino acid sequence of psLAAO are linked by disulfide bonds, contributing to the maintenance of the psLAAO structure, and thus are presumed to contribute to its antibacterial activity. 【0157】 Based on this information, Figure 13 shows the estimated three-dimensional structures of psLAAO1(29-522) with the sequences at positions 1 to 27 and leucine (L28) at position 28 excluded, and psLAAO1(35-505) with the sequences at positions 1 to 34 and 506 to 522 excluded. As shown in Figure 13, psLAAO1(29-522) well preserves the structure of psLAAO1(1-522) which has the complete sequence. 【0158】 Furthermore, the antimicrobial activity of isolated and purified psLAAO1 (29-522) and psLAAO1 (35-505) was evaluated in the same manner as described in (6-1) above. As a result, inhibition zones were confirmed in both cases, indicating that the recombinant psLAAO1 possesses antimicrobial activity. The sequence identity between psLAAO1 (35-505) and the wild-type psLAAO1 (1-522) was 90.2%. 【0159】 From these results, it was found that recombinant psLAAO, which possesses C37 and C200 of wild-type psLAAO1 and closely conserves the structure of wild-type psLAAO1 (except for the N-terminal signal peptide sequence and the C-terminal α-helix structure sequence), has antibacterial activity. Structural similarity can be quantified using structural alignment with PyMOL 3.0 (open source). For example, using PyMOL 3.0, the RMSD (root mean square deviation) of the predicted structures of psLAAO1 (35-505) and wild-type psLAAO1 (1-522) was 0.18, indicating high homology. 【0160】 [10. Evaluation of Sensitization Potency of Purified Recombinant psLAAO] To evaluate the sensitization potency of recombinant psLAAO1, the effective concentration of CD54 expression (EC) was determined by the method described in <Examples> of Patent Document 1. ２００ ) and effective concentration of CD86 expression (EC １５０ The following values ​​were calculated for each of them. 【0161】 As a result, (1) the effective concentration of the cell surface marker CD54 expression in the human monocytic leukemia cell line (THP-1) (EC ２００ (2) The effective concentration (EC) of the cell surface marker CD86 expression in human monocytic leukemia cell line (THP-1) is 4.88 μg / mL. １５０ The concentration was 9.70 μg / mL. Recombinant psLAAO derived from transformed rice callus had significantly lower human sensitization potential than recombinant psLAAO derived from transformed yeast. 【0162】 [11. Sequence Listing] The sequences listed in the sequence listing are shown in Tables 2A to 2C below. 【0163】 【0164】 【0165】 Cross-reference of related applications 【0166】 This application claims priority to Japanese Patent Application No. 2024-218818, filed on 13 December 2024, the entire contents of which are incorporated herein by reference. Furthermore, the entire contents of all documents referenced in the detailed description of the invention of this application, including Patent Document 1 and Non-Patent Documents 1-2, are incorporated herein by reference.

Claims

1. A transgenic plant in which the soluble flounder (Platicithys stellatus) L-amino acid oxidase (psLAAO) gene and the viral RNA silencing repressor gene have been introduced as foreign genes, and which expresses the introduced genes.

2. The transformed plant according to claim 1, wherein the soluble psLAAO is psLAAO having an endoplasmic reticulum anchoring signal peptide or psLAAO from which the secretion signal peptide sequence has been removed.

3. The transformed plant according to claim 1, wherein the viral RNA silencing inhibitor is at least one protein selected from the group consisting of Cucumber mosaic virus 2b protein, Tombus virus p19 protein, Turnip crinkle virus p38 protein, and Flockhouse virus B2 protein.

4. The transformed plant according to claim 1, wherein the transformed plant is transformed rice, transformed strawberry, or transformed soybean.

5. Seeds or callus of a transformed plant according to any one of claims 1 to 4.

6. A method for producing soluble psLAAO, comprising the step of obtaining soluble psLAAO from the seeds or callus of a transformed plant described in claim 5.

7. A DNA fragment comprising, or a combination of, a first DNA fragment containing a soluble psLAAO gene and a second DNA fragment containing a viral RNA silencing suppressor gene.

8. The DNA fragment according to claim 7, wherein the soluble psLAAO is a psLAAO having an endoplasmic reticulum anchoring signal peptide or a psLAAO from which a secretion signal peptide sequence has been removed.

9. The DNA fragment according to claim 7, wherein the viral RNA silencing inhibitor is at least one protein selected from the group consisting of cucumber mosaic virus 2b protein, tombus virus p19 protein, turnip crinkle virus p38 protein, and flockhouse virus B2 protein.

10. A method for producing a transformed plant that expresses soluble psLAAO, comprising the step of obtaining a transformed plant that expresses soluble psLAAO by introducing a DNA fragment described in any one of claims 7 to 9 as an exogenous gene into a plant.

11. An antimicrobial composition comprising soluble psLAAO obtained from the seeds or callus of the transformed plant described in claim 5 as an active ingredient.

12. The antimicrobial composition according to claim 11, wherein the antimicrobial composition is used for antimicrobial action against at least one selected from the group consisting of Staphylococcus epidermidis, Staphylococcus aureus, Vibrio parahaemolyticus, methicillin-resistant Staphylococcus aureus, Pseudomonas aeruginosa, Streptococcus pyogenes, Propionibacterium acnes, Acinetobacter, Trichophyton, Yersinia cyanocarcinoma, Escherichia coli, and Enterococcus.

13. The antimicrobial composition according to claim 11, wherein the antimicrobial composition is used as a topical agent applied to the skin or eyes.

14. A method for preventing or treating an infectious disease, comprising administering a soluble psLAAO obtained from a transformed plant according to any one of claims 1 to 4 to a target organism.

15. An antimicrobial composition having antimicrobial activity, comprising soluble Platicitys stellatus L-amino acid oxidase (psLAAO) as an active ingredient, wherein the antimicrobial activity is antimicrobial activity against at least one selected from the group consisting of Staphylococcus epidermidis, Staphylococcus aureus, Vibrio parahaemolyticus, methicillin-resistant Staphylococcus aureus, Pseudomonas aeruginosa, Streptococcus pyogenes, Propionibacterium acnes, Acinetobacter, Trichophyton, Yersinia cyanobacteria, Escherichia coli, and Enterococcus.

16. The antimicrobial composition according to claim 15, wherein the antimicrobial activity is antimicrobial activity against at least one selected from the group consisting of Staphylococcus aureus, Pseudomonas aeruginosa, Streptococcus pyogenes, Propionibacterium acnes, Acinetobacter, Trichophyton, Yersinia cyanocarcinoma, Escherichia coli, and Enterococcus.

17. The antimicrobial composition according to claim 15, wherein the soluble psLAAO is psLAAO having an endoplasmic reticulum anchoring signal peptide or psLAAO from which the secretion signal peptide sequence has been removed.

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