KR102098808B1 - Composition of synbiotics for suppressing constipation and diarrhea induction comprising Lactobacillus plantarum VH13 and Undaria pinnatifida Sporophyll hot-water extract - Google Patents

Abstract

The present invention relates to synbiotics comprising Lactobacillus plantarum VH13 and a hot-water extract of a head part of Undaria pinnatifida Sporophyll. The synbiotics according to the present invention can effectively suppress the induction of constipation and diarrhea, and then actually improve intestinal environment, thereby being beneficial for intestinal health.

Description

Composition of synbiotics for suppressing constipation and diarrhea induction comprising Lactobacillus plantarum VH13 and Undaria pinnatifida Sporophyll hot-water extract} Lactobacillus plantarum VH13 and seaweed return hydrothermal extract

The present invention relates to a new biotics composition for inhibiting constipation and diarrhea, and specifically, a new biotics composition for suppressing constipation and diarrhea including Lactobacillus plantarum VH13 and seaweed hydrothermal extract.

Lactobacillus is one of the most commonly used health functional foods in the world as it is in the spotlight as a probiotic that helps the host's health when taken as a probiotic. In Korea, the Ministry of Food and Drug Safety has also recognized suiting as a function, and sales are being made with the effect of improving constipation and suppressing diarrhea. However, there has been a recent report that lactobacillus injected from the outside has a very small effect on the intestinal microflora pattern because it is difficult to occupy the habitat in competition with bacteria in the intestine (Zmore et al, 2018). It is being questioned that it shows the functionality of. This seems to be consistent with the market's assessment that functionalities such as improving constipation and suppressing diarrhea in the intestine do not appear collectively. This is analyzed because the probiotics injected from the outside do not adhere to the intestines and are excreted as they are. The proof of this is that the probiotics are supposed to take more than a certain number of live bacteria every day, even though they are probiotics.

In order to solve this contradiction, prebiotics, which are food sources of lactic acid bacteria, are formulated with probiotics in order to help the lactic acid bacteria occupy the intestinal habitat. However, it has been found that the prebiotics, which are widely introduced in general, have little effect on the attachment of lactic acid bacteria to the intestine, so it is expected that the effect on improving the intestinal environment is minimal.

On the other hand, prior art related to this includes the registered patent No. 10-1381794 new biotic food composition containing tagatose and probiotic lactic acid bacteria, Patent Publication No. 10-2018-0116093 yacon extract and probiotic lactic acid bacteria Synthetic bio-compositions, etc., but these are prebiotics each containing tagatose and yacon extracts, and the method for preparing the new bio-fermented fermentation product of Patent No. 10-1805863 and the new bio-fermented product produced thereby The technical feature is that the mixture of soybean and oat is inoculated and fermented with lactic acid bacteria culture. In addition, Patent Publication No. 10-2016-0114800, a prebiotic composition comprising a zucchini, and a new probiotic composition comprising the same, an antimicrobial agent, a food and pharmaceutical composition are known to be related to a neobiotic composition comprising a zucchini, , Means for solving the above problems are not disclosed.

Therefore, there is a need to develop a new prebiotic that can actually help to improve the intestinal environment and suppress constipation and diarrhea.

Registered Patent No. 10-1381794 Patent Publication No. 10-2018-0116093 Registered Patent No. 10-1805863 Patent Publication No. 10-2016-0114800

Zmora N, Zilberman-Schapira G, Sues J, More U, Dor-Bachaxh M et al, Personalized but mucosal colonization resistance to empiric probiotics is associated with unique host and microbiome features. Cell 174: 1388-1405, 2018. Nair AB, Jacob S, A simple practice guide for dose conversion between animals and human. J basic Clin Pharma 7: 27-31, 2016.

In order to solve the above problems, the present inventors intend to provide a new biotics composition comprising Lactobacillus plantarum VH13 (KFCC11759P) and seaweed hydrothermal extract.

In addition, to provide a health functional food containing the composition.

In order to achieve the above object, the present invention provides a new biotics composition comprising Lactobacillus plantarum VH13 (KFCC11759P) and seaweed hydrothermal extract.

In addition, it provides a health functional food composition comprising the new biotics composition.

When the seaweed extract contained in the composition is extracted using water as a solvent, it may be any one selected from the group consisting of hot water extraction, cold immersion extraction, reflux cooling extraction, ultrasonic extraction and steam extraction, and preferably hot water extract. have.

When extracting by hot water extraction, the extraction temperature can be extracted at 25 to 90 ° C.

The extraction time may be 1 to 3 hours, preferably 1 to 2 hours, more preferably 1.5 hours.

The final property of the seaweed hydrothermal water extract may be 0.2 to 1 mg / ml on a reducing sugar basis, preferably 0.5 mg / ml, and more preferably 1 mg / ml.

The reducing sugar is a generic term for the aldehyde group or ketone group of a sugar molecule in a free or hemiacetal form, and the reducing sugar as used in the present invention may be glucose.

When converting the appropriate concentration of the reducing sugar standard of the present invention to the concentration applied to the human body, it can be converted by the following Table 1 (Nair and Jacob, 2016).

If the optimal concentration is calculated after experimenting with rats according to Table 1 above and applying it to the human body,

Optimal conversion concentration X without side effects (mg / kg) * 0.16 / kg

It can be calculated by calculating.

The composition is characterized by suppression of constipation and diarrhea.

In addition, the health functional food comprising the new biotics composition according to the present invention may additionally include a food additive or an additive.

The additive or auxiliary material may include sugar, sweetener, milk cream, fragrance, and the like. Specific examples of the auxiliary raw material may include crystalline glucose, crystalline cellulose, maltodextrin, fructooligosaccharide, galactooligosaccharide, crystalline fructose, lactose mixed powder, vegetable cream powder, bromelain, isomalt, xylitol, and the like.

The new biotics composition of the present invention can be prepared as a health functional food in various forms by a conventionally known method, for example, liquids (drinks, pouches), tablets, pills, capsules, sticks, films, jelly And the like.

When formulating the composition of the present invention, it may further include suitable carriers, excipients, lubricants and diluents commonly used in the manufacture of pharmaceutical compositions. Carriers, excipients, lubricants and diluents that may be included in the compositions of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, cellulose, methylcellulose, microcrystalline cellulose, magnesium stearate, Silicon dioxide, etc. may be included, but is not limited thereto.

The present invention relates to neobiotics comprising Lactobacillus plantarum VH13 and seaweed hydrothermal extract, and the new biotics according to the present invention can effectively suppress constipation and diarrhea inducing and thus intestinal health. helpful.

1 is a graph showing the absorption spectrum measured at a wavelength of 550 mm to measure the concentration of reducing sugar.
Figure 2a shows the supernatant salinity concentration eluted from the seaweed according to the room temperature water immersion time.
Figure 2b shows the supernatant reducing sugar concentration eluted from the seaweed according to the room temperature water immersion time.
Figure 3 shows the degree of elution of reducing sugar, A is The concentration of reducing sugar in the seaweed, detected in the supernatant, B represents the concentration of reducing sugar detected in the supernatant of alginic acid exposed at 90 ° C for 1.5 hours.
Figure 4 shows the adhesion rate of Lactobacillus plantarum VH13 lactic acid bacteria according to the concentration of seaweed reflux water extract.
Fig. 5 Lactobacillus plantarum VH13 + The design of animal experiments for functional testing by new biotics of seaweed reflux extract.
Figure 6 shows the results of inhibiting constipation in rats caused by oral administration of Lactobacillus plantarum VH13 + seaweed reflux extract neobiotics.
Figure 7 shows the relative expression of the CxCl1 (IL-8) gene in descending colon mesothelial epithelial cells by oral administration of Lactobacillus plantarum VH13 + seaweed hydrothermal extract synbiotics.
Figure 8 shows the effect of inhibiting diarrhea in rats caused by oral administration of Lactobacillus plantarum VH13 + seaweed reflux extract neobiotics.
Figure 9 shows the relative expression of IL-10 and Muc2 gene in descending colon mesothelial epithelial cells by oral administration of Lactobacillus plantarum VH13 + seaweed hydrothermal extract Sinbiotics.

Hereinafter, the present invention will be described in detail by examples and experimental examples.

However, the following examples and experimental examples are merely illustrative of the present invention, and the contents of the present invention are not limited to the following examples and experimental examples.

<Example> Production of seaweed reflux extract

1.1 Standardization of Seaweed Return Water Extracts

Since the representative material of seaweed is a complex polysaccharide and the functional component in vitro is water-soluble, the total reducing sugar in the hot-water extract was designated as an indicator for standardization of the water-soluble fraction. The concentration was estimated by applying a colorimetric method using dinitrosalicylic acid (DNS) to measure the absorption at 550 nm with a UV-Vis Spectrophotometer (Shimadzu, UV-1800, JP) and substituting it into a standard graph. The standard graph for concentration prediction was introduced in the seaweed regression and introduced Glucose represented by reducing sugar (FIG. 1).

1.2. Sea salt removal and calculation of extraction time

In order to calculate the minimum time for removing the salts inherent in the seaweeds, the concentration of the salts flowing into the supernatant was measured over time after immersing 10 g of seaweeds in 100 g of water. The salt eluted with water became the maximum value after 60 minutes, and it was seen that 2 g of the 10 g of seaweed that was introduced, and a total of 20% escaped with salt (FIG. 2A). However, since the component exhibiting the functionality of seaweed was predicted to be water-soluble, there was a possibility that it would be lost when immersed, and accordingly, a reducing sugar represented by water-soluble was measured to find out how much it was released for 60 minutes (FIG. 2B).

As a result of the measurement, the reducing sugar eluted with the supernatant was about 60 mg in 10 g of seaweed, and was stopped after the maximum loss after 60 minutes. It is thought that the monosaccharides of seaweed, which were decomposed according to external environmental conditions when the seaweed is harvested, are attached to the outer surface and eluted. Therefore, the condition for desalination was set to 60 minutes in 10 times water.

1.3. Establishment of seaweed return hydrothermal extract production conditions

From the above, 10 times more water was added to 10 g of seaweed, from which salt was removed, and the elution degree of the reducing sugar with time at 90 ° C or 25 ° C was measured (FIG. 3). In the seaweed retrieval extracted at 25 ° C, the amount of the reducing sugar eluted with the supernatant was 0.01%, and there was no significant change for 3 hours (A in FIG. 3), whereas in the extract with heat applied at 90 ° C, the extract was added up to 0.15% after 1.5 hours after extraction. Of reducing sugar was detected, which is the result of obtaining 150 mg of reducing sugar at 10 g of seaweed. When the time was applied for a longer time, the eluted reducing sugar decreased rather, which is thought to be a decomposition effect due to prolonged exposure to high temperature. Therefore, the production conditions of seaweed reflux water extract were set at 90 ° C for 1.5 hours. Alginic acid contained in a large amount of water-soluble components of seaweed is not detected as a reducing sugar under the hot water extraction conditions of 90 ° C 1.5 hours (Fig. 3B), it was found that the reducing sugar detected in Fig. 1 is not a byproduct of alginic acid. The produced seaweed hydrothermal water extract had a liquid phase, and the concentration of reducing sugar by the colorimetric method was 0.2 to 1 mg / ml.

1.4 Seaweed reflux extract in vitro  Intestinal attachment effect test

Before the in vivo functional assay for the seaweed hydrothermal extract produced in 1.2, an experiment was first conducted to test whether the extract exhibits in vitro function, and the specific method is as follows.

After incubating the HT-29 cell line in a 24-well plate with 90% confluency, and after treating Lactobacillus plantarum VH13 Lactobacillus 10 ⁶ cfu / ml and seaweed reflux export, it was allowed to stand at 37 ° C for a total of 3 hours. After removing the medium solution, washing twice with PBS solution, and removing the epithelial cells with trypsin, the number of lactic acid bacteria attached to the epithelial cells was counted by dilution plating method. The ratio of the lactic acid bacteria attached to the total administered lactic acid bacteria was obtained, and the adhesion rate according to the concentration of seaweed reflux extract was shown, and the results are shown in FIG. 4 (C in FIG. 4 contains only lactic acid bacteria, 5 is lactic acid bacteria + Seaweed reflux extract (including 5 μg per reduction), 25 means lactic acid bacteria + seaweed reflux extract (including 25 μg per reduction), 50 means lactic acid bacteria + seaweed reflux extract (including 50 μg per reduction).

As a result of the experiment, it was found that when 5 μg of total reducing sugar was applied, the adhesion rate increased by 44% compared to the control and 133% by 50 μg. Therefore, based on the maximum value of this experiment, 50 μg, the volume of the seaweed retrieval extract that can be physically introduced into the final product is calculated, and the animal dose concentration of the seaweed reciprocating hydrothermal extract for in vivo functional assay is 100 μg / rat (0.2 mg per reduction) / kg).

<Experimental Example 1> According to the invention Lactobacillus planta room ( Lactobacillus plantarum ) Suppression of constipation caused by taking new biotics of VH13 + seaweed reflux extract

1-1. Lactobacillus planta room ( Lactobacillus plantarum ) VH13 + Seaweed Hydrothermal Water Extract Inhibition of Constipation / Diarrhea Induced Functionality by Taking New Biotics Animal Experiment Design

Constipation (A) induces symptoms by oral administration of Loperamide 5mg / kg twice a day for 5 days (9am, 6pm), and diarrhea (B; inflammatory growth disease, IBD) is 3% Dextran Sulfate Sodium was diluted in drinking water and administered for 7 days to induce symptoms. Lactobacillus plantarum VH13 + seaweed reflux water extract new biotics ( Lactobacillus plantarum VH13 10 ⁸ cfu + seaweed reflux water extract) according to the present invention is administered simultaneously with the drug for 1 week, in the case of constipation 1 week, Diarrhea collected feces two weeks later. Genetic analysis was performed by collecting 30-40 mg tissue directly above the rectum to identify the total weight and moisture by collecting feces for 24 hours immediately before collecting feces, and to view the health status of the descending colon, and the animal experiment design is shown in FIG. 5. Did.

1-2. According to the invention Lactobacillus planta room ( Lactobacillus plantarum ) Evaluation of constipation inhibition by taking new biotics of VH13 + seaweed reflux extract

The amount of feces collected for 1 to 24 hours in 1-1 was found to be reduced by about 35% in the drug administration, Loperamide, compared to the control, but the lactic acid bacteria + seaweed reflux extract new biotics administration (Lop + Lp + Uex) It can be seen that the action is canceled and restored to the control level (Fig. 6 a. Fig. 6C is a control; Lop, Loperamide drug sphere; Lp, Lactobacillus plantarum ) VH13; Uex, seaweed reflux extract; Fos, Fructo-oligosaccharides). On the other hand, the comparative lactic acid bacteria + fructooligosaccharide new biotics administration group (Lop + Lp + Fos) was significantly increased than the drug administration, but did not reach the level of lactic acid bacteria + seaweed reflux extract new biotics administration.

The difference in the effect of inhibiting constipation caused by administration of seaweed hydrothermal extract and fructooligosaccharide synbiotics can be confirmed in FIG.

In other words, it was found that the content of water did not increase in the new biotics-administered formulations formulated with drugs or fructooligosaccharides, but increased significantly in the new biotics-administered lactic acid bacteria + seaweed hydrothermal extract, effectively suppressing constipation.

However, in FIG. 6B, the moisture content of the drug sphere is not significantly reduced compared to the control group, which is considered to be because the rodent's feces themselves had little moisture, and thus the width of water loss due to the drug was not large. On the other hand, it was interpreted that the effect of containing water in the feces was higher in the group administered with new biotics (LP + Lop + Uex) containing seaweed hydrothermal extract, which was higher than the control level. Since these effects cannot be seen with new biotics (LP + Lop + Fos) formulated with fructooligosaccharides, it is thought that the effect of seaweed hydrothermal water extract had a significant positive effect on suppression of constipation. Since human stool has a relatively larger variation in moisture content than rodents, it is predicted that the amount of change in moisture content in feces due to the presence or absence of constipation will be greater, so that the effect of suppressing constipation caused by taking new biotics of seaweed hydrothermal extract will be greater than that in rodents. It is expected.

In conclusion, constipation inhibition by fructooligosaccharide neobiotics could not be seen at the introduction concentration, but it was found that taking the seaweed hydrothermal water extract neobiotics has a large effect of suppressing constipation.

1-3. Evaluation of constipation-inducing inhibition according to inflammatory factor expression

Improvement of the intestinal environment, such as suppressing constipation, can also be seen through the frequency of expression of inflammatory substances that inform the health of the intestine, so real-time RT-PCR using the primers mentioned in Table 2 is an indicator inflammation factor for colorectal cancer, Cxcl1 ( In the case of humans, IL-8) gene expression was analyzed (the expression level of the intrinsic house keeping gene was measured by β-actin ).

Sequence number gene Reference Primer sequence Size
(bp) Tm (℃) / GC (%) One Cxcl1 NCBI Forward: CAG ACA GTG GCA GGG ATT CAC 254 59/57 2 Reverse: GGA CAA TCT TCT GAA CCA TGG 55/48 3 β-actin NCBI Forward: GTC ACC AAC TGG GAC GAC AT 210 58/55 4 Reverse: GAG GCG TAC AGG GAT AGC AC 58/60

Referring to FIG. 7, which shows the relative dielectric constant compared to the control, the Cxcl1 gene expression in the group fed the seaweed hydrothermal extract neobiotics (LP + Lop + Uex) was reduced by 20% in the descending colon compared to the control and drug groups. This indicates that the inflammation of the colon was well controlled by the seaweed hydrothermal extract neobiotics, and the overall intestinal environment was improved in a healthy direction even when compared to the control group fed only with water as well as with the drug. On the other hand, fructooligosaccharide neobiotics also show an effect in inflammation control, but in FIG. 7, it was shown that they did not show constipation-inhibiting effect. Therefore, the direct cause of inflammation control is present in neobiotics, not prebiotics, according to the results of previous studies. It is interpreted as the action of lactic acid bacteria.

However, since fructooligosaccharides do not help lactic acid bacteria adhere to intestinal cells, it is predicted to be more disadvantageous in the long-term inflammation control than seaweed hydrothermal extract, which increased the adhesion rate of lactic acid bacteria.

In conclusion, even if there are occasional causes of constipation, it is expected that if you are taking seaweed hydrothermal extract neobiotics, you can effectively control constipation. In addition, it is thought to be effective in maintaining overall intestinal health due to long-term inflammation control.

<Experimental Example 2> Lactobacillus planta room ( Lactobacillus plantarum ) VH13 + Seaweed Hydrothermal Water Extract Suppression of diarrhea caused by taking new biotics

2-1. Inhibition experiment of diarrhea inducement by measuring bowel movement

The effect of inhibiting diarrhea was evaluated by measuring the amount of feces collected for 24 hours immediately before the sacrifice of the experimental animal. C: Control, DSS: Dextran Sulfate Sodum, Lp: Lactobacillus plantarum VH13, Uex: Seaweed hydrothermal extract, Fos: Each of the experimental groups ingested fructooligosaccharide, a in FIG. 8 is the weight of the feces, and b is the moisture content of the feces.

As a result of the experiment, the drug administration (DSS), lactic acid bacteria + seaweed reflux extract new biotics (DSS + Lop + Uex), lactic acid bacteria + fructooligosaccharide new biotics (DSS + Lop + Fos) administration increased than the control (Fig. 8, a). In order to confirm the presence or absence of diarrhea, the moisture content in the feces was compared (b in FIG. 8), but the drug administration was caused by diarrhea caused by the drug, so that the water content increased compared to the control, but the water content of the seaweed reflux extract new biotics administration Showed that it was maintained at the control level. Contrary to this, the administration of fructooligosaccharide synbiotics contained water at the level of drug. Therefore, in FIG. 8A, the amount of increased stool of the seaweed reflux extract neobiotics administration increased the amount of defecation, and the amount of increased stool of the fructooligosaccharide neobiotics administration was from moisture increase. In other words, it can be seen that diarrhea is not controlled in the drug sphere and in the administration of fructooligosaccharide synbiotics.

2-2. Evaluation of constipation-inducing inhibition according to inflammatory factor expression

Inflammatory growth disease is caused by diarrhea caused by destruction of the mucous membrane protecting the intestine as the production and recovery of inflammation in the intestinal epithelial cells is repeated by a specific cause, and when severe, bloody stools are seen. Therefore, diarrhea regulated by taking seaweed reflux extract neobiotics is due to the control of inflammatory factors in the intestinal epithelial tissue, or anti-inflammatory, an indicator marker to see how much the action of mucosal production factors for the recovery of intestinal epithelial cells is healthy. The expression level of factor IL-10 and muc2 , a major gene for mesothelial membrane production, was measured by real time RT-PCR using the primer pair in Table 3 (the expression level of the intrinsic house keeping gene was measured by β-actin ).

Sequence number gene Reference Primer sequence Size (bp) Tm (℃) / GC (%) 5 Muc2 NCBI Forward: GGC ACC TAC CTT GTT GTG GAG 193 59/57 6 Reverse: GCA TTC TCG GTG CAC GCA CA 62/60 7 IL-10 NCBI Forward: GCT CAG CAC TGC TAT GTT GCC 207 60/57 8 Reverse: GCA GTA AGG AAT CTG TCA GC 54/50

It was found that the seaweed reflux extract sinbiotics (DSS + Lop + Uex) administration increased by 43% compared to the drug sphere, indicating that inflammation was effectively controlled (FIG. 9). In addition, when looking at the expression level of the muc2 gene, it shows that the intestinal epithelium is well protected by an increase of 80% or more compared to the drug sphere. On the other hand, when the administration of fructooligosaccharide new biotics (DSS + Lop + Fos) was also used as a genetic index, the expression of anti-inflammatory and mucosal production factors increased compared to drug administration, indicating that the action of lactic acid bacteria by short-term administration occurred. You can. However, since fructooligosaccharides do not support the attachment of lactic acid bacteria to the intestinal mucosa, it is judged that the lactic acid bacteria effect is difficult to last for a long time. Furthermore, even though fructooligosaccharide new biotics has a short-term inflammation control effect, it has no visible diarrhea inhibition Because it did not appear as (Fig. 8), it seems to be an inefficient combination for short-term diarrhea control.

In conclusion, lactic acid bacteria + seaweed reflux extract biobiotics is considered to have a great effect in controlling diarrhea caused by drugs in a short period of time, and to play a positive role in controlling inflammation and forming membranous membrane. This is, even if there are occasional causes of diarrhea, if you are taking seaweed hydrothermal extract sinbiotics as usual, it can effectively suppress the induction of diarrhea, and is also expected to be beneficial for maintaining long-term intestinal health.

3. Conversion of human body value

In the present invention, the reference concentration of reducing sugar in the seaweed hydrothermal extract that can be administered to humans is the rat test effective concentration of 0.01mg / day / 0.5kg rat, based on Table 1, based on 60kg of human standard 0.2 (mg / kg) * 0.16 * 60kg = 1.92 mg or 2 mg / day. Since the weight of a person varies and the degree of constipation or diarrhea that appears varies, it is desirable to determine a range rather than a specific value, so a minimum of 0.1 mg / day is administered by applying the minimum value of 0.005 mg / day / 0.5 kg rat shown in FIG. Or, you can take 10mg / day, which is 5 times higher than the applied 2mg / day calculated in animal experiments.

Therefore, in the present invention, the reference concentration of reducing sugar in the seaweed hydrothermal water extract that can be administered to a person weighing 60 kg is preferably 0.2 (mg / kg) * 0.16 * 60kg = 1.92, about 2 mg / day, or 0.01 (mg / kg) * 0.16 * 60kg = 0.092 to 1 * 0.16 * 60kg = 9.6, that is, about 0.1 mg / day to 10 mg / day.

Korea Microbial Conservation Center (KCCM) KFCC11759P 20180104

<110> VITAMIN HOUSE INC. <120> Composition of synbiotics for suppressing constipation and          diarrhea induction comprising Lactobacillus plantarum VH13 and          Undaria pinnatifida Sporophyll hot-water extract <130> 18-11224 <160> 8 <170> KoPatentIn 3.0 <210> 1 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Forward primer of Cxcl1 <400> 1 cagacagtgg cagggattca c 21 <210> 2 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Reverse primer of Cxcl1 <400> 2 ggacaatctt ctgaaccatg g 21 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Forward primer of beta-actin <400> 3 gtcaccaact gggacgacat 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Reverse primer of beta-actin <400> 4 gaggcgtaca gggatagcac 20 <210> 5 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Forward primer of Muc2 <400> 5 ggcacctacc ttgttgtgga g 21 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Reverse primer of Muc2 <400> 6 gcattctcgg tgcacgcaca 20 <210> 7 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Forward primer of IL-10 <400> 7 gctcagcact gctatgttgc c 21 <210> 8 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Reverse primer of IL-10 <400> 8 gcagtaagga atctgtcagc 20

Claims (6)

delete delete As a new biotics composition for inhibiting constipation and diarrhea, including seaweed reciprocal hydrothermal extract and Lactobacillus plantarum VH13 (KFCC11759P),
The seaweed reciprocating hydrothermal extract is extracted at 90 ° C. for 1.5 hours, and the new biotics composition for suppressing constipation and diarrhea is characterized by comprising reducing sugars at a concentration of 0.2 to 1 mg / ml. delete delete A health functional food composition comprising the new biotics composition for suppressing constipation and diarrhea of claim 3.

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