TW202319062A - Probiotic composition and its use - Google Patents

Abstract

A probiotic composition comprising kiwi fruit ferment, inulin and fructooligosaccharides, and the weight ratio of kiwi fruit ferment, inulin and fructooligosaccharides is 3-4:2.5-4:2.5-4. Based on this, the probiotic composition prepared by kiwifruit and sugars can be used to improve the intestinal tract of the subject.

Description

Prebiotic compositions and uses thereof

The invention relates to the use of a prebiotic composition, which uses fermented kiwi fruit and sugars to prepare the prebiotic composition, and uses the prebiotic composition to improve the intestinal tract of a recipient.

Prebiotics, also known as prebiotics and probiotics, are polysaccharides in natural foods that are not easily digested by human enzymes, but they can be used by probiotics in the digestive system (mainly the large intestine) to form a flora Growth, expansion and metabolism to produce short-chain fatty acids (Short Chain Fatty Acids, SFCAs).

According to the joint statement of the International Scientific Association for Probiotics and Prebiotics (ISAPP) on "Nature report" in 2017 on prebiotics, it is defined as "the ability to be selectively utilized by microorganisms that are symbiotic with the host. , and thus substances that promote the health of the host".

Specifically, prebiotics help good bacteria grow and help keep bad bacteria in your gut. The probiotics in the gut will also metabolize the prebiotics into short-chain fatty acids, which are then provided to the probiotics and the host as an energy source.

In view of this, the present invention provides a prebiotic composition, which includes fermented kiwi fruit and sugars, and can be used to improve the intestinal tract of the recipient, thereby assisting the recipient to lose weight.

In some embodiments, a prebiotic composition includes fermented kiwifruit, inulin, and fructooligosaccharides, wherein the weight ratio of fermented kiwifruit, inulin, and fructooligosaccharides is 3-4: 2.5-4: 2.5-4.

In some embodiments, a kiwifruit fermented product and sugars are used to prepare a prebiotic composition for improving the intestinal tract of a recipient, wherein the prebiotic composition includes kiwifruit fermented product, inulin and fructooligosaccharides, and The weight ratio of the kiwifruit fermented product, inulin and fructooligosaccharide is 3-4:2.5-4:2.5-4.

In some embodiments, the weight ratio of the kiwifruit fermented product, inulin and fructooligosaccharide contained in the prebiotic composition is 4:3:3.

In some embodiments, improving the intestinal tract of the recipient includes improving the flora of the intestinal tract, improving the defecation status of the recipient, improving the intestinal barrier of the recipient, or a combination thereof.

In some embodiments, the bacterial phase includes Akkermansia muciniphila , Parabacteroides goldsteini and Bifidobacterium .

In some embodiments, the prebiotic composition is used to promote the growth ability of Parabacteroides gordii.

In some embodiments, improving the recipient's defecation status refers to improving the recipient's defecation difficulty, promoting the recipient's intestinal peristalsis frequency, reducing the recipient's unclean defecation, reducing the recipient's defecation time, or a combination thereof.

In some embodiments, the prebiotic composition is used to increase the concentration of compact structure proteins in the recipient's blood.

In some embodiments, the compact structure protein is claudin 3 (CLDN3).

In some embodiments, a prebiotic composition is used to reduce a subject's body weight, waist circumference, body fat percentage, or a combination thereof.

In summary, the prebiotic composition of any embodiment includes fermented kiwifruit, inulin and fructooligosaccharides. That is to say, the kiwi fruit fermented product and polysaccharide can be used to prepare a prebiotic composition for improving the intestinal tract of the recipient. In some embodiments, the prebiotic composition can be used to improve the intestinal flora, improve the defecation status of the subject, improve the intestinal barrier of the subject, or a combination thereof. For example, the prebiotic composition can increase the growth ability of probiotics such as Kremniphila, Parabacteroides gordii and Bifidobacterium and promote their content in the intestinal tract of the recipient. In some embodiments, the prebiotic composition can improve the subject's defecation difficulty, promote the subject's intestinal peristalsis frequency, reduce the subject's unclean defecation, reduce the subject's defecation time, or a combination thereof. In some embodiments, the prebiotic composition increases the concentration of a compact structure protein (eg, Claudin 3 (CLDN3)) in the recipient's blood. In some embodiments, the prebiotic composition is used to reduce the subject's body weight, waist circumference, body fat percentage or a combination thereof, thereby achieving a slimming effect.

In the description of the following embodiments, unless otherwise stated, the symbol "%" refers to weight percentage and the symbol "vol%" usually refers to volume percent concentration.

The prebiotic composition includes fermented kiwi fruit and sugar. Among them, the sugars are polysaccharides such as oligosaccharides and dietary fiber, such as fructooligosaccharides and inulin. Moreover, the kiwifruit fermented product is prepared by fermenting the fruit juice of kiwifruit ( Actinidia deliciosa ) through various bacterial strains.

In some embodiments, the prebiotic composition includes kiwifruit fermented product, inulin and fructooligosaccharide, and the weight ratio of kiwifruit fermented product, inulin and fructooligosaccharide is 3-4:2.5-4:2.5-4 . For example, the prebiotic composition includes kiwifruit fermented product, inulin and fructooligosaccharide in a weight ratio of 4:3:3.

In some embodiments, the preparation process of the kiwi fruit fermented product is as follows: glucose and kiwi fruit are mixed with water to obtain a culture solution, and the weight of the water is 3-6 times the total weight of the kiwi fruit. Next, ferment the culture solution and the plurality of strains for 1-4 days to obtain kiwifruit fermented juice. Here, the plural strains include yeast and lactic acid bacteria. Here, "kiwifruit fruit" may include its pulp/fruit, pulp/fruit containing pericarp, or pulp/fruit containing seeds.

Wherein, the yeast used may be commercially available brewer's yeast ( Saccharomyces cerevisiae ). For example, it may be brewer's yeast of the strain with deposit number BCRC20271 (international deposit number ATCC26602) procured from the Institute of Food Work Development.

The lactic acid bacteria used may be commercially available Lactobacillus helveticus , Streptococcus thermophiles or Lactobacillus plantarum . For example, Streptococcus thermophilus TCI028 and BCRT910760 (International Deposit No. Streptococcus thermophilus strain TCI378 under deposit number DSM32451) or Streptococcus thermophilus TCI633 under deposit number BCRC910636 (international deposit number DSM28121).

For example, first mix the fruit of kiwifruit with water, and extract at 80-100° C. for 0.5-1.5 hours to obtain a kiwifruit extract; then add glucose to the kiwifruit extract to obtain a Fermented broth. Wherein, the added amount of glucose is 8-10% of the total weight of kiwi fruit and water. Moreover, by adding glucose, the culture solution has sufficient sugar content to ensure that the bacteria have sufficient nutrients during the subsequent fermentation and ensure that the subsequent fermentation can proceed smoothly. Next, add yeast to the culture medium to ferment for 0.5-2 days to form a primary fermentation liquid, then add lactic acid bacteria to the primary fermentation liquid to ferment for 0.5-2 days to form a kiwi fruit fermentation liquid, and filter the kiwi fruit fermentation liquid , concentrated, freeze-dried and pulverized to obtain the kiwifruit fermented product. In some embodiments, the total polyphenol content of the kiwi fruit fermentation broth is 200-220 μg/ml.

Based on this, the prebiotic composition can be obtained by mixing the fermented kiwifruit obtained through a specific process with inulin and fructooligosaccharides in a specific ratio. Wherein, the weight ratio of the kiwifruit fermented product, inulin and fructooligosaccharide is 3-4:2.5-4:2.5-4. Moreover, the prebiotic composition can increase the amount of probiotic bacteria in the recipient's intestinal tract by at least 3 times.

In some embodiments, a prebiotic composition improves the microbiota of the intestinal tract. For example, the prebiotic composition can increase the bacterial count of Akkermansia muciniphila in the recipient's intestine by at least 3 times, and the prebiotic composition can increase the amount of Parabacteroides gordii in the recipient's intestine. goldsteini i) at least 1.5 times, and the prebiotic composition can increase the bacteria of the genus Bifidobacterium in the intestinal tract of the recipient at least 3 times.

In some embodiments, the prebiotic composition improves the bowel movement of the subject. For example, by taking a prebiotic composition, the recipient can reduce the frequency of defecation and the defecation time of the recipient to improve the difficulty of defecation, reduce the frequency of unclean defecation, and promote the frequency of intestinal peristalsis, Thereby improving your own defecation status.

In some embodiments, the prebiotic composition increases the concentration of compact structural proteins in the blood of the recipient, thereby improving the intestinal barrier of the recipient. For example, the compact structure protein may be claudin 3 (CLDN3).

Thus, it can be seen that the prebiotic composition can improve the intestinal tract of the recipient. Here, the subject is a human.

In some embodiments, the prebiotic composition can improve the intestinal tract of the recipient, thereby achieving a slimming effect. For example, by taking the prebiotic composition, the subject can reduce body weight, waist circumference, body fat percentage, or a combination thereof. Among them, the body fat percentage refers to the body fat percentage of the whole body and the body fat percentage of the trunk.

In some embodiments, the prebiotic composition can be in solid form, eg, powder, lozenge, capsule, and the like.

In some embodiments, the dosage of the prebiotic composition is 800 mg/day. For example, the prebiotic composition is mainly composed of fermented kiwifruit, inulin and fructooligosaccharides, and the daily dose of 800 mg of the prebiotic composition refers to the amount of fermented kiwifruit, inulin and fructooligosaccharides The total amount is 800 mg.

Any of the aforementioned prebiotic compositions can be pharmaceuticals. In other words, the medicinal product contains kiwifruit fermented product, inulin and fructooligosaccharide in specific proportions in effective content.

In some embodiments, the foregoing pharmaceutical products may be formulated for parenteral, parenteral, oral, or topical administration using techniques well known to those skilled in the art. Dosage form.

In some embodiments, the dosage form for enteral or oral administration can be, but not limited to, tablet, troche, lozenge, pill, capsule , dispersible powder or granule, solution, suspension, emulsion, syrup, elixir, slurry or the like. In some embodiments, the dosage form for parenteral or topical administration may be, but not limited to, injection, sterile powder, external preparation, or the like. In some embodiments, the injection can be administered by subcutaneous injection, intraepidermal injection, intradermal injection or intralesional injection.

In some embodiments, the above-mentioned pharmaceuticals may contain pharmaceutically acceptable carriers (pharmaceutically acceptable carriers) which are widely used in pharmaceutical manufacturing technology. In some embodiments, the pharmaceutically acceptable carrier can be one or more of the following carriers: solvent, buffer, emulsifier, suspending agent, disintegrating agent ( decomposer), disintegrating agent, dispersing agent, binding agent, excipient, stabilizing agent, chelating agent, diluent ), gelling agents, preservatives, wetting agents, lubricants, absorption delaying agents, liposomes, and the like. The type and amount of carrier to be used is within the expertise and routine skill 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 aqueous solution containing alcohol (aqueous solution containing alcohol).

In some embodiments, any of the aforementioned prebiotic compositions can be edible products. In other words, the edible product contains a specific content of fermented kiwifruit, inulin and fructooligosaccharides in a specific ratio. In some embodiments, the edible product can be general food, health food or dietary supplement.

In some embodiments, the aforementioned edible products can be manufactured into dosage forms suitable for oral administration using techniques well known to those skilled in the art. In some embodiments, the aforementioned general food may be the edible product itself. In some embodiments, general foods may be, but not limited to: beverages, fermented foods, bakery products or seasonings.

In some embodiments, the obtained prebiotic composition can be further used as a food additive to obtain a prebiotic composition containing a specific ratio of fermented kiwifruit, inulin, and fructooligosaccharides. food composition. Here, the prebiotic composition of any embodiment can be added during the preparation of raw materials by conventional methods, or the prebiotic composition of any embodiment can be added during the production of food, and can be combined with any edible material Edible products (i.e. food compositions) formulated for ingestion by humans and non-human animals.

example 1 : Preparation of kiwifruit fermented product

First, the kiwi fruit ( Actinidia deliciosa ) is thoroughly washed, and the whole fruit is broken into kiwi fruit particles with a particle size of less than 12mm. Mix the kiwi fruit particles and water evenly at a ratio of 1:5 to obtain a raw material mixture. Then, according to the total weight of the raw material mixed liquid, 7.5% glucose was added to the raw material mixed liquid, and the raw material mixed liquid was leached at 95° C. for 0.5 hours to obtain a kiwi fruit culture liquid. Here, the sugar content of the culture solution at this time is 8.0°Bx.

After the culture solution was cooled to room temperature, the fermentation procedure was carried out. First, 0.1% of Saccharomyces cerevisiae (purchased from the Bioresource Conservation and Research Center (BCRC) of the Food Industry Development Institute, relative to the kiwi fruit culture solution) was added. No. BCRC20271) in the kiwi fruit culture medium, fermented at 30°C for 1 day to form the primary fermentation liquid, and then added 0.05% Streptococcus thermophilus (purchased from BCRC, deposit No. BCRC910636) was fermented at 30° C. for 1 day in the primary fermentation liquid to obtain the kiwi fruit fermentation liquid. Here, the specifications set for the kiwi fruit fermentation broth are pH 3.5±0.3 and sugar content less than 4.0°Bx. When these specifications are met, the fermentation can be judged to be complete. At this point, most of the sugar content in the kiwi fruit fermentation broth has been exhausted.

Next, the kiwifruit fermented liquid is filtered with a 200-mesh sieve to remove fruit residues in the fermented juice, and then freeze-dried, dried and pulverized to obtain a kiwifruit fermented product.

example 2 : single ingredient Effect on the growth of probiotics

Here, the single component refers to the fermented kiwi fruit prepared in Example 1, fructooligosaccharide (purchased from Meiji), and inulin (purchased from Cosucra). The probiotic refers to Parabacteroides goldsteini (purchased from ATCC). The liquid medium used was tryptone soybean broth (Tryptone Soy Broth, TSB; hereinafter referred to as TSB medium) added with 5% sheep blood (purchased from BD).

Divide the groups into a control group and three experimental groups. The experimental medium of the control group is a simple liquid medium (95% TSB medium and 5% sheep blood), while the three experimental groups are added with a single component to be tested. The kiwi fruit fermentation group, inulin group and fructooligosaccharide group. Among them, the experimental medium used in the kiwifruit fermented product group was 94% TSB medium, 5% sheep blood and 1% kiwifruit fermented product prepared in Example 1. The experimental medium used in the inulin group was 94% TSB medium, 5% sheep blood and 1% inulin. The experimental medium used in the fructooligosaccharide group was 94% TSB medium, 5% sheep blood and 1% fructooligosaccharide.

Take 1% of the activated Parabacteroides gordii into 15mL test tubes containing 5mL of each group of medium, and culture them anaerobically at 37°C for 48 hours. Then, after culturing for 48 hours, 100 μL of each group of bacterial liquid was placed on solid TSB medium containing 5% sheep blood, and the colony-forming unit (CFU) of each group was calculated. Herein, the CFU of the control group was regarded as 100% growth rate, and the growth rate (%) of other groups was correspondingly converted.

Please refer to Figure 1, the growth rate of the control group without adding the single ingredient to be tested is 100%, while in the other three experimental groups, the growth rate of the kiwifruit fermented product group is 106%, and the growth rate of the inulin group is 107% and the growth rate of the fructooligosaccharide group was 100%. It can be seen that for Parabacteroides gordii, 1% fructo-oligosaccharides cannot promote its growth, but 1% inulin and kiwi fruit fermentation can increase the number of bacteria and improve its growth ability.

example 3 : different ingredients Effect on the growth of probiotics

Here, the prebiotic ingredients used include inulin (purchased from Cosucra), xylooligosaccharide (purchased from Shandong Longli Biotechnology Co., Ltd.), fructooligosaccharide (purchased from Meiji), pomegranate enzyme, Example 1 The prepared kiwifruit fermented product. The liquid medium used was tryptone soybean broth (Tryptone Soy Broth, TSB; hereinafter referred to as TSB medium) added with 5% sheep blood (purchased from BD).

Among them, the production process of pomegranate enzyme is: thoroughly wash the pomegranate ( Punica granatum ), and break the whole fruit into pomegranate particles with a particle size of less than 12mm. Mix pomegranate particles and water evenly at a ratio of 1:5 to obtain a raw material mixture. Then, according to the total weight of the raw material mixed liquid, 7.5% glucose was added to the raw material mixed liquid, and the raw material mixed liquid was leached at 95° C. for 0.5 hours to obtain a pomegranate culture liquid. Here, the sugar content of the culture solution at this time is 8.0°Bx. After the culture solution was cooled to room temperature, the fermentation procedure was carried out, first adding 0.1% of Saccharomyces cerevisiae relative to the pomegranate culture solution (purchased from the Bioresource Conservation and Research Center (BCRC) of the Food Industry Development Institute, deposit number BCRC20271) in pomegranate culture broth, fermented at 30°C for 1 day to form a primary fermentation broth, and then added 0.05% Streptococcus thermophilus (purchased from BCRC, deposit number BCRC910636) relative to the pomegranate culture broth In the primary fermentation broth, ferment at 30° C. for 1 day to obtain a pomegranate fermentation broth. Here, the pomegranate fermentation broth is set with a pH value of 3.5±0.3 and a sugar content of less than 4.0°Bx. When these specifications are met, the fermentation can be judged to be complete. At this point, most of the sugar in the pomegranate broth has been consumed. Next, the pomegranate fermented liquid is filtered with a 200-mesh screen to remove fruit residues in the fermented juice, freeze-dried, dried and pulverized to obtain the pomegranate enzyme.

The groups were divided into control group, experimental group and 7 control groups (control group A~G). The composition of the medium used in each group is shown in Table 1:

Table 1 group Medium composition Components of each group of prebiotic combinations control group 95% TSB medium + 5% sheep blood none test group 94% TSB medium + 5% sheep blood + 1% experimental group prebiotic combination (ie, prebiotic composition) 40% kiwifruit fermented product + 30% inulin + 30% fructooligosaccharide control group A 94% TSB medium + 5% sheep blood + 1% prebiotic combination A 40% pomegranate enzyme + 30% inulin + 30% xylooligosaccharide control group B 94% TSB medium + 5% sheep blood + 1% prebiotic combination B 40% pomegranate enzyme + 30% fructooligosaccharide + 30% xylooligosaccharide Control group C 94% TSB medium + 5% sheep blood + 1% prebiotic combination C 40% pomegranate enzyme + 30% inulin + 30% fructooligosaccharide Control group D 94% TSB medium + 5% sheep blood + 1% prebiotic combination D 25% pomegranate enzyme + 25% inulin + 25% xylooligosaccharide + 25% fructooligosaccharide Control group E 94% TSB medium + 5% sheep blood + 1% prebiotic combination E 40% kiwifruit fermented product + 30% inulin + 30% xylooligosaccharide control group F 94% TSB medium + 5% sheep blood + 1% prebiotic combination F 40% kiwifruit fermented product + 30% fructooligosaccharide + 30% xylooligosaccharide Control group G 94% TSB medium + 5% sheep blood + 1% prebiotic combination G 25% kiwifruit fermented product + 25% inulin + 25% xylooligosaccharide + 25% fructooligosaccharide

It can be seen from the above table that, except for the control group which is pure liquid medium (95% TSB medium + 5% sheep blood), the medium of the other experimental groups and control groups (control groups A~G) is 94% TSB medium + 5% sheep blood + 1% prebiotic combination (experimental group prebiotic combination (ie, prebiotic composition), prebiotic combination A~G). Taking the experimental medium of the experimental group as an example, 100mL of the experimental medium contains 99mL of liquid medium (94mL of TSB medium and 5mL of sheep blood) and 1mL of the experimental group's prebiotic combination, while 1mL of the experimental group's prebiotic combination is composed of 400 μL of 10% kiwifruit fermented product, 300 μL of 10% fructooligosaccharide solution and 300 μL of 10% inulin solution, and so on for the other groups.

Take 1% of the activated Parabacteroides gordii into 15mL test tubes containing 5mL of each group of medium, and culture them anaerobically at 37°C for 48 hours. Then, after culturing for 48 hours, 100 μL of each group of bacterial liquid was placed on solid TSB medium containing 5% sheep blood, and the colony-forming unit (CFU) of each group was calculated. Herein, the CFU of the control group was regarded as 100% growth rate, and the growth rate (%) of other groups was correspondingly converted.

Please refer to Figure 2, the growth rate of the control group without adding probiotic combination was 100%, while in the other groups: the growth rate of the experimental group was 409.8% (nearly 4.1 times), the growth rate of the control group A was 207.7%, The growth magnification of control group B was 72.7%, the growth magnification of control group C was 80.8%, the growth magnification of control group D was 35.3%, the growth magnification of control group E was 124.1%, and the growth magnification of control group F was 209.4%. The growth magnification of control group G was 227.3%. It can be seen that the prebiotic composition composed of 40% kiwifruit fermented product, 30% inulin and 30% fructooligosaccharide can significantly improve the growth ability of Parabacteroides gordii and increase the number of bacteria.

Based on this, certain kinds of prebiotic combinations can effectively and significantly improve the growth ability of probiotics (such as Parabacteroides gordii), and not all common prebiotics (such as inulin, fructooligosaccharides, ) can effectively improve the growth ability of any probiotic bacteria (eg, Parabacteroides gordii).

example 4 : Different proportions of prebiotic composition Effect on the growth of probiotics

Here, the prebiotic combination used is inulin (purchased from Cosucra), fructooligosaccharide (purchased from Meiji) and the kiwifruit fermented product prepared in Example 1. The probiotic refers to Parabacteroides goldsteini (purchased from ATCC). The liquid medium used was Tryptone Soy Broth (TSB; hereinafter referred to as TSB medium) added with 5% sheep blood (purchased from BD).

The groups were divided into control group and four experimental groups (experimental groups a~d), and the composition of the medium used in each group is shown in Table 2:

Table 2 group Medium composition Composition ratio of prebiotic composition Kiwi fruit ferment FOS Inulin Experimental group a 94% TSB medium + 5% sheep blood + 1% prebiotic composition a 30% 30% 40% Experimental group b 94% TSB medium + 5% sheep blood + 1% prebiotic composition b 30% 40% 30% Experimental group c 94% TSB medium + 5% sheep blood + 1% prebiotic composition c 40% 30% 30% Experimental group d 94% TSB medium + 5% sheep blood + 1% prebiotic composition d 50% 25% 25% control group 95% TSB medium + 5% sheep blood none

It can be seen from the above table that the weight ratio of kiwifruit fermented product, fructooligosaccharide and inulin in each group is 3-4:2.5-4:2.5-4.

Take 1% of the activated Parabacteroides gordii into 15mL test tubes containing 5mL of each group of medium, and culture them anaerobically at 37°C for 48 hours. Then, after culturing for 48 hours, 100 μL of each group of bacterial liquid was placed on solid TSB medium containing 5% sheep blood, and the colony-forming unit (CFU) of each group was calculated. Herein, the CFU of the control group was regarded as 100% growth rate, and the growth rate (%) of other groups was correspondingly converted.

Please refer to Figure 3, the growth rate of the control group without the combination of probiotics was 100%, while in the other groups: the growth rate of the experimental group a was 312.0% (nearly 3.12 times), and the growth rate of the experimental group b was 354.0% (nearly 3.54 times), the growth rate of experimental group c was 409.8% (nearly 4.1 times) and the growth rate of experimental group d was 388.0% (nearly 3.88 times). In other words, the prebiotic composition composed of fermented kiwi fruit, fructooligosaccharides and inulin can effectively increase the growth ability of Parabacteroides gordii by at least 3 times. Moreover, when the ratio of the prebiotic composition is 40% kiwifruit fermented product, 30% inulin and 30% fructooligosaccharide, the growth ability of Parabacteroides gordii can be significantly increased by at least 4 times.

Based on this, a combination of certain proportions and specific types of probiotics can effectively and significantly improve the growth ability of probiotics (eg, Parabacteroides gordii).

example 5 : human experiment

In order to further confirm the impact of specific types of prebiotic compositions on the human body. Each capsule contains 400 mg of the kiwi fruit fermented product prepared in Example 1 with a weight ratio of 4:3:3, inulin (purchased from Cosucra) and fructooligosaccharides (purchased from Meiji) The capsules were provided to 8 subjects, and each person took two capsules a day for 4 consecutive weeks. In other words. The daily dose per person is 800 mg of the prebiotic composition. In addition, at week 0 (before taking), week 2 (after taking for 2 weeks), and week 4 (after taking for 4 weeks), test items such as blood drawing, stool sampling, questionnaire feedback and body composition determination were performed.

Among them, the blood test is to measure the intestinal barrier protein content. The basic structure of the intestinal barrier is that intestinal epithelial cells are co-constructed by tight structural proteins, and this connection is also called tight junction TJ (tight junction). Therefore, tightly structured proteins (such as CLDN3 protein, OCLN protein, etc.) are also called intestinal shielding proteins. The index protein of this blood test is CLDN3 protein. It contains four transmembrane structures that are the main components of tight junctions.

Among them, fecal sampling is to determine the growth ability of intestinal bacteria. The strains detected in this study were Kemansia muciniphila, Parabacteroides gordii and Bifidobacterium.

Among them, the questionnaire feedback is the feedback of defecation bacteria.

Among them, the determination of body composition includes weight, body fat percentage, waist circumference and so on.

example 5-1. in the subject's blood CLDN3 Protein Content Analysis

The venous blood of 8 subjects was collected before taking the prebiotic composition (that is, week 0) and after taking the prebiotic composition (that is, the second and fourth weeks) respectively, using purple-headed blood collection tubes containing EDTA anticoagulant. 6mL, and entrusted TCI GENE to perform performance analysis of intestinal shielding protein content in blood.

Here, the intestinal barrier protein detected is CLDN3 protein.

Please refer to Figure 4. The average CLDN3 protein concentration in the blood of the 8 subjects at week 0 was 8.9 ng/mL, and the average CLDN3 protein concentration in the blood increased to 9.96 ng/mL after taking the prebiotic composition for 2 weeks. mL, and the average concentration of CLDN3 protein in blood increased to 12.70 ng/mL after taking the prebiotic composition for 4 weeks. That is to say, after taking the prebiotic composition for 2 weeks, the average CLDN3 protein concentration in the blood increased by 1.06 ng/mL, and after taking the prebiotic composition for 4 weeks, the average CLDN3 protein concentration in the blood increased by 3.8 ng/mL . It can be known that the concentration of CLDN3 protein can be effectively increased by taking the prebiotic composition, thereby improving the intestinal barrier ability.

example 5-2. Subject's intestinal flora analyze

The stools of 8 subjects were collected before taking the prebiotic composition (that is, week 0) and after taking the prebiotic composition (that is, the second and fourth weeks), and entrusted BIOTOOLS Perform NGS analysis of fecal bacteria (sequencing position: 16S rRNA V3-V4). Moreover, the analysis results of the 8 subjects are organized as shown in Figures 5 to 7.

Here, the strains of the bacterial phase analyzed are Akkermansia muciniphila (hereinafter referred to as AKK bacteria), Parabacteroides goldsteini (hereinafter referred to as PG bacteria) and Bifidobacterium ( Bifidobacterium ; Hereinafter referred to as BF bacteria).

See Figure 5. The average number of AKK bacteria in the 8 subjects at the 0th week was regarded as 1, and the ratio of the average AKK bacteria in the 2nd and 4th weeks of the 8 subjects was converted based on this. Here, the average number ratio of AKK bacteria in the 2nd week of 8 subjects was 3.44, and the average number ratio of AKK bacteria in the 4th week was 4.87. That is to say, after 8 subjects took the prebiotic composition continuously for 4 weeks, the amount of AKK bacteria in their intestines increased by 4.87 times, and the metabolites of AKK bacteria can suppress the host's appetite and induce the host's FIAF (Fasting-Induced Adipose Factor) gene expression, thereby reducing the ability of the host to store fat. In addition, when the amount of AKK bacteria in the intestine is increased, it can reverse the obesity caused by high-fat diet, reduce the concentration of lipopolysaccharide of harmful bacteria in the blood, reduce chronic inflammation, and reduce insulin resistance. Therefore, by taking a prebiotic composition composed of fermented kiwifruit, inulin and fructooligosaccharides, it can effectively increase the amount of AKK bacteria in the user's intestinal tract, thereby achieving weight loss, reducing chronic inflammation, and reducing insulin resistance and many other effects.

See Figure 6. The average bacterial count of PG bacteria in the 8 subjects at the 0th week was regarded as 1, and the ratio of the average PG bacterial counts in the 2nd and 4th weeks of the 8 subjects was converted based on this. Here, the average number ratio of PG bacteria in the eight subjects was 3.76 at the second week, and the average number ratio of PG bacteria at the fourth week was 1.93. That is to say, after 8 subjects took the prebiotic composition continuously for 4 weeks, the amount of PG bacteria in their intestines increased by 1.93 times. Moreover, the increase of PG bacteria can help improve metabolic syndrome, reduce intestinal leakage, and improve intestinal inflammation. Specifically, when the amount of PG bacteria in the intestine increases, it can reduce the host's body weight, slow down the host's insulin resistance, increase the host's fat metabolism and activate the brown cells in the host's body, thereby achieving the effect of weight loss. Therefore, by taking a prebiotic composition composed of fermented kiwifruit, inulin and fructooligosaccharides, the amount of PG bacteria in the user's intestinal tract can be effectively increased, thereby achieving weight loss, reducing intestinal leakage, and improving the intestinal tract Various effects such as inflammation state.

See Figure 7. The average bacterial count of BF bacteria in the 8 subjects at the 0th week was regarded as 1, and the ratio of the average BF bacterial counts of the 8 subjects in the 2nd and 4th weeks was basically converted. Here, the average bacterial count ratio of BF bacteria in the 2nd week was 3.33, and the average bacterial count ratio of BF bacteria in the 4th week was 3.27. That is to say, after 8 subjects took the prebiotic composition continuously for 4 weeks, the amount of BF bacteria in their intestines increased by 3.27 times. Moreover, BF bacteria, as a physiologically beneficial bacterium, has various important physiological functions such as improving the biological barrier, improving nutrition, anti-tumor effect, immune enhancement, improving gastrointestinal function, and anti-aging to the health of the host. Therefore, by taking a prebiotic composition composed of fermented kiwifruit, inulin and fructooligosaccharides, the amount of BF bacteria in the user's intestinal tract can be effectively increased, thereby achieving improved biological barrier capabilities, improved nutrient absorption, Anti-tumor, enhance immunity, improve gastrointestinal tract and other effects.

example 5-3. Analysis of Subject Defecation Questionnaire

The defecation status of the 8 subjects was investigated by questionnaires before taking the prebiotic composition (that is, week 0) and after taking the prebiotic composition (that is, the second and fourth weeks). Moreover, the questionnaire feedback results of the 8 subjects are organized as shown in Figure 8 to Figure 11 .

Here, the questionnaire items analyzed include: degree of difficulty in defecation (such as the frequency of defecation, the time required for defecation, whether the defecation is clean) and the frequency of intestinal peristalsis, etc., as shown in Table 3.

table 3 Comprehensive assessment of defecation status Defecation status/Bowel condition options 1. How often do you have a bowel movement? □ once a day □More than twice a day □ once every two days 2. How long does it take to defecate in the toilet? □Less than 5 minutes □5~10 minutes □10~20 minutes 3. In the past period of time, did you feel the frequency of defecation is not clean? □ no □ very little □Sometimes □Often 4. In the past period of time, how often have you felt bowel movements? □ no □ very little □Sometimes □Often

See Figure 8. Before taking the prebiotic composition at week 0, 4 of the 8 subjects (i.e. 50% of the subjects) defecated more than once in two days, but after taking the prebiotic composition for 2 weeks, more than two days The number of subjects who defecated once dropped to 1, and 4 subjects defecated once a day, and 3 subjects defecated more than twice a day. After taking the prebiotic composition for 4 weeks, all the subjects (i.e. 100% of the subjects) defecated every day, among which 5 subjects defecated once a day, and 3 subjects defecated twice a day above. It can be known that taking the prebiotic composition can improve the recipient's defecation frequency (at least once a day), thereby making it easier for the recipient to clear the stool.

See Figure 9. Before taking the prebiotic composition at week 0, 1 of the 8 subjects had a defecation time of 10-20 minutes, and 4 subjects had a defecation time of 5-10 minutes , while the defecation time required for 3 subjects was less than 5 minutes. However, after taking the prebiotic composition for 2 weeks, none of the subjects needed to defecate for more than 10 minutes, and there were 5 subjects who needed to defecate for 5 to 10 minutes, and 3 subjects who needed to defecate. The required bowel movement time is less than 5 minutes. And, after taking the prebiotic composition for 4 weeks, as with the results of the 2nd week survey, none of the subjects needed to defecate for more than 10 minutes, and among the 8 subjects, the defecation time required by the subjects was 10 minutes. The number of people who are 5-10 minutes and less than 5 minutes are 4. In other words, 50% of the subjects need to defecate for less than 5 minutes, and 100% of the subjects need to defecate. 10 minutes or less. It can be seen that taking the prebiotic composition can improve the defecation time of the receptors, thereby avoiding problems caused by sitting on the toilet for a long time (such as foot numbness and nerve compression), and can reduce the time of using the toilet.

See Figure 10. Before taking the prebiotic composition at week 0, 50% of the 8 subjects felt "sometimes" that their bowel movements were not clean, and 12.5% of the subjects "often" felt that their bowel movements were not clean, Only 25% of the subjects "rarely" felt unclean bowel movements and 12.5% of the subjects "never" felt unclean bowel movements. After taking the prebiotic composition for 2 weeks, 50% of the subjects felt "rarely" unclean bowel movements, 12.5% felt "no" unclean bowel movements, and no subjects felt "often" Dirty bowel movements. Moreover, only 37.5% of the subjects "sometimes" feel that their bowel movements are not clean. After taking the prebiotic composition for 4 weeks, 62.5% of the subjects felt "rarely" unclean bowel movements, 25% felt "no" unclean bowel movements, and none of the subjects felt "often" Dirty bowel movements. Moreover, only 12.5% of the subjects "sometimes" feel that their bowel movements are not clean. In other words, taking the prebiotic composition can reduce the feeling of unclean defecation, so that the recipient can feel smooth defecation.

See Figure 11. Before taking the prebiotic composition at week 0, none of the 8 subjects felt "often" intestinal peristalsis, while 1 subject "did not" feel intestinal peristalsis. In addition, 2 subjects "rarely" felt intestinal peristalsis, and 5 subjects "sometimes" felt intestinal peristalsis. After taking the prebiotic composition for 2 weeks, there were 2 people who "often" felt the situation of intestinal peristalsis, and no subject "did not" feel the situation of intestinal peristalsis, while "seldom" and "some "Time" felt the situation of intestinal peristalsis, a total of 6 people. After taking the prebiotic composition for 4 weeks, the number of people who "frequently" felt the situation of intestinal peristalsis reached 3, and no subject "did not" feel the situation of intestinal peristalsis, while 3 subjects " "Sometimes" felt intestinal peristalsis and 2 subjects "rarely" felt intestinal peristalsis. Therefore, after taking the prebiotic composition for 4 weeks, the 8 subjects felt that the frequency of intestinal peristalsis increased. It can be known that taking the prebiotic composition can promote the frequency of intestinal peristalsis of the recipient, thereby improving the intestinal tract and improving the defecation situation, so that the recipient can defecate smoothly.

example 5-4. Subject body composition analysis

The 8 subjects were measured by body fat (brand: TANITA BC-601FS) before taking the prebiotic composition (that is, week 0) and after taking the prebiotic composition (that is, the second and fourth weeks). Measure its body composition. In addition, the body composition results of the 8 subjects are arranged as shown in Figure 12 to Figure 15 .

Here, the body composition items analyzed include: body weight, total body fat percentage, trunk body fat percentage and waist circumference.

See Figure 12. Before taking the prebiotic composition at week 0, the average body weight of the 8 subjects was 73.6 kg, but after taking the prebiotic composition for 2 weeks, the average body weight of the 8 subjects decreased to 72.7 kg. Moreover, after taking the prebiotic composition for 4 weeks, the average body weight of the 8 subjects dropped to 72.5 kg. In other words, the average body weight of the subjects decreased by 0.9 kg after taking the prebiotic composition for 2 weeks, and the average body weight of the subjects decreased by 1.1 kg after taking the prebiotic composition for 4 weeks. It can be known that by taking the prebiotic composition composed of fermented kiwi fruit, inulin and fructooligosaccharides, the body weight of the recipient can be effectively reduced and the effect of weight loss can be achieved.

See Figure 13. Before taking the prebiotic composition in the 0th week, the average body fat rate of the 8 subjects was 32%, and after taking the prebiotic composition for 2 weeks, the average body fat rate of the 8 subjects decreased to 31.5%. Moreover, after taking the prebiotic composition for 4 weeks, the average body fat rate of the 8 subjects dropped to 31.2%. In other words, the average body fat percentage of the subjects decreased by 0.5% after taking the prebiotic composition for 2 weeks, and the average body fat percentage of the subjects decreased by 0.8% after taking the prebiotic composition for 4 weeks. It can be known that by taking the prebiotic composition composed of fermented kiwifruit, inulin and fructooligosaccharides, it can effectively reduce the body fat rate of the recipient and achieve a slimming effect.

See Figure 14. Before taking the prebiotic composition in the 0th week, the average trunk body fat rate of the 8 subjects was 33.6%, and after taking the prebiotic composition for 2 weeks, the average trunk body fat rate of the 8 subjects decreased to 32.9%. Moreover, after taking the prebiotic composition for 4 weeks, the average trunk body fat rate of the 8 subjects dropped to 32.7%. In other words, the average trunk body fat percentage of the subjects decreased by 0.7% after taking the prebiotic composition for 2 weeks, and the average trunk body fat percentage of the subjects decreased by 0.9% after taking the prebiotic composition for 4 weeks. It can be seen that by taking the prebiotic composition composed of fermented kiwi fruit, inulin and fructooligosaccharides, the body fat percentage of the recipient's trunk can be effectively reduced to achieve a slimming effect.

See Figure 15. Before taking the prebiotic composition at week 0, the average waist circumference of the 8 subjects was 91.9 cm, and after taking the prebiotic composition for 2 weeks, the average waist circumference of the 8 subjects decreased to 89.4 cm. Moreover, after taking the prebiotic composition for 4 weeks, the average waist circumference of the 8 subjects dropped to 87.9 cm. In other words, the average waist circumference of the subjects decreased by 1.7 cm after taking the prebiotic composition for 2 weeks, and the average waist circumference of the subjects decreased by 3.2 cm after taking the prebiotic composition for 4 weeks. It can be known that by taking the prebiotic composition composed of fermented kiwi fruit, inulin and fructooligosaccharides, the waist circumference of the recipient can be effectively reduced, thereby achieving a slimming effect.

In addition, one of the subjects detected the change of abdominal fat with a bone density and body composition analyzer (DXA) at week 0 and week 4, as shown in Figure 16 . FA0 is the subject's abdominal fat at week 0 (the white area above the spine in the left figure of Figure 16), and FA4 is the abdominal fat of the subject at week 4 (the white area above the spine in the right figure of Figure 16 area), and comparing the size of FA0 and FA4, it can be found that the subject's abdominal fat was significantly reduced. In other words, by taking the prebiotic composition composed of fermented kiwi fruit, inulin and fructooligosaccharides, the abdominal fat of the recipient can be effectively reduced, thereby achieving a slimming effect.

To sum up, the prebiotic composition comprising fermented kiwi fruit, inulin and fructooligosaccharide according to any embodiment of the present invention can be used to improve the intestinal tract of the recipient. Wherein, the weight ratio of kiwifruit fermented product, inulin and fructooligosaccharide contained in the prebiotic composition is 3-4:2.5-4:2.5-4. In some embodiments, a prebiotic composition comprising fermented kiwifruit, inulin, and fructooligosaccharides can be used to improve the intestinal flora (such as Klemensia muciniphila, Parabacteroides gordii, and Bifidobacterium sp. ), improving the recipient's defecation status (such as improving the recipient's difficulty in defecation, promoting the recipient's intestinal peristalsis frequency, reducing the recipient's unclean defecation, reducing the recipient's defecation time or a combination), increasing the recipient's Gut barrier (eg, increased concentration of tightly-structured proteins in the recipient) or a combination thereof. Moreover, in some embodiments, the prebiotic composition including fermented kiwi fruit, inulin and fructooligosaccharides can be used to reduce the recipient's body weight, waist circumference, body fat percentage or a combination thereof, thereby achieving the effect of weight loss.

Although the technical content of the present invention has been disclosed above with preferred embodiments, it is not intended to limit the present invention. Any modification and modification made by those skilled in the art without departing from the spirit of the present invention should be covered by the present invention. Therefore, the scope of protection of the present invention should be defined by the scope of the appended patent application.

FA0: Belly Fat Week 0 FA4: Belly Fat Week 4

Fig. 1 is the result figure of the analysis experiment of the effect of a single component on the growth of probiotics; Fig. 2 is the result figure of the analysis experiment of multiple groups of different ingredients affecting the growth of probiotics; Fig. 3 is the analysis experiment result diagram of the effect of multiple groups of different ratios of prebiotic combinations on the growth of probiotics; Fig. 4 is a graph showing the experimental results of the prebiotic composition on the expression of compact structure proteins in the recipient's blood; Fig. 5 is a graph showing the experimental results of the growth ability of the prebiotic composition for Kreman mucinophilus; Fig. 6 is the experimental result diagram of the growth ability of the prebiotic composition for Parabacteroides gordii; Figure 7 is a graph showing the experimental results of the growth ability of the prebiotic composition for Bifidobacteria; Figure 8 is a graph showing the analysis results of the prebiotic composition for the average number of bowel movements of the subjects; Figure 9 is a graph of the analysis results of the prebiotic composition for the average defecation time of the subject; Figure 10 is a graph showing the analysis results of the prebiotic composition on the frequency of unclean defecation of the subject; Figure 11 is a diagram of the analysis results of the frequency of intestinal peristalsis of the subject for the prebiotic composition; Figure 12 is an analysis chart of the average body weight of the subjects in the 0th week, the 2nd week and the 4th week; Figure 13 is an analysis chart of the average body fat percentage of the subjects in the 0th week, the 2nd week and the 4th week; Figure 14 is an analysis chart of the average trunk body fat percentage of the subjects in the 0th week, the 2nd week and the 4th week; Figure 15 is an analysis chart of the average waist circumference of the subjects in the 0th week, the 2nd week and the 4th week; and Fig. 16 is an analysis photograph of the abdominal fat mass of the test subjects at the 0th week and the 4th week.

Claims (11)

A prebiotic composition comprising kiwifruit fermented product, inulin and fructooligosaccharide, wherein the weight ratio of the kiwifruit fermented product, the inulin and the fructooligosaccharide is 3-4:2.5-4:2.5-4. The prebiotic composition according to claim 1, wherein the weight ratio of the kiwifruit fermented product, the inulin and the fructooligosaccharide is 4:3:3. The prebiotic composition as described in claim 1, used to promote the growth ability of Parabacteroides goldsteinii. A use of kiwifruit fermented product and sugars for preparing a prebiotic composition for improving the intestinal tract of a recipient, wherein the prebiotic composition includes the kiwifruit fermented product, inulin and fructooligosaccharides, and the kiwifruit The weight ratio of the fermented product, the inulin and the fructooligosaccharide is 3-4:2.5-4:2.5-4. The use according to claim 4, wherein the weight ratio of the kiwi fruit fermented product, the inulin and the fructooligosaccharide is 4:3:3. The use according to claim 4 or 5, wherein improving the intestinal tract of the recipient includes improving the bacterial phase of the intestinal tract, improving the defecation status of the recipient, improving the intestinal barrier of the recipient or a combination thereof. The use as described in Claim 6, wherein the bacterial phase includes Akkermansia muciniphila , Parabacteroides goldsteinii and Bifidobacterium . The use as described in claim 6, wherein improving the defecation status of the recipient refers to improving the difficulty of defecation of the recipient, promoting the frequency of intestinal peristalsis of the recipient, reducing the unclean defecation of the recipient, reducing the The body's defecation time or a combination thereof. The use as claimed in claim 6, wherein the prebiotic composition is used to increase the concentration of the compact structure protein in the blood of the recipient. The use as described in Claim 9, wherein the compact structure protein is claudin 3 (Claudin 3, CLDN3). The use according to claim 4, wherein the prebiotic composition is used to reduce the subject's body weight, waist circumference, body fat percentage or a combination thereof.

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