Saccharomyces cerevisiae strain exhibiting acid resistance, bile resistance and adhesion to microplastics, and use thereof

Abstract

The present invention relates to a Saccharomyces cerevisiae strain, which meets the conditions of a prebiotic, such as acid resistance and bile resistance, and exhibits excellent adhesion to microplastics, thereby having the effect of removing microplastics in the environment and microplastics in living organisms.

Description

Saccharomyces cerevisiae strains having acid resistance, bile resistance, and microplastic adhesion and their uses

[0001] The present invention relates to a Saccharomyces cerevisiae strain, and more particularly, to a Saccharomyces cerevisiae strain having acid resistance, bile resistance, and microplastic adhesion ability.

[0002]

[0003] Plastics, which have excellent properties such as plasticity, flexibility, heat and electrical insulation, and corrosion resistance, are also inexpensive to produce, so their use is gradually increasing across all aspects of life, and as a result, the amount of plastic being discarded is also increasing.

[0004] Discarded plastics break down into small pieces due to ultraviolet rays, weathering, and other factors, decomposing to the point where they are invisible to the naked eye. They persist widely in nature, including soil, air, and the ocean. They are absorbed by plants and animals, and instead of decomposing, they accumulate in their bodies, significantly impacting marine life, humans, and the Earth's ecosystem. Microplastics, in particular, can combine with various toxins (heavy metals, PAHs, PCBs, OCPs, PBDEs, etc.) and harmful microorganisms (Vibrio parahaemolyticus, etc.), posing a risk of toxicity in living organisms.

[0005] The need to control these microplastics continues to be raised, but their extremely small size makes detection and separation difficult. Therefore, recent efforts have focused on developing new biodegradable microplastics or discovering new microorganisms capable of decomposing microplastics.

[0006] Patent Document 1 (Publication No. 10-2023-0012307) discloses an Enterobacter spp. strain with polyethylene-decomposing activity, but its effectiveness is not at a level that is commercially successful, and there is a problem that it cannot be applied to microplastics that have already entered the body from landfills or are present in food.

[0007] Therefore, continuous research is needed to identify new strains that are stable in vivo and can inhibit the absorption of microplastics.

[0008]

[0009] Considering the above problems, the purpose of the present invention is to provide a Saccahromyces cerevisiae strain having acid resistance, bile resistance, and microplastic adhesion ability.

[0010] More specifically, another object of the present invention is to provide a food composition for inhibiting microplastic absorption containing a Saccahromyces cerevisiae strain as an active ingredient.

[0011] More specifically, another object of the present invention is to provide a composition for adsorbing microplastics comprising a Saccahromyces cerevisiae strain.

[0012]

[0013] In order to achieve the above object, the present invention provides a Saccahromyces cerevisiae strain having acid resistance, bile resistance and microplastic adhesion ability, characterized in that it is any one selected from the group consisting of Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P. Provides.

[0014] The above strain may have a portion of the sequence deleted or substituted in at least one gene among the FLO11 gene and the FLO10 gene.

[0015] By the above deletion or substitution, the Saccahromyces cerevisiae strain may have a reduced biofilm forming ability.

[0016] The above Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, the above Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P and the above Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P may have a portion of the FLO11 gene and FLO10 gene sequence deleted or substituted, and the above Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P may have a portion of the FLO11 gene sequence deleted or substituted.

[0017] The above strain may be a strain in which the base sequence from the 390973rd base to the 391033rd or 391204th base in the FLO11 gene sequence of Saccahromyces cerevisiae S288C chromosome IX (position NC_001141.2) is deleted and replaced with T.

[0018] Among the above strains, the Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P is characterized by a deletion of the base sequence from the 390973rd base to the 391033rd base in the FLO11 gene sequence of S288C chromosome IX (position NC_001141.2) and a substitution with T, and the Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P may be a deletion of the base sequence from the 390973rd base to the 391204th base in the FLO11 gene sequence of S288C chromosome IX (position NC_001141.2) and a substitution with T.

[0019] The above strain may be a strain in which the sequence from the 647797th base to the 647906th base in the FLO10 gene sequence of Saccahromyces cerevisiae S288C chromosome IX (position NC_001143.9) is deleted and replaced with CC.

[0020] The above microplastics may be 1 to 500 nm.

[0021] In order to achieve the above other objects, the present invention provides a food composition for inhibiting microplastic absorption, comprising as an active ingredient any one Saccahromyces cerevisiae strain selected from the group consisting of Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P, and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P.

[0022] In order to achieve the above other objects, the present invention provides a composition for adsorbing microplastics, comprising any one Saccahromyces cerevisiae strain selected from the group consisting of Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P, and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P.

[0023]

[0024] The Saccahromyces cerevisiae strain of the present invention has the conditions as a prebiotic, such as acid resistance and bile resistance, and has excellent adhesion to microplastics, so it is effective in removing microplastics in the environment and in living organisms.

[0025] In addition, the Saccahromyces cerevisiae strain has a gene that forms biofilms deleted or replaced, so it does not form colonies (biofilms) in living organ tissues and is easily excreted, so it can be used as a functional strain that is biocompatible and removes intestinal microplastics.

[0026]

[0027] Figure 1 is a graph showing the polystyrene (PS) adhesion ability of 36 acid-resistant yeast strains selected in the first round. S. cerevisiae S288c (indicated as S288c in the figure) was used as a negative control group.

[0028] Figure 2 is a graph showing the polystyrene (PS) adhesion ability of four Saccharomyces cerevisiae strains according to the present invention and the negative control S. cerevisiae S288c strain analyzed by viable cell count. Figure 2a is a graph for S. cerevisiae ReY29-4_P16, Figure 2b is a graph for S. cerevisiae ReY29-5_P17, Figure 2c is a graph for S. cerevisiae ReY43-1_P18, and Figure 2d is a graph for S. cerevisiae ReY43-2_P19.

[0029] Figure 3 is a graph showing the polypropylene (PP) adhesion ability of four Saccharomyces cerevisiae strains according to the present invention and the negative control S. cerevisiae S288c strain analyzed by viable cell count. Figure 3a is a graph for S. cerevisiae ReY29-4_P16, Figure 3b is a graph for S. cerevisiae ReY29-5_P17, Figure 3c is a graph for S. cerevisiae ReY43-1_P18, and Figure 3d is a graph for S. cerevisiae ReY43-2_P19.

[0030] Figure 4 is a graph showing the adhesion ability of four strains of Saccharomyces cerevisiae and the negative control strain S. cerevisiae S288c to fluorescently labeled polystyrene (PS) beads (ultrafine plastics) according to size. Figure 4a is a graph for S. cerevisiaeReY29-4_P16, Figure 4b is a graph for S. cerevisiaeReY29-5_P17, Figure 4c is a graph for S. cerevisiaeReY43-1_P18, and Figure 4d is a graph for S. cerevisiaeReY43-2_P19.

[0031] Figure 5a is a graph showing the results of analyzing the efficacy of inhibiting in vivo microplastic absorption when the S. cerevisiae ReY29-5_P17 strain was orally administered. Figure 5b is a graph showing the results of analyzing the efficacy of inhibiting in vivo microplastic absorption when the S. cerevisiae ReY43-1_P18 strain was orally administered. Figure 5c is a graph showing the results of analyzing the efficacy of inhibiting in vivo microplastic absorption when the S. cerevisiae ReY43-2_P19 strain was orally administered.

[0032] Figure 6 is a graph showing the results of confirming the FLO11 gene expression level of Saccharomyces cerevisiae strains (S. cerevisiaeReY29-4_P16, S. cerevisiaeReY29-5_P17, S. cerevisiaeReY43-1_P18 and S. cerevisiaeReY43-2_P19) according to the present invention by RT-PCR, and the gene expression level is expressed as a relative gene expression level compared to the FLO11 gene expression level of the S. cerevisiaeS288c strain.

[0033]

[0034] Below, various aspects and various implementation examples of the present invention will be examined in more detail.

[0035] The purposes, other objectives, features, and advantages of the present invention will be readily understood through the following preferred embodiments, illustrated in the accompanying drawings. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. Rather, the embodiments presented herein are provided to ensure that the disclosure is thorough and complete, and to ensure that the spirit of the present invention is fully conveyed to those skilled in the art.

[0036] In this specification, terms such as “include” or “have” are intended to specify the presence of a feature, number, step, operation, component, part, or combination thereof described in the specification, but should be understood not to exclude in advance the possibility of the presence or addition of one or more other features, numbers, steps, operations, components, parts, or combinations thereof.

[0037] In this specification, when a range is described for a variable, the variable will be understood to include all values ​​within the described range including the described endpoints of the range. For example, the range "5 to 10" will be understood to include the values ​​5, 6, 7, 8, 9, and 10, as well as any subranges such as 6 to 10, 7 to 10, 6 to 9, 7 to 9, etc., and any value between integers that fall within the described range, such as 5.5, 6.5, 7.5, 5.5 to 8.5, and 6.5 to 9. Also, for example, a range of "10% to 30%" would be understood to include all integers up to and including 30%, as well as any subranges such as 10% to 15%, 12% to 18%, 20% to 30%, etc., and any value between reasonable integers within the stated range, such as 10.5%, 15.5%, 25.5%, etc.

[0038] In this specification, “A and / or B” means “A and B, or A or B.”

[0039]

[0040] Hereinafter, the present invention will be described in detail.

[0041]

[0042] One aspect of the present invention relates to a Saccahromyces cerevisiae strain having acid resistance, bile resistance, and microplastic adhesion, characterized in that the strain is any one selected from the group consisting of Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P, and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P.

[0043] The present invention is a strain isolated from traditional nuruk, which is safe for ingestion. Among the strains, four Saccahromyces cerevisiae strains were identified, which exhibit bile and acid resistance, as well as excellent adhesion to plastics such as polystyrene and polypropylene.

[0044] The four strains of Saccahromyces cerevisiae were identified as Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P, and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P, respectively, and deposited with the Korea Center for Microorganism Conservation (KCCM) on March 30, 2023, and designated as "KCCM They were assigned the accession numbers "13340P", "KCCM 13341P", "KCCM 13342P", and "KCCM 13343P".

[0045] The above Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340 contains a gene base sequence encoding 18S rRNA of sequence number 1. Specifically, it was identified as 18S NS1 and NS24.

[0046] [Sequence number 1]

[0047] CAGGAGTGGAGCCTGCGGCTTAATTTGACTCAACACGGGGAAACTCACCAGGTCCAGACACAATAAGGATTGACAGATTGAGAGCTCTTTCTTGATTTTGTGGTGGTGGTGCATGGCCGTTCTTAGTT GGTGGAGTGATTTGTCTGCTTAATTGCGATAACGAACGAGACCTTAACCTACTAAATAGTGGTGCTGCATTTGCTGGTTATCCACTTCTTAGAGGGACTATCGGTTTCAAGCCGATGGAAGTTTGAGGC AATAACAGGTCTGTGATGCCCTTAGACGTTCTGGGCCGCACGCGCGCTACACTGACGGAGCCAGCGAGTCTAACCTTGGCCGAGAGGTCTTGGTAATCTTGTGAAACTCCGTCGTGCTGGGGATAGAGCA TTGTAATTATTGCTCTTCAACGAGGAATTCCTAGTAAGCGCAAGTCATCAGCTTGCGTTGATTACGTCCCTGCCCTTTGTACACACCGCCCGTCGCTAGTACCGATTGAATGGCTTAGTGAGGCCTCAGG

[0048] The above Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P contains a gene base sequence encoding 18S rRNA of sequence number 2. Specifically, since S. cerevisiae is a fungi, it was identified as 18S NS1 and NS24.

[0049] [Sequence number 2]

[0050] GGAGCCTGCGGCTTAATTTGACTCAACACGGGGAAACTCACCAGGTCCAGACACAATAAGGATTGACAGATTGAGAGCTCTTTCTTGATTTTGTGGGGTGGTGGTGCATGGCCGTTCTTAGTTGGTGGAG TGATTTGTCTGCTTAATTGCGATAACGAACGAGACCTTAACCTACTAAATAGTGGTGGCTAGCATTTGCTGGTTATCCACTTCTTAGAGGGACTATCGGTTTCAAGCCGATGGAAGTTTGAGGCAATAAC AGGTCTGTGATGCCCTTAGACGTTCTGGGCCGCACGCGCGCTACACTGACGGAGCCAGCGAGTCTAACCTTGGCCGAGAGGTCTTGGTAATCTTGTGAAACTCCGTCGTGCTGGGGATAGAGCATTGTA ATTATTGCTCTTCAACGAGGAATTCCTAGTAAGCGCAAGTCATCAGCTTGCGTTGATTACGTCCCTGCCCTTTGTACACACCGCCCGTCGCTAGTACCGATTGAATGGCTTAGTGAGGCCTCAGGATCT

[0051] The above Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P contains a gene base sequence encoding 18S rRNA of sequence number 3. Specifically, since S. cerevisiae is a fungi, it was identified as 18S NS1 and NS24.

[0052] [Sequence number 3]

[0053] AGCCTGCGGCTTAATTTGACTCAACACGGGGAAACTCACCAGGTCCAGACACAATAAGGATTGACAGATTGAGAGCTCTTTCTTGATTTTGTGGGGTGGTGGTGCATGGCCGTTCTTAGTTGGTGGAGTGATTTGTCTG CTTAATTGCGATAACGAACGAGACCTTAACCTACTAAATAGTGGTTGCTAGCATTTGCTGGTTATCCACTTCTTAGAGGGACTATCGGTTTCAAGCCGATGGAAGTTTGAGGCAATAACAGGTCTGTGATGCCCTTAGA CGTTCTGGGCCGCACGCGCGCTACACTGACGGAGCCAGCGAGTCTAACCTTGGCCGAGAGGTCTTGGTAATCTTGTGAAACTCCGTCGTGCTGGGGATAGAGCATTGTAATTATTGCTCTTCAACGAGGAATTCCTAG TAAGCGCAAGTCATCAGCTTGCGTTGATTACGTCCCTGCCCTTTGTACACACCGCCCGTCGCTAGTACCGATTGAATGGCTTAGTGAGGCCTCAGGATCTGCTTAGAGAAGGGGGCAACTCCATCTCAGAGCGGAGAA

[0054] The above strain contains a gene sequence encoding 18S rRNA of sequence number 4. Specifically, since S. cerevisiae is a fungi, it was identified as 18S NS1 and NS24.

[0055] [Sequence number 4]

[0056] AGCCTGCGGCTTAATTTGACTCAACACGGGGAAACTCACCAGGTCCAGACACAATAAGGATTGACAGATTGAGAGCTCTTTCTTGATTTTGTGGGGTGGTGGTGCATGGCCGTTCTTAGTTGGTGGAGTG ATTTGTCTGCTTAATTGCGATAACGAACGAGACCTTAACCTACTAAATAGTGGTGGCTAGCATTTGCTGGTTATCCACTTCTTAGAGGGACTATCGGTTTCAAGCCGATGGAAGTTTGAGGCAATAACAGG TCTGTGATGCCCTTAGACGTTCTGGGCCGCACGCGCGCTACACTGACGGAGCCAGCGAGTCTAACCTTGGCCGAGAGGTCTTGGTAATCTTGTGAAACTCCGTCGTGCTGGGGATAGAGCATTGTAATT ATTGCTCTTCAACGAGGAATTCCTAGTAAGCGCAAGTCATCAGCTTGCGTTGATTACGTCCCTGCCCTTTGTACACACCGCCCGTCGCTAGTACCGATTGAATGGCTTAGTGAGGCCTCAGGATCTGCTT

[0057] All of the Saccahromyces cerevisiae strains according to the present invention may have acid resistance and bile resistance.

[0058] All of the Saccahromyces cerevisiae strains according to the present invention may have the ability to adhere to microplastics.

[0059] In this specification, 'acid resistance' means that it can exhibit a survival rate of 50% or more for 3 days even under conditions of pH 2 to 3, and 'bile resistance' means that it can exhibit a survival rate of 50% or more for 24 hours in an environment similar to human bile.

[0060] In this specification, "microplastics" refers to plastic particles (or fragments) measuring 5 mm or less in length or diameter, created during the degradation of plastic products by factors such as ultraviolet rays, weathering, and waves. Due to their small size, these microplastics are difficult to recover from the natural environment, gradually accumulating in the natural world (marine and soil ecosystems). Microplastics not only enter plants and animals through the food chain and reach humans, but can also be exposed to them through drinking water, food, and air. As they absorb various toxic elements during this process, concerns about the adverse effects of microplastics have arisen. Recently, it has been revealed that microplastics accumulate in the fetal brain through the maternal mammary glands, causing abnormal behaviors such as anxiety and depression, and lack of social skills. This finding confirms that microplastics pose a risk to brain health not only during neural development but also throughout life, and concerns about microplastics are growing.

[0061] In this specification, 'microplastic adhesion' refers to the ability to stick to microplastics and not fall off, and is also called microplastic adsorption ability. When the strain according to the present invention (any one or more selected from the group consisting of Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P, and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P) stably proliferates in the internal digestive organ, it adsorbs and attaches to microplastics entering the digestive organ, thereby preventing the microplastics from being absorbed into the body through the blood vessels in the digestive organ. In addition, the strain of the present invention forms less biofilm and is thus easily discharged from the inside of the body to the outside, thereby easily removing the adsorbed microplastics from the body.

[0062] In other words, microplastic adhesion can stick to any plastic material regardless of charge type or surface functional group, thereby preventing absorption into the body, enabling excellent microplastic removal.

[0063] The above microplastics are not particularly limited as long as they are created by wear and tear from general plastics due to physical, chemical, biological actions and weathering, or are generated during an industrial processing process, and specifically, they may be at least one selected from the group consisting of polyethylene (PE), polypropylene (PP), polystyrene (PS), polyethylene terephthalate (PET), polyvinyl chloride (PVC), polyvinyl alcohol (PVA), and mixtures thereof.

[0064] The size of the above microplastics is not particularly limited, but may be 5 mm or less, preferably have an average diameter of 1 to 1000 nm, and more preferably have an average diameter of 1 to 500 nm.

[0065] Any one or more strains selected from the group consisting of Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P according to the present invention may be used without limitation in fields requiring adsorption and / or removal of microplastics, such as soil, rivers, and seas contaminated with microplastics, but may preferably include a food composition, a feed composition, or a pharmaceutical composition.

[0066] At this time, the Saccahromyces cerevisiae strain having acid resistance, bile resistance and microplastic adhesion according to the present invention has a part of the sequence of a gene (FLO11 and / or FLO10) that forms a biofilm that inhibits the immune function in the body deleted or substituted, so that the overall biofilm forming ability is reduced and lowered. Preferably, among the strains, Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P may have a portion of the FLO11 gene and FLO10 gene sequence deleted or substituted, and Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P may have a portion of the FLO11 gene sequence deleted or substituted.

[0067] Among the technologies for removing microplastics, those that utilize microbial biofilms have been mentioned. However, these biofilms can interfere with the action of immune cells and antibiotics within the body, delaying the healing process and increasing susceptibility to bacterial and fungal infections. Furthermore, since microplastics are adsorbed to the biofilm after microbial colonies form within the body's tissues, they are not excreted from the body and remain in the tissues, leading to the persistent and prolonged accumulation of microplastics.

[0068] On the other hand, Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P according to the present invention have excellent adhesion to microplastics, but have reduced and decreased ability to form biofilms, so that they are easily discharged from the body, and thus can effectively prevent and block microplastics from being absorbed into the body.

[0069] According to one embodiment of the present invention, the strain may be one in which the base sequence from the 390973rd base to the 391033rd or 391204th base in the FLO11 gene sequence of Saccahromyces cerevisiae S288C chromosome IX (position NC_001141.2) is deleted and replaced with T. Preferably, among the above strains, the Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P has a base sequence deleted from the 390973rd base to the 391033rd base in the FLO11 gene sequence of S288C chromosome IX (position NC_001141.2) and substituted with T, and the Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P may be a deletion of the base sequence from the 390973rd base to the 391204th base in the FLO11 gene sequence of S288C chromosome IX (position NC_001141.2) and a substitution with T.

[0070] According to one embodiment of the present invention, the strain may be one in which the 647797th base sequence to the 647906th base sequence of the FLO10 gene sequence of Saccahromyces cerevisiae S288C chromosome IX (position NC_001143.9) is deleted and replaced with CC. Preferably, among the above strains, Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P may be those in which the 647797th base sequence to the 647906th base sequence in the FLO10 gene sequence of S288C chromosome Ⅸ (position NC_001143.9) is deleted and replaced with CC.

[0071]

[0072] Another aspect of the present invention relates to a food composition for inhibiting microplastic absorption, comprising as an active ingredient any one Saccahromyces cerevisiae strain selected from the group consisting of Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P, and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P.

[0073] The above food composition utilizes probiotic properties such as microplastic adhesion properties, acid resistance, and bile resistance of any one Saccahromyces cerevisiae strain selected from the group consisting of Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P, and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P.

[0074] With respect to the food composition of the present invention, the microplastics may be microplastics that enter or exist in the body through the intake of drinking water, food, food ingredients, etc., and may preferably be microplastics that enter or exist in the digestive organs, and most preferably, may be microplastics that enter or exist in the intestines.

[0075] Any one Saccahromyces cerevisiae strain selected from the group consisting of Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P is the same as described above. At this time, the strain may be a live cell or a dead cell, and preferably may be a live cell. Additionally, the Saccahromyces cerevisiae strain may or may not contain a culture.

[0076] The active ingredient, Saccahromyces cerevisiae strain, is included in a therapeutically effective amount or nutritionally effective concentration relative to the total weight of the above composition, which is 10 4 10 inland 16 CFU / g, preferably 10 6 10 inland 12 Contains a content of CFU / g or a culture containing an equivalent number of viable cells. Typically, for adult patients, 1×10 6 Viable bacteria greater than CFU / g, preferably 1×10 8 1×10 12 CFU / g of live bacteria can be administered once or in multiple doses.

[0077] In this specification, 'microplastic absorption' may refer to microplastics being absorbed and accumulated in the body through the digestive system through the mouth, or to microplastics accumulating in the body, causing human health problems (inflammation, absorption of toxic substances, metabolic disorders, endocrine disorders, decreased function of organs or tissues, respiratory problems, etc.) or causing damage to cells, tissues, or organs in the body.

[0078] In this specification, "inhibiting microplastic absorption" may mean protecting one or more cells or tissues selected from the brain, heart, lungs, liver, and kidneys from damage caused by microplastics being absorbed or accumulated in the digestive system when they enter the body through oral ingestion. Furthermore, it may mean inhibiting the accumulation of microplastics in one or more cells or tissues selected from the brain, heart, lungs, liver, and kidneys.

[0079] According to one embodiment of the present invention, the strain can inhibit orally administered microplastics from being absorbed from the digestive tract into the blood vessels and accumulating in the tissues of internal organs. Specifically, in one embodiment of the present invention, when microplastics were orally administered to a mouse model that had ingested the strain, the concentration of microplastics entering the blood vessels of the mouse model significantly decreased. On the other hand, when microplastics were orally administered to a mouse model that had not ingested anything, the concentration of microplastics in the blood vessels of the mouse model doubled. Through this, it was confirmed that the strain inhibits the accumulation of microplastics in the body.

[0080] The above 'food composition' comprises, as an active ingredient, any one Saccahromyces cerevisiae strain selected from the group consisting of Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P, in addition to food raw materials that can be used as food described in the standards and specifications for food commonly used in food manufacturing ('Food Code'), Includes food additives listed in the Food Additives Codex.

[0081] Although not particularly limited, examples thereof include proteins, carbohydrates, fats, nutrients, seasonings, and flavoring agents. The carbohydrates may include monosaccharides such as glucose, fructose, etc.; disaccharides such as maltose, sucrose, lactose, etc.; oligosaccharides or polysaccharides such as dextrin, corn syrup, cyclodextrin, etc.; and sugar alcohols such as xylitol, sorbitol, erythritol, etc. The flavoring agent may include natural flavoring agents [thaumatin, stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.]) and synthetic flavoring agents (saccharin, aspartame, etc.).

[0082] In the case of manufacturing a food composition using any one Saccahromyces cerevisiae strain selected from the group consisting of Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P as an active ingredient, the Saccahromyces cerevisiae strain absorbs microplastics. There is no need to specifically limit the content as long as it exhibits the effect of suppressing or absorbing microplastics, but it may be included in amounts of, for example, 0.1 to 99 wt%, 0.5 to 95 wt%, 1 to 90 wt%, 2 to 80 wt%, 3 to 70 wt%, 4 to 60 wt%, and 5 to 50 wt%.

[0083] In the above food composition, any one Saccahromyces cerevisiae strain selected from the group consisting of Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P, and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P, which is an effective ingredient, may be appropriately selected by a person skilled in the art, depending on the condition, weight, presence or absence of a disease, degree, and duration of the ingestion. For example, the daily dosage may be 1 to 5,000 mg, preferably 5 to 2,000 mg, more preferably 10 to 1,000 mg, even more preferably 20 to 800 mg, and most preferably 50 to 500 mg. The number of administrations need not be particularly limited, but can be adjusted by a person skilled in the art within the range of 3 times a day to once a week. In the case of long-term intake for the purpose of health and hygiene or health control, the dosage may be below the above range.

[0084] The above food composition need not be particularly limited, but may be, for example, a powder, granule, tablet, capsule, pill, extract, jelly formulation, tea bag formulation, or beverage formulation.

[0085] In addition, in order to suppress the absorption of microplastics in general foods, any one Saccahromyces cerevisiae strain selected from the group consisting of Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P may be added, and foods to which the addition is possible are not particularly limited, but for example, those stipulated in Article 7 of the Food Sanitation Act. It can be added to confectionery, bread or rice cakes, cocoa products or chocolates, meat or egg products, fish products, tofu or jelly products, noodles, tea, coffee, beverages, special-purpose foods, soy sauce, seasonings, dressings, kimchi, salted seafood, pickled foods, stewed foods, alcoholic beverages, dried products, and other foods as exemplified in the Food Standards and Ingredients Specifications for Livestock Products ('Food Code'). It can also be added to dairy products, meat products, packaged meat, and egg products as exemplified in the Livestock Products Sanitation Management Act's Processing Standards and Ingredients Specifications for Livestock Products ('Livestock Products Code').

[0086] Meanwhile, a food composition comprising any one Saccahromyces cerevisiae strain selected from the group consisting of Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P as an active ingredient can be used alone as a "health functional food for inhibiting absorption of microplastics" and "in vivo absorption of microplastics" Or it can be used as a "health functional food to reduce accumulation."

[0087] The above "health functional food" refers to a food manufactured (including processed) in accordance with legal standards using raw materials or ingredients with beneficial functions for the human body (Article 3, Paragraph 1 of the Health Functional Food Act). While the terminology and scope of the above "health functional food" may vary by country, it can be defined as a "dietary supplement" in the United States, a "food supplement" in Europe, a "health functional food" or "food for special health use (FoSHU)" in Japan, or a "health food" in China.

[0088] The above food composition or health functional food may additionally contain food additives, and its suitability as a food additive shall be determined in accordance with the specifications and standards for the relevant item in accordance with the general provisions and general test methods of the ‘Food Additives Code’, unless otherwise provided.

[0089] In addition, the health functional food may be used in combination with a health functional food material that can promote absorption of microplastics from a living body or suppress absorption of microplastics into a living body, together with any one Saccahromyces cerevisiae strain selected from the group consisting of Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P, and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P. Can be.

[0090]

[0091] Another aspect of the present invention provides a feed composition for inhibiting microplastic absorption or absorbing microplastics, comprising any one Saccahromyces cerevisiae strain selected from the group consisting of Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P, and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P.

[0092] Any one Saccahromyces cerevisiae strain selected from the group consisting of Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P is the same as described above.

[0093] The above feed composition for inhibiting microplastic absorption or absorbing microplastics can be prepared by adding any one Saccahromyces cerevisiae strain selected from the group consisting of Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P, and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P in an appropriate effective concentration range according to various feed preparation methods known in the art.

[0094]

[0095] Another aspect of the present invention provides a composition for preventing or treating a microplastic-related disease of any one Saccahromyces cerevisiae strain selected from the group consisting of Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P, and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P.

[0096] At this time, any one Saccahromyces cerevisiae strain selected from the group consisting of Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P is the same as described above.

[0097] When microplastics are absorbed orally, they travel through the intestinal cells into the bloodstream, and then through the bloodstream to various organs and tissues, including the lymphatic and hepatobiliary systems. These microplastics can cause one or more diseases selected from the group consisting of cardiovascular disease, endocrine disease, inflammatory disease, obesity, and metabolic disease.

[0098] The composition of the present invention can prevent or treat diseases related to microplastics by adsorbing or attaching microplastics and preventing the introduced microplastics from being absorbed into blood vessels.

[0099] The above cardiovascular disease may be at least one selected from the group consisting of myocardial infarction, atherosclerosis, atherothrombosis, coronary artery disease, stable and unstable angina, stroke, vascular stenosis, vascular restenosis, aortic aneurysm, and acute ischemic arteriovascular event.

[0100] The above endocrine disease may be any one selected from the group consisting of diabetes, thyroid disease (hypothyroidism and hyperthyroidism, thyroiditis, thyroid nodules), Cushing's syndrome, insulin resistance, and growth hormone deficiency.

[0101] The inflammatory disease may be any one selected from the group consisting of sepsis, septic shock, inflammatory bowel disease (IBD), peritonitis, nephritis, acute bronchitis, chronic bronchitis, osteoarthritis, enteropathic spondylitis, chronic obstructive pulmonary disease (COPD), rheumatoid arthritis, acute lung injury, and broncho-pulmonary dysplasia.

[0102] The above inflammatory bowel disease (IBD) may be ulcerative colitis (UC) or Crohn's disease.

[0103] The above metabolic disease may be any one selected from the group consisting of obesity, hypertension, arteriosclerosis, hyperlipidemia, fatty liver, non-alcoholic fatty liver disease, hyperinsulinemia, diabetes, and insulin resistance syndrome.

[0104] The pharmaceutical composition may further comprise a suitable carrier, excipient or diluent according to a conventional method. Carriers, excipients and diluents that may be included in the pharmaceutical composition of the present invention include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

[0105] In addition, the pharmaceutical composition according to the present invention can be formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories, or sterile injection solutions, respectively, according to conventional methods. Specifically, when formulating, it can be prepared using diluents or excipients such as commonly used fillers, bulking agents, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and such solid preparations can be prepared by mixing the pharmaceutical composition of the present invention with at least one excipient, such as starch, calcium carbonate, sucrose, lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc can also be used. Liquid preparations for oral administration include suspensions, solutions, emulsions, and syrups. In addition to commonly used simple diluents such as water and liquid paraffin, they may contain various excipients such as wetting agents, sweeteners, flavoring agents, and preservatives. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions can include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases can include witepsol, macrogol, Tween 61, cacao butter, laurin butter, and glycerogelatin.

[0106] The pharmaceutical composition of the present invention can be administered to mammals such as rats, mice, livestock, and humans through various routes.

[0107] The pharmaceutical composition of the present invention may be in any form suitable for the intended administration method. For the pharmaceutical composition of the present invention, administration refers to introducing a given substance into a patient through any suitable method. The pharmaceutical composition may be administered via any conventional route, as long as the drug can reach the target tissue.

[0108] The route of administration of the pharmaceutical composition according to the present invention may be oral or parenteral, but is not limited thereto, and the parenteral administration may include oral, intravenous, intramuscular, intraarterial, intramedullary, intraarticular, intrasynovial, intrasternal, intrathecal, intracardiac, transdermal, subcutaneous, intradermal, intraperitoneal, intranasal, enteral, topical, intracranial, intracerebroventricular, intrauterine, intrauterine epidural, sublingual, or rectal. The pharmaceutical composition of the present invention may be administered by any device capable of transporting the active ingredient to the target site, but is preferably administered orally.

[0109] The content of the active ingredient in the pharmaceutical composition can be appropriately adjusted depending on the purpose of use of the pharmaceutical composition, the form of the formulation, etc., and may be, for example, 0.001 to 99 wt%, 0.001 to 90 wt%, 0.001 to 50 wt%, 0.01 to 50 wt%, 0.1 to 50 wt%, or 1 to 50 wt% based on the total weight of the pharmaceutical composition, but is not limited thereto.

[0110] The pharmaceutical composition of the present invention may vary depending on various factors including the activity of the active ingredient used, age, body weight, general health, sex, diet, administration time, administration route, excretion rate, drug combination, and the severity of a specific disease to be prevented or treated, and the dosage of the pharmaceutical composition may vary depending on the patient's condition, body weight, degree of disease, drug form, administration route, and period, but may be appropriately selected by those skilled in the art, and may be administered at 0.0001 to 50 mg / kg or 0.001 to 50 mg / kg per day. Administration may be administered once a day or divided into several times. The dosage does not limit the scope of the present invention in any way. The pharmaceutical composition according to the present invention may be formulated as a pill, a dragee, a capsule, a liquid, a gel, a syrup, a slurry, or a suspension.

[0111]

[0112] Another aspect of the present invention relates to a composition for adsorbing microplastics, comprising any one Saccahromyces cerevisiae strain selected from the group consisting of Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P, and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P.

[0113] The composition may adsorb microplastics present in the natural environment. The natural environment is not particularly limited as long as it is contaminated with microplastics, but preferably includes at least one selected from the group consisting of soil, seawater, drinking water, and groundwater.

[0114] To determine whether the strain exhibited microplastic adhesion, the strain was incubated with microplastics. The strain was found to adhere to microplastics, confirming that the strain of the present invention possesses microplastic adhesion or adsorption. Furthermore, this microplastic adhesion was shown to exhibit excellent adsorption efficiency in live bacteria.

[0115]

[0116] Hereinafter, the present invention will be described in more detail through examples and the like. However, the scope and content of the present invention should not be construed as being limited or reduced by the examples and the like. Furthermore, based on the disclosure of the present invention, including the following examples, it is clear that those skilled in the art can easily implement the present invention, even though specific experimental results are not presented. It is also natural that such variations and modifications fall within the scope of the appended claims.

[0117]

[0118] <Experimental Example 1> Selection of a strain with excellent acid resistance and PS adhesion from traditional foods

[0119] Isolation and cultivation of yeast strains

[0120] Approximately 200 Saccharomyces genus colonies were isolated from 60 kinds of traditional nuruk (Lee JE, Lee AR, Kim HR, Lee E, Kim TW, Shin WC, et al. Restoration of traditional Korean nuruk and analysis of the brewing characteristics. J Microbiol Biotechnol. 2017. 27:896-908) manufactured and collected according to the manufacturing method of ancient literature. The 200 isolated yeast colonies were stored on YM agar (yeast malt extract agar) and cultured on PDB (potato dextrose agar, Becton Dickinson and Company, Sparks, MD, USA) medium at 25°C for 48 h, and used in a screening analysis for yeast selection.

[0121] 1st selection (acid-resistant strain)

[0122] The growth of isolated Saccharomyces colonies at pH 2.0 was investigated. To check acid tolerance, 100 μl of the selected yeast colonies were inoculated into YM medium adjusted to pH 2.0 with 0.1 M HCl, and cultured at 37°C for 3 days. Then, the strains were spread on PDA agar medium to check for colony formation. Through the above process, 36 candidate strains with excellent acid tolerance were initially selected.

[0123] Second selection (adhesion to plastic surface)

[0124] A secondary selection was conducted to select candidate strains with excellent adhesion to polystyrene. For this purpose, 36 strains selected in the first round were prepared and confirmed through crystal violet staining assay (see Reynolds, Todd B., and Gerald R. Fink. "Bakers' yeast, a model for fungal biofilm formation." Science 291.5505 (2001): 878-881). In brief, each yeast strain was cultured in SC medium containing 2% (w / v) glucose (Synthetic complete media and plates and YPD plates were made as described [C. Guthrie, G. R. Fink, Methods Enzymol. 194, 12 (1991)] with the exception of the altered agar or glucose concentrations, which are specified in the text.), and OD 600 The cells were harvested at 1.5 days. Then, the harvested cells were washed with H2O and cultured in SC medium containing 2% glucose at OD 600 After resuspending to 1.0, the cells were transferred to each well of a 96-well polystyrene plate (Falcon Microtest flat bottom plate, 35-1172; Becton-Dickinson Lab-ware) using a 100 μl pipette. Cells attached to the polystyrene were identified by staining with crystal violet. Staining with crystal violet was performed to visualize the cells attached to each well of the polystyrene plate (GA O'Toole et al. [Methods Enzymol. 310, 91 (1999)]). Crystal violet solution was added to each well, left for 15 minutes, washed with H2O, and then the absorbance (Abs) was measured at 595 nm (Fig. 1).

[0125] S. cerevisiae S288c (strain ATCC 204508) was selected as a negative control to compare plastic adhesion (polystyrene (PS) adhesion). S. cerevisiae S288c (strain ATCC 204508) is a standard strain that has microplastic adhesion despite not expressing the FLO11 gene (Literature: The Flo11p-deficient Saccharomyces cerevisiae strain background S288c can adhere to plastic surfaces. Colloids and Surfaces B: Biointerfaces Volume 60, Issue 1, 15 October 2007, Pages 131-134).

[0126] Statistical analysis

[0127] The experimental results were calculated as the mean ± standard deviation, and the analyzed experimental data were subjected to one-way ANOVA from the experimental data obtained from the negative control group and each sample, and significance was verified at the *** p<0.001 level.

[0128] Experimental results

[0129] Figure 1 is a graph showing the polystyrene (PS) adhesion ability of 36 acid-resistant yeast strains selected in the first round. S. cerevisiae S288c (indicated as S288c in the figure) was used as a negative control group.

[0130] As shown in Fig. 1, four strains (P16, P17, P18, and P19) were identified as having significantly higher plastic adhesion ability compared to the negative control, S. cerevisiae S288c. Four strains (P16, P17, P18, and P19) were finally selected.

[0131] To identify the four finally selected strains, 18S rRNA gene sequence analysis was performed. The universal bacterial primer pair NS1 (sequence 5: 5'-GTAGTCATATGCTTGTCTC-3') and NS24 (sequence 6: 5'-AAACCTTGTTACGACTTTTA-3') were used to amplify the genes. The gene sequences were then confirmed using BLAST (Basic Local Alignment Search Tool) of the National Center for Biotechnology Information (NCBI; http: / www.ncbi.nlm.nih.gov / BLAST / ). 18S rRNA gene sequence analysis confirmed that all four strains were Saccahromyces cerevisiae.

[0132] The four finally selected strains were named Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P, and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P, respectively, and deposited with the Korea Center for Microorganisms (KCCM), an internationally recognized microorganism depository under the Budapest Treaty, and assigned the deposit number "KCCM" on March 30, 2023. 13340P", "KCCM 13341P", "KCCM 13342P" and "KCCM 13343P" were awarded.

[0133]

[0134] <Experimental Example 1-2> Acid and bile resistance analysis of the finally selected S. cerevisiae strains

[0135] The acid resistance and bile resistance of the four finally selected strains (Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P, and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P) were examined. To confirm acid resistance, 100 μl of the selected yeast colonies were inoculated onto YM medium adjusted to pH 2.0 with the addition of 0.1 M HCl, cultured at 37°C for 3 days, and then plated onto PDA agar medium to confirm colony formation of the strain. In addition, to confirm bile resistance, 100 μl of the selected yeast colonies were inoculated onto YM medium containing 1% bile salts, cultured at 37°C for 24 hours, and then plated onto PDA agar medium to confirm colony formation of the strain. Comparison was made with three commercially available strains of S. boulardii, which are acid-resistant yeasts sold as positive controls.

[0136] StrainResistance to acid (pH2.0)Resistance to bile acid (1%)S.cerevisiaeReY29-4_P16++S.cerevisiaeReY29-5_P17++++S.cerevisiaeReY29-5_P18++++S.cerevisiaeReY29-5_P19+++S.boulardii1+++S.boulardii2++S.boulardii3+++S.cerevisiaeS288c--

[0137] -, no survival rate; +, low-survival rate; ++, high-survival rate

[0138] According to Table 1, Saccharomyces cerevisiae, present in traditional nuruk, is a safe microorganism that has been used for food and beverages for a long time. All four strains finally selected through the first and second selection processes in the present invention have excellent characteristics not only of acid resistance but also of microplastic adhesion. Therefore, when administered orally, they survive until the intestines without losing activity. Therefore, by adsorbing microplastics present in the intestines, they have the effect of preventing microplastics from being absorbed into the body, and thus can exhibit significant effects.

[0139]

[0140] <Experimental Example 2> Adhesion Analysis for Polystyrene and Polypropylene

[0141] Verification of plastic adhesion ability by viable cell count of the four finally selected Saccharomyces cerevisiae strains

[0142] The plastic adhesion ability of the four finally selected Saccharomyces cerevisiae strains (S. cerevisiaeReY29-4_P16, S. cerevisiaeReY29-5_P17, S. cerevisiaeReY43-1_P18, and S. cerevisiaeReY43-2_P19) was confirmed using a crystal violet staining assay (see Reynolds, Todd B., and Gerald R. Fink. "Bakers' yeast, a model for fungal biofilm formation." Science 291.5505 (2001): 878-881).

[0143] Saccharomyces cerevisiae strains (S. cerevisiaeReY29-4_P16, S. cerevisiaeReY29-5_P17, S. cerevisiaeReY43-1_P18, and S. cerevisiaeReY43-2_P19) were seeded at 2.5 × 10 in 96-well polystyrene plates (Falcon Microtest flat bottom plate, 35-1172; Becton-Dickinson Lab-ware) or 96-well polypropylene plates (96 well plate, deep, plate-dome, 90063; Bioneer). 5 cells / well (≥98% viable cell count), 5.0 × 10 5 cells / well (≥98% viable cell count), 1.0 × 10 6 cells / well (≥98% viable cell count) and 2.0 × 10 6 Each well was inoculated at a concentration of 10 cells / well (≥98% viable cell count) and cultured for 1 hour. Cells attached to polystyrene were identified by staining with crystal violet (GA O'Toole et al. [Methods Enzymol. 310, 91 (1999)]). Crystal violet solution was added to each well, left to stand for 15 minutes, washed with H2O, and then the absorbance (Abs) was measured at 595 nm. S. cerevisiae S288c was used as a negative control (Figs. 2, 3).

[0144] Statistical analysis

[0145] The experimental results were calculated as the mean ± standard deviation, and the analyzed experimental data were subjected to one-way ANOVA from the experimental data obtained from the negative control group and each sample, and significance was verified at the *** p<0.001 level.

[0146]

[0147] Figure 2 is a graph showing the polystyrene (PS) adhesion ability of four Saccharomyces cerevisiae strains according to the present invention and the negative control S. cerevisiae S288c strain analyzed by viable cell count. Figure 2a is a graph for S. cerevisiae ReY29-4_P16, Figure 2b is a graph for S. cerevisiae ReY29-5_P17, Figure 2c is a graph for S. cerevisiae ReY43-1_P18, and Figure 2d is a graph for S. cerevisiae ReY43-2_P19.

[0148] As shown in Fig. 2, the adhesion ability to plastic (polystyrene, PS) of four Saccharomyces cerevisiae strains (S. cerevisiaeReY29-4_P16, S. cerevisiaeReY29-5_P17, S. cerevisiaeReY43-1_P18, and S. cerevisiaeReY43-2_P19) significantly increased as the number of viable cells increased, and it was confirmed that the adhesion ability for each viable cell number was significantly superior to that of S. cerevisiaeS288c.

[0149]

[0150] Figure 3 is a graph showing the polypropylene (PP) adhesion ability of four Saccharomyces cerevisiae strains according to the present invention and the negative control S. cerevisiae S288c strain analyzed by viable cell count. Figure 3a is a graph for S. cerevisiae ReY29-4_P16, Figure 3b is a graph for S. cerevisiae ReY29-5_P17, Figure 3c is a graph for S. cerevisiae ReY43-1_P18, and Figure 3d is a graph for S. cerevisiae ReY43-2_P19.

[0151] As shown in Fig. 3, it was confirmed that the four Saccharomyces cerevisiae strains (S. cerevisiaeReY29-4_P16, S. cerevisiaeReY29-5_P17, S. cerevisiaeReY43-1_P18 and S. cerevisiaeReY43-2_P19) according to the present invention have excellent adhesion not only to polystyrene but also to polypropylene, a representative non-degradable plastic. In particular, it was confirmed that they have superior adhesion than the negative control strain S. cerevisiaeS288c.

[0152]

[0153] <Experimental Example 3> Adhesion by Microplastic Size

[0154] The purpose of this study was to investigate the adhesion ability of the four finally selected Saccharomyces cerevisiae strains (S. cerevisiaeReY29-4_P16, S. cerevisiaeReY29-5_P17, S. cerevisiaeReY43-1_P18, and S. cerevisiaeReY43-2_P19) to microplastics of different sizes (less than ~1,000 nm).

[0155] Four sizes of polystyrene beads (PS beads) (20, 50, 500, and 1000 nm) labeled with FITC, Rhodamine, and Cy5 fluorescence were prepared. Four strains of Saccharomyces cerevisiae (S. cerevisiaeReY29-4_P16, S. cerevisiaeReY29-5_P17, S. cerevisiaeReY43-1_P18, and S. cerevisiaeReY43-2_P19) and the negative control, S. cerevisiaeS288c strain was cultured on SC medium containing 2% (w / v) glucose (Synthetic complete media and plates and YPD plates were made as described [C. Guthrie, GR Fink, Methods Enzymol. 194, 12 (1991)] with the exception of the altered agar or glucose concentrations, which are specified in the text.) and 2 × 10 7 A culture medium was prepared at a concentration of cells / ml. 100 μg of polystyrene beads (PS beads) were added to each culture medium and cultured for 3 hours. Each cell culture medium was centrifuged to remove the supernatant, collect the pellet, add PBS to resuspend, and measure fluorescence (FITC, 495 / 519; Rhodamine, 546 / 568; Cy5, 651 / 670). For accurate comparison, Comparative Group 1 (only beads), which contained only fluorescently labeled PS beads, and Comparative Group 2 (only yeast), which contained the same number of yeast without PS beads, were prepared and measured using the same process as described above.

[0156]

[0157] Figure 4 is a graph showing the adhesion ability of four strains of Saccharomyces cerevisiae and the negative control strain S. cerevisiae S288c to fluorescently labeled polystyrene (PS) beads (ultrafine plastics) according to size. Figure 4a is a graph for S. cerevisiaeReY29-4_P16, Figure 4b is a graph for S. cerevisiaeReY29-5_P17, Figure 4c is a graph for S. cerevisiaeReY43-1_P18, and Figure 4d is a graph for S. cerevisiaeReY43-2_P19.

[0158] As shown in Fig. 4, four strains of Saccharomyces cerevisiae (S. cerevisiaeReY29-4_P16, S. cerevisiaeReY29-5_P17, S. cerevisiaeReY43-1_P18, and S. cerevisiaeReY43-2_P19) did not show significant adhesion to plastics larger than 1000 nm, but were found to have significantly better adsorption / binding ability than the control strain, S. cerevisiaeS288c, to ultrafine plastics smaller than 500 nm.

[0159]

[0160] <Experimental Example 4> Evaluation of the efficacy of inhibiting in vivo microplastic absorption.

[0161] It was intended to confirm whether the Saccharomyces cerevisiae strains (S. cerevisiaeReY29-5_P17, S. cerevisiaeReY43-1_P18 and S. cerevisiaeReY43-2_P19) according to the present invention adsorb microplastics such as polystyrene and polypropylene in vivo and inhibit their bioabsorption.

[0162] C57BL / 6 male mice (6 weeks old) were purchased from Orient Bio. They were housed in a controlled-conditioning room (Korea Food Research Institute Animal Laboratory) at 22±1℃, 55±1% humidity, and a 12-h light / dark cycle, with free access to food and water. After acclimation for 7 days, they were used in the experiment.

[0163] The experimental group was divided into four groups of 10 animals each, evenly distributed by weight, and each group was treated as follows.

[0164] Group 1 (normal group, control): 0.2 ml / day of physiological saline solution was administered orally for 3 weeks, and 0.3 ml of physiological saline solution was administered orally on the last day (21st day).

[0165] Group 2 (control group, Plastic): 0.2 ml / day of saline solution was administered orally for 3 weeks, and on the last day (21st day), 50 mg / kg (in 0.3 ml) of fluorescently labeled PS beads 20, 50, and 500 nm were administered orally, respectively.

[0166] Group 3 (negative control group, S288C): Oral administration of S. cerevisiae S288c strain culture daily for 3 weeks (1 × 10 9 cells / day in 0.2 ml), and on the last day (21st day), 50 mg / kg (in 0.3 ml) of fluorescently labeled PS beads 20, 50, and 500 nm were orally administered, respectively.

[0167] Group 4 (experimental group, 17): Oral administration of S. cerevisiae ReY29-5_P17 strain culture solution (1 × 10 9 cells / day in 0.2 ml), and on the last day (21st day), 50 mg / kg (in 0.3 ml) of fluorescently labeled PS beads 20, 50, and 500 nm were administered orally, respectively.

[0168] Group 5 (experimental group, 18): Oral administration of S. cerevisiae ReY43-1_P18 strain culture solution (1 × 10 9 cells / day in 0.2 ml), and on the last day (21st day), 50 mg / kg (in 0.3 ml) of fluorescently labeled PS beads 20, 50, and 500 nm were administered orally, respectively.

[0169] Group 6 (experimental group, 19): Orally administered S. cerevisiae ReY43-2_P19 strain culture solution daily for 3 weeks (1 × 10 9 cells / day in 0.2 ml), and on the last day (21st day), 50 mg / kg (in 0.3 ml) of fluorescently labeled PS beads 20, 50, and 500 nm were administered orally, respectively.

[0170] Blood was collected from each group 0, 1, 3, 6, 12, and 24 hours later, and the blood was fluorescence-measured to confirm whether the single-administered PS bead was absorbed into the body (existed in the blood).

[0171]

[0172] Figure 5a is a graph showing the results of analyzing the efficacy of inhibiting in vivo microplastic absorption when the S. cerevisiae ReY29-5_P17 strain was orally administered. Figure 5b is a graph showing the results of analyzing the efficacy of inhibiting in vivo microplastic absorption when the S. cerevisiae ReY43-1_P18 strain was orally administered. Figure 5c is a graph showing the results of analyzing the efficacy of inhibiting in vivo microplastic absorption when the S. cerevisiae ReY43-2_P19 strain was orally administered.

[0173] As shown in Fig. 5, groups 4 (17), 5 (18), and 6 (19) orally administered Saccharomyces cerevisiae strains (S. cerevisiaeReY29-5_P17, S. cerevisiaeReY43-1_P18, and S. cerevisiaeReY43-2_P19) were confirmed to inhibit orally administered microplastics from being absorbed into the blood (absorbed in vivo). Specifically, group 2 (control group, plastic) was orally administered microplastics and 1 hour later, they were absorbed into the blood at a concentration of 80-100 ㎍ / ml, and over time, they were transferred and accumulated in other organ tissues from the blood (accumulated at 60-80 ㎍ / ml), and the concentration of microplastics in the blood decreased to 20 ㎍ / ml.

[0174] Group 3 (S288c) also confirmed that microplastics were orally administered and absorbed into the blood at a concentration of 80 ㎍ / ml 1 hour later, and that they were transferred and accumulated (60 ㎍ / ml) from the blood to other organ tissues over time, reducing the concentration of microplastics in the blood to 20 ㎍ / ml.

[0175] On the other hand, when microplastics were orally administered to groups 4 (17), 5 (18), and 6 (19) orally administered Saccharomyces cerevisiae strains (S. cerevisiaeReY29-5_P17, S. cerevisiaeReY43-1_P18, and S. cerevisiaeReY43-2_P19), it was confirmed that only 40-60 ㎍ / ml of microplastics were absorbed into the blood after 1 hour (a value 1.5-2 times lower than groups 2 and 3). It was confirmed that Saccharomyces cerevisiae strains (S. cerevisiaeReY29-5_P17, S. cerevisiaeReY43-1_P18, and S. cerevisiaeReY43-2_P19) significantly inhibited the absorption of orally administered microplastics into the body.

[0176] In addition, groups 4 (17), 5 (18), and 6 (19) that were orally administered Saccharomyces cerevisiae strains (S. cerevisiaeReY29-5_P17, S. cerevisiaeReY43-1_P18, and S. cerevisiaeReY43-2_P19) also showed a decrease in blood concentration over time as they moved and accumulated in organ tissues. However, in groups 4, 5, and 6, when orally administered microplastics were absorbed into the blood, they were initially absorbed at a very reduced concentration, so the amount of microplastics accumulated in the body (20-40 ㎍ / ml) was significantly lower by 1.5 to 4 times than in group 2 (60-80 ㎍ / ml accumulated) and group 3 (60 ㎍ / ml).

[0177]

[0178] <Experimental Example 5> Presence of biofilm formation genes

[0179] Summary of the experiment

[0180] It is known that a protein called flocculin, which belongs to the yeast cell membrane glycoprotein, is involved in plastic adhesion. Flocculin is encoded by the FLO11 gene (Reynolds, Todd B., and Gerald R. Fink. "Bakers' yeast, a model for fungal biofilm formation." Science 291.5505 (2001): 878-881).

[0181] The transcriptional level of the FLO11 gene is related to biofilm formation in yeast, specifically, the higher the transcriptional level of the FLO11 gene, the higher the possibility of forming biofilm in yeast (Zara, Giacomo, et al. "FLO11gene length and transcriptional level affect biofilm-forming ability of wild flor strains of Saccharomyces cerevisiae." Microbiology 155.12 (2009): 3838-3846).

[0182] Biofilms are three-dimensional structures formed by microorganisms within a multimeric substrate secreted by themselves, forming in the form of a film on solid surfaces and living biological tissues. Therefore, the excellent biofilm-forming ability of yeast gives it an advantage in terms of environmental resistance, but it can also interfere with the action of immune cells or antibiotics in the body, delaying the healing process and making it vulnerable to bacterial and fungal infections. In particular, if it has the ability to adhere to plastic, it can form biofilm colonies in tissues without being excreted from the body, and continue to accumulate plastic on the walls of the digestive organs, which can cause organ damage caused by plastic.

[0183] That is, the expression level of the FLO11 gene of four Saccharomyces cerevisiae strains (S. cerevisiaeReY29-4_P16, S. cerevisiaeReY29-5_P17, S. cerevisiaeReY43-1_P18 and S. cerevisiaeReY43-2_P19) according to the present invention was confirmed using real-time PCR and FLO11gene specific primers.

[0184] Isolation of genomic DNA and amplification of the FLO11 gene sequence

[0185] Four strains of Saccharomyces cerevisiae (S. cerevisiaeReY29-4_P16, S. cerevisiaeReY29-5_P17, S. cerevisiaeReY43-1_P18, and S. cerevisiaeReY43-2_P19) and S. cerevisiaeS288c strain were cultured in SC medium containing 2% (w / v) glucose, respectively, and the OD 600 The culture was continued until it reached 1.5, and the pellet was collected by centrifugation. Then, total RNA was extracted using an RNA extraction kit (RNeasy, Qiagen).

[0186] SuperScript to synthesize the above total RNA into cDNA TM Ⅲ Forst-strand (Invitrogen, USA) was used and performed according to the manufacturer's manual. cDNA was stored frozen at -20°C until RT-PCR.

[0187] The above cDNA was diluted 10-fold with DEPC-water and used. The diluted cDNA, Fast SYBER® Green Supermix (Thermo Fisher, USA), forward primer and reverse primer diluted to a concentration of 10 pM, and DEPC-water were mixed to make a reaction volume of 20 μl. Real-time PCR was performed using StepOnePlus (Thermo Fisher, USA) by repeating 40 cycles of 95°C for 3 sec and 60°C for 30 sec.

[0188] FLO11 primer Forward sequence 75'-CCTCCGAAGGAACTAGCTGTAATT-3' Reverse sequence 85'-AGTCACATCCAAAGTATACTGCATGAT-3'

[0189]

[0190] Figure 6 is a graph showing the results of confirming the FLO11 gene expression level of Saccharomyces cerevisiae strains (S. cerevisiaeReY29-4_P16, S. cerevisiaeReY29-5_P17, S. cerevisiaeReY43-1_P18 and S. cerevisiaeReY43-2_P19) according to the present invention by RT-PCR, and the gene expression level is expressed as a relative gene expression level compared to the FLO11 gene expression level of the S. cerevisiaeS288c strain.

[0191] As shown in Fig. 6, it was confirmed that the four Saccharomyces cerevisiae strains (S. cerevisiaeReY29-4_P16, S. cerevisiaeReY29-5_P17, S. cerevisiaeReY43-1_P18, and S. cerevisiaeReY43-2_P19) did not have a significant difference in the FLO11 gene expression level from the negative control strain, S. cerevisiaeS288c.

[0192]

[0193] <Experimental Example 6> Genetic Analysis of Four Saccharomyces cerevisiae Strains

[0194] Saccharomyces cerevisiae has genetic diversity among strains within the species, and even within the same Saccahromyces cerevisiae, there are significant differences in activities such as acid resistance and plastic adhesion.

[0195] Based on the experimental results described above, the four strains finally selected (Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P, and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P) were subjected to whole genome resequencing to identify S. cerevisiae, which is known to have plastic adhesion ability among Saccharomyces cerevisiae. The genetic differences with cerevisiaeS288c were analyzed, and the results are presented as follows.

[0196] Genetic analysis results of S. cerevisiaeReY29-4_P16 strain

[0197] Among the four strains selected in the present invention, genes with mutations were selected and analyzed based on the gene sequences of the S. cerevisiaeReY29-4_P16 strain and the standard strain S. cerevisiaeS288C, and the results are shown in Table 3.

[0198] ChromosomeSV startSV endSV typeRef (in S288c)Alt(in sample yeast)Gene NameNC_001141.2390973391033DeletionTGATGGAGGTGACAGTAGTGCCAGTAGAAAAGCTTTCAGTAGTAGAGCTGAATGGAATTGAA: Sequence 9TFLO11NC_001143.9647797647906DeletionCTAGCTCTTCCTCGTCGTCCAGTGAAGTCTGTACAGAGTGCACCGAGACCGAGTCTACCAGTTATGTGACACCATATGTCAGCTCGTCTACTGCTGCCGCAAACTACACT: SEQ ID NO: 10CCFLO10

[0199] As shown in Table 3, the S. cerevisiaeReY29-4_P16 strain was confirmed to have the sequence from base 390973 to base 391033 (61 bp) of position NC_001141.2 (S288C chromosome IX) encoding the FLO11 gene deleted and substituted with T.

[0200] In addition, the S. cerevisiaeReY29-4_P16 strain was confirmed to have a deletion in which the sequence (110 bp) from the 647797th base to the 647906th base of position NC_001143.9 (S288C chromosome IX) encoding the FLO10 gene was substituted with CC.

[0201] That is, the S. cerevisiaeReY29-4_P16 strain exhibits acid resistance at pH 2.0, so it can safely settle in the digestive tract when administered orally, and has excellent adhesion to the surface of microplastics less than 1000 nm that are administered orally in vivo, effectively inhibiting the absorption of microplastics into the body.

[0202] Furthermore, the S. cerevisiaeReY29-4_P16 strain has excellent adhesion to microplastics despite not forming expression of FLO11, a gene related to biofilm formation. If it is used as an active ingredient, it can be used to inhibit microplastic bioabsorption by consuming it as a probiotic composition or food, and can also be used to adsorb and remove microplastics in the environment.

[0203] Genetic analysis results of S. cerevisiaeReY29-5_P17 strain

[0204] Among the four strains selected in the present invention, genes with mutations were selected and analyzed based on the gene sequences of the S. cerevisiaeReY29-5_P17 strain and the standard strain S. cerevisiaeS288C, and the results are shown in Table 4.

[0205] ChromosomeSV startSV endSV typeRef (in S288c)Alt(in sample yeast)Gene NameNC_001141.2390973391204DeletionTGATGGAGTGACAGTAGTGCCAGTAGAAAAGCTTTCAGTAGTAGAGCTGAATGGAATTGAAGATGGAGCGGAGGAAGTGATGTTGCTAGAGGAAGATG GGGTTGGTACTGGGTGCTACAGAGCTTTCAGTGGTGGAGCTGGATACTGGAGCAGAAGAGCTTTCAGTAGTAGAGCTTGATGGGGTTGGTACTGGAACAGAAGAGCTTTCAGTGCTAGAGCTGAATGGGGTTGAA : sequence 11TFLO11NC_001143.9647797647906DeletionCTAGCTCTTCCTCGTCGTCCAGTGAAGTCTGTACAGAGTGCACCGAGACCGAGTCTACCAGTTATGTGACACCATATGTCAGCTCGTCTACTGCTGCCGCAAACTACACT: SEQ ID NO: 10CCFLO10

[0206] As shown in Table 4, the S. cerevisiaeReY29-5_P17 strain was confirmed to have a deletion of the 390973rd to 391204th base sequence (232 bp) of the NC_001141.2 position (S288C chromosome IX) encoding the FLO11 gene and a replacement with T.

[0207] In addition, the S. cerevisiaeReY29-5_P17 strain was confirmed to have a deletion in which the sequence (110 bp) from the 647797th base to the 647906th base of position NC_001143.9 (S288C chromosome IX) encoding the FLO10 gene was substituted with CC.

[0208] That is, the S. cerevisiaeReY29-5_P17 strain exhibits acid resistance at pH 2.0, so it can safely settle in the digestive tract when administered orally, and it has excellent adhesion to the surface of microplastics less than 1000 nm that are administered orally in vivo, so it was confirmed that it effectively inhibits microplastics from being absorbed into the body.

[0209] Furthermore, the S. cerevisiaeReY29-5_P17 strain has excellent adhesion to microplastics despite not forming expression of FLO11, a gene related to biofilm formation. If it is used as an active ingredient, it can be used to inhibit bioabsorption of microplastics by consuming it as a probiotic composition or food, and can also be used to adsorb and remove microplastics in the environment.

[0210] Genetic analysis results of S. cerevisiaeReY43-1_P18 strain

[0211] Among the four strains selected in the present invention, genes with mutations were selected and analyzed based on the gene sequences of the S. cerevisiaeReY43-1_P18 strain and the standard strain S. cerevisiaeS288C, and the results are shown in Table 5.

[0212] ChromosomeSV startSV endSV typeRef (in S288c)Alt(in sample yeast)Gene NameNC_001141.2390973391033DeletionTGATGGAGGTGACAGTAGTGCCAGTAGAAAAGCTTTCAGTAGTAGAGCTGAATGGAATTGAA: Sequence 9TFLO11

[0213] As shown in Table 5, it was confirmed that the S. cerevisiaeReY43-1_P18 strain had a deletion of the 390973rd base to the 391033rd base sequence (61 bp) of the NC_001141.2 position (S288C chromosome IX) encoding the FLO11 gene and a replacement with T.

[0214] That is, the S. cerevisiaeReY43-1_P18 strain exhibits acid resistance at pH 2.0, so it can safely settle in the digestive tract when administered orally, and it has excellent adhesion to the surface of microplastics less than 1000 nm that are administered orally in vivo, so it was confirmed that it effectively inhibits microplastics from being absorbed into the body.

[0215] Furthermore, the S. cerevisiaeReY43-1_P18 strain has excellent adhesion to microplastics despite not forming expression of FLO11, a gene related to biofilm formation, and when it is used as an active ingredient, it can be used to inhibit bioabsorption of microplastics by consuming it as a probiotic composition or food, and can also be used to adsorb and remove microplastics in the environment.

[0216] Genetic analysis results of S. cerevisiaeReY43-2_P19 strain

[0217] Among the four strains selected in the present invention, genes with mutations were selected and analyzed based on the gene sequences of the S. cerevisiaeReY43-2_P19 strain and the standard strain S. cerevisiaeS288C, and the results are shown in Table 6.

[0218] ChromosomeSV startSV endSV typeRef (in S288c)Alt(in sample yeast)Gene NameNC_001141.2390973391033DeletionTGATGGAGGTGACAGTAGTGCCAGTAGAAAAGCTTTCAGTAGTAGAGCTGAATGGAATTGAA: Sequence 9TFLO11NC_001143.9647797647906DeletionCTAGCTCTTCCTCGTCGTCCAGTGAAGTCTGTACAGAGTGCACCGAGACCGAGTCTACCAGTTATGTGACACCATATGTCAGCTCGTCTACTGCTGCCGCAAACTACACT: SEQ ID NO: 10CCFLO10

[0219] As shown in Table 6, it was confirmed that the S. cerevisiaeReY43-2_P19 strain had a deletion of the 390973rd base to the 391033rd base sequence (61 bp) of the NC_001141.2 position (S288C chromosome IX) encoding the FLO11 gene and a replacement with T.

[0220] In addition, it was confirmed that the S. cerevisiaeReY43-2_P19 strain had a deletion of the 647797th base to the 647906th base sequence (110 bp) of position NC_001143.9 (S288C chromosome IX) encoding the FLO11 gene and a replacement with CC.

[0221] That is, the S. cerevisiaeReY43-2_P19 strain exhibits acid resistance at pH 2.0, so it can safely settle in the digestive tract when administered orally, and it has excellent adhesion to the surface of microplastics less than 1000 nm that are administered orally in vivo, so it was confirmed that it effectively inhibits microplastics from being absorbed into the body.

[0222] Furthermore, the S. cerevisiaeReY43-2_P19 strain has excellent adhesion to microplastics despite not forming expression of FLO11 and FLO10, genes related to biofilm formation, and when made into an active ingredient, it can be used to inhibit bioabsorption of microplastics by consuming it as a probiotic composition or food, and can also be used to adsorb and remove microplastics in the environment.

[0223]

[0224] [Accession number]

[0225] Name of depositor: Korea Center for Microbiological Conservation (KCCM)

[0226] Accession number: KCCM13340P

[0227] Date of acceptance: 20230330

[0228] Name of depositor: Korea Center for Microbiological Conservation (KCCM)

[0229] Accession number: KCCM13341P

[0230] Date of acceptance: 20230330

[0231] Name of depositor: Korea Center for Microbiological Conservation (KCCM)

[0232] Accession number: KCCM13342P

[0233] Date of acceptance: 20230330

[0234] Name of depositor: Korea Center for Microbiological Conservation (KCCM)

[0235] Accession number: KCCM13343P

[0236] Date of acceptance: 20230330

[0237]

[0238]

[0239]

[0240]

Claims

1. A Saccahromyces cerevisiae strain that has acid resistance, bile resistance, and microplastic adhesion. A Saccahromyces cerevisiae strain, characterized in that it is any one selected from the group consisting of Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P, and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P.

2. In paragraph 1, The above strain is a Saccharomyces cerevisiae strain characterized in that a portion of the sequence of at least one gene among the FLO11 gene and the FLO10 gene is deleted or substituted.

3. In paragraph 2, A Saccahromyces cerevisiae strain characterized in that the biofilm forming ability is reduced by the deletion or substitution described above.

4. In paragraph 1, The above Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, the above Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P and the Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P have a part of the FLO11 gene and FLO10 gene sequence deleted or substituted, The above Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P is a Saccahromyces cerevisiae strain characterized by a deletion or substitution of a portion of the FLO11 gene sequence.

5. In paragraph 1, The above strain is a Saccharomyces cerevisiae strain characterized in that the base sequence from the 390973rd base to the 391033rd or 391204th base in the FLO11 gene sequence of the S288C chromosome IX of Saccahromyces cerevisiae (position NC_001141.2) is deleted and substituted with T.

6. In paragraph 5, Among the above strains, The above Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P is characterized by a deletion of the base sequence from the 390973rd base to the 391033rd base in the FLO11 gene sequence of S288C chromosome IX (position NC_001141.2) and a substitution with T. The above Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P are Saccharomyces cerevisiae strains characterized in that the base sequence from the 390973rd base to the 391204th base in the FLO11 gene sequence of S288C chromosome Ⅸ (position NC_001141.2) is deleted and substituted with T.

7. In paragraph 1, The above strain is a Saccharomyces cerevisiae strain characterized in that the sequence from the 647797th base to the 647906th base in the FLO10 gene sequence of the S288C chromosome IX of Saccahromyces cerevisiae (position NC_001143.9) is deleted and replaced with CC.

8. In paragraph 1, A strain of Saccharomyces cerevisiae, characterized in that the above microplastics have a size of 1 to 500 nm.

9. A food composition for inhibiting absorption of microplastics, comprising as an active ingredient any one Saccahromyces cerevisiae strain selected from the group consisting of Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P, and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P.

10. A composition for adsorbing microplastics, comprising any one Saccahromyces cerevisiae strain selected from the group consisting of Saccahromyces cerevisiae ReY29-4_P16 KCCM 13340P, Saccahromyces cerevisiae ReY29-5_P17 KCCM 13341P, Saccahromyces cerevisiae ReY43-1_P18 KCCM 13342P, and Saccahromyces cerevisiae ReY43-2_P19 KCCM 13343P.

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