TWI767377B - Composition for promoting growth of akkermansia muciniphila and uses thereof - Google Patents

Abstract

The present disclosure provides a composition for promoting growth ofAkkermansia muciniphila . Wherein the composition comprises Pachyrhizus erosus ferment. The composition further promotes the growth ofParabacteroides goldsteinii andBifidobacterium in the user, and can modulate the user’s weight, body fat, waist circumference, and skin condition.

Description

Composition for promoting the growth of Ekman Muxophila and use thereof

The present invention relates to a composition, particularly a composition for promoting the growth of Akkermansia muciniphila .

According to research, more than 70% of microorganisms symbiotically exist in the human digestive system, especially in the gastrointestinal tract to build a rich and complex ecosystem, called the intestinal flora, which affects many physiological states and regulation of the human body, and can reflect the human body's Various physical states have an important impact on human beings.

Akkermansia muciniphila is a non-spore-forming, non-motile, obligate anaerobe (Obligate anaerobe) gram-negative bacterium, usually present in the intestines of most mammals.

Recent studies have pointed out that obese or diabetic people have less Ekman mucophilus in their intestines. In addition, oral ingestion of Ekmanmucinophilus was found to reverse high-fat diet-induced obesity in a mouse model, possibly due to the outer membrane protein (Amuc_1100 ) reduces fat accumulation in mice and stabilizes blood sugar.

However, due to the obligate anaerobic characteristics of Ekman Muxophila, it is not easy to grow, and requires a special medium derived from animals. Therefore, it is relatively difficult to supplement the bacteria orally after culturing the bacteria externally. Therefore, it is necessary to develop a natural composition that can promote the number of Ekmanmucinophilus in the gastrointestinal tract.

In view of this, there is a need for research or development of a natural plant-based composition for promoting Ekman Muxophila to effectively improve the health of the organism.

An object of the present invention is to provide a composition for promoting the growth of Akkermansia muciniphila , which comprises a cold potato fermented product.

In some embodiments, the cold potato fermented product is obtained by fermenting a culture solution of a cold potato and water through a plurality of strains.

In some embodiments, the culture solution is obtained by mixing jicama and water in a weight ratio of 1:3 to 1:6.

In some embodiments, the plurality of species consists of a Saccharomyces cerevisiae , a Lactobacillus plantarum and an Acetobacter aceti .

In some embodiments, the composition is capable of promoting the growth of Ekmanyl Muxophila more than 3-fold.

In some embodiments, the compositions are further capable of promoting the growth of Parabacteroides goldsteinii and Bifidobacterium bacteria.

According to some embodiments, the present invention provides a use of a composition for reducing body weight, body fat and waist circumference in a subject.

According to some embodiments, the present invention provides a use of a composition for improving the condition of a subject's skin.

In some embodiments, the composition has at least two of the following functions: increasing skin hydration, increasing skin elasticity, reducing skin wrinkles, reducing skin UV spots, and enhancing skin radiance.

In some embodiments, the individual is an individual with a Body Mass Index (BMI) between 24 and 27 or a body fat percentage greater than 25%.

In some embodiments, the composition further comprises xylo-oligosaccharides and inulin.

In some embodiments, the weight ratio of jicama fermented product, xylo-oligosaccharide and inulin ranges from 3 to 5: 1 to 3: 1 to 3.

The invention provides a cold potato fermented product and a composition thereof for promoting the growth of Ekman Mucophilus. The jicama fermented product can be used as a component in a composition for effectively promoting the growth of Ekman Muxophila, and the composition can be further used for reducing the body weight, body fat and waist circumference of an individual, or for improving the skin of an individual situation. The composition can be in the form of powder, granule, solution, or capsule, and can be made into food, drink, medicine, or nutritional supplement, and administered to an individual orally.

The embodiments of the present invention will be further described below in conjunction with the drawings. The following examples are used to illustrate the inventive features and applications of the present invention, but not to limit the scope of the present invention. Within the spirit and scope of the present invention, some changes and modifications can be made, so the protection scope of the present invention should be determined by the scope of the appended patent application.

Some specific implementation aspects of this case will be described below. Without departing from the spirit of this case, this case can still be practiced in many different forms, and the scope of protection should not be limited to the conditions specifically stated in the description.

The case used Excel software for statistical analysis. Data are presented as mean ± standard deviation (SD), and differences between groups were analyzed by Student's t -test .

Numerical values used herein are approximations and all experimental data are expressed within 20%, preferably within 10%, and most preferably within 5%.

The so-called "Brix" in this article refers to Brix (Degrees Brix, symbol °Bx), which is a unit for measuring sugar content, representing the number of grams of sucrose dissolved in 100 grams of aqueous solution at 20 °C, which can be used to judge fermentation. The degree to which sugar is converted in the process.

The term "individual" as used herein refers to a human or non-human mammal, preferably a human.

The term "skin" herein refers to the skin of a human or non-human mammal, preferably the skin of a human.

In this article, the so-called "cold potato" refers to the perennial entwining herbaceous vines of the Fabaceae ( Fabaceae ) genus Pachyrhizus , also known as jicama, sand kudzu and other aliases.

The so-called "inulin" in this article, also called inulin, is a type of fructan, which is formed by the polymerization of fructose molecules.

The term "xylo-oligosaccharides" herein refers to oligosaccharides containing 2 to 7 xylose units, which can be derived from plants, or obtained by microbial fermentation, enzymatic transformation, or chemical synthesis.

As used herein, the terms " Saccharomyces cerevisiae ,"" Lactobacillus plantarum ," and " Acetobacter aceti ," respectively, are intended to encompass those readily available to those skilled in the art Saccharomyces cerevisiae, Lactobacillus endospermum and Acetobacter (for example, those that can be purchased from domestic or foreign depository institutions), or Saccharomyces cerevisiae isolated and purified from natural sources using conventional microbial isolation methods in the art , Plasmodium and Acetobacter strains.

According to the present invention, types of food products include, but are not limited to, beverages, fermented foods, bakery products, health foods, and dietary supplements.

Unless otherwise stated, the experimental procedures in the following examples were carried out at room temperature (25±5°C) and normal pressure (1 atm).

Example 1 : solid state Preparation of cold potato fermented product

Cold potato ( Pachyrhizus erosus ) is a perennial vine-like herb of the genus Pachyrhizus (Legumen), also known as jicama, kudzu, and kudzu. Its agricultural cultivation is mainly distributed in tropical regions, including Ecuador in South America. , Peru, Taiwan, Guangdong, Sichuan, Ryukyu, Indonesia, Vietnam, Philippines, Malaysia in Asia. The origin of the jicama used in the present invention is not limited. Cold potato is named after the root is like potato pieces. The root of cold potato can be eaten raw or cooked, with a diameter of about 20-30 cm, and can be processed into sand arrowroot powder, which has the effect of cooling and removing heat. In the present invention, jicama is fermented to obtain a fermented product.

( 1-1 ): Preparation example of solid-state jicama fermented product 1

First, thoroughly clean the root parts of the jicama, and break them into jicama root granules with a particle size of less than 12 mm. The jicama root granules and water are uniformly mixed in a weight ratio of 1:3-6 to obtain a raw material mixed solution. Then, the raw material mixed solution is heated at 80°C-100°C for 0.5-1 hour to obtain a culture solution of jicama. Next, according to the total weight of the jicama culture solution, 9.0 wt% (weight percent) of glucose was added to the jicama culture solution, and the sugar content of the jicama culture solution at this time was 9.0°Bx.

After the cold potato culture solution was cooled to room temperature, the fermentation procedure was carried out, and 0.05-0.2 wt% of Saccharomyces cerevisiae (purchased from the Biological Resource Conservation and Research Center of the Food Industry Development Institute (BCRC) was added relative to the culture solution. ), deposit number BCRC20271) (international deposit ATCC26602) in the culture medium, fermented for 0.5-2 days to form the first primary fermentation product, and then added 0.02-0.1 wt% of Lactobacillus plantarum relative to the culture medium ) (purchased from BCRC, deposit number BCRC910805) (international deposit DSM33108) in the first primary fermentation product, fermented for 0.5-2 days to obtain the second primary fermentation product, and then added 3.0-7.0 wt% relative to the culture medium Acetobacter aceti (purchased from BCRC, deposit number BCRC11688) (international deposit ATCC15973) was fermented in the second primary fermentation for 2-4 days. Finally, without removing these three kinds of bacteria, use the set sugar content range of 4.0°Bx ± 1.0, pH value of 3.4 ± 0.3 and other specifications, if the inspection meets the specifications, it is determined that the fermentation is completed and the cold potato fermented product is obtained.

In an embodiment of the present invention, the strains used in the fermentation can also be other yeast strains of the same genus and different species other than Saccharomyces cerevisiae, and other yeast strains of the same genus and different species other than Lactobacillus sp. Bacillus strains and other Acetobacter strains of the same genus and different species other than acetic acid bacteria, the present invention is not limited to this.

Then, the fermented jicama is prepared into a solid jicama fermented product, and the preparation method thereof is briefly described as follows. The fermented jicama is subjected to a drying procedure (eg, hot air drying, freeze drying, spray drying) to remove moisture. When carrying out the drying procedure, first add 30-50wt% maltodextrin to the jicama fermented product as an excipient. After stirring evenly, confirm that the maltodextrin is completely dissolved, and the sugar content of the jicama fermented product at this time is If the value is greater than 23°Bx, the jicama fermented product is dried by hot air, freeze drying or spray drying to remove water to obtain a solid jicama fermented product. The specification set for the solid-state jicama fermentation product is moisture <5%.

( 1-2 ): Preparation example of solid-state jicama fermented product 2

The preparation method of the solid-state jicama fermented product used in subsequent Examples 2, 3 and 4 of the present invention is briefly described as follows. First, thoroughly clean the root parts of the jicama, and break them into jicama root granules with a particle size of less than 12 mm. The jicama root granules and water are uniformly mixed at a weight ratio of 1:5 to obtain a raw material mixed solution. Then, the raw material mixed solution was heated at 95° C. for 0.5 hours to obtain a cultured solution of jicama. Next, according to the total weight of the jicama culture solution, 9.0 wt% (weight percent) of glucose was added to the jicama culture solution, and the sugar content of the jicama culture solution at this time was 9.0°Bx.

After the cold potato culture solution was cooled to room temperature, the fermentation procedure was carried out, and 0.1 wt% of Saccharomyces cerevisiae (purchased from the Biological Resource Conservation and Research Center (BCRC) of the Institute of Food Industry Development) was added relative to the culture solution. The accession number BCRC20271) (international deposit ATCC26602) was fermented for 1 day in the culture medium to form the first primary fermentation product, and then 0.05 wt% of Lactobacillus plantarum (purchased from BCRC, relative to the culture medium) was added. The accession number BCRC910805) (international deposit DSM33108) was fermented in the first primary fermentation product for 1 day to obtain the second primary fermentation product, and then 5.0 wt% acetic acid bacteria ( Acetobacter aceti ) (purchased from BCRC, deposit number BCRC11688) (international deposit ATCC15973) was fermented for 3 days in the second primary fermentation. Finally, without removing these three kinds of bacteria, use the set sugar content range of 4.0°Bx ± 1.0, pH value of 3.4 ± 0.3 and other specifications, if the inspection meets the specifications, it is determined that the fermentation is completed and the cold potato fermented product is obtained, Then, the jicama fermented product is prepared into a solid jicama fermented product. First add 30wt% maltodextrin (as an excipient) to the cold potato fermented product, stir evenly, confirm that the maltodextrin is completely dissolved, and the sugar content of the cold potato fermented product is greater than 23°Bx. The potato fermented product is freeze-dried to remove moisture to obtain a solid-state jicama fermented product. The specification that the solid-state sweet potato fermented product is set to is moisture＜5%, and if the inspection meets the specification, it is determined that the preparation is completed and the solid-state sweet potato fermented product is obtained, which is also the used solid-state sweet potato fermented product of Example 2, Example 3 and Example 4. .

Example 2 : Probiotic test of Ekman Mucophilus

Source of xylo-oligosaccharide, fructooligosaccharide, inulin, lactitol

The xylo-oligosaccharides used in the examples herein were purchased from Shandong Longli Biotechnology Co., Ltd., the fructooligosaccharides were purchased from Meiji Co., Ltd., the inulin was purchased from Cosucra Groupe Warcoing, and the lactitol was purchased from Hongwei Biotechnology Co., Ltd. . However, the source of these sugars is not limited.

Source and sample preparation of Ekman Mucophilus

The source of Ekman Muxophila can be isolated from the intestines of human volunteers or directly purchased from the Food Industry Development Research Institute. In the following examples, Ekmanella muciniphila was purchased from the Bioresource Collection and Research Center (BCRC), number BCRC81048.

An activation procedure was carried out to activate Ekman Muxophila. The Ekman Mucophilus was inoculated into a brain-heart medium (Brain Heart Infusion, also known as BHI medium) with 10% of the bacteria in a cryovial and cultured anaerobically at 37°C for 48 hours to activate Ekman. Mucinophilus, follow-up probiotic experiments can be carried out.

In order to identify the nutrients that are beneficial to the growth of Eikeman Muxophila, the pre-cultured Eikeman Muxophila was added to a 3% inoculum (about 1x10 ⁴ CFU/mL) containing 1% (w/w) The candidate probiotics were cultured in the brain-heart medium (volume 5mL) in an anaerobic environment at 37°C for 48 hours. The candidate prebiotics are xylo-oligosaccharide (experimental group 1), lactitol (experimental group 2), inulin (experimental group 3), fructooligosaccharide (experimental group 4), or solid-state jicama fermented product (experimental group 5 ). As a control, Ekmannii mucinophilus was additionally cultured in brain-heart medium without candidate probiotics (control group).

Figure 1 shows the effects of the aforementioned various candidate probiotics on the growth of Ekman Muxophila. In this experiment, the dilution smear method was used to calculate the bacterial count of Ekman Muxophila after 48 hours of culture. According to Figure 1, the bacterial count of the control group was 6.50 log CFU/ml, equivalent to 1x10 ^6.5 CFU/ml; the bacterial count of Eikmanthophilus in experimental group 1 was 6.75 log CFU/ml, equivalent to 1x10 ^6.75 CFU /ml; the bacterial count of Ekman Muxophila in experimental group 2 was 6.67 log CFU/ml, which is equivalent to 1×10 ^6.67 CFU/ml; the bacterial count of Ekman Muxophila in experimental group 3 was 6.53 log CFU /ml, equivalent to 1x10 ^6.53 CFU/ml; the bacterial count of Ekmanmucinophilus in experimental group 4 was 6.53 log CFU/ml, equivalent to 1x10 ^6.53 CFU/ml; The bacterial count was 6.73 log CFU/ml, equivalent to 1x10 ^6.73 CFU/ml.

Then, the number of bacteria in each experimental group was calculated by the formula: the number of bacteria in the experimental group (CFU/ml)/the number of bacteria in the control group (CFU/ml). According to this formula, compared with the control group, the candidate probiotics that can increase the bacterial count include xylo-oligosaccharide (experimental group 1), lactitol (experimental group 2), inulin (experimental group 3), fructooligosaccharide (experimental group 3) Group 4) and solid-state cold potato fermented product (experimental group 5), the bacterial counts were increased by 1.8 times, 1.6 times, 1.1 times, 1.1 times and 1.7 times, respectively. This result indicated that any of xylo-oligosaccharide, lactose, inulin, fructooligosaccharide, and solid Jicama fermented products could promote the growth of Ekman Mucophilus. However, the effects of xylo-oligosaccharide, lactose and solid jicama fermented products were more obvious.

Example 3 : Composition for promoting the growth of Ekman Muxophilus

In order to evaluate the promotion effect of different combinations of the aforementioned Ekmania muciniphila probiotics on the growth of Ekmania muciniphila, the precultured Ekmanella muciniphila was adjusted to 3% (about 1×10 ⁴ CFU/mL) Add the brain heart medium (volume 5 mL) containing 1% (w/w) composition (experimental group 6) to the inoculum, and culture at 37°C under anaerobic conditions for 48 hours. Bacteria amount. The composition in the brain-heart medium had the following composition: 0.5% solid jicama ferment, 0.25% xylo-oligosaccharide, and 0.25% inulin. As a control, Ekmannii Muxophila was additionally cultured in brain-heart medium without the aforementioned composition (control group).

Figure 1 shows the effect of the aforementioned compositions on the growth of Ekman Muxophila. According to Figure 1, the bacterial count of the control group was 6.50 log CFU/ml, equivalent to 1x10 ^6.5 CFU/ml; the bacterial count of experimental group 6 was 7.02 log CFU/ml, equivalent to 1x10 ^7.02 CFU/ml; compared with the control group, The addition of the composition significantly increased the bacterial count of Ekman Muxophila by 3.3 times. Therefore, it can be known that the combination of solid-state jicama fermented product, xylo-oligosaccharide and inulin can most effectively promote Ekman mucoprotein when the weight ratio is 3~5:1~3:1~3 The growth of bacteria, even inulin showed the lowest growth-promoting effect shown in Figure 1, only 1.1 times.

Since this composition had the best ability to enhance the growth of Ekman Muxophila, subsequent human experiments were carried out with this composition.

Example 4 : Physiological effect of the composition

This example investigates the physiological effects of continuously taking a composition capsule containing 200 mg solid-state jicama fermented product, 100 mg xylo-oligosaccharide, and 100 mg inulin on subjects who meet these conditions, (1 ) in the age range of 25-60 years old, (2) mildly obese groups or groups with excessive body fat. Among them, the mildly obese group refers to the group with a body mass index (BMI) between 24 and 27, and the high body fat refers to the body fat rate of more than 25% (men) or more than 30% (women). .

Eight subjects were orally administered the composition daily for four consecutive weeks at a dose of 400 mg/person/day, and the subjects were evaluated for changes in gastrointestinal flora, body posture, and skin before and after administration for four weeks. Numerical values such as status, and the detailed detection methods of each item are as follows.

( 4-1 ) Phase change detection method of gastrointestinal bacteria

In this experiment, Ekmannii, Parabacteroides gordonii, and Bifidobacterium were selected as the targets for the detection of bacterial phase changes. The following describes the physiological significance reflected by these species.

Akkermansia muciniphila , a bacterium belonging to the genus Akkermansia , is an intestinal bacterium that exists in the mucus layer of the human intestine, and was proposed by Muriel Derrien et al in 2004 A new Ekmania bacterium.

Ekmannii is known to be abundantly present in non-obese humans and only in small amounts in the guts of obese humans. In addition, it has been reported that, compared with normal mice, Ekmannii mucinophilus is also less in obese model mice and type 2 diabetes model mice. See, eg, Proc Natl Acad Sci USA. 2013 May 28;110(22):9066-71., which is hereby incorporated by reference in its entirety.

It has been reported that the metabolite propionate of Ekmannii can regulate fasting-induced adipose factor (FIAF), G Protein Coupled Receptor 43 (G Protein Coupled Receptor 43). , GPR43), histone deacetylases, and peroxisome proliferator-activated receptor gamma (Peroxisome proliferator-activated receptor gamma), which are involved in transcription factor regulation, cell cycle control, lipid Important regulators of breakdown resistance and satiety. See, eg, mBio. 2014 Aug 12;5(4):e01438-14., which is hereby incorporated by reference in its entirety.

Parabacteroides goldsteinii is a bacterium belonging to the genus Parabacteroides and is a Gram-negative obligate anaerobic bacterium.

According to reports, supplementation with this bacteria can make the intestinal cells of mice more compact, reduce the phenomenon of intestinal leakage, and improve the intestinal inflammation state. And can reduce the body weight of mice, slow down insulin resistance, and increase fat metabolism, activation of brown fat. From this, it is known that this bacteria can be used as a research target for improving metabolic syndrome. See e.g.: Gut. 2019 Feb;68(2):248-262, which is hereby incorporated by reference in its entirety.

Bifidobacterium ( Bifidobacterium ) is a Gram-positive anaerobic bacteria, and Bifidobacterium is an important intestinal beneficial microorganism. As a kind of physiological beneficial bacteria, Bifidobacterium has biological barrier, nutritional effect, anti-tumor effect, immune enhancement effect, improvement of gastrointestinal tract function, anti-aging and other important physiological functions to human health.

Methods of conducting phase change experiments of gastrointestinal bacteria

The subjects were tested for intestinal flora in the 0th week (before taking the above composition capsules) and the 4th week (after taking the above composition capsules).

The detection method can be to extract DNA from the stool after collecting the subject's stool sample, and perform quantitative real-time polymerase chain reaction (qRT-PCR) for the 16S ribosomal gene (16S ribosomal gene). , by using the primer sequences of known bacteria to detect the bacterial count ratio of common intestinal bacteria. The detection method can also perform DNA extraction from the stool, and then perform 16S Amplicon Sequencing on the specimen, especially for the V3-V4 region of the 16S DNA, and then sequence the next generation. (Next Generation Sequencing (NGS)) method, and perform subsequent sequencing analysis.

In this example, the next-generation sequencing method was used. The 16S Amplicon Sequencing of microorganisms was carried out by Toulsi Biotechnology Co., Ltd., and DNA was extracted from the stool samples. The 16S metagenomic sequencing library preparation protocol developed by San Diego, USA) was used for 16S rRNA gene amplicon sequencing. Briefly, fecal-extracted DNA was amplified using primers targeting the V3/V4 variable regions of the 16S rRNA gene: 16S amplicon PCR forward primer (V3 region) TCGTCGGCAGCGTCAGATGTGTATAAGAGACAGCCTACGGGNGGCWGCAG (SEQUENCE ID NO: 1); 16S amplification Amplifier PCR reverse primer (V4 region) GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAGGACTACHVGGGTATCTAATCC (SEQUENCE ID NO: 2) PCR was performed to amplify the variable region V3-V4 of the 16S rRNA gene (positions 341 to 805 in the rRNA gene) (refer to Herlemann et al., ISME J. 2011 Oct;5(10):1571-9.; where W=A/T, H=A/C/T, V=G/C/A, N=G/A/T/C). Complete the sequence and library construction of the amplified DNA, build a pair-end library according to MiSeq Reagent Kit v3 (Illumina, Wilmington, DE, USA), and perform high-throughput sequencing on the Illumina system. The raw data (Raw Data) obtained after the sequencing is completed will undergo paired sequence splicing (Raw Tags) and filtering to obtain Clean Tags, and then remove chimeras to obtain effective data (Effective Tags) that can be used for subsequent analysis. Then, based on the valid data, the similarity (greater than 97%) OTUs (Operational Taxonomic Units) clustering and species classification analysis were performed. Species annotation was performed on the representative sequences according to the OTUs clustering results to obtain the corresponding species information and species-based abundance (Abundance) distribution.

Among them, the change of the proportion of a specific bacterial species before taking (week 0) and after taking it (week 4) is calculated by the following formula: the proportion of a certain bacterial species to all bacterial counts in the specimen (week 4) )/the proportion of a certain bacterial count to all bacterial counts in the specimen (week 0). If the value is greater than 1, it means that the proportion of a certain bacteria in a certain organism has increased, which can also be called an increase in the relative abundance of a certain bacteria in a certain organism.

Referring to Fig. 2, taking the ratio of Akkermansia muciniphila strains before taking (week 0) as 1, after taking the capsules containing the composition of the present invention for 4 weeks, Akkermansia muciniphila The proportion of proteobacteria was 2.13 (week 4), which was equivalent to a 2.13-fold increase in the proportion of bacteria.

Referring to FIG. 3 , taking the ratio of Parabacteroides goldsteinii strains before taking (week 0) as 1, after taking the capsules containing the composition of the present invention for 4 weeks, the strains of Parabacteroides goldsteinii The ratio was 5.72 (week 4), which was equivalent to a 5.72-fold increase in the bacterial species ratio.

Referring to FIG. 4 , taking the Bifidobacterium strain ratio before taking (week 0) as 1, after taking the capsules containing the composition of the present invention for 4 weeks, the Bifidobacterium strain ratio was 16.87 (4th week), which is equivalent to an increase of 16.87 times the proportion of bacteria.

The ability of the composition of the present invention to regulate the bacterial phase of the human body, especially the ability of the gastrointestinal bacterial phase of the subjects with mild obesity or excessive body fat, was analyzed through the sequencing detection of the fecal samples of the subjects. From the above results It can be seen that after taking the composition of the present invention containing 200 mg of solid-state jicama fermented product, 100 mg of inulin and 100 mg of xylo-oligosaccharide, the gastrointestinal tract of the subjects was Ekman Mucophilus, Parabacteroides gordonii, Bifidobacterium The proportion of bacilli increased significantly.

( 4-2 ) Body posture change detection

Again, the subjects for the body composition change detection here are the same batch as the subjects for the aforementioned (3-1) Gastrointestinal bacteria phase change.

Subjects were tested for body weight, total body fat percentage, body fat percentage on the trunk, waist circumference in the 0th week (before taking the above composition capsules) and the 4th week (after taking the above composition capsules), and the measurement of each value The way is as follows.

Body weight, total body fat percentage and trunk body fat percentage were measured using a body weight and body fat meter (TANITA limb and trunk body composition meter, model BC-545F), while waist circumference was measured by a tape measure. The subject's waist is covered with clothing, the subject keeps standing relaxed, and his hands hang down naturally. The tester passes the tape measure around the subject's waist, adjusts the height, and takes the navel as the horizontal measurement point. , Without squeezing the skin, the subjects maintained normal breathing. At the end of the exhale, measure the waist circumference.

It should be noted that the statistically significant difference between the measurement results in the 0th week and the measurement results in the 4th week was statistically analyzed by the student t-test in the Excel software.

See Figure 5. Before the test (ie, week 0 in Figure 5), the average weight of all subjects was 74.0 kg, and after the test (ie, week 4 in Figure 5), the average weight of all subjects was 73.6 kg. In other words, after taking one tablet of the composition of the present invention containing 200 mg of solid jicama fermented product, 100 mg of inulin and 100 mg of xylo-oligosaccharide daily for 4 weeks, the average body weight of the subjects decreased by 0.6 kg. Based on this, the compositions of the present invention are capable of reducing body weight, particularly in subjects who are mildly obese or have excess body fat.

See Figure 6. Before the test (ie, week 0 in Figure 6), the average total body fat rate of all subjects was 28.2%, and after the test (ie, week 4 in Figure 6), the average total body fat of all subjects was The rate was 27.5%. In other words, after 4 weeks of daily consumption of a composition of the present invention containing 200 mg of solid jicama fermented product, 100 mg of inulin and 100 mg of xylo-oligosaccharide, the subjects' average total body fat rate decreased by 0.7%. Based on this, the composition of the present invention can reduce the total body fat rate, especially the total body fat rate of subjects with mild obesity or excessive body fat.

See Figure 7. Before the test (ie, week 0 in Figure 7), the average trunk body fat rate of all subjects was 23.6%, and after the test (ie, week 4 in Figure 7), the average trunk body fat of all subjects. The rate was 22.7%. In other words, after taking one tablet of the present invention containing 200 mg of solid jicama fermented product, 100 mg of inulin and 100 mg of xylo-oligosaccharide daily for 4 weeks, the subjects' average trunk body fat percentage decreased by 0.9%. Based on this, the composition of the present invention can reduce the trunk body fat rate, especially the trunk body fat rate in subjects with mild obesity or excessive body fat.

See Figure 8. Before the test (ie, week 0 in Figure 8), the average waist circumference of all subjects was 88.4 cm, and after the test (ie, week 4 in Figure 8), the average waist circumference of all subjects was 85.5 cm. In other words, after 4 weeks of daily consumption of a composition of the present invention containing 200 mg of jicama fermented product, 100 mg of inulin and 100 mg of xylo-oligosaccharide, the subjects' average waist circumference decreased by 2.9 cm, equivalent to a decrease of 1.14 inches. Based on this, the composition of the present invention can reduce the average waist circumference, especially the mean waist circumference of subjects who are mildly obese or have excess body fat.

Therefore, the composition of the present invention containing jicama fermented product, inulin and xylo-oligosaccharide has the effects of reducing body weight, reducing body fat rate, reducing body fat rate and waist circumference, especially reducing mild obesity or excessive body fat. Subject's body weight, total body fat percentage, trunk body fat percentage and waist circumference.

( 4-3 ) Detection of changes in various values of the skin

Once again, the subjects who were tested for the changes in various skin values were the same batch as the subjects for the aforementioned (3-1) Gastrointestinal bacteria phase changes.

Experiment method: After the above subjects took the 0th week (before taking the capsules of the above composition) and the 4th week (after taking the capsules of the above composition), the corresponding instruments and measurements were used according to different test items. way to record the value of facial skin.

Before and after taking the test, the subject will clean the face first, and the temperature and humidity of the test area where the subject is located are consistent to reduce the impact of external temperature and humidity on the skin.

Test items: skin moisture content, skin elasticity, skin wrinkles, skin UV spots and skin luster will be tested separately.

skin moisture test

Using Corneometer ^® CM825 (C+K Multi Probe Adapter System, Germany) purchased from Courage+Khazaka electronic company in Germany, the same subjects before taking (week 0) and after taking (week 4) facial skin for testing. The detection probe is based on the principle of capacitance for measurement. When the water content changes, the capacitance value of the skin also changes, so the water content on the skin surface can be analyzed by measuring the skin capacitance value.

Please refer to FIG. 9 , which shows the improvement of the moisture content of the skin (upper cheek) of the subject after taking the composition of the present invention (containing 200 mg of solid jicama fermented product, 100 mg of inulin and 100 mg of xylo-oligosaccharide) for 4 weeks, Specifically, the skin moisture content before taking the composition of the present invention is regarded as 1 (ie 100%), and after taking the composition of the present invention (test group), the skin moisture content of the subject is 1.066 (ie 106.6%), That is an average significant increase of 6.6%.

It can be seen that the composition of the present invention can improve the moisture content of the skin, and thus has the effect of improving the moisture content of the skin.

skin elasticity ( elasticity ) detection method

Using the skin elasticity detection probe Cutometer® MPA580 (C+K Multi Probe Adapter System, Germany) purchased from Courage+Khazaka electronic company in Germany, the same subjects before taking (week 0) and after taking (week 4) were tested. Facial skin test. The test principle is based on the principle of suction and stretching. A negative pressure is generated on the surface of the tested skin to suck the skin into a test probe, and the depth of the skin being sucked into the probe is detected through the optical test system, and the built-in software analyzes and calculates. Skin elasticity.

Please refer to FIG. 10 , which shows the improvement of skin elasticity state of the subject after taking the composition of the present invention (containing 200 mg of solid jicama fermented product, 100 mg of inulin and 100 mg of xylo-oligosaccharide) for 4 weeks. The skin elasticity before taking the composition of the present case was regarded as 1 (ie 100%), and after taking the composition of the present case, the skin elasticity of the subjects was 1.128 (ie 112.8%), that is, an average significant increase of 12.8%.

Thus, it can be seen that the composition of the present invention has the effect of enhancing skin elasticity.

skin wrinkles ( wrinkle ) detection method

Use the VISIA high-end digital skin detector sold by Canfield Company in the United States to conduct the test. The facial skin of the same subject before and after the test is photographed through a high-resolution camera lens. The change of the skin shadow can detect the texture position and get the value, which can represent the wrinkle degree of the skin.

Please refer to FIG. 11 , which shows the improvement in the number of skin wrinkles after taking the composition of the present invention (containing 200 mg of solid jicama fermented product, 100 mg of inulin and 100 mg of xylo-oligosaccharide) for 4 weeks. The number of skin wrinkles before taking the composition of the present case was regarded as 1 (ie 100%), and after taking the composition of the present case, the number of skin wrinkles of the subjects was 0.775 (ie 77.5%), that is, an average significant reduction of 22.5%.

It can be seen that the composition of the present invention has the effect of reducing the number of wrinkles.

Skin Ultraviolet Spot Detection

The facial skin of the same subject was tested before and after taking the VISIA high-end digital skin tester sold by Canfield in the United States. This instrument uses UV light for facial skin photography. Ultraviolet rays can be absorbed by melanin, improve the visibility of pigment spots, and detect melanin spots in the epidermis that are invisible to the naked eye. The higher the measured value, the more UV stains.

Please refer to FIG. 12 , which shows the improvement of skin ultraviolet pigmentation after taking the composition of the present invention (containing 200 mg of solid jicama fermented product, 100 mg of inulin and 100 mg of xylo-oligosaccharide) for 4 weeks. Specifically, Taking the number of skin UV spots before taking the composition of this case as 1 (ie 100%), after taking the composition of this case, the number of skin UV spots of the subjects was 0.963 (ie 96.3%), that is, an average significant reduction of 3.7%.

Thus, it can be seen that the composition of the present invention has the effect of reducing skin ultraviolet pigmentation.

Skin gloss detection method

Using the Glossymeter GL200 (C+K Multi Probe Adapter System, Germany) of the German Courage+Khazaka electronic company to detect the facial skin of the same subject before and after using the mask, the instrument is irradiated to the skin surface. The direct reflection and scattered reflection of the light are tested and calculated. The higher the measured value, the more radiant the skin is.

Please refer to FIG. 13 , which shows the improvement of skin luster of the subject after taking the composition of the present invention (containing 200 mg of solid jicama fermented product, 100 mg of inulin and 100 mg of xylo-oligosaccharide) for 4 weeks. The skin luster before the composition of the present case was regarded as 1 (ie 100%), and after taking the composition of the present case, the skin luster of the subjects was 1.073 (ie 107.3%), that is, an average significant increase of 7.3%.

From this, it can be seen that the composition of the present invention has an effect of enhancing skin luster.

To sum up, the present invention discloses various probiotics and their combinations that are beneficial to the growth of Ekman Muxophila. These prebiotics can be used as components in compositions effective for promoting the growth of Ekmanxomyxa, which can be used, for example, as a medium supplement for in vitro bacterial cultures, or for promoting the intestinal tract of an individual (eg, a human) A product of the proliferation of Eikmann's Muxophila. The composition can be in the form of powder, granule, solution, or capsule, and can be made into food, drink, medicine, or nutritional supplement, and administered to an individual orally.

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 person who is familiar with the art, makes some changes and modifications without departing from the spirit of the present invention, should be included in the present invention. Therefore, the protection scope of the present invention shall be determined according to the scope of the appended patent application.

none

Fig. 1 is the result of the detection of the effect of various probiotics on the growth of Akkermansia muciniphila in the test tube experiment; Fig. 2 is the detection result of Akkermansia muciniphila in the human experiment Figure 3 is the result of the detection of Parabacteroides goldsteinii in the human experiment; Figure 4 is the result of the detection of Bifidobacterium in the human experiment; Figure 5 is the result of the human experiment Fig. 6 is the result diagram of the whole body fat detection in the human experiment; Fig. 7 is the result diagram of the trunk body fat detection in the human experiment; Fig. 8 is the result diagram of the waist circumference detection in the human experiment; Fig. 9 is the result chart of skin moisture content detection in human experiment; Fig. 10 is the result chart of skin elasticity detection in human experiment; Fig. 11 is the result chart of skin wrinkle detection in human experiment; Fig. 12 is the result chart of human experiment Figure 13 shows the results of skin gloss detection in human experiments; in the diagram, "*" represents p-value less than 0.05, "**" represents p-value less than 0.01, and "***"" means p-value less than 0.001. The more "*", the more significant the statistical difference.

Claims (6)

The use of a jicama fermented product for preparing a composition for promoting the growth of Akkermansia muciniphila , the composition comprising the jicama fermented product, a xylo-oligosaccharide and an inulin, wherein the jicama The weight ratio of the fermented product, the xylo-oligosaccharide and the inulin ranges from 3 to 5: 1 to 3: 1 to 3, wherein the cold potato fermented product is a culture solution of a cold potato and water and is fermented through a plurality of bacterial species As a result, these strains are composed of a Saccharomyces cerevisiae , a Lactobacillus plantarum and an Acetobacter aceti , and the Saccharomyces cerevisiae is added at 0.1 wt% relative to the culture solution Bacteria fermented for 1 day, then added 0.05wt% of the Lactobacillus embryonic bacteria relative to the culture solution for 1 day, and finally added 5.0wt% of the acetic acid bacteria relative to the culture solution and fermented for 3 days to obtain the cold potato fermentation thing. The use according to claim 1, wherein the culture solution is obtained by mixing the jicama and the water in a weight ratio of 1:3 to 1:6. The use as claimed in claim 1, wherein the composition promotes the growth of Ekman Muxophila more than 3 times. The use of claim 1, wherein the composition is further capable of promoting the growth of Parabacteroides goldsteinii and Bifidobacterium bacteria. Use of a composition for reducing body weight, body fat and waist circumference of an individual, the composition comprising a jicama fermented product, a xylo-oligosaccharide and an inulin, wherein the jicama fermented product, the xylo-oligosaccharide and the The weight ratio of inulin ranges from 3 to 5: 1 to 3: 1 to 3, wherein the individual is an individual with a body mass index (BMI) between 24 and 27, wherein the jicama fermented product is a jicama and A culture solution of the water is obtained by fermenting a plurality of bacterial species, and the bacterial species are composed of a Saccharomyces cerevisiae , a Lactobacillus plantarum and an Acetobacter aceti . The Saccharomyces cerevisiae was added at 0.1wt% relative to the culture solution for 1 day, and then added at 0.05wt% relative to the culture solution for 1 day of fermentation, and finally 5.0wt% relative to the culture solution was added. % of the acetic bacteria was fermented for 3 days to obtain the cold potato fermented product. The use of claim 5, wherein the individual is an individual with a body fat percentage greater than 25%.

Images