Prebiotic compositions and uses thereof

Abstract

The present application discloses a probiotic composition, which comprises grape ferment, a first ingredient and a second ingredient. The first ingredient is lactitol or xylo-oligosaccharide, and the second ingredient is fructo-oligosaccharide, xylo-oligosaccharide or inulin. The first ingredient is different from the second ingredient, and the weight ratio of the grape ferment, the first ingredient and the second ingredient is 1-2:1-2:1-2. Therefore, the probiotic composition prepared by the grape ferment can be used to promote the intestinal health of the recipient and improve the antioxidant capacity of the recipient.

Description

TECHNICAL FIELD

[0001] The present disclosure relates to a use of a prebiotic composition, a grape ferment for preparing a prebiotic composition, and a prebiotic composition for promoting intestinal health and enhancing antioxidant capacity in a subject. BACKGROUND

[0002] Prebiotics, also known as prebiotic fibers, are polysaccharide components in natural foods that are not easily digested by human enzymes, but can be utilized by probiotics in the digestive system (mainly the large intestine) for growth, expansion, and metabolism to generate short-chain fatty acids (SFCAs).

[0003] According to the common statement of International Scientific Association for Probiotics and Prebiotics (ISAPP) in 2017 Nature report, prebiotics are defined as "substances that are selectively utilized by host-associated microorganisms, thereby promoting the health of the host".

[0004] Specifically, prebiotics can help probiotics grow and inhibit harmful bacteria in the intestine. Probiotics in the intestine also metabolize prebiotics to generate short-chain fatty acids, thereby providing energy sources for probiotics and the host. SUMMARY

[0005] Therefore, the present disclosure provides a prebiotic composition including grape ferment, which can be used to promote the growth of Akkermansia muciniphila, and can promote the intestinal health and / or enhance the antioxidant capacity of a subject.

[0006] In some embodiments, a prebiotic composition includes grape ferment, a first ingredient, and a second ingredient. The first ingredient is lactitol or xylo-oligosaccharide, and the second ingredient is fructo-oligosaccharide, xylo-oligosaccharide, or inulin. The first ingredient is different from the second ingredient. The weight ratio of grape ferment, the first ingredient, and the second ingredient is 1-2: 1-2: 1-2.

[0007] In some embodiments, the weight ratio of grape ferment, the first ingredient, and the second ingredient in the prebiotic composition is 2: 1: 1.

[0008] In some embodiments, the prebiotic composition is used to promote the growth of Akkermansia muciniphila.

[0009] In some embodiments, a prebiotic composition includes grape ferment, lactitol as a first ingredient, and fructo-oligosaccharide as a second ingredient.

[0010] In some embodiments, a grape ferment is used for preparing a prebiotic composition for promoting gut health of a subject, wherein the prebiotic composition includes grape ferment, lactitol, and fructo-oligosaccharide, and the weight ratio of grape ferment, lactitol, and fructo-oligosaccharide is 1-2: 1-2: 1-2.

[0011] In some embodiments, promoting gut health of a subject includes promoting growth of probiotics, and the probiotics include Akkermansia muciniphila and Bifidobacterium.

[0012] In some embodiments, the prebiotic composition is used to promote growth capacity of Akkermansia muciniphila.

[0013] In some embodiments, a grape ferment is used for preparing a prebiotic composition for enhancing antioxidant capacity of a subject, wherein the prebiotic composition includes grape ferment, lactitol, and fructo-oligosaccharide, and the weight ratio of grape ferment, lactitol, and fructo-oligosaccharide is 1-2: 1-2: 1-2.

[0014] In some embodiments, the prebiotic composition is used to increase sulfur-containing compounds and total Glutathione content in a subject.

[0015] In some embodiments, the sulfur-containing compounds are Thiols compounds.

[0016] In some embodiments, the prebiotic composition includes grape ferment, lactitol, and fructo-oligosaccharide in a weight ratio of 2: 1: 1.

[0017] In summary, the probiotic composition of any embodiment, which includes grape ferment, a first ingredient and a second ingredient, and the first ingredient is lactitol or xylo-oligosaccharide, and the second ingredient is fructo-oligosaccharide, xylo-oligosaccharide or inulin. Also, the probiotic composition has the ability to promote the growth of Akkermansia muciniphila. In some embodiments, the grape ferment can be used to prepare a probiotic composition that promotes the gut health of a subject and / or enhances the antioxidant capacity of a subject, and the probiotic composition includes grape ferment, lactitol and fructo-oligosaccharide. In some embodiments, the probiotic composition can promote the growth of probiotic bacteria. For example, the probiotic bacteria include Akkermansia muciniphila and Bifidobacterium. In some embodiments, the probiotic composition can increase the level of sulfur-containing compounds (e.g., Thiols) and total Glutathione in a subject. BRIEF DESCRIPTION OF DRAWINGS

[0018] Figure 1 is a graph of the results of an analysis experiment of the effect of different groups of ingredients on probiotic bacteria;

[0019] Figure 2 is a graph of the results of an analysis experiment of the effect of different groups of ratios of probiotic compositions on probiotic bacteria;

[0020] Figure 3 is a graph of the results of an experiment of the effect of probiotic compositions on the growth ability of Akkermansia muciniphila;

[0021] Figure 4 is a graph of the results of an experiment of the effect of probiotic compositions on the growth ability of Bifidobacterium;

[0022] Figure 5 is a graph of the results of an analysis experiment of the effect of probiotic compositions on the level of f-Thiols in the blood of a subject; and

[0023] Figure 6 is a graph of the results of an analysis experiment of the effect of probiotic compositions on the level of t-Glutathione in the blood of a subject. DETAILED DESCRIPTION

[0024] The prebiotic composition includes grape ferment, sugar alcohol as a first component, and saccharide as a second component, and the first component is different from the second component. The sugar alcohol can be lactitol, and the saccharide can be oligosaccharide, dietary fiber, or other polysaccharide, such as xylo-oligosaccharide, fructo-oligosaccharide, and inulin. The grape ferment is prepared by fermenting grape extract prepared from red grape fruit by yeast and lactobacillus.

[0025] In some embodiments, the prebiotic composition includes grape ferment, lactitol, and polysaccharide, and the weight ratio of grape ferment, lactitol, and polysaccharide is 1-2:1-2:1-2. In this case, the polysaccharide can be fructo-oligosaccharide, xylo-oligosaccharide, or inulin. For example, the weight ratio of grape ferment, lactitol, and polysaccharide is 2:1:1.

[0026] In some embodiments, the prebiotic composition includes grape ferment, xylo-oligosaccharide, and polysaccharide, and the weight ratio of grape ferment, xylo-oligosaccharide, and polysaccharide is 1-2:1-2:1-2. In this case, the polysaccharide can be fructo-oligosaccharide or inulin. For example, the weight ratio of grape ferment, xylo-oligosaccharide, and polysaccharide is 2:1:1.

[0027] In some embodiments, the prebiotic composition includes grape ferment, lactitol, and fructo-oligosaccharide, and the weight ratio of grape ferment, lactitol, and fructo-oligosaccharide is 1-2:1-2:1-2. For example, the prebiotic composition includes grape ferment, lactitol, and fructo-oligosaccharide with a weight ratio of 2:1:1.

[0028] In some embodiments, the grape ferment preparation process includes preparing grape extract, and preparing grape ferment by implanting multiple strains in the grape extract and then culturing.

[0029] In this case, the grape extract can be red grape juice obtained by squeezing red grape (Vits vinifera) fruit, or concentrated grape juice obtained by concentrating red grape juice, or grape juice diluent obtained by diluting red grape juice or concentrated grape juice, or grape juice obtained by extracting red grape fruit with a solvent.

[0030] For example, red grape fruits are directly crushed and filtered to obtain grape extract. In some embodiments, the red grape fruits include skins, pulp, and seeds. In some embodiments, the grape extract can be prepared by mixing commercially available concentrated grape juice with water. In some embodiments, the grape extract can be prepared by mixing concentrated grape juice, water, and glucose. In some embodiments, the grape extract can be prepared by heating concentrated grape juice, water, and glucose to 95°C or above for 30 minutes. The volume ratio of the concentrated grape juice to water can be 1:8. The concentration of the glucose can be 3% (W / V), or an amount of glucose is added to make the Brix of the grape extract greater than or equal to 8. That is, the Brix of the solution is measured simultaneously during the addition of the glucose, and the addition of the glucose is stopped when the Brix of the grape extract reaches 8 or above. In some embodiments, the red grape can be Sangiovese, Lambrusco, or a mixture thereof.

[0031] In this regard, the plurality of bacteria includes yeast and Lactobacillus.

[0032] In some embodiments, the grape fermentation product can be prepared by inoculating 0.1% (W / V) of yeast and 0.05% (W / V) of Lactobacillus into the grape extract and then incubating the mixture. In some embodiments, the grape fermentation product can be prepared by mixing 0.1% (W / V) of yeast and 0.05% (W / V) of Lactobacillus into the grape extract, incubating the mixture, and then concentrating and / or filtering the mixture. In other words, the grape fermentation product can be the initial mixture of the grape extract and the plurality of bacteria, the concentrated mixture of the initial mixture, the filtered mixture of the initial mixture, or the concentrated and filtered mixture of the initial mixture.

[0033] In some embodiments, the incubation time of the grape extract and the plurality of bacteria can be 72 hours. In some embodiments, the incubation temperature of the grape extract and the plurality of bacteria can be 30°C.

[0034] In some embodiments, the concentration temperature can be 55°C to 65°C.

[0035] In some embodiments, the yeast can be Saccharomyces cerevisiae. For example, the yeast can be the Saccharomyces cerevisiae strain deposited in the BCRC (Biological Resource Center, Taiwan Food Industry Development Institute) with the accession number BCRC20271 (also deposited in the DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH) with the accession number ATCC33107) or other commercially available Saccharomyces cerevisiae.

[0036] The lactic acid bacteria can be Streptococcus thermophilus, Lactobacillus helveticus, or Lactobacillus plantarum. For example, the lactic acid bacteria can be Streptococcus thermophilus TCI633 (BCRC 910636, also DSM 28121), Streptococcus thermophilus TCI357 (BCRC 910846, also DSM 33107), Streptococcus thermophilus TCI028 (BCRC 910805, also DSM 33108), Streptococcus thermophilus TCI378 (BCRC 910760, also DSM 32451), other commercially available Streptococcus thermophilus, other commercially available Lactobacillus helveticus, or other commercially available Lactobacillus plantarum.

[0037] For example, a concentrated grape juice is mixed with water to obtain a grape juice solution. The volume ratio of the concentrated grape juice to water is 1:8. Then, 3% (W / V) of glucose is added to the grape juice solution and left at 95°C for 0.5 hours to obtain a grape extract. Then, 0.1% (W / V) of brewer's yeast and 0.05% (W / V) of Streptococcus thermophilus TCI633 are inoculated into the grape extract, and the mixture is left at 30°C for 72 hours to obtain a grape fermentation broth. Then, the grape fermentation broth is concentrated under reduced pressure at 60°C, and the concentrated grape fermentation broth is filtered through a filter with a pore size of 400 mesh to obtain a grape ferment.

[0038] Accordingly, the grape ferment, the first component, and the second component obtained by the specific process are mixed in a specific ratio to obtain a prebiotic composition. The weight ratio of the grape ferment, the first component, and the second component is 1-2: 1-2: 1-2. In addition, the prebiotic composition can increase the amount of probiotics (e.g., Akkermansia muciniphila) in the intestine of the subject.

[0039] In some embodiments, the prebiotic composition including the grape ferment, lactulose, and fructooligosaccharide can be used to promote the health of the intestine of the subject. For example, the prebiotic composition can increase the amount of Akkermansia muciniphila in the intestine of the subject by at least 2.5 times and increase the amount of Bifidobacterium in the intestine of the subject by at least 3.8 times. Thus, the prebiotic composition can improve the intestinal flora, thereby achieving the effect of promoting the health of the intestine of the subject.

[0040] In some embodiments, the prebiotic composition comprising grape ferment, lactitol, and fructooligosaccharide increases the antioxidant capacity of the subject. For example, the prebiotic composition can increase the level of sulfur-containing compounds, the level of total Glutathione, or a combination thereof, in the subject. In some embodiments, the sulfur-containing compounds are Thiols compounds. By increasing the level of sulfur-containing compounds and / or total Glutathione, the ability of the subject to scavenge free radicals can be increased, thereby strengthening the defense against oxidative stress to the cells.

[0041] In some embodiments, the prebiotic composition can be in a solid form, such as a powder, a tablet, a capsule, and the like.

[0042] In some embodiments, the dosage of the prebiotic composition is 400 mg per day. For example, the prebiotic composition is mainly composed of grape ferment, lactitol, and fructooligosaccharide, and the daily dosage of 400 mg of the prebiotic composition means that the total amount of grape ferment, lactitol, and fructooligosaccharide is 400 mg.

[0043] Any of the prebiotic compositions described above can be a pharmaceutical product. In other words, the pharmaceutical product comprises an effective amount of grape ferment, lactitol or xylo-oligosaccharide as a first component, and fructooligosaccharide, xylo-oligosaccharide, or inulin as a second component, in a specific ratio.

[0044] In some embodiments, the pharmaceutical product described above can be manufactured into a dosage form suitable for enteral, parenteral, oral, or topical administration using techniques well known to those skilled in the art.

[0045] In some embodiments, the enterally or orally administered dosage form can be, but is not limited to, a tablet, a troche, a lozenge, a pill, a capsule, a dispersible powder or a granule, a solution, a suspension, an emulsion, a syrup, an elixir, a slurry, or the like. In some embodiments, the parenterally or topically administered dosage form can be, but is not limited to, an injection, a sterile powder, an external preparation, or the like. In some embodiments, the injection can be administered subcutaneously, intraepidermally, intradermally, or intralesionally.

[0046] In some embodiments, the aforementioned pharmaceutical product can comprise a pharmaceutically acceptable carrier that is widely used in pharmaceutical manufacturing technology. In some embodiments, the pharmaceutically acceptable carrier can be one or more of the following: a solvent, a buffer, an emulsifier, a suspending agent, a decomposer, a disintegrating agent, a dispersing agent, a binding agent, an excipient, a stabilizing agent, a chelating agent, a diluent, a gelling agent, a preservative, a wetting agent, a lubricant, an absorption delaying agent, a liposome, and the like. The kind and amount of the carrier to be selected are within the scope of the professional knowledge and routine skills of those skilled in the art. In some embodiments, the solvent as the pharmaceutically acceptable carrier can be water, normal saline, phosphate buffered saline (PBS), or an aqueous solution containing alcohol.

[0047] In some embodiments, any of the aforementioned prebiotic compositions can be a consumable product. In other words, the consumable product comprises a specific content of grape ferment, lactulose or xylo-oligosaccharide as a first ingredient, and fructo-oligosaccharide, xylo-oligosaccharide or inulin as a second ingredient, in a specific ratio. In some embodiments, the consumable product can be a general food, a health food, or a dietary supplement.

[0048] In some embodiments, the aforementioned consumable product can be manufactured into a dosage form suitable for oral administration using techniques well known to those skilled in the art. In some embodiments, the aforementioned general food can be the consumable product itself. In some embodiments, the general food can be, but is not limited to, beverages, fermented foods, bakery products, or condiments.

[0049] In some embodiments, the resulting prebiotic composition can be further used as a food additive to prepare a food composition containing a grape ferment, lactitol or xylooligosaccharide as a first ingredient, and fructooligosaccharides, xylooligosaccharide or inulin as a second ingredient in a specific ratio. In this regard, any of the embodiments of the prebiotic composition can be added to the raw materials during the preparation of the raw materials by existing methods, or added to the food during the preparation of the food, and formulated with any edible material into an edible product (i.e., a food composition) for human and non-human animals to consume.

[0050] Example 1: Preparation of grape ferment

[0051] First, a concentrated grape juice from red grapes of Sangiovese variety from Italy was mixed with water to obtain a grape juice solution. The volume ratio of the concentrated grape juice to water was 1 :8. In this regard, the concentrated grape juice was purchased from supplier Diana Food with product number CC01460001.

[0052] Next, 3% (W / V) of glucose was added to the grape juice solution and incubated at 95 °C for 0.5 hours to obtain a grape extract. Then, 0.1% (W / V) of Saccharomyces cerevisiae with accession number BCRC 20271 and 0.05% (W / V) of Streptococcus thermophilus TCI633 with accession number BCRC 910636 were inoculated into the grape extract and incubated at 30 °C for 72 hours to obtain a grape fermentate. In this regard, the grape fermentate had a Brix of 4.0 ± 0.5 (20 °C) and a pH of 3.0 ± 0.5.

[0053] Then, the grape fermentate was concentrated under reduced pressure at about 60 °C, and then filtered through a filter with a pore size of 400 mesh to obtain a grape ferment.

[0054] Example 2: Effect of different compositions of prebiotic on probiotics

[0055] In this regard, the xylooligosaccharide used was purchased from Shandong Longli Biological Technology Co., Ltd. The lactitol used was purchased from Hongwei Biological Technology. The inulin used was purchased from Cosucra. The fructooligosaccharides used were purchased from Meiji. The grape ferment used was the grape ferment prepared in Example 1. The liquid medium used was Tryptone Soy Broth (TSB; hereinafter referred to as TSB medium) with 5% sheep blood (purchased from BD).

[0056] The groups were divided into a blank group, 5 experimental groups (experimental groups A-E) and 5 control groups (control groups A-E), as shown in Table 1.

[0057] Table 1

[0058]

[0059]

[0060] As shown in Table 1, the blank group used a simple liquid culture medium (i.e., 95% TSB medium and 5% sheep blood). The control groups A-E used a 94% TSB medium, 5% sheep blood, and 1% of a single-component probiotic combination A-E, and the probiotic combinations A-E were xylo-oligosaccharides, lactitol, inulin, fructo-oligosaccharides, and grape ferment, respectively. The experimental groups A-E used a 94% TSB medium, 5% sheep blood, and 1% of a different composition of probiotic combination F-J. Here, the probiotic combination F was grape ferment, xylo-oligosaccharides, and lactitol in a weight ratio of 2:1:1. The probiotic combination G was grape ferment, xylo-oligosaccharides, and fructo-oligosaccharides in a weight ratio of 2:1:1. The probiotic combination H was grape ferment, xylo-oligosaccharides, and inulin in a weight ratio of 2:1:1. The probiotic combination I was grape ferment, lactitol, and fructo-oligosaccharides in a weight ratio of 2:1:1. The probiotic combination J was grape ferment, lactitol, and inulin in a weight ratio of 2:1:1.

[0061] 1% of activated Akkermansia muciniphila was added to 15 mL test tubes containing 5 mL of each group of culture medium, and anaerobic culture was carried out at 37°C for 48 hours. After 48 hours of culture, 100 μL of bacterial solution was taken from each group of test tube on solid TSB medium containing 5% sheep blood, and after 72 hours of anaerobic culture at 37°C, the bacterial amount of each group was calculated. Here, the bacterial amount was expressed in colony-forming units (CFU).

[0062] See Table 2 Figure 1 The bacterial amount of the blank group was 6.50 log CFU, the bacterial amount of the control group A was 6.75 log CFU, the bacterial amount of the control group B was 6.67 log CFU, the bacterial amount of the control group C was 6.53 log CFU, the bacterial amount of the control group D was 6.53 log CFU, the bacterial amount of the control group E was 6.88 log CFU, the bacterial amount of the experimental group A was 7.00 log CFU, the bacterial amount of the experimental group B was 6.96 log CFU, the bacterial amount of the experimental group C was 6.95 log CFU, the bacterial amount of the experimental group D was 7.11 log CFU, and the bacterial amount of the experimental group E was 7.00 log CFU.

[0063] Therefore, compared with the blank group, the single-component probiotic composition of grape ferment, xylo-oligosaccharide and lactitol can increase the amount of Akkermansia, while inulin and fructo-oligosaccharide have no significant effect on promoting the growth of Akkermansia.

[0064] In addition, after combining these probiotic components based on grape ferment, compared with the blank group, the probiotic composition composed of multiple components can promote the growth of Akkermansia and increase the amount of Akkermansia. Moreover, the probiotic composition with the best effect on increasing the amount of Akkermansia is the probiotic composition D composed of grape ferment, lactitol and fructo-oligosaccharide.

[0065] Therefore, when the probiotic composition is composed of grape ferment, lactitol or xylo-oligosaccharide as the first component, and fructo-oligosaccharide, xylo-fructose or inulin as the second component, the probiotic composition can promote the growth of probiotics (such as Akkermansia) and increase the amount of probiotics.

[0066] Example 3: Effect of different proportions of probiotic composition on probiotics

[0067] Here, the composition of the probiotic composition used is grape ferment prepared in Example 1, lactitol (purchased from Hongwei Biotechnology) and fructo-oligosaccharide (purchased from Meiji). The liquid medium used is Tryptone Soy Broth (TSB; hereinafter referred to as TSB medium) with 5% sheep blood (purchased from BD).

[0068] The groups are divided into a blank group and 4 experimental groups (experimental groups A-D), as shown in Table 2.

[0069] Table 2

[0070]

[0071]

[0072] From the above table, the weight ratio of grape ferment, lactitol and fructo-oligosaccharide in probiotic composition a is 1:1:1, the weight ratio of grape ferment, lactitol and fructo-oligosaccharide in probiotic composition b is 2:1:1, the weight ratio of grape ferment, lactitol and fructo-oligosaccharide in probiotic composition c is 1:2:1, and the weight ratio of grape ferment, lactitol and fructo-oligosaccharide in probiotic composition d is 1:1:2.

[0073] Example 3: Animal Experiment

[0074] Please refer to Figure 2 The blank group had a bacterial amount of 6.50 log CFU, the experimental group A had a bacterial amount of 6.92 log CFU, the experimental group B had a bacterial amount of 7.11 log CFU, the experimental group C had a bacterial amount of 6.77 log CFU, and the experimental group d had a bacterial amount of 6.66 log CFU.

[0075] Therefore, compared to the blank group, the probiotic composition a-d composed of grape ferment, lactosaccharose, and fructo-oligosaccharide can effectively promote the growth ability of Akkermansia muciniphila and increase the bacterial amount of Akkermansia muciniphila. Moreover, when the probiotic composition is 50% grape ferment, 25% lactosaccharose, and 25% fructo-oligosaccharide, this probiotic composition can significantly promote the growth ability of Akkermansia muciniphila by at least 4.1 times.

[0076] Therefore, grape ferment, lactosaccharose, and fructo-oligosaccharide can effectively improve the growth ability of probiotics (such as Akkermansia muciniphila) regardless of the proportion of the probiotic composition. Among them, compared to other proportions, when the proportion of grape ferment, lactosaccharose, and fructo-oligosaccharide is 2:1:1, the probiotic composition can significantly improve the growth ability of probiotics (such as Akkermansia muciniphila).

[0077] Example 4: Human Experiment

[0078] 4-1, Experimental Design

[0079] Each of the 7 subjects was given a capsule containing 400 mg of the probiotic composition composed of grape ferment, lactosaccharose (purchased from Hongwei Biotechnology), and fructo-oligosaccharide (purchased from Meiji) in a weight ratio of 2:1:1 prepared in Example 1, and each person took one capsule per day, for a continuous period of 2 weeks. In other words, the daily dose of each person was 400 mg of probiotic composition. Moreover, each subject was tested at week 0 (i.e., before taking) and week 2 (i.e., after taking for 2 weeks).

[0080] 4-2, Intestinal flora analysis of subjects

[0081] 7 subjects were collected feces before taking the prebiotic composition (i.e. at week 0) and after taking the prebiotic composition (i.e. at week 2), and commissioned BIOTOOLS to conduct fecal flora NGS analysis (sequencing position: 16S rRNA V3-V4) to determine the growth ability of the intestinal flora.

[0082] Here, the analyzed flora species are Akkermansia muciniphila (hereinafter referred to as AKK bacteria) and Bifidobacterium.

[0083] See Figure 3 The average AKK bacteria count of the 7 subjects at week 0 was taken as 1, and the average AKK bacteria count ratio of the 7 subjects at week 2 was calculated based thereon. Here, the average AKK bacteria count ratio of the 7 subjects at week 2 was 2.56. That is, the AKK bacteria count in the intestines of the 7 subjects was increased by 2.56 times after taking the prebiotic composition for 2 weeks, and the metabolites of the AKK bacteria can suppress the appetite of the host and induce the expression of the FIAF (Fasting-Induced Adipose Factor) gene of the host, thereby reducing the ability of the host to store fat. In addition, when the AKK bacteria count in the intestines is increased, obesity caused by a high-fat diet can be reversed, the concentration of lipopolysaccharides of harmful bacteria in the blood can be reduced to reduce chronic inflammation, and insulin resistance can be reduced. Therefore, by taking the prebiotic composition composed of grape ferment, lactitol, and fructooligosaccharide, the AKK bacteria count in the intestines of the user can be effectively increased, thereby achieving various effects such as slimming, reducing chronic inflammation, and reducing insulin resistance.

[0084] See Figure 4 The average Bifidobacterium count of the 7 subjects at week 0 was taken as 1, and the average Bifidobacterium count ratio of the 7 subjects at week 2 was calculated based thereon. Here, the average Bifidobacterium count ratio of the 7 subjects at week 2 was 3.84. That is, the Bifidobacterium count in the intestines of the 7 subjects was increased by 3.84 times after taking the prebiotic composition for 2 weeks. In addition, when the Bifidobacterium count in the intestines of the human body is increased, the Bifidobacterium, which is a physiologically beneficial bacteria, can improve the biological barrier of the human body, provide nutrition to the human body, enhance the immunity of the human body, improve the health of the gastrointestinal tract of the human body, and improve the anti-aging ability of the human body, thereby enabling the human body to be healthy. Therefore, by taking the prebiotic composition composed of grape ferment, lactitol, and fructooligosaccharide, the Bifidobacterium count in the intestines of the user can be effectively increased, thereby increasing the amount of probiotic bacteria in the intestines of the recipient, and achieving various physiological functions beneficial to health, such as improving the biological barrier of the recipient, providing nutrition to the recipient, enhancing the immunity of the recipient, improving the health of the gastrointestinal tract of the recipient, and improving the anti-aging ability of the recipient.

[0085] 4-3, Analysis of sulfur-containing compounds and total glutathione content in blood of the subject

[0086] 7 subjects were collected 6 mL of venous blood each using EDTA-containing anticoagulant purple head blood collection tubes before taking the probiotic composition (i.e. at week 0) and after taking the probiotic composition (i.e. at week 2), and entrusted TCI GENE to analyze the expression of sulfur-containing compounds and total glutathione (total-Glutathione; GSH) in blood. At this point, the detected sulfur-containing compounds are f-thiols. f-thiols and total glutathione are indicators of the body's antioxidant capacity. When the content of f-thiols or glutathione increases, it represents an increase in the body's ability to scavenge free radicals, thereby enhancing the ability to resist the damage of oxidative stress to cells.

[0087] Please refer to Figure 5 The average f-thiol content in the blood of 7 subjects at week 0 was 322.29 μg / mL, and the average f-thiol content in the blood after taking the probiotic composition for 2 weeks was increased to 377.86 μg / mL. That is, after taking the probiotic composition for 2 weeks, the average f-thiol content in the blood increased by 55.57 μg / mL (equivalent to an increase of 17.2%). Therefore, by taking the probiotic composition, the content of f-thiols can be effectively increased, thereby improving the body's ability to scavenge free radicals and improving antioxidant capacity.

[0088] Please refer to Figure 6 The average total glutathione content in the blood of 7 subjects at week 0 was 1380 μmol / L, and the average total glutathione content in the blood after taking the probiotic composition for 2 weeks was increased to 1449 μmol / L. That is, after taking the probiotic composition for 2 weeks, the average total glutathione content in the blood increased by 69 μmol / L (equivalent to an increase of 5%). Therefore, by taking the probiotic composition, the content of total glutathione can be effectively increased, thereby improving the body's ability to scavenge free radicals and improving antioxidant capacity.

[0089] In summary, the probiotic composition prepared from the grape ferment according to any of the embodiments of the present application can be used to promote the growth of Akkermansia, wherein the probiotic composition comprises grape ferment, lactulose or xylo-oligosaccharide as the first ingredient, and fructo-oligosaccharide, xylo-oligosaccharide or inulin as the second ingredient, and the weight ratio of grape ferment, the first ingredient and the second ingredient is 1-2: 1-2: 1-2. In some embodiments, the probiotic composition prepared from grape ferment, lactulose and fructo-oligosaccharide can be used to promote the intestinal health of the subject and / or improve the antioxidant capacity of the subject. The weight ratio of grape ferment, lactulose and fructo-oligosaccharide contained in the probiotic composition is 1-2: 1-2: 1-2. In some embodiments, the probiotic composition can promote the intestinal health of the subject by promoting the growth of probiotics such as Akkermansia and Bifidobacterium. In some embodiments, the probiotic composition can improve the antioxidant capacity of the subject by increasing the content of sulfur-containing compounds such as thiol compounds and total glutathione in the body of the subject.

[0090] Of course, the present application can have other various embodiments, and those skilled in the art can make various corresponding changes and modifications according to the present application without departing from the spirit and essence of the present application. However, these corresponding changes and modifications should all belong to the protection scope of the claims of the present application.

Claims

1. A prebiotic composition, characterized in that, To promote the growth of Akkermansia muciniphila, the following are included: Grape fermentation products; The first component, wherein the first component is lactitol; and The second component is fructooligosaccharides; The weight ratio of the grape ferment, the first component, and the second component is 2:1:1; The grape fermentation product is prepared by inoculating red grape extract with 0.1% W / V yeast and 0.05% W / V lactic acid bacteria and then allowing it to stand for culture.

2. The use of a composition comprising grape fermentation for preparing a probiotic composition that promotes the growth of Akkermansia muciniphila, wherein the grape fermentation is prepared by inoculating an extract of red grapes with 0.1% w / v yeast and 0.05% w / v lactic acid bacteria and then allowing it to stand for culture; the composition comprising grape fermentation includes the grape fermentation, a first component, and a second component; wherein the first component is lactitol, the second component is fructooligosaccharide, and the weight ratio of the grape fermentation, the first component, and the second component is 2:1:1; wherein the dosage of the probiotic composition is 400 mg / day.

3. Use according to claim 2, characterized in that, The yeast is brewer's yeast, and the lactic acid bacteria is Streptococcus thermophilus TCI633.

4. The use according to claim 2, wherein the prebiotic composition has the ability to promote the growth of Bifidobacterium.

Images