Pediococcus pentosaceus HNTP and application thereof

Abstract

The invention relates to the technical field of microorganisms, and particularly discloses pediococcus pentosaceus HNTP and application thereof. The preservation number of the Pediococcus pentosaceus HNTP is CGMCC No.29067, the Pediococcus pentosaceus HNTP is high in acid resistance and fast in growth, when the Pediococcus pentosaceus HNTP is applied to fermentation of high-fiber matrix grains, the relative abundance of various bacteria can be adjusted, various volatile gases can be generated, the content of beneficial components in a fermentation product can be increased, traditional fermented foods and beverages can be regulated, controlled and improved, and the effects of improving the quality of the high-fiber matrix grains are achieved. And a new choice is provided for whole grain plant-based fermentation.

Description

Technical Field

[0001] This invention relates to the field of microbial technology, and more specifically, to a strain of Pediococcus pentosaceus HNTP and its applications. Background Technology

[0002] Pediococcus pentosaceus possesses various beneficial functions for the human body. According to existing research, the reported functions of Pediococcus pentosaceus mainly include: promoting digestion and gastrointestinal motility, improving symptoms of indigestion; enhancing immunity: Pediococcus pentosaceus, containing polysaccharide components, can stimulate immune cells to produce antibodies, enhancing the body's immunity and preventing disease. In agriculture and animal husbandry, as a growth stimulant and preservative, Pediococcus pentosaceus can extend the shelf life of forage during low-temperature storage. Despite these functions, Pediococcus pentosaceus still has some shortcomings in practical applications, such as limited adaptability to the environment, potentially less effective inhibition of certain pathogens than expected, and issues with stability and efficacy in food processing.

[0003] In the field of grain fermentation, commonly used microbial strains include lactic acid bacteria and yeast. These strains can produce specific enzymes during fermentation, which help reduce the content of anti-nutritional factors in grains and improve their nutritional value and digestibility. However, these traditional fermentation strains also have some drawbacks, such as the need to improve their effectiveness and the limited number of strains suitable for plant-based beverages such as grains. Therefore, it is necessary to research and develop new strains. Summary of the Invention

[0004] One of the objectives of this invention is to provide a new strain of Pediococcus pentosaceus suitable for grain fermentation and its applications.

[0005] To achieve this objective, the technical solution of the present invention is as follows:

[0006] This invention provides a Pediococcus pentosaceus HNTP strain, with accession number CGMCC No. 29067.

[0007] The present invention also provides a microbial agent containing the above-mentioned Pediococcus pentosaceus HNTP.

[0008] The microbial agent of the present invention contains Pediococcus pentosaceus HNTP, which can be a solid or liquid agent. Those skilled in the art can prepare the agent using methods known in the art. Preferably, the agent comprises a fermentation broth of Pediococcus pentosaceus HNTP.

[0009] The present invention also provides the use of the above-mentioned Pediococcus pentosaceus HNTP or bacterial agent in fermenting high-fiber matrix grains or in the preparation of high-fiber matrix grain beverages or in antibacterial applications.

[0010] In the application of this invention, the high-fiber matrix grain is oats.

[0011] The present invention, Pediococcus pentosaceus HNTP, can increase the content of various beneficial components in fermentation products during oat fermentation, thereby enhancing the efficacy of oat fermentation products.

[0012] The Pentosaccharide Pediococcus HNTP of the present invention has excellent inhibitory effects on Escherichia coli and Staphylococcus aureus.

[0013] The present invention also provides the application of the above-mentioned Pediococcus pentosaceus HNTP or bacterial agent in increasing the content of lactic acid, flavonoids, amino acids, β-glucan and / or reducing sugar in the products after oat fermentation.

[0014] The present invention also provides the application of the above-mentioned Pediococcus pentosaceus HNTP or bacterial agent in altering the relative abundance of one or more of the genera Weissella, Rhodococcus, Lactobacillus, Gordonia, and Bacteroides during oat fermentation.

[0015] The present invention further provides the application of the above-mentioned Pediococcus pentosaceus HNTP or bacterial agent in altering the relative abundance of one or more of the genera Weissella, Rhodococcus, Pseudomonas, Lactobacillus, Gordonia, and Bacteroides during oat fermentation.

[0016] In the application of the present invention, the above-mentioned Pediococcus pentosaceus is used. When using pentosaceus HNTP or microbial agents for oat fermentation, the relative abundance of *Weissella*, *Rhodococcus*, *Pseudomonas*, and *Lactobacillus* is increased before 24 hours; from 24 to 48 hours, the relative abundance of *Weissella*, *Pseudomonas*, *Gordonia*, and *Bacteroides* is increased, while the relative abundance of *Rhodococcus* and *Lactobacillus* is decreased; from 48 to 72 hours, the relative abundance of *Lactobacillus* is increased, while the relative abundance of *Weissella*, *Rhodococcus*, *Pseudomonas*, *Gordonia*, and *Bacteroides* is decreased.

[0017] The Pentosacchariphyte HNTP of this invention can adjust the relative abundance of various bacteria during oat fermentation, thereby adjusting the composition of fermentation products and the generation of volatile gases, providing a new approach for regulating and improving traditional fermented foods and beverages.

[0018] This invention also provides the application of the above-mentioned Pediococcus pentosaceus HNTP or bacterial agent in the fermentation enhancement of methyl acetate, butyric acid, propionic acid, amyl acetate, 1-propanol, isoamyl acetate, ethyl butyrate, ethyl propionate, butyl acetate, propyl acetate, isobutyl acetate, 3-methyl-2-butenal, 3-methyl-3-buten-1-ol, cis-2-penten-1-ol, butyl 3-methylbutyrate, ethyl acetate, propional, 1-penten-3-ol, ethyl isobutyrate, isobutanol, acetic acid, and 3-methylbutanol. Most of these substances have fruity, floral, herbal, or cheese-like aromas, and therefore, HNTP fermentation can produce and control unique product flavors. Preferably, the fermentation substrate is oats.

[0019] The present invention also provides an antibacterial agent comprising the above-mentioned Pediococcus pentosaceus HNTP or bacterial agent.

[0020] The present invention also provides an oat beverage or sweetened mash, which is prepared by fermenting oats with the above-mentioned Pediococcus pentosaceus HNTP or bacterial agent. Preferably, in the preparation of the oat beverage, the oats are first subjected to high-pressure jet homogenization before fermentation.

[0021] The Pediococcus pentosaceus HNTP of the present invention can be used to ferment solid oat grains to prepare sweet mash. Compared with other microbial agents, the present invention can obtain products with increased content of beneficial components.

[0022] The *Pediococcus pentosaceus* HNTP of this invention can also be used to ferment oat pulp to prepare beverages, resulting in products with stable texture and resistance to spoilage. High-pressure jet homogenization can thoroughly break down the fermentation substrate, thereby releasing more substances for fermentation bacteria and adjusting the composition of the final product.

[0023] The beneficial effects of this invention are at least as follows:

[0024] This invention provides a novel Pediococcus pentosacchariformis HNTP strain, which is a Gram-positive bacterium that undergoes glucose homofermentation, is highly acid-resistant, can grow at around pH 2, can grow and reproduce at 10-50℃, has a rapid growth rate, can inhibit the growth of pathogens, and is low in cost, safe, reliable, and easy to use.

[0025] The *Pediococcus pentosaceus* HNTP strain of this invention, when used in grain fermentation, has a good effect on regulating and improving traditional fermented foods and beverages. It can increase the content of beneficial components such as lactic acid, flavonoids, amino acids, β-glucan, and reducing sugars, and has a good promoting effect on the fermentation of new whole grain beverages. It is simple to operate, inexpensive, and makes up for the current lack of suitable plant-based fermentation strains for whole grains. It can improve the efficiency and effect of grain fermentation, and at the same time help promote the application of microbial technology in the food industry and other fields. Attached Figure Description

[0026] Figure 1 For the phylogenetic tree of the family gene.

[0027] Figure 2 The images show the colony morphology of Pediococcus pentosaceus HNTP in LB and MRS media. The left image shows the colony morphology in LB medium, the middle image shows the colony morphology in MRS medium, and the right image shows the colony morphology in LB liquid medium.

[0028] Figure 3 The growth curve of Pediococcus pentosaceus HNTP is shown.

[0029] Figure 4 The growth of Pediococcus pentosaceus HNTP at different pH values.

[0030] Figure 5 The growth of Pediococcus pentosaceus HNTP at different temperatures.

[0031] Figure 6 The inhibition results of Pediococcus pentosus HNTP on Escherichia coli (left image, MRS plate) and Staphylococcus aureus (right image, LB plate).

[0032] Figure 7 This describes the growth of Pediococcus pentosaceus HNTP on GSP medium, producing viscous polysaccharides.

[0033] Figure 8 This describes the reduction of Cr(VI) to Cr(III) by *Pediococcus pentosaceus* HNTP on potassium chromate medium.

[0034] Figure 9 This describes the hydrolysis of skim milk powder by Pediococcus pentosaceus HNTP.

[0035] Figure 10 This describes the formation of a transparent hydrolysis zone (2 cm radius) by Pediococcus pentosaceus HNTP on carboxymethyl cellulose sodium (CMC) medium.

[0036] Figure 11 This diagram shows the composition of sweet mash bacteria during oat fermentation in Example 3 of the present invention. A represents the phylum level, B represents the genus level, C represents the Venn diagram at each time point, and D represents the PCA diagram of sweet mash bacteria during fermentation.

[0037] Figure 12 The image shows the composition of bacteria in the sweet mash during the oat fermentation process of the control bacteria in Example 3 of this invention, where A represents the phylum level and B represents the genus level.

[0038] Figure 13 The lactic acid content at different stages during oat fermentation in Example 3 of this invention.

[0039] Figure 14 The content of flavonoids at different stages during oat fermentation in Example 3 of the present invention.

[0040] Figure 15 The amino acid content at different stages during oat fermentation in Example 3 of this invention.

[0041] Figure 16 The content of β-glucan at different time points during oat fermentation in Example 3 of the present invention.

[0042] Figure 17 The reducing sugar content at different stages during oat fermentation in Example 3 of this invention.

[0043] Figure 18 This is a db-RDA analysis diagram of bacterial composition and sweet mash properties during fermentation, where LA—lactic acid, F—flavonoids, AA—amino acids, S—starch, RS—reducing sugars, and β-glucan—β-glucan.

[0044] Figure 19 This is a fingerprint spectrum of volatile components.

[0045] Figure 20The results of the Circos genome circle map analysis.

[0046] Figure 21 The results are from the analysis of carbohydrate-active enzymes.

[0047] Figure 22 This is a linear map of gene clusters for the synthesis of secondary metabolites. Detailed Implementation

[0048] The preferred embodiments of the present invention will be described in detail below with reference to examples. It should be understood that the following examples are given for illustrative purposes only and are not intended to limit the scope of the invention. Those skilled in the art can make various modifications and substitutions to the present invention without departing from its spirit and intent. Unless otherwise specified, the experimental methods used in the following examples are conventional methods. Unless otherwise specified, the materials and reagents used in the following examples can be obtained commercially or prepared according to conventional methods in the art. Example 1: Isolation and Screening of Pediococcus pentosaceus

[0049] In this embodiment, a strain of *Pediococcus pentosaceus* was obtained by screening using the dilution plate method, as follows: An Erlenmeyer flask containing 500 mL of distilled water, a test tube containing 4.5 mL of distilled water, MRS solid medium, and plates were sterilized. After sterilization, the MRS medium was poured into the plates. Under aseptic conditions, 10 g of traditionally fermented whole-grain sweet mash was mixed thoroughly, and then 90 mL of sterile distilled water was added and shaken until a final concentration of 10% was achieved. -1 Then, take 500 μL of this liquid and add it to a test tube containing 4500 μL of sterile distilled water. Shake to mix thoroughly, and then continuously dilute to achieve concentrations of 10 μL and 10 μL respectively. -2 10 -3 10 -4 and 10 -5 , 10 -3 and 10 -5 Repeatedly streak plate separation with 20 μL of liquid until a single colony is obtained. Inoculate the single colony into a 2 mL cryovial containing 20% ​​glycerol using a sterile inoculation needle and store at -80°C for later use.

[0050] MRS medium consisted of: 10.0 g peptone; 10.0 g beef extract; 5.0 g yeast extract; 20.0 g dextrose; 1.0 g polysorbate 80; 2.0 g ammonium citrate; 5.0 g sodium acetate; 0.1 g magnesium sulfate; 0.05 g manganese sulfate; 2.0 g dipotassium phosphate; and 1000 mL distilled water. Solid medium was added with 15 g / L agar and sterilized at 121°C for 15 min.

[0051] A fast-growing, acid-resistant, and wide-temperature-range lactic acid bacterium, *Pediococcus pentosaceus* HNTP, was selected for biopreservation. This strain was deposited on November 20, 2023, at the China General Microbiological Culture Collection Center (CGMCC, address: No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, 100101, China), and classified as *Pediococcus pentosaceus*, with accession number CGMCC No. 29067.

[0052] The full-length HNTP gene of *Pediococcus pentosaceus* was extracted, and then the 16S rRNA gene was amplified using PCR primers 27f (5'-AGAGTTTGATCCTGGCTC-3', SEQ ID No. 1) and 1492r (5'-TACGGCTACCTTGTTACGACT-3', SEQ ID No. 2). BLAST (http: / / www.ncbi.nlm.nih.gov / BLAST) was used to identify the species with the highest homology to the gene sequence of the target lactic acid bacteria strain among those of known taxonomic positions. Gene sequences of known lactic acid bacteria strains were selected from GenBank and compared with the 16S rRNA gene sequence of the target lactic acid bacteria strain to determine their similar species and taxonomic position.

[0053] Specifically, based on 31 housekeeping genes (dnaG, frr, infC, nusA, pgk, pyrG, rplA, rplB, rplC, rplD, rplE, rplF, rplK, rplL, rplM, rplN, rplP, rplS, rplT, rpmA, rpoB, rpsB, rpsC, rpsE, rpsI, rpsJ, rpsK, rpsM, rpsS, smpB, tsf), 19 strains most closely related at the species level were selected, and a phylogenetic tree was constructed using the Neighbor-Joining (NJ) method with MEGA 6.0 software. Results are shown below. Figure 1 .

[0054] Example 2: Cultivation and observation of Pediococcus pentosaceus

[0055] This embodiment studies the growth characteristics of the Pediococcus pentosacchari HNTP obtained in Example 1.

[0056] 1. *Pediococcus pentosaceus* HNTP was cultured in MRS and LB media, and its colony morphology was observed. See the attached table for details. Figure 2 .

[0057] MRS medium consisted of: 10.0 g peptone; 10.0 g beef extract; 5.0 g yeast extract; 20.0 g glucose; 1 ml polysorbate 80; 2.0 g ammonium citrate; 5.0 g sodium acetate; 0.2 g magnesium sulfate; 0.05 g manganese sulfate; 2.0 g dipotassium phosphate; and 1000 mL distilled water. Solid medium was added with 15 g / L agar and sterilized at 121°C for 15 min.

[0058] LB medium: 10g tryptone, 10g NaCl, 5g yeast extract, 1000mL distilled water; add 15g / L agar to solid medium, sterilize at 121℃ for 15min.

[0059] 2. Pediococcus pentosaceus HNTP and Lactobacillus plantarum L1 (described in: Ren Fei, Liu Yuchun, Wang Chao, et al. Screening and growth characteristics of highly efficient probiotics for degrading corn gluten powder [J]. Grain and Oil Food Science and Technology, 2021, 29(3):183-191, in which Lactobacillus plantarum L1 was named CGM57) were cultured in LB medium (pH 7.0) at 37℃ for 48 h, and the growth was recorded. The specific results are shown in [link to results]. Figure 3 .

[0060] 3. Following the culture conditions described in section 2 above, but changing the pH of the LB medium to 2-10, culture *Pediococcus pentosaceus* HNTP and *Lactobacillus plantarum* L1 was performed. See the results below. Figure 4 .

[0061] 4. Under the same culture conditions as in section 2 above, but with the culture temperature changed to 10℃, 20℃, 30℃, 40℃, and 50℃ respectively, *Pediococcus pentosaceus* HNTP and *Lactobacillus plantarum* L1 were cultured. See the results below. Figure 5 .

[0062] 5. The antibacterial activity, extracellular mucopolysaccharide production, Cr reduction, protease production, and cellulose decomposition of *Pediococcus pentosaceus* HNTPs were detected. Results are shown below. Figures 6 to 10 .

[0063] Detection method: In a sterile operating table, culture 10 μL of bacterial solution (under the same culture conditions as in section 2 above, with a bacterial concentration of 1×10⁻⁶) 6 The bacterial culture medium (CFU / mL) was inoculated onto sterile filter paper on a bacterial agar plate evenly coated with pathogens, and incubated in a 30°C incubator for 48 hours. The inhibition zones of the colonies were then observed and measured.

[0064] Specifically, the HNTP of *Pediococcus pentosaceus* exhibits antibacterial activity against *Escherichia coli* and *Staphylococcus aureus*, with clear zone diameter to colony diameter ratios of 1.9 (against *Escherichia coli*) and 2.4 (against *Staphylococcus aureus*). The same antibacterial experiment was conducted using the control strain CICC23190 (a related species of *Pediococcus pentosaceus*), with clear zone diameter to colony diameter ratios of 1.2 (against *Escherichia coli*) and 1.3 (against *Staphylococcus aureus*).

[0065] 6. The sensitivity of Pediococcus pentosaceus HNTP to different carbon sources and chemical sensitivities was tested. Results: After testing with the Biolog method, the carbon sources available were: dextrin, D-trehalose, D-cellobiose, gentiobiose, D-minobiose, stachyose, raffinose, β-formyl-D-glucoside, D-salicylic acid, α-D-glucose, D-mannose, D-galactose, inosine, L-malic acid, and Tween 40.

[0066] Chemically sensitive substances: lincomycin, guanidine hydrochloride, sodium tetradecanoate sulfate, sodium butyrate, sodium bromate, potassium tellurite. Example 3: Preparation of sweet fermented whole-grain oats using Pediococcus pentosaceus HNTP.

[0067] In this embodiment, HNTP bacterial culture was added to cooked oats at a ratio of 1% (v / w, mL / g) and fermented at 25°C. Illumina sequencing was used to analyze the succession of the bacterial community and changes in nutrients during fermentation. The HNTP bacterial culture was obtained by culturing *Pediococcus pentosaceus* HNTP in LB medium (pH 7.0) at 37°C for 48 hours, with a bacterial concentration of 1×10⁻⁶. 6 CFU / mL.

[0068] To investigate the succession of the microbial community in the samples, a related species, CICC 23190, was used as a control under the same conditions. To investigate the changes in nutrient content, Pediococcus pentosaceus CICC 23190 (DZJ) and Lactobacillus plantarum L1 were used as controls fermented in oats.

[0069] When using HNTPs for fermentation, the composition of bacteria in the sweet mash during oat fermentation is shown in [reference needed]. Figure 11 The changes in various nutrients are shown in the table below. Figures 13 to 17 The bacterial composition and properties of sweet mash are shown in the db-RDA analysis diagram. Figure 18 .

[0070] When using the control strain CICC 23190 for fermentation, the bacterial composition of the sweet mash during oat fermentation is shown in the figure. Figure 12 .

[0071] When fermented with HNTP, at the bacterial phylum level, the abundance of all four phyla (Firmwallis, Actinobacteria, Proteobacteria, and Bacteroidetes) in the sweet mash samples was greater than 1%, with Firmicutes being the absolute dominant phylum. The relative abundance increased from 68.84% at 24 h of fermentation to 69.49% at 48 h, reaching a peak of 94.81% at 72 h. At the genus level, during the fermentation of oat sweet mash samples by *Pediococcus pentosaceus*, the initial inoculum *Pediococcus pentosaceus* was significantly dominant throughout the fermentation process. Its relative abundance decreased slightly from 34.96% at 24 h to 28.83% at 48 h, reaching a peak of 59.69% at 72 h. Using Pediococcus pentosaceus HNTP as a fermentation strain can regulate and alter the relative abundance of genera such as Weissella, Rhodococcus, Pseudomonas, Lactobacillus, Gordonia, and Bacteroides during oat fermentation. Specifically, before 24 hours, HNTPs can increase the relative abundance of *Weissella*, *Rhodococcus*, *Pseudomonas*, and *Lactobacillus*; from 24 to 48 hours, they increase the relative abundance of *Weissella*, *Pseudomonas*, *Gordonia*, and *Bacteroides*, while decreasing the relative abundance of *Rhodococcus* and *Lactobacillus*; from 48 hours to 72 hours, they increase the relative abundance of *Lactobacillus*, while decreasing the relative abundance of *Weissella*, *Rhodococcus*, *Pseudomonas*, *Gordonia*, and *Bacteroides*.

[0072] When fermentation was carried out using control bacteria, the changes in the bacterial community were significantly different from those during HNTP fermentation.

[0073] Principal component analysis (PCA) revealed that the sweet mash samples formed distinct microbial communities at 24h, 48h, and 72h during fermentation, exhibiting different microbial community compositions and succession processes at these times. From 0h to 72h, the contents of lactic acid, flavonoids, amino acids, β-glucan, and reducing sugars showed a continuous increase, reaching their highest values ​​at 72h, while starch content showed the opposite trend. The bacterial composition during fermentation was significantly correlated with the properties of the sweet mash (LA—lactic acid, F—flavonoids, AA—amino acids, S—starch, RS—reducing sugars, β-glucan—β-glucan). This analysis provides a theoretical foundation and reference for the development of microbial starter cultures for traditional grain fermentation foods, the improvement of oat product quality, and industrial production.

[0074] When HNTPs are fermented at 30°C, they reach the level achieved after 72 hours of fermentation at 25°C in just 48 hours. Pediococcus pentosaceus HNTPs can be used to ferment oats to produce sweet mash samples and to regulate the microbial community and nutrients.

[0075] pass Figures 13 to 17 It is known that using the Pentosacchariformis HNTP of this invention for oat fermentation can increase the content of various active ingredients.

[0076] Example 4: Study on volatile organic compounds of HNTP strain during oat fermentation in culture medium

[0077] Pediococcus pentosaceus HNTP was cultured in LB medium (pH 7) at 30°C for 48 h, and volatile organic compounds (VOCs) were analyzed using GC-IMS (three biological replicates, HNTP group). Pediococcus pentosaceus HNTP was then used to ferment oats according to the method described in Example 3, and VOCs were analyzed using GC-IMS after 48 h (three biological replicates, HNTPYM group). The control strain, Pediococcus pentosaceus CICC 23190, was cultured in LB medium under the same conditions (DZJ group) and fermented oats (23TP group), and the results were analyzed under the same conditions (three biological replicates). A detailed list of VOCs from the four experiments is shown in Table 1. The results of the comparative analysis of VOC fingerprint spectra are shown in... Figure 19 .

[0078] The results showed that fermentation of Pediococcus pentosus HNTP in LB medium could produce the following substances: butyric acid, propionic acid, acetic acid, ethyl lactate, etc.

[0079] The fermentation of Pediococcus pentosus HNTP in oats can produce the following substances: methyl acetate, butyric acid, propionic acid, amyl acetate, 1-propanol, isoamyl acetate, ethyl butyrate, ethyl propionate, butyl acetate, etc.

[0080] Significant differences were observed between *Pediococcus pentosaceus* HNTP and the control strain CICC 23190 during oat fermentation in LB medium. During oat fermentation, the group fermented with *Pediococcus pentosaceus* HNTP (HNTPYM group) exhibited significantly higher levels and types of volatile organic compounds than the group fermented with the control strain CICC 23190 (23TP group). *Pediococcus pentosaceus* HNTP fermentation of oats produced large amounts of esters, acids, and alcohols; ethanol and furfural levels were higher in the DZJ and 23TP groups. After fermentation with the control strain (23TP group), the difference between the control group and the unfermented group (DZJ group) was small, and no new substances were produced during fermentation.

[0081] Specifically comparing the two groups: Compared to the DZJ group, HNTP fermentation in LB medium produces higher levels of substances including butyraldehyde, 3-methylbutyraldehyde, and octanal; compared to the 23TP group, HNTP fermentation in oats produces higher levels of substances including methyl acetate, butyric acid, propionic acid, amyl acetate, 1-propanol, isoamyl acetate, ethyl butyrate, ethyl propionate, butyl acetate, propyl acetate, isobutyl acetate, 3-methyl-2-butenal, 3-methyl-3-buten-1-ol, cis-2-penten-1-ol, butyl 3-methylbutyrate, ethyl acetate, propionaldehyde, 1-penten-3-ol, ethyl isobutyrate, isobutanol, acetic acid, and 3-methylbutanol. Most of these substances have fruity, floral, herbal, and cheese-like aromas, thus HNTP fermentation produces unique flavors compared to other bacteria of the same genus.

[0082] Table 1. List of Volatile Components

[0083]

[0084]

[0085]

[0086]

[0087]

[0088]

[0089] Example 5: High-pressure jet coupled bacterial fermentation to produce whole grain beverage

[0090] In this embodiment, roasted oats (10% w) were mixed with water and treated with a high-pressure jet (Beijing Collaborative Innovation Food Technology Co., Ltd., FJ-3037D) at 120 MPa. Then, Pediococcus pentosaceus HNTP bacterial solution (see Example 3) was added at a ratio of 0.5% (v / w, mL / g) and fermented at 25°C for 8 hours to obtain a whole-grain oat beverage. Quality was tested (average of 3 tests). Pediococcus pentosaceus CICC 23190 and Lactobacillus plantarum L1 were used as controls. The comparison of test results is shown in Table 2. Comparison photos of HNTP and L1 are shown below. Figure 20 .

[0091] Table 2. Effects of high-pressure jet-coupled bacterial strains on the quality of whole-grain beverages.

[0092]

[0093] When whole-grain oat beverages are prepared using HNTP fermentation, the levels of bioactive substances (dry basis), dietary fiber, folic acid, and stability are significantly enhanced, making it suitable for fermenting high-fiber matrix whole-grain beverages. No obvious spoilage or mold is observed after two weeks of storage at room temperature following fermentation. If prepared and then sterilized and bottled using UHT, it can be stored intact for over one year.

[0094] Example 6: HNTP Genome Analysis

[0095] This embodiment analyzed the genome of HNTPs, and the results are shown in Table 3. Figures 20 to 22 .

[0096] Table 3

[0097]

[0098] Results of Circos genome mapping analysis are shown below Figure 20 The results of carbohydrate activity enzyme analysis are shown in [the table below]. Figure 21 The linear map of the gene clusters for secondary metabolite synthesis is shown below. Figure 22 .

[0099] Although the present invention has been described in detail above with general descriptions and specific embodiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention fall within the scope of protection claimed by the present invention.

Claims

1. A Pediococcus pentosaceus HNTP, characterized in that, The accession number is CGMCCNo.29067.

2. A microbial agent, characterized in that, Contains the Pediococcus pentosaceus HNTP as described in claim 1; preferably, the inoculum is a fermentation broth of Pediococcus pentosaceus HNTP.

3. The application of the Pediococcus pentosaceus HNTP as described in claim 1, or the bacterial agent as described in claim 2, in the fermentation of high-fiber substrate grains, the preparation of high-fiber substrate grain beverages, or in antibacterial activities.

4. The application according to claim 3, characterized in that, The high-fiber matrix grain is oats.

5. The application of the Pediococcus pentosaceus HNTP as described in claim 1, or the inoculant as described in claim 2, in increasing the content of lactic acid, flavonoids, amino acids, β-glucan, and / or reducing sugars in the products after oat fermentation.

6. The application of the Pediococcus pentosaceus HNTP of claim 1, or the inoculum of claim 2, in adjusting the relative abundance of one or more of the genera Weissella, Rhodococcus, Pseudomonas, Lactobacillus, Gordonia, and Bacteroides during oat fermentation.

7. The application according to claim 6, characterized in that, In the Pediococcus pentosaceus as described in claim 1 When using the *P. pentosaceus* HNTP or the microbial agent of claim 2 for oat fermentation, the relative abundance of *Weissella*, *Rhodococcus*, *Pseudomonas*, and *Lactobacillus* is increased before 24 hours; from 24 to 48 hours, the relative abundance of *Weissella*, *Pseudomonas*, *Gordonia*, and *Bacteroides* is increased, while the relative abundance of *Rhodococcus* and *Lactobacillus* is decreased; from 48 to 72 hours, the relative abundance of *Lactobacillus* is increased, while the relative abundance of *Weissella*, *Rhodococcus*, *Pseudomonas*, *Gordonia*, and *Bacteroides* is decreased.

8. The application of the Pediococcus pentosaceus HNTP according to claim 1, or the inoculum according to claim 2, in the fermentation of methyl acetate, butyric acid, propionic acid, amyl acetate, 1-propanol, isoamyl acetate, ethyl butyrate, ethyl propionate, butyl acetate, propyl acetate, isobutyl acetate, 3-methyl-2-butenal, 3-methyl-3-buten-1-ol, cis-2-penten-1-ol, butyl 3-methylbutyrate, ethyl acetate, propional, 1-penten-3-ol, ethyl isobutyrate, isobutanol, acetic acid, and 3-methylbutanol, preferably, wherein the substrate for fermentation is oats.

9. An antibacterial agent, characterized in that, It contains Pediococcus pentosaceus HNTP as described in claim 1 or the bacterial agent as described in claim 2.

10. An oatmeal beverage or sweetened cereal, characterized in that, It is prepared by fermenting oats with Pediococcus pentosaceus HNTP as described in claim 1 or the bacterial agent as described in claim 2. Preferably, in the preparation of oat beverage, the oats are first homogenized by high-pressure jet before fermentation.

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