Pediococcus pentosaceus strains, fermentation broth of Porphyra yezoensis-Pediococcus pentosaceus and their application in enhancing intestinal barrier function
Patent Information
- Application Number
- CN202510770141.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2025-06-10
- Publication Date
- 2026-09-01
- Estimated Expiration
- 2045-06-10
AI Technical Summary
目前，我国坛紫菜加工业整体处于加工品种单一，开发利用程度较低，坛紫菜的精深加工产品较为缺乏，市场上主要以烘干制品(紫菜盘)、烘焙食品(烤海苔、夹心海苔)、速食汤品(紫菜汤)等形式流通，产品附加值低
[0028]本发明筛选得到一株戊糖片球菌，将其与坛紫菜共同进行发酵，可以得到具有增强肠道屏障功能的发酵物，具体可以上调ZO-1、occludin、claudin-1及claudin-4紧密连接蛋白相关基因的表达水平，以起到保护肠屏障的功能。本发明提供的发酵菌株以及发酵方法可以提高坛紫菜原有的生物活性与营养价值，开发具有增强肠道屏障活性的海藻健康食品，促进海藻深加工产品的多元化和加工技术的多层次化，推动我国海藻精深产业的快速发展。
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Abstract
Description
Technical Field
[0001] This invention belongs to the field of food fermentation, specifically relating to Pediococcus pentosaceus strains, Porphyra yezoensis-Pediococcus pentosaceus fermentation broth, and their application in enhancing intestinal barrier function. Background Technology
[0002] As the largest immune organ and a crucial component of the digestive system, the gut's health is closely linked to overall health. Intestinal barrier function, a core mechanism for maintaining gut homeostasis, encompasses multiple levels including physical barriers, chemical barriers, and immune regulation, playing a key role in resisting pathogen invasion, regulating nutrient absorption, and maintaining gut microbiota balance. With changing modern lifestyles, the incidence of intestinal diseases is on the rise, such as inflammatory bowel disease and irritable bowel syndrome. These diseases are often closely related to impaired intestinal barrier function. Therefore, finding safe and effective strategies to enhance intestinal barrier function has significant clinical and social value.
[0003] Marine biological resources, as one of the richest biological resource banks on Earth, contain a large number of substances with unique biological activity. Porphyra yezoensis, a common marine red algae, is rich in nutrients such as protein, dietary fiber, amino acids, and minerals. Long-term consumption has antioxidant, lipid-lowering, blood pressure-lowering, and immune-regulating effects. Currently, my country's Porphyra yezoensis processing industry is characterized by a limited range of processed products and a low level of development and utilization. Deep-processed products of Porphyra yezoensis are scarce, and the market mainly circulates in the form of dried products (seaweed trays), baked goods (roasted seaweed, stuffed seaweed), and instant soups (seaweed soup), resulting in low added value. Therefore, developing deep-processing technologies for Porphyra yezoensis to fully utilize its nutritional and bioactive value is an important industrial issue that urgently needs to be addressed.
[0004] Lactic acid bacteria, as a typical representative of probiotics, play a vital role in regulating the balance of intestinal flora and enhancing intestinal barrier function. They maintain the balance of the intestinal microecology by competitively inhibiting the growth of harmful bacteria, producing antibacterial substances, and promoting the proliferation of beneficial bacteria. Furthermore, lactic acid bacteria strains such as *Lactobacillus plantarum*, *Lactobacillus reuteri*, and *Lactobacillus paracasei* have been widely used in food processing due to their excellent fermentation characteristics. Studies have shown that lactic acid bacteria fermentation can promote the release of plant-based bioactive substances, thereby improving their bioactivity and enhancing their nutritional properties. Therefore, lactic acid bacteria fermentation is an important processing technology for improving the nutritional quality and bioactivity of food, with broad prospects for development and application. Summary of the Invention
[0005] To address the need for effective utilization of the nutritional value of *Porphyra yezoensis* in existing technologies, this invention provides a *Pediococcus pentosaceus* strain, a *Porphyra yezoensis*-*Pediococcus pentosaceus* fermentation broth, and its application in enhancing intestinal barrier function. The fermentation broth obtained by fermenting *Porphyra yezoensis* using the *Pediococcus pentosaceus* strain provided by this invention significantly enhances intestinal barrier function. The specific technical solution is as follows:
[0006] In a first aspect, the present invention provides a strain of Pediococcus pentosaceus, characterized in that the Pediococcus pentosaceus strain is classified as Pediococcus pentosaceus, the depositary institution is the China Center for Type Culture Collection, the accession number is CCTCC M 20251312, and the deposit date is June 9, 2025.
[0007] In a second aspect, the present invention provides a Pediococcus pentosacchari bacterial suspension, characterized in that the active ingredient of the Pediococcus pentosacchari bacterial suspension is the Pediococcus pentosacchari strain as described in claim 1.
[0008] Furthermore, the concentration of Pediococcus pentosaceus in the bacterial solution is 1.0 × 10⁻⁶. 8 cfu / mL.
[0009] Thirdly, the present invention provides a method for preparing a fermentation broth of *Porphyra yezoensis*-*Pediococcus pentosaceus*, characterized in that it comprises:
[0010] Disperse the laver in water, sterilize it, add the Pediococcus pentosacchari bacterial solution as described in claim 2, and ferment it to obtain laver-Pediococcus pentosacchari fermentation broth.
[0011] Furthermore, the dispersion operation involves adding laver to water to form a uniform slurry.
[0012] Furthermore, the dispersion ratio of the laver to water is 1g: 20-100mL.
[0013] Furthermore, the dispersion ratio of the laver to water is 1g:50mL.
[0014] Furthermore, the amount of Pediococcus pentosaceus bacterial solution added is 1-10% of the fermentation broth after addition.
[0015] Furthermore, the amount of Pediococcus pentosaceus bacterial solution added is 5% of the fermentation broth after addition.
[0016] Furthermore, the fermentation time is 1 to 12 days.
[0017] Furthermore, the fermentation time is 72 hours.
[0018] Furthermore, the sterilization method is high-temperature sterilization.
[0019] Furthermore, the sterilization method is as follows: sterilize the laver dispersion in a high-pressure steam sterilizer at 121°C for 15 minutes.
[0020] Fourthly, the present invention provides a fermentation broth of *Porphyra yezoensis*-Pediococcus pentosacchari prepared by the above preparation method.
[0021] Fifthly, the present invention provides the above-mentioned Pediococcus pentosacchari strain, or the above-mentioned Pediococcus pentosacchari bacterial solution, or the fermentation broth of Porphyra yezoensis-Pediococcus pentosacchari prepared by the above-mentioned preparation method, or the application of the above-mentioned Porphyra yezoensis-Pediococcus pentosacchari fermentation broth in the preparation of products that enhance intestinal barrier function.
[0022] Furthermore, in the aforementioned application, the supernatant of the fermentation broth of *Porphyra yezoensis*-*Pediococcus pentosaceus* is taken.
[0023] Furthermore, in the aforementioned application, the supernatant of the *Porphyra yezoensis*-*Pediococcus pentosacchari* fermentation broth is diluted.
[0024] Furthermore, the supernatant of the *Porphyra yezoensis*-*Pediococcus pentosacchari* fermentation broth was diluted to 1 / 10 to 2 / 5 of its original volume.
[0025] Furthermore, the supernatant of the *Porphyra yezoensis*-*Pediococcus pentosacchari* fermentation broth was diluted to 1 / 5 of its original volume.
[0026] Furthermore, the enhancement of intestinal barrier function is manifested by upregulating the expression levels of ZO-1, occludin, claudin-1, and / or claudin-4 tight junction protein genes.
[0027] Compared with the prior art, the present invention has the following beneficial effects:
[0028] This invention screened a strain of *Pediococcus pentosaceus*, which, when co-fermented with *Porphyra yezoensis*, yielded a fermented product that enhances intestinal barrier function. Specifically, it upregulates the expression levels of genes related to ZO-1, occludin, claudin-1, and claudin-4 tight junction proteins, thereby protecting the intestinal barrier. The fermentation strain and method provided by this invention can improve the original bioactivity and nutritional value of *Porphyra yezoensis*, develop seaweed health foods with enhanced intestinal barrier activity, promote the diversification of deep-processed seaweed products and the multi-level development of processing technologies, and drive the rapid development of my country's deep-processed seaweed industry. Attached Figure Description
[0029] Figure 1 Microscopic image of Pediococcus pentosaceus HYZ2-1.
[0030] Figure 2 Microscopic image of Pediococcus pentosaceus 3-5-2.
[0031] Figure 3 This is a schematic diagram of the phylogenetic tree of Pediococcus pentosaceus HYZ2-1 based on 16S rRNA.
[0032] Figure 4 The graph shows the pH changes during the fermentation of laver by three strains of Pediococcus pentosaceus; where Control represents the laver slurry group in the unfermented jar.
[0033] Figure 5 This is a schematic diagram illustrating the effect of different concentrations of fermentation broth on the activity of Caco-2 cells.
[0034] Figure 6 This diagram illustrates the effects of different strains of *Porphyra yezoensis* fermentation broth on LPS-induced tight junction proteins in Caco-2 cells; where Control represents the normal control group, LPS represents the model group, and Untreated represents the untreated *Porphyra yezoensis* slurry intervention group.
[0035] Figure 7 This diagram illustrates the effect of Porphyra yezoensis-Pediococcus pentosaceus fermentation broth on LPS-induced tight junction proteins in Caco-2 cells; where Control represents the normal control group, Untreated represents the unfermented Porphyra yezoensis slurry intervention group, and LPS represents the model group. Detailed Implementation
[0036] To enable those skilled in the art to better understand the present invention, the technical solution of the present invention will be clearly and completely described below with reference to specific embodiments. It should be noted that the following detailed descriptions are exemplary and are only some embodiments of the present invention, not all embodiments.
[0037] Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort should fall within the scope of protection of the present invention.
[0038] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains. The experimental materials used in the embodiments of this invention are all conventional experimental materials in the art and are commercially available. Experimental methods not specifying detailed conditions are performed according to conventional experimental methods or the operating instructions recommended by the supplier.
[0039] In this invention, the source of the laver is not required; commercially available laver is sufficient. In one specific embodiment of this invention, laver from Xiapu, Ningde is selected.
[0040] The *Pediococcus pentosaceus* 3-5-2 used in this invention is derived from marine algae, with accession number CCTCC AB 2025091, and is deposited at the China Center for Type Culture Collection. The *Pediococcus pentosaceus* PP is derived from sourdough, with accession number CCTCC AB2025090, and is also deposited at the China Center for Type Culture Collection.
[0041] Example 1: Screening and Identification of Pediococcus pentosaceus
[0042] Lactic acid bacteria were isolated using the streak plating method: Marine samples were added to MRS liquid medium and incubated at 37°C for 24 hours for bacterial enrichment. 1 mL of the marine sample was added to 9 mL of sterile physiological saline (0.90% by mass) to prepare a 10% concentration. -1 The bacterial suspension was diluted in a 10-fold series until the concentration was 10. -1 -10 -4 Each gradient was replicated three times. After vortexing to mix, 200 μL of the bacterial suspension with the appropriate gradient was spread onto MRS liquid medium and incubated at 37°C for 48 h. Representative strains were selected based on colony color, elevation, and edge shape, and streaked onto MRS agar medium, incubated at 37°C for 48 h. After multiple subcultures, strains with uniform morphology observed by the naked eye were Gram-stained and examined under a microscope. Strains with uniform microscopic examination were confirmed as purified and stored at -80°C (25% glycerol) for later use.
[0043] The purified bacterial culture was obtained using the above method. The sequence obtained from sequencing was compared with the nucleic acid sequence in NCBI BLAST. The results showed that the strain was Pediococcus pentosaceus, named HYZ2-1 and 3-5-2. Figure 1 A microscopic photograph of Gram-stained Pediococcus pentosaceus HYZ2-1. Figure 2 Microscopic photograph of Pediococcus pentosaceus 3-5-2 Gram staining. Figure 3 The phylogenetic tree of Pediococcus pentosacchari HYZ2-1, obtained based on 16S rRNA results, shows that HYZ2-1 has high homology with Pediococcus pentosacchari DSM 20336.
[0044] The 16S rRNA sequence of Pediococcus pentosaceus HYZ2-1:
[0045]
[0046] The 16S rRNA sequence of Pediococcus pentosaceus 3-5-2:
[0047]
[0048] Example 2: Preparation of Porphyra yezoensis-Marine Lactic Acid Bacteria Fermentation Broth
[0049] Porphyra yezoensis from Xiapu, Ningde was selected, and the Porphyra yezoensis was mixed with purified water at a ratio of 1:50 (g / mL) to obtain a Porphyra yezoensis dispersion.
[0050] Pediococcus pentosaceus bacterial suspension was prepared according to the method in Example 1, with a bacterial suspension concentration of 1.0 × 10⁻⁶. 8 cfu / mL.
[0051] The laver solution was sterilized in a high-pressure steam sterilizer at 121°C for 15 minutes; then, the laver dispersion was cooled to below 37°C; 5% Pediococcus pentosaceus culture was added to the laver dispersion as a starter culture, and the mixture was allowed to ferment at 37°C for 72 hours to obtain the laver-Pediococcus pentosaceus fermentation broth.
[0052] Example 3: pH changes during the preparation of Porphyra yezoensis-marine lactic acid bacteria fermentation broth
[0053] Porphyra yezoensis from Xiapu, Ningde was selected, and the Porphyra yezoensis was mixed with purified water at a ratio of 1:50 (g / mL) to obtain a Porphyra yezoensis dispersion.
[0054] Pediococcus pentosaceus bacterial suspension was prepared according to the method in Example 1, with a bacterial suspension concentration of 1.0 × 10⁻⁶. 8 cfu / mL.
[0055] The laver dispersion was sterilized in a high-pressure steam sterilizer at 121°C for 15 minutes. Then, the laver dispersion was cooled to below 37°C. 5% Pediococcus pentosaceus culture was added to the laver dispersion as a starter culture, and the mixture was allowed to ferment at 37°C for 12 days. During the fermentation process, the pH of the fermentation broth was measured every 3 days using a pH meter.
[0056] The Control group was treated by adding unfermented laver slurry.
[0057] Depend on Figure 4 It can be seen that during the 12 days of fermentation, the pH of the control group (unfermented group) remained basically unchanged. The pH of the fermentation broth prepared from *Pediococcus pentosaceus* strains 3-5-2 and PP changed slightly, but the pH of the fermentation broth in the HYZ2-1 group decreased the most, dropping sharply in the early stage of fermentation (days 0-6) and remaining basically unchanged in the later stage (days 6-12). These results indicate that the *Porphyra yezoensis* dispersion inoculated with *Pediococcus pentosaceus* strains 3-5-2 and PP underwent almost no fermentation, while the *Porphyra yezoensis* dispersion inoculated with strain HYZ2-1 fermented for at least six days.
[0058] Example 4: Determination of Cell Concentration in Porphyra yezoensis-Marine Lactic Acid Bacteria Fermentation Broth
[0059] Porphyra yezoensis from Xiapu, Ningde was selected, and the Porphyra yezoensis was mixed with purified water at a ratio of 1:50 (g / mL) to obtain a Porphyra yezoensis dispersion.
[0060] Pediococcus pentosaceus bacterial suspension was prepared according to the method in Example 1, with a bacterial suspension concentration of 1.0 × 10⁻⁶. 8 cfu / mL.
[0061] The *Porphyra yezoensis* dispersion was sterilized in a high-pressure steam sterilizer at 121°C for 15 minutes. Then, the *Porphyra yezoensis* dispersion was cooled to below 37°C. 5% *Pediococcus pentosaceus* culture was added to the *Porphyra yezoensis* solution as a starter culture, and the mixture was allowed to ferment at 37°C for 72 hours to obtain the *Porphyra yezoensis*-*Pediococcus pentosaceus* fermentation broth.
[0062] After centrifuging the fermentation broth of *Porphyra yezoensis* and *Pediococcus pentosacchari* at 6000 rpm for 20 min to remove algal residue and *Lactobacillus* cells, the broth was filtered through a 0.22 μm membrane to obtain the fermentation supernatant of *Porphyra yezoensis* and *Pediococcus pentosacchari*.
[0063] Caco-2 cells were grown at a rate of 1.0 × 10⁻⁶. 4 Cells were seeded at a density of 100 μL / well in 96-well plates, with three replicates per group. Cells were cultured for 24 hours, with medium changed every other day. When cells reached 80-90% confluence, the 96-well plates were removed, washed once with PBS, and then 100 μL of different concentrations (2%, 4%, 10%, 20%, 40%, 100%) of fermentation broth were added to each well. The plates were then co-cultured with Caco-2 cells at 37°C and 5% CO2 for 24 hours. Then, 10 μL of CCK-8 reagent was added to each well, and absorbance was measured at 450 nm to calculate cell viability.
[0064] Control group: Caco-2 cells were 1.0 × 10⁻⁶ 4 Cells were seeded at a density of 100 μL / well in 96-well plates, with three replicates per group. Cells were cultured for 24 h, with medium changed every other day. When cells reached 80-90% confluence, the 96-well plates were removed, washed once with PBS, and then 100 μL of different concentrations (2%, 4%, 10%, 20%, 40%, 100%) of unfermented *Porphyra yezoensis* dispersion was added to each well. The cells were then co-cultured with Caco-2 cells at 37°C and 5% CO2 for 24 h. Then, 10 μL of LCK-8 reagent was added to each well, and absorbance was measured at 450 nm to calculate cell viability.
[0065] Depend on Figure 5It can be seen that, in both the control group and the experimental group, the toxicity to cells was relatively low at a fermentation broth concentration of 20% (fermentation broth diluted 5 times). Therefore, subsequent cell experiments will use a 20% concentration of fermentation broth.
[0066] Example 5: Effects of different Pediococcus pentosaceus-Porphyra yezoensis fermentation broths on the expression of intestinal barrier-related genes in Caco-2 cells.
[0067] Porphyra yezoensis from Xiapu, Ningde was selected, and the Porphyra yezoensis was mixed with purified water at a ratio of 1:50 (g / mL) to obtain a Porphyra yezoensis solution.
[0068] Pediococcus pentosaceus bacterial suspension was prepared according to the method in Example 1, with a bacterial suspension concentration of 1.0 × 10⁻⁶. 8 cfu / mL.
[0069] The laver solution was sterilized in a high-pressure steam sterilizer at 121°C for 15 minutes; then, the laver solution was cooled to below 37°C; 5% Pediococcus pentosaceus bacterial solution was added to the laver solution as a fermentation agent, and the solution was allowed to ferment statically at 37°C for 72 hours to obtain laver-Pediococcus pentosaceus fermentation broth.
[0070] After centrifuging the fermentation broth of *Porphyra yezoensis* and *Pediococcus pentosacchari* at 6000 rpm for 20 min to remove algal residue and *Lactobacillus* cells, the broth was filtered through a 0.22 μm membrane to obtain the fermentation supernatant of *Porphyra yezoensis* and *Pediococcus pentosacchari*.
[0071] qRT-PCR experiment: Caco-2 cells were cultured in DMEM medium supplemented with 10% (v / v) heat-inactivated fetal bovine serum (FBS) and 1% (v / v) penicillin-streptomycin solution at 37°C and 5% CO2. Cells were then cultured in 12-well plates (Greiner, Shanghai, China) at a rate of 1×10⁻⁶ cells / well. 5 Cells were seeded at a rate of 1 cell per well until the cells reached 80-90% confluence, at which point experimental treatment began. The groups were as follows: Experimental treatment group: Received fermentation broth containing one of the following: *Porphyra yezoensis*-HYZ2-1, *Porphyra yezoensis*-3-5-2, or *Porphyra yezoensis*-PP, along with DMEM medium containing LPS; Model group (LPS): Received DMEM medium containing LPS; Untreated group: Received unfermented *Porphyra yezoensis* slurry for intervention; Normal control group (Control): Received only DMEM medium. After a period of cell intervention, Caco-2 cells were washed with PBS, and RNA was extracted from the cells according to the instructions of the RNA extraction kit (R1200-100, Solarbio, Beijing, China). The All-in-One qRT Mix with dsDNase kit (RTQ-204, TianSai) reverse transcribed RNA into cDNA. qPCR analysis was performed on a BioRad-CFX96Touch instrument (Bio-Rad, California, USA). qPCR was performed using the qPCRMaster Mix (QST-100, TianSai) kit. The expression levels of the target gene were normalized to the expression levels of β-actin in the same samples. Data were analyzed using a 2-... ΔΔct The method is used for analysis.
[0072] Depend on Figure 6 It can be seen that the fermentation broth of *Porphyra yezoensis*-HYZ2-1 can specifically upregulate the expression levels of occludin and claudin-2 genes, and is significantly higher than that of *Porphyra yezoensis*-3-5-2 fermentation broth and *Porphyra yezoensis*-PP fermentation broth. The fermentation broth of *Porphyra yezoensis*-HYZ2-1 shows the ability to upregulate the expression levels of genes related to intestinal barrier function.
[0073] Example 6: Effects of *Porphyra yezoensis*-marine lactic acid bacteria fermentation broth, which has the potential to enhance intestinal barrier function, on the expression of intestinal barrier-related genes in Caco-2 cells.
[0074] Porphyra yezoensis from Xiapu, Ningde was selected, and the Porphyra yezoensis was mixed with purified water at a ratio of 1:50 (g / mL) to obtain a Porphyra yezoensis dispersion.
[0075] Pediococcus pentosaceus bacterial suspension was prepared according to the method in Example 1, with a bacterial suspension concentration of 1.0 × 10⁻⁶. 8 cfu / mL.
[0076] The *Porphyra yezoensis* dispersion was sterilized in a high-pressure steam sterilizer at 121°C for 15 minutes. Then, the *Porphyra yezoensis* dispersion was cooled to below 37°C. 5% *Pediococcus pentosaceus* culture was added to the *Porphyra yezoensis* dispersion as a starter culture, and the mixture was allowed to ferment at 37°C for 72 hours. This yielded the *Porphyra yezoensis*-*Pediococcus pentosaceus* fermentation broth.
[0077] After centrifuging the fermentation broth of *Porphyra yezoensis* and *Pediococcus pentosacchari* at 6000 rpm for 20 min to remove algal residue and *Lactobacillus* cells, the broth was filtered through a 0.22 μm membrane to obtain the fermentation supernatant of *Porphyra yezoensis* and *Pediococcus pentosacchari*.
[0078] qRT-PCR experiment: Caco-2 cells were cultured in DMEM medium supplemented with 10% (v / v) heat-inactivated fetal bovine serum (FBS) and 1% (v / v) penicillin-streptomycin solution at 37°C and 5% CO2. To determine the expression of ZO-1, occludin, claudin-1, and claudin-4 tight junction protein-related genes, qRT-PCR was performed on 12-well cell culture plates (Greiner, Shanghai, China) at a rate of 1 × 10⁻⁶. 5 Cells were seeded at a rate of [number] cells / well until 80-90% confluence was achieved, at which point experimental treatment began. The groups were as follows: the experimental treatment group received DMEM medium containing *Porphyra yezoensis*-HYZ2-1 fermentation broth and LPS; the model group received DMEM medium containing LPS; the untreated group received unfermented *Porphyra yezoensis* slurry; and the normal control group received only DMEM medium. After a period of time, Caco-2 cells were washed with PBS, and RNA was extracted from the cells according to the instructions of the RNA extraction kit (R1200-100, Solarbio, Beijing, China). The All-in-One qRT Mix with dsDNase kit (RTQ-204, TianSai) reverse transcribed RNA into cDNA. qPCR analysis was performed on a BioRad-CFX96 Touch instrument (Bio-Rad, California, USA). qPCR was performed using the qPCR Master Mix (QST-100, TianSai) kit. The expression levels of the target gene were normalized to the expression levels of β-actin in the same samples. Data were analyzed using a 2-... ΔΔct The method is used for analysis.
[0079] Depend on Figure 7 It can be seen that the fermentation broth of *Porphyra yezoensis*-HYZ2-1 can specifically upregulate the expression levels of ZO-1, occludin, claudin-1, and claudin-4 genes. Compared with the untreated group, the HYZ2-1 group showed an upregulation of 1.427-fold, 2.031-fold, 1.36-fold, and 1.948-fold, respectively. The fermentation broth of *Porphyra yezoensis*-HYZ2-1 demonstrates the ability to upregulate the expression levels of genes related to intestinal barrier function.
Claims
1. A strain of Pediococcus pentosaceus, characterized in that, The Pediococcus pentosaceus strain is classified as follows: Pediococcus pentosaceus The depositary institution is the China Center for Type Culture Collection, the accession number is CCTCC M 20251312, and the deposit date is June 9, 2025.
2. A Pediococcus pentosaceus bacterial suspension, characterized in that, The active ingredient in the Pediococcus pentosaccharis bacterial solution is the Pediococcus pentosaccharis strain as described in claim 1.
3. The Pediococcus pentosaceus bacterial suspension according to claim 2, characterized in that, The concentration of Pediococcus pentosaceus in the bacterial solution was 1.
0. 10 8 cfu / mL.
4. A method for preparing a fermentation broth of *Porphyra yezoensis*-*Pediococcus pentosaceus*, characterized in that, include: Disperse the laver in water, sterilize it, add the Pediococcus pentosacchari bacterial solution as described in claim 2, and ferment it to obtain laver-Pediococcus pentosacchari fermentation broth.
5. The preparation method according to claim 4, characterized in that, The dispersion ratio of the laver to water is 1g: 20~100mL.
6. The preparation method according to claim 4, characterized in that, The amount of Pediococcus pentosaceus bacterial solution added is 1-10% of the volume of the fermentation liquid after addition.
7. The preparation method according to claim 4, characterized in that, Fermentation time is 1 to 12 days.
8. The fermentation broth of *Porphyra yezoensis*-Pediococcus pentosaceus prepared by the method described in claim 4.
9. The application of the *Porphyra yezoensis*-*Pediococcus pentosacchari* fermentation broth prepared by the preparation method of claim 4, or the *Porphyra yezoensis*-*Pediococcus pentosacchari* fermentation broth of claim 8, in the preparation of products that enhance intestinal barrier function.
10. The application according to claim 9, characterized in that, The enhancement of intestinal barrier function is manifested by upregulating the expression levels of ZO-1, occludin, claudin-1 and / or claudin-4 tight junction protein genes.