KR102520354B1 - Composition for improving stomach function comprising fermented kiwi - Google Patents

Abstract

The present invention relates to a composition for improving gastric function, comprising, as an active ingredient, fermented kiwifruit which is fermented with lactic acid bacteria. According to the present invention, the composition containing the fermented kiwifruit of the present invention can improve gastric function by improving the digestive function of a stomach, alleviating damage to the gastric mucosa, and inhibiting the secretion of gastric acid.

Description

Composition for improving gastric function containing fermented kiwifruit

The present invention relates to a composition for improving gastric function containing fermented kiwifruit, and more particularly, to a composition for improving gastric function containing fermented kiwifruit fermented with lactic acid bacteria as an active ingredient.

The stomach is part of the digestive tract, an organ in the form of a hollow sac that connects the esophagus to the small intestine. The stomach is an organ that stores food entering through the esophagus for a while and controls the delivery of food to the duodenum through partial digestion to ensure efficient digestion and absorption in harmony with the secretion of pancreatic enzymes. When food comes in, the stomach secretes gastric acid to digest it. At this time, the gastric mucosal protective layer acts to prevent damage to the gastric mucosa by strong acidic gastric acid.

There are various factors that negatively affect human gastric function, such as genetic, physiological, environmental and mental factors. When the exercise capacity of the stomach is reduced due to these various factors, symptoms such as indigestion, heartburn, heavy bloating, and vomiting may appear, and when these symptoms persist for more than 3 months, it is called functional gastrointestinal disorder. In addition, when gastric acid is secreted excessively, the gastric mucosal protective layer is damaged by gastric acid, and at this time, inflammation occurs in the stomach, and when this inflammation is continuously repeated, gastritis and gastric ulcer may be caused. Gastritis refers to a case in which an inflammatory lesion is limited to the gastric mucosa, and gastric ulcer refers to a case in which an ulcer has penetrated the gastric mucosa and reached the submucosa and muscle layer. Moreover, when chronic gastritis is repeated and worsened, intestinal metaplasia changes may occur in the mucous membrane, and these changes may develop into gastric cancer, so it is necessary to maintain gastric health by improving gastric function. .

The kiwi fruit is a dioecious vine deciduous tree belonging to the genus Actinidia of the family Acinidiaceae and grows mainly in temperate regions. It is said that the fruit shape is covered with brown hairs and resembles a bird called 'Kiwi' that lives in New Zealand, so it was named kiwi. Kiwi contains phenolic compounds known to contribute to various physiological activities as well as the taste and flavor of food, is rich in vitamins C and E, and contains a large amount of minerals such as folic acid, potassium, calcium, and phosphorus. It contains physiologically active substances beneficial to health, such as chlorophyll and carotenoids. As such, since kiwifruit contains various physiologically active substances, various studies using kiwifruit are continuously being conducted. Specifically, Korean Patent Registration No. 1081910 relates to a cosmetic composition containing a kiwi extract that has skin tone improvement and skin aging prevention effects, and the kiwi extract effectively inhibits the glycation reaction of the skin to improve skin tone and relieve skin wrinkles. It has been disclosed that Korean Registered Patent No. 2027798 relates to an antioxidant composition containing fermented gold kiwi lactic acid bacteria as an active ingredient . It is disclosed that the content of total phenols and flavonoids is significantly increased compared to the antioxidant activity. In addition, actinidin contained in kiwi is an enzyme that decomposes protein and is effective in promoting digestion, so it is known as a fruit that is helpful for stomach health. However, in the prior art, there is no mention of the effect of fermented kiwi to improve digestive function and gastric mucosal damage, and to improve gastric function by inhibiting the secretion of gastric acid.

Accordingly, the inventors of the present invention have made diligent research efforts to overcome the problems of the prior art, and as a result, in the case of a composition for improving gastric function containing as an active ingredient a fermented product of kiwifruit fermented with lactic acid bacteria, the digestive function of the stomach and damage to the gastric mucosa. and it was confirmed that gastric function could be improved by inhibiting the secretion of gastric acid, and the present invention was completed.

KR 10-1081910 B1 KR 10-2027798 B1

Accordingly, the main object of the present invention is to provide a composition for improving gastric function containing a fermented kiwi fruit capable of improving gastric function by improving digestive function and gastric mucosal damage and inhibiting secretion of gastric acid.

According to one aspect of the present invention, the present invention provides a composition for improving gastric function comprising, as an active ingredient, a fermented product of kiwifruit fermented with lactic acid bacteria.

The term 'improvement of gastric function' in the present invention refers to improving and enhancing the function of the stomach to maintain and promote the health of the stomach, specifically, by increasing the activity of digestive enzymes such as amylase, lipase and protease, and It means promoting and improving digestive function by improving motility, preventing and improving gastric mucosal damage by preventing gastritis and improving damaged gastric mucosa, and protecting the stomach by inhibiting gastric acid secretion.

Kiwi in the present invention may be a kiwi of the genus Actinidia chinensis (gold kiwi), and detailed varieties include Jessie Gold (Actinidia chinensis Planch var. chinensis 'Jecy Gold'), Halla Gold (Actinidia chinensis Planch var. chinensis 'Halla Gold') '), Haegeum (Actinidia chinensis Planch var. chinensis 'Haegum'), Zespri®Gold (Actinidia chinensis Planch var. chinensis 'Hort16A'), Zespri®SunGold (Actinidia chinensis Planch var. chinensis 'Zesy002') , Zespri®Zesy003 (Actinidia chinensis Planch var. chinensis 'Zesy003'), Zespri®ZESH004 (Actinidia chinensis Planch var. chinensis 'Zesh004'), Consorzio Dori Europe®Dori (Actinidia chinensis Planch var. chinensis 'AC 1536') and Jingold® (Actinidia chinensis Planch 'Jintao'), but is not limited thereto.

In the composition for improving gastric function of the present invention, the lactic acid bacteria are kiwi-derived lactic acid bacteria, and the kiwi-derived lactic acid bacteria are Lactococcus Lactis VI-01 KCTC 14351BP and Lactobacillus paracasei VI-02 Characterized in that one or more of KCTC 14352BP.

The present inventors isolated lactic acid bacteria from strains isolated from kiwi and decoded the genome sequences of the selected strains through 16s rRNA analysis. In addition, the strains selected through genome sequencing were named “ Lactococcus Lactis VI-01” and “ Lactobacillus paracasei VI-02”, and the Bioscience and Biotechnology Research Center (KCTC) on November 3, 2020, and were given accession numbers KCTC14351BP and KCTC14352BP, respectively.

In the composition for improving gastric function of the present invention, the lactic acid bacteria may be lactic acid bacteria commonly used for fermentation in the art, preferably Lactobacillus paracasei, Lactobacillus acidophilus , Lactobacillus casei, Lactobacillus helveticus , Lactobacillus gasseri, Lactobacillus delbrueckii ssp. bulgaricus, Lactobacillus fermentum (Lactobacillus fermentum), Lactobacillus plantarum , Lactobacillus reuteri, Lactobacillus rhamnosus and At least one strain selected from the group consisting of Lactobacillus salivarius, more preferably Lactobacillus acidophilus, Lactobacillus casei and Lactobacillus Helveticus ) It is characterized by comprising one or more strains selected from the group consisting of.

In the fermented product of kiwifruit in the present invention, each strain is cultured (seed culture) in a first medium to prepare a first culture product (seed culture medium), and the first culture product is inoculated and cultured in a second medium (mass culture) After preparing the second culture product (mass culture medium), it can be obtained by centrifuging it to remove the supernatant, and then mixing and fermenting the obtained culture medium with kiwi puree. More specifically, the first culture product (species culture medium) is prepared by inoculating the strain with 0.01 to 0.5% of each medium and culturing at 35 ° C to 37 ° C for 18 to 28 hours, and the second culture product (Massive culture) can be prepared by inoculating the strain at 0.01 to 1% of the medium and culturing at 35 ° C to 37 ° C for 7 to 10 hours. In addition, the mixing ratio of kiwi puree and culture medium may be 7.5 to 8.5:2.5 to 1.5, preferably 8:2.

In the composition for improving gastric function of the present invention, the fermented kiwi product is characterized in that it is included in 30 to 80% by weight, preferably 30 to 60% by weight, based on the total weight of the composition. If the content of the fermented kiwi product is less than 30% by weight, it is difficult to show sufficient gastric function improvement effect, and if it exceeds 80% by weight, the effect of improving gastric function compared to the content is not sufficient, which is not preferable from an economic point of view.

In the composition for improving gastric function of the present invention, the composition is characterized in that it is a health functional food composition.

In the composition for improving gastric function of the present invention, the health functional food composition may have a convenient dosage form for ingestion and utilization, preferably capsules, tablets, powders, granules, liquids, pills, flakes, pastes, syrups, It is characterized in that it is at least one formulation selected from the group consisting of gels, beverages, jellies and bars. In order to prepare such a formulation, conventional excipients, stabilizers, thickeners, and the like may be further included. In addition, food additives may be additionally included, and the suitability as a “food additive” shall be determined according to the general rules of the Food Additive Code and general test methods approved by the Ministry of Food and Drug Safety, unless otherwise specified. judged by

When the health functional food composition of the present invention is formulated in powder form, the fermented product of kiwi is stored at -180 ° C to -195 ° C, preferably at -195 ° C, using a liquid nitrogen freezer (LNF). After quick-freezing, the quick-frozen fermented product can be formulated by grinding the dried raw material obtained by lyophilization at -35°C to -45°C, preferably at -40°C.

In the composition for improving gastric function of the present invention, the composition is characterized in that it is a food composition.

In the composition for improving gastric function of the present invention, the food composition may be formulated as a food that can use fermented kiwi as a main component, preferably one selected from the group consisting of dairy products, beverages, sauces, jams and baked goods It is characterized by the above formulation.

According to another aspect of the present invention, the present invention provides a composition for improving digestive function comprising, as an active ingredient, a fermented product of kiwifruit fermented with lactic acid bacteria.

According to an experimental example of the present invention, the fermented kiwi product according to the present invention has excellent digestive enzymes such as amylase, lipase and protease activity (see Experimental Example 5), gastric emptying ability (see Experimental Example 6) and gastrointestinal motility. (See Experimental Example 7) was confirmed to be excellent. These results suggest that the fermented kiwi product according to the present invention can improve digestive function, and the improvement of this digestive function can ultimately improve gastric health by improving gastric function.

According to another aspect of the present invention, the present invention provides a composition for improving gastric mucosal damage comprising, as an active ingredient, a fermented product of kiwifruit fermented with lactic acid bacteria.

According to an experimental example of the present invention, the fermented kiwi product according to the present invention has an excellent antioxidant effect (see Experimental Example 4), restores the damaged gastric mucosa (see Experimental Example 8), and reduces inflammation in the gastric mucosa (see Experimental Example 8). Experimental Example 9) was confirmed. These results suggest that the fermented kiwi product according to the present invention can prevent and improve gastric mucosal damage, and that the prevention and improvement of gastric mucosal damage can ultimately improve gastric health by improving gastric function. .

According to another aspect of the present invention, the present invention provides a composition for inhibiting gastric acid secretion comprising, as an active ingredient, a fermented product of kiwifruit fermented with lactic acid bacteria.

According to one experimental example of the present invention, it was confirmed that the fermented kiwi product according to the present invention inhibits gastric acid secretion (see Experimental Example 11). These results suggest that the fermented kiwi product according to the present invention can protect the gastric mucosa by inhibiting gastric acid secretion, and that the inhibition of gastric acid secretion can ultimately improve gastric health by improving gastric function.

As described above, the composition containing the fermented kiwifruit according to the present invention can improve gastric function by improving digestive function of the stomach and gastric mucosal damage, and inhibiting the secretion of gastric acid.

1 is a diagram showing Gram staining of deposited strains according to the present invention.
Figure 2 is a view showing the formation of a transparent ring according to the calcium carbonate decomposition of the deposited strain according to the present invention.
Figure 3 is a view of observing the morphology of the deposited strain Lactococcus Lactis VI-01 KCTC 14351BP according to the present invention.
Figure 4 is a view of observing the morphology of the deposited strain Lactobacillus paracasei VI-02 KCTC 14352BP according to the present invention.
5 is a phylogenetic tree of the deposited strain Lactococcus Lactis VI-01 KCTC 14351BP according to the present invention.
Figure 6 is a phylogenetic tree of the deposited strain Lactobacillus paracasei VI-02 KCTC 14352BP according to the present invention.
7 and 8 are results confirming the antioxidant effect of the fermented kiwi product according to the present invention.
9 is a result of measuring the gastric emptying ability of the fermented kiwi product according to the present invention.
10 is a result of gastrointestinal motility evaluation of fermented kiwifruit according to the present invention.
11 and 12 are results confirming the degree of inhibition of gastric mucosal damage of the fermented kiwi product according to the present invention.
13 to 18 show the results confirming the improvement of the gastric mucosa of the fermented kiwifruit according to the present invention (FIGS. 13 to 15; western blotting, FIGS. 16 to 18; ELISA).
19 and 20 are the results of measuring the length of the gastric line.
21 to 24 are the results of confirming the degree of gastric acid secretion.
25 is the result of measuring pepsin activity.

Hereinafter, the present invention will be described in more detail through examples. Since these examples are intended to illustrate the present invention only, the scope of the present invention is not to be construed as being limited by these examples.

Preparation Example 1: Isolation and identification of microorganisms

1) Separation Source and Sample Pretreatment

Gold kiwi fruit and puree were used to separate lactic acid bacteria from gold kiwi. Gold kiwi fruit was purchased from Zespri Gold Kiwi imported from New Zealand at a mart in September 2020, washed with tertiary distilled water to remove foreign substances, and then cut into small pieces, including the peel, and used. Gold kiwi puree was purchased from Namyang Frozen Food Co., Ltd. in May 2020, stored at -20 ° C, completely melted at room temperature and mixed evenly before use in the experiment.

2) Natural fermentation

Chopped gold kiwi and puree were fermented under salting and degreasing conditions. For salting conditions, each sample was salted for 3 hours by adding 8% of cheil salt, and then sterilized 1% fructooligosaccharide solution was mixed. Oligosaccharide solutions were mixed. All mixed samples were corrected to pH 6.0 or higher using sodium hydroxide solution, and fermented for about 3 days in a 37°C incubator.

3) Selection and identification of lactic acid bacteria

After mixing the fermented sample evenly, a portion of the fermentation broth was inoculated using BCP plate count agar (EIKEN, Japan) and incubated at 37 ° C for 48 hours. Afterwards, the medium was yellow and the phenotypes were purified using MRS agar (Difco, USA) plate medium for colony. For the pure isolated strain, the characteristics of the colony were confirmed, the cell morphology was confirmed through microscopic observation (×1000), and only the strains showing Gram-positive were first screened through Gram staining (FIG. 1). Thereafter, it was inoculated into MRS agar medium containing 1% calcium carbonate (DAEJUNG, Korea) and cultured at 37 ° C for 48 hours to decompose calcium carbonate with organic acid to finally select two strains with large clear zones. (Fig. 2).

The two separated strains were arbitrarily named VI-01 and VI-02, and were prepared as stocks using 30% glycerol and stored at -80 ° C. To confirm the biochemical characteristics of the two strains, such as glycolysis, arginin and esculin resolution, API 50 CHL kit (Biomerieux, France) was used to change the color of the medium using the apiweb program ( http://apiweb.biomerieux.com ). I sympathized with it.

For 16s rRNA analysis, PCR was performed using 27F and 1492R primers and sequencing analysis was performed by requesting Biofact Co., Ltd. The nucleotide sequence of each identified strain was compared for homology with the 16S ribosomal RNA gene sequence registered in GenBank using the blast program of NCBI, and aligned using the ClustalX 2.1 program using the neighbor-joining method. The phylogenetic tree used the bootstrap N-J tree method of ClustalX 2.1 and was confirmed with the NJplot program.

4) Genome sequence decoding

As a result of confirming the nucleotide sequence of 1,414 bp through 16S rRNA gene analysis of strain VI-01 strain, it showed 100% homology with Lactococcus lactis NBRC 100933 strain. Therefore, strain VI-01 was named Lactococcus lactis VI-01 (SEQ ID NO: 1).

As a result of confirming the nucleotide sequence of 1,441 bp through 16S rRNA gene analysis of strain VI-02 strain, it showed 99% homology with Lactobacillus paracasei R094 strain. Therefore, the VI-02 strain was named Lactobacillus paracasei VI-02 (SEQ ID NO: 2).

5) Identification through glucose availability survey

As a result of strain identification through sugar utilization investigation using API 50 CHL kit of the isolated strain, strain VI-01 was confirmed to be 98.4% similar to Lactococcus lactis , and strain VI-02 was confirmed to be 99.6% similar to Lactobacillus paracasei .

Preparation Example 2: Biochemical and Morphological Characteristics of Strains

1) Biochemical characteristics

Biochemical characteristics of the isolated strains were measured using API 50 CHL kit. The pure cultured colony on the MRS agar medium was diluted in API 50 CHL medium to an appropriate concentration and inoculated into the API 50 CHL kit, then cultured at 37 ° C for 24 to 48 hours and the color change of the inoculated medium was observed.

Lc. lactis VI-01 strain used a total of 19 sugars, including galactose, D-glucose, D-fructose, D-mannose, mannitol, maltose, and lactose among 49 sugars, but it was confirmed that sorbitol or xylitol could not be used ( Table 1).

No. sugar type usability No. sugar type usability One Glycerol - 26 Salicine + 2 Erythritol - 27 Cellobiose + 3 D-Arabinose - 28 Maltose + 4 L-Arabionse - 29 Lactose + 5 Ribose + 30 Melibiose - 6 D-Xylose - 31 Sanccharose + 7 L-Xylose - 32 Trehalose + 8 Adonitol - 33 Inuline - 9 β-Methl-xyloside - 34 Melezitose - 10 Galactose + 35 D-Raffinose - 11 D-Glucose + 36 Amidon + 12 D-Fructose + 37 Glycogen - 13 D-Mannose + 38 Xylitol - 14 L-Sorbose - 39 β-Gentiobiose + 15 Rhamnose - 40 D-Turanose - 16 Dulicitol - 41 D-Lyxose - 17 Inositol - 42 D-Tagatose - 18 Mannitol + 43 D-Fucose - 19 Sorbitol - 44 L-Fucose - 20 α Methyl-D-mannoside - 45 D-Arabitol - 21 α Methyl-D-glucoside - 46 L-Arabitol - 22 N-acetyl glucosamine + 47 Gluconate + 23 Amygdaline + 48 2-ceto-gluconate - 24 Arbutine + 49 5-ceto-gluconate - 25 Esculin +

L. paracasei VI-02 strain used 22 sugars, including galactose, D-glucose, D-fructose, D-mannose, mannitol, and sorbitol among 49 sugars, but lactose and xylitol were not used ( Table 2).

No. sugar type usability No. sugar type usability One Glycerol - 26 Salicine + 2 Erythritol - 27 Cellobiose + 3 D-Arabinose - 28 Maltose + 4 L-Arabionse - 29 Lactose - 5 Ribose + 30 Melibiose - 6 D-Xylose - 31 Sanccharose + 7 L-Xylose - 32 Trehalose + 8 Adonitol - 33 Inuline - 9 β-Methl-xyloside - 34 Melezitose + 10 Galactose + 35 D-Raffinose - 11 D-Glucose + 36 Amidon - 12 D-Fructose + 37 Glycogen - 13 D-Mannose + 38 Xylitol - 14 L-Sorbose - 39 β-Gentiobiose + 15 Rhamnose - 40 D-Turanose + 16 Dulicitol - 41 D-Lyxose - 17 Inositol - 42 D-Tagatose + 18 Mannitol + 43 D-Fucose - 19 Sorbitol + 44 L-Fucose - 20 α Methyl-D-mannoside - 45 D-Arabitol - 21 α Methyl-D-glucoside - 46 L-Arabitol + 22 N-acetyl glucosamine + 47 Gluconate + 23 Amygdaline + 48 2-ceto-gluconate - 24 Arbutine + 49 5-ceto-gluconate - 25 Esculin +

2) Morphological characteristics

As for the morphological characteristics, the shape and color of the colonies of the strain cultured in the MRS agar medium were confirmed, and the strain shape was observed under a microscope.

The colony of strain Lc.lactis VI-01 is small, round, 0.5-1.5 mm, and glossy white. As a result of observing the morphology of the strain under a microscope, it was confirmed as cocci (FIG. 3).

The colony of strain L. paracasei VI-02 is 1.5-2.5 mm round and convex, and has white or creamy luster. As a result of microscopic observation, long chains were formed as bacilli (FIG. 4).

Preparation Example 3: Deposit of Strains

After confirming the 16S RNA sequence analysis and morphological characteristics, and finally confirming the strain, the deposit procedure was performed at the Center for Biological Resources and a deposit number was assigned (Table 3).

strain name deposit number Lactococcus Lactis VI-01 KCTC 14351BP Lactobacillus paracasei VI-02 KCTC 14352BP

In order to conduct an experiment to confirm the activity of the deposited strain isolated as described above, the composition of the test group was set as shown in Table 4 below, and an acid resistance test, an artificial gastric juice resistance test, and an artificial bile acid resistance test were performed. Deposited strains 1 and 2 are novel strains isolated from kiwifruit in the present invention, and general strains 1 and 2 are commercially available strains.

strain name deposited strain 1 Lactobacillus paracasei VI-02 deposited strain 2 Lactococcus lactis VI-01 Common strain 1 Lactobacillus paracasei Common strain 2 Lactococcus lactis

Experimental Example 1: Acid resistance test

After transferring 30mL of sterilized MRS broth to a 50mL conical tube, 1% of the bacterial solution was inoculated and cultured at 37°C for 18 hours. The culture solution was centrifuged at 4,500 rpm for more than 10 minutes to completely settle the cells, and then only the supernatant was removed. The supernatant was removed, and the same amount of the supernatant with pH 2.0 and pH 3.0 PBS buffer removed was added to a 50mL conical tube containing only cells, and mixed evenly. After incubation at 37 ° C for 2 hours, a portion of the sample was diluted to an appropriate concentration using 0.85% NaCl solution, inoculated into MRS agar medium, and cultured at 37 ° C for 48 hours. The MRS agar plate in which 15 to 300 colonies were distributed was selected, the colonies were counted, and the results were confirmed. The results are shown in Table 5 below.

division survival rate pH 2 survival rate pH 3 deposited strain 1 75.3% 91.5% deposited strain 2 69.4% 90.1% Common strain 1 34.2% 69.4% Common strain 2 31.3% 65.3%

As a result, as shown in Table 5, it can be confirmed that the deposited strains isolated from kiwifruit in the present invention have excellent acid resistance from a high survival rate compared to general strains.

Experimental Example 2: artificial gastric juice resistance test

After preparing 1% pepsin solution and transferring 30ml of sterilized MRS Broth to a 50ml conical tube, 1% of the bacterial solution of each experimental sample was inoculated and incubated at 37℃ for 18 hours. After adding 10 mL of 1% pepsin solution to 89 mL of pH 2.5 MRS broth, 1% culture medium was mixed, incubated at 37 ° C for 2 hours, and a portion of the sample was taken at 0, 1, and 2 hours of culture and used in 0.85% NaCl solution. After diluting to an appropriate concentration, it was inoculated into MRS agar medium and incubated at 37°C for 48 hours. After completion of the culture, the survival rate was measured through the confirmation of the number of viable cells to confirm the result, and the results are shown in Table 6 below.

division survival rate deposited strain 1 67.2% deposited strain 2 61.4% Common strain 1 27.4% Common strain 2 20.7%

As a result, as shown in Table 6, the deposited strains isolated from kiwifruit in the present invention showed a higher survival rate in acidic conditions of artificial gastric juice than the general strains. It was found that the acid resistance of the deposited strain of the present invention isolated from kiwi at a pH level of 3 to 4 was remarkably excellent.

Experimental Example 3: Artificial bile acid resistance test

In the case of the internal bile acid test, as an experimental method to confirm the viability and activity maintenance due to bile in the duodenum after passing through the stomach, the survival rate was confirmed by measuring the number of viable cells after treatment with artificial bile.

Specifically, a sample inoculated with 1% of the cells in a sterilized MRS borth and cultured at 37 ° C for 18 hours was pretreated with artificial gastric juice for 2 hours, and then mixed with 87ml of MRS broth and 3ml of 10% Oxgall solution and artificial gastric juice pretreated sample 10 ml of the mixture was cultured at 37 ° C. for 4 hours, and the treated sample was spread on a plate, and the survival rate was calculated by checking the number of colonies generated. The results are shown in Table 7 below. As a result of the test, in the case of the bile acid test, artificial gastric juice was treated and then artificial bile acid was treated to confirm the final result in order to correct for the digestive process of the human body.

division survival rate deposited strain 1 58.2% deposited strain 2 51.4% Common strain 1 13.7% Common strain 2 10.1%

As a result, as shown in Table 7 above, when treated with artificial bile acid in a state where the survival rate was reduced by pre-treatment with artificial gastric juice, all deposited strains isolated from kiwi in the present invention showed a survival rate of 50% or more, compared to the general strains. showed only a survival rate of 30%, indicating that the deposited strains were highly active against digestive enzymes such as gastric juice or bile acid in the digestive tract.

Example: Preparation of kiwi fermented product using lactic acid bacteria

1) wearing kiwi

Although various types of kiwifruit can be used, in the present invention, kiwifruit of the genus Actinidia chinensis (gold kiwifruit) was used.

2) Pretreatment process (manufacturing of kiwi puree)

Kiwi fruit is washed several times, and fruit kiwi fruit that can be used as a raw material is selected. The selection criteria are to select fruits that have no damage when observed with the naked eye, that are not rotten or contaminated with bacteria, and that have a sugar content of 13 Brix% or more (if not exceeded, go through a post-ripening process). Peel the skin of the selected fruit and cut the flesh.

3) Grinding and seed separation process (making kiwi puree)

In trituration and seed separation, the cut flesh is put into a centrifugal separator and ground, and the seeds filtered through a sieve are removed and stirred to homogenize.

4) Lactic acid bacteria cultivation process (species culture - tank culture - centrifugation)

4-1) Lactic acid bacteria species

The lactic acid bacteria used to prepare the fermented kiwifruit were L. paracasei VI-02 KCTC14352BP, Lc. lactis VI-01 KCTC14351BP strain was mixed with Lactobacillus acidophilus , Lactobacillus casei , and Lactobacillus Helveticus for fermentation. In the case of three species of Lactobacillus strains, products from Sacco were used.

4-2) Cultivation process

4-2-1) Seed culture

Seed culture was cultured with MRS Broth. In the specific culture method, each strain was inoculated with 0.1% of each medium and then cultured at 37° C. for 24 hours to obtain a first culture product (strain). Sterilization was performed at 121° C. for 15 minutes.

4-2-2) Mass culture

Mass culture was carried out using the first culture product. After inoculating the first culture product in the MRS medium, the second culture product was obtained by culturing at 37° C. for 9 hours. Sterilization was performed at 121° C. for 20 minutes.

4-2-3) Centrifugation and concentration

The cultured lactic acid bacteria are centrifuged to concentrate the culture medium and lactic acid bacteria strain.

5) mix

Mix the raw material (kiwi puree) and the lactobacillus culture medium at a ratio of 8:2. Specific composition and inoculation amount are shown in Table 8 below.

Composition for preparing kiwi fermented product Inoculation amount (% by weight) of the strain (first culture product) during mass culture Kiwi Puree + Lb. paracasei VI-02 + Lc. lactis VI-01 + Lb. casei + Lb. acidophillus + Lb. Helveticus 25:25:16.5:16.5:16.5

6) fermentation

The mixed raw materials are incubated at 37°C for 8 to 12 hours.

7) Powdering

For use in experiments, the prepared fermented product was rapidly frozen and freeze-dried, and then pulverized and powdered to prepare a fermented product powder (FGKP) of kiwifruit.

Preparation Example 4: Preparation of animal model

After purchasing a 6-week-old male Sprague-Dawley (SD) rat, it was put into the experiment after a 7-day acclimatization period in a breeding facility (temperature: 22±2°C, humidity: 40-60%, light/dark cycle: 12 hours). During the breeding period, the temperature and humidity of the breeding room were checked every 8 hours.

Experimental Example 4: Confirmation of antioxidant effect

1) Measurement of DPPH radical scavenging activity

The fermented kiwi powder (FGKP) prepared in Example was suspended at room temperature for 3 hours or more at a concentration of 100 mg/ml, diluted to an appropriate concentration, and used in the experiment. L-ascorbic acid was used as a positive control, and each sample was mixed with a diluted solution of each concentration and DPPH at a concentration of 6.3mg/50ml at a ratio of 1:9, reacted in the dark for 10 minutes, and then measured absorbance at 517 nm. And the results are shown in Table 9 and FIG. 7 below.

2) Measurement of ABTS radical scavenging activity

Powder of fermented kiwifruit (FGKP; Example) suspended at room temperature for 3 hours or more at a concentration of 100 mg/ml was diluted to an appropriate concentration. As a positive control, L-ascorbic acid was used, and a mixture of 7 mM ABTS solution and 2.4 mM potassium persulfate was left in the dark for more than 16 hours. After forming ABTS+ from the left ABTS stock, it was diluted with absolute ethanol so that the absorbance value at 734 nm was 0.70 ± 0.02, and the diluted solution and sample were left for 1 minute at a ratio of 9: 1, and then the absorbance was measured. The results are shown in Table 9 and FIG. 8 below.

AA100 FGKP 1.25 FGKP 2.5 FGKP 5 FGKP 10 DPPH (%) 96.88±0.30 6.35±0.65 16.67±1.45 39.73±2.87 90.23±2.01 ABTS (%) 92.33±2.52 31.67±2.08 41.00±1.00 65.00±2.00 94.33±2.52

As a result, as shown in Table 9 and FIGS. 7 and 8, the antioxidant effect was 96% at 100 μg/ml of L-ascorbic acid, and the fermented kiwi product (FGKP) according to the present invention has an antioxidant effect as the concentration increases. increased.

Experimental Example 5: Measuring Digestive Enzyme Activity

1) Measurement of amylase activity

10 ml (1% solution) of distilled water was added to 0.1 g of fermented kiwifruit powder (FGKP) prepared in Example, and the supernatant obtained by centrifugation after shaking at room temperature for 4 hours after sealing was used as an enzyme solution. 1.0 ml of the previously prepared enzyme solution was added to 1.0 ml of 1% starch solution, reacted at 40 ° C for 30 minutes, 10 ml of 1 M acetic acid was added to stop the reaction, 10 ml of 0.00025 N iodine solution was added to develop color, and Absorbance was measured. At this time, after reacting in the same way using distilled water instead of the enzyme solution as a blank test (blank), the absorbance was measured at 660 nm. The absorbance measurement value of the sample reaction solution was expressed as 10% reduction based on the absorbance value of the blank test, converted into 1.0 g (dry weight base) of the sample. The results are shown in Table 10.

2) Measurement of lipase activity

For the measurement of α-Amylase activity, a prepared enzyme solution (a solution diluted to a concentration of 1% in distilled water) was used. Lipase activity was measured by slightly modifying the method of Lee et al. (1999) and Cao et al. (2015). A substrate in which a solution of 4-nitrophenyl acetate dissolved in acetonitrile to a concentration of 10 mM, ethanol and 100 mM sodium phosphate buffer solution (pH 7.0) were mixed at a ratio of 1:4:95 (v/v/v) before the experiment. 200 ul of the enzyme solution was added to 1.8 mL of the solution, reacted at 37° C. for 20 minutes, and then absorbance was measured at 405 nm. At this time, as a blank test solution, distilled water was used instead of the enzyme solution and the test was performed under the same conditions. Separately, the activity of lipase was calculated based on the calibration curve using p-nitrophenol standards, and 1 U of lipase is the amount of enzyme that liberates 1 uM of p-nitrophenol per minute per 1 g of sample (dry weight base). It was expressed in terms of . The results are shown in Table 10.

3) Measurement of protease activity

0.10 g of fermented kiwi powder (FGKP) prepared in Example was added to 10 mL of 0.1 M sodium phosphate buffer (pH 7.5), followed by voltexing (about 10 seconds) until the powder was completely dissolved. Subsequently, after shaking for about 10 minutes using a reciprocating shaker and centrifuging (4500 rpm, 7 minutes), the supernatant was used as a test solution, and all samples were stored in a refrigerator at 4 ° C or less until used in the experiment. 0.1 mL of test solution was added to 1.5 mL of 0.1 M sodium phosphate buffer (pH 7.5), equilibrated in a water bath at 37 °C for about 1 minute, and then 0.1 mL of substrate solution was added and reacted for 10 minutes. Subsequently, 0.4 mL of a 50% acetic acid solution was added to the reaction solution to stop the enzyme reaction, and then the absorbance was measured at 405 nm using a spectrophotometer. At this time, the blank test was corrected after measuring the absorbance using 0.1 ml of 0.1 M sodium phosphate buffer instead of 0.1 ml of the test solution. The amount of p-nitroaniline released from the substrate by the action of the protease was quantified using a calibration curve prepared with p-nitroaniline standards, and 1 unit (U) of enzyme activity is the amount of enzyme required to release 1 uM of p-nitroaniline per minute. It was expressed in terms of conversion per 1 g (dry weight base) of one sample. It is shown in Table 10 below.

α-Amylase Lipase Protease Enzymes (U/g) 121.5 ± 1.98 4.42 ± 0.01 21.15 ± 0.07

※ All experiments are expressed as average and standard deviation after repeated experiments 3 times.

Enzyme foods contain enzymes (α-Amylase, Protease) and are approved as enzyme foods (products containing grain enzymes, products containing embryo enzymes, etc.) It should have an enzyme titer. As shown in Table 10, α-Amylase, Lipase, and Protease are 121.5 ± 1.98 U / g, 4.42 ± 0.01 U / g, and 21.15 ± 0.07 U / g, respectively. A similar level of enzyme titer was confirmed.

Experimental Example 6: Measurement of gastric emptying capacity

1) Animal model

Experimental animals were divided into 6 groups with 8 animals per group. After acclimatization was completed, the experimental animals were fasted for 48 hours, and then each test substance was orally administered. HPMC was used as an excipient, and 2 ml of 3% HPMC was orally administered to the normal group (NOR) and the control group (CON), and to the experimental group, the powder (FGKP) of the fermented kiwifruit prepared in Example was given by concentration (FGKP 50 mg/kg, 125 mg/kg, 250 mg/kg) was mixed with 3% HPMC and then administered orally. In the positive control group, 30 mg/kg of itopride was mixed with 3% HPMC and then administered orally. At the same time, distilled water was intraperitoneally administered to the normal group (NOR), and cisplatin was intraperitoneally administered at a concentration of 10 mg/kg to the control group (CON), experimental group (FGKP), and positive control group (itopride). After 1 hour, 0.5% phenol red was used as a liquid meal and mixed with 1.5% HPMC was orally administered to all groups.

2) Experiment method

In order to measure the amount of phenol red remaining in the stomach, gastric emptying was delayed with cisplatin, a substance that induces gastric emptying, and 20 minutes later, euthanasia was performed with isoflurane under inhalational anesthesia. . The extracted stomach was placed in a sterilized beaker and homogenized for 20 seconds with 100 ml of 0.1N NaOH. The completely ground stomach and stomach contents were allowed to stand at room temperature for 1 hour, and then 5 ml of the contents were taken and mixed with 0.5 ml of 20% TCA solution. After sufficiently mixing, centrifuge at 2500xg for 20 minutes, separate 2 ml of the supernatant into a tube, and mix with 2 ml of 0.5 N NaOH. After sufficient reaction, absorbance was measured at a wavelength of 560 nm using a microplate spectrophotometer. The result of measuring the change in gastric emptying capacity was converted using Equation 1 (Gastric emptying rate (%)), and the results are shown in Table 11 and FIG. 9.

[Equation 1]

NOR CON FGKP 50 FGKP 125 FGKP 250 Itopride 30 Gastric emptying
(%) 67.7±5.3 20.1±10.9 42.1±7.3 43.6±5.7 58.6±8.8 59.1±4.4

As a result, as shown in Table 11 and FIG. 9, the control group (CON) showed a significantly lower value as 20.1 ± 10.9 compared to the normal group (NOR) 67.7 ± 5.3, and the FGKP experimental group was concentration dependent compared to the control group. It was confirmed that the gastric emptying capacity was significantly increased. Itopride, a positive control group, also showed higher significance in gastric emptying capacity compared to the control group.

Experimental Example 7: Gastrointestinal motility evaluation

1) Animal model

Experimental animals were divided into 6 groups with 8 animals per group. After acclimatization was completed, the experimental animals were fasted for 48 hours, and then each test substance was orally administered. Using HPMC as an excipient, 2 ml of 3% HPMC was orally administered to the normal group (NOR) and the control group (CON), and to the experimental group, powder (FGKP) of the fermented kiwifruit prepared in Example was given at different concentrations (FGKP 50 mg/kg). , 125 mg/kg, 250 mg/kg) was mixed with 3% HPMC and then administered orally. In the positive control group, 10 mg/kg of mosapride was mixed with 3% HPMC and then orally administered. At the same time, distilled water was intraperitoneally administered to the normal group (NOR), and atropine was intraperitoneally administered at a concentration of 1 mg/kg to the control group (CON), experimental group (FGKP), and positive control group (mosapride). After 1 hour, FITC-dextran was orally administered at a concentration of 6.25 mg/ml by 0.1 ml each using an evaluation diet.

2) Experiment method

After 15 minutes of oral administration of FITC-dextran, anesthetize and euthanize the small intestine, and then separate only the small intestine again and tie the end of the small intestine with a thread to fix it so that the contents of the small intestine do not flow. The isolated small intestine was regularly divided into 10 segments, and the amount of FITC-dextran in each segment was measured using fluorescence to calculate a geometric center (G.C). The G.C calculation method contained each segment in a tube, put 3 ml of 0.05M Tris buffer into each tube, stirred, and centrifuged at 1200 rpm for 5 minutes. After centrifugation, the supernatant was taken and the fluorescent signal was measured at 490-520 nm using a microplate spectrophotometer. In order to confirm the change in gastrointestinal metastasis, it was converted using Equation 2 (Geometric center), and the results are shown in Table 12 and FIG. 10.

[Equation 2]

NOR CON FGKP 50 FGKP 125 FGKP 250 Mosapride 10 Geometric center
(cm) 6.55 ±
0.58 3.77±
0.57 4.98±0.38 5.81±0.42 5.12±0.43 5.58±0.42

As a result, as shown in Table 12 and FIG. 10, the control group (CON) significantly decreased to 3.77 ± 0.57 compared to the normal group (NOR) 6.55 ± 0.58, and the experimental group FGKP group was at low, medium, and high concentrations, respectively. It was measured as 4.98±0.38, 5.81±0.42, and 5.12±0.43, and showed high significance together with the positive control Mosapride group, and the highest value was shown in the FGKP group (125 mg/kg).

Experimental Example 8: Confirmation of inhibition of gastric mucosal damage

1) Animal model

Experimental animals were divided into 6 groups, 8 animals per group, and the experiment was conducted. They were fasted for 24 hours before inducing acute gastritis, and drinking water was provided ad libitum. On the day of the experiment, the experimental group (FGKP) was orally administered the fermented kiwifruit powder (FGKP) prepared in Example at concentrations of 50 mg/kg, 125 mg/kg, and 250 mg/kg, respectively, and the positive control group (Sucralfate) was administered to the gastric mucosa. Sucralfate, which has a protective effect, was orally administered at a concentration of 50 mg/kg. One hour after the sample and sucralfate were administered, distilled water was orally administered to the normal group (NOR), and 2 ml of a mixture of 150 mM HCl and 60% ethanol was orally administered to the experimental group (FGKP) and the positive control group (sucralfate).

2) Experiment method

The excised gastric tissue was fixed with pins, fixed in 2% PFA for 30 minutes, and images were taken using an optical digital camera (DSC-HX50V, Sony, Tokyo, Japan). For the measurement of the damaged gastric mucosa, the area of the actual damaged area (Equation 3) was measured using the image-J program, and then it was expressed as a damage ratio (Equation 4) compared to the total area of the stomach. The results are shown in Table 14 and FIG. 11 and shown in FIG. 12 .

[Equation 3]

[Equation 4]

NOR CON FGKP 50 FGKP 125 FGKP 250 Sucralfate 50 Gastric lesion area % (AU) 0 18.15±
1.05 10.36±0.61 7.87±1.53 2.46±1.01 4.05±0.76

As a result, as shown in Table 13 and FIGS. 11 and 12, as a result of visual observation of the gastric mucosa in the animal experimental model for acute gastritis induced by HCL/ethanol, the gastric mucosa showed no lesions in the normal group (NOR). Compared to the control group (CON), gastric mucosa was damaged due to HCL/ethanol induction, and redness, hemorrhage, and edema were evident. On the other hand, it was observed that hemorrhagic mucosal damage was significantly reduced in both the positive control group (Sucralfate) and the sample administration group (FGKP). In addition, when the degree of gastric mucosal damage was measured using the i-solution program, no gastric mucosal lesions were seen in the normal group (NOR), but the damaged area was as high as 18.15±1.05 in the control group (CON). On the other hand, the positive control group, Sucralfate, was 4.05±0.76, which was significantly reduced compared to the control group, and the experimental group, FGKP, showed significant decreases to 10.35±0.61, 7.87±1.52, and 2.45±1.00.

Experimental Example 9: Confirmation of gastric mucosal improvement

1) Animal model

Experimental animals were divided into 6 groups, 8 animals per group, and the experiment was conducted. They were fasted for 24 hours before inducing acute gastritis, and drinking water was provided ad libitum. On the day of the experiment, the experimental group (FGKP) was orally administered the fermented kiwifruit powder (FGKP) prepared in Example at concentrations of 50 mg/kg, 125 mg/kg, and 250 mg/kg, respectively, and the positive control group (Sucralfate) was administered to the gastric mucosa. Sucralfate, which has a protective effect, was orally administered at a concentration of 50 mg/kg. One hour after the sample and sucralfate were administered, distilled water was orally administered to the normal group (NOR), and 2 ml of a mixture of 150 mM HCl and 60% ethanol was orally administered to the experimental group (FGKP) and the positive control group (sucralfate).

2) Experiment method

To obtain the cytoplasm of the stomach, add protease inhibitors and RIPA buffer [140 mM, Tris-HCl (25 mM, pH 7.4), 0.1% SDS, 1% Triton X-100], grind with a tissue grinder, and place on ice for 15 minutes. After leaving it on, it was centrifuged at 12,000 rpm for 20 minutes. iNOS and COX-2 protein expressions were measured by western blotting with gastric tissue proteins, and PGE ₂ , TNF-α and IL-6 contents were measured using an ELISA kit to confirm the amount of change. Western blotting results are shown in Table 14 and FIGS. 13 to 15, and ELISA results are shown in Table 15 and FIGS. 16 to 18.

NOR CON FGKP 50 FGKP 125 FGKP 250 Sucralfate 50 iNOS
(% of NOR) 100.0±
13.5 249.7±
12.1 134.7±13.7 123.1±8.6 120.1±8.2 77.9±5.2 COX-2
(% of NOR) 100.0±
9.8 156.8±
9.2 138.3±11.6 126.8±7.7 105.3±4.4 120.7±9.4

As a result of western blotting, the expression of COX-2 was significantly increased in the control group compared to the normal group, and decreased in the FGKP experimental group and positive control group compared to the control group. The expression of iNOS showed a significant decrease in both the FGKP experimental group and the positive control group compared to the control group.

NOR CON FGKP 50 FGKP 125 FGKP 250 Sucralfate 50 _PGE2 (ng/mg) 43.3 ±
2.1 19.4±
0.6 21.6±1.9 24.3±2.8 36.0±2.0 25.6±1.7 TNF-α (pg/mg) 372.6±
6.9 811.3±
37.2 485.9±5.0 456.6±47.0 400.6±13.1 402.6±28.2 IL-6 (ng/mg) 24.5 ±
1.3 47.2±
4.9 30.4±2.1 29.5±1.8 27.7±2.2 28.1±1.6

As a result of ELISA, the control group significantly increased the expression of both TNF-α and IL-6 compared to the normal group. The FGKP-administered group showed a tendency to significantly decrease compared to the control group. In addition, as a result of confirming the concentration of PGE ₂ present in the stomach tissue, the control group was 19.43±1.85ng/mg, which was significantly reduced compared to the normal group of 43.26±2.12ng/mg. The positive control group increased to 25.59±1.68ng/mg compared to the control group, and the FGKP experimental group increased to 21.56±1.85, 24.34±2.81, and 36.01±2.02, respectively.

Experimental Example 10: Pseudolinear Length Measurement

1) Animal model

Experimental animals were divided into 6 groups, 8 animals per group, and the experiment was conducted. They were fasted for 24 hours before inducing acute gastritis, and drinking water was provided ad libitum. On the day of the experiment, the experimental group (FGKP) was orally administered the fermented kiwifruit powder (FGKP) prepared in Example at concentrations of 50 mg/kg, 125 mg/kg, and 250 mg/kg, respectively, and the positive control group (Sucralfate) was administered to the gastric mucosa. Sucralfate, which has a protective effect, was orally administered at a concentration of 50 mg/kg. One hour after the sample and sucralfate were administered, distilled water was orally administered to the normal group (NOR), and 2 ml of a mixture of 150 mM HCl and 60% ethanol was orally administered to the experimental group (FGKP) and the positive control group (sucralfate).

2) Experiment method

In order to examine the degree of gastric damage in experimental animals with acute gastritis induced by HCL/ethanol, the extracted stomach was fixed in 4% PFA for 24 hours, and then dehydrated using xylen to prepare a paraffin tissue specimen. Subsequently, 4 μm-thick slices were prepared using a cutting machine and attached to slide glass. After removing the paraffin, the tissue sections were stained using hematoxylin and eosin (H&E), and the stained tissue sections were observed at x100 magnification using an optical microscope (zeiss, Germany), and the gastric glands were analyzed using an i-solution image analysis program. gastric gland) was measured, and the results are shown in Table 16 and FIGS. 19 and 20.

NOR CON FGKP 50 FGKP 125 FGKP 250 Sucralfate 50 mucosa thickness (μm) 1181±279 491±96 660±159 722±143 812±145 737±177

As a result, as shown in Table 16 and FIGS. 19 and 20, no significant lesions were observed in the gastric mucosa of all experimental animals in the normal group not treated with HCL/ethanol, and in the control group, the length of the gastric gland compared to the normal group. was observed short, confirming the induction of gastric mucosal damage, and in the positive control group treated with HCL/ethanol and Sucralfate, the gastric gland length was longer than that of the control group, confirming the inhibitory effect of Sucralfate on gastric mucosal damage. Also, in the experimental group treated with FGKP and then with HCL/ethanol, the gastric gland length was longer than that of the control group.

Experimental Example 11: Confirmation of gastric acid secretion

1) Animal model

Experimental animals were divided into 5 groups with 8 animals per group. After acclimatization, the experimental animals were fasted for 24 hours, and then each test substance was orally administered. Distilled water was orally administered to the control group (CON), and the experimental group was orally administered with the fermented kiwifruit powder (FGKP) prepared in Example at concentrations of 50 mg/kg, 125 mg/kg, and 250 mg/kg. To the positive control group (Sucralfate), 50 mg/kg of Sucralfate was orally administered. After 30 minutes of sample administration, each group was anesthetized with isoflurane, and then the abdominal cavity was opened about 2 cm, and the pyloric region connecting the stomach and duodenum was ligated. The incised abdominal cavity was sutured, and then moved to a cage after staying in a recovery chamber.

2) Experiment method

After 6 hours of pyloric ligation, they were euthanized with anesthesia, and the stomach was removed and gastric juice was collected and used for the experiment. The total volume of the obtained gastric juice is measured, and then the pH is measured using a pH meter. After transferring 1ml of gastric juice to the tube, add 2 to 3 drops of phenolphtalein solution. Then add 0.05N NaOH sodium hydroxide until it turns red. Total acidity was calculated according to Equation 5 below, and the results are shown in Table 17 and Figures 21 to 24.

[Equation 5]

CON FGKP 50 FGKP 125 FGKP 250 Sucralfate 50 Gastric juice vol. (ml/6h) 10.13±1.22 8.30±1.13 6.63±1.02 4.53±0.95 5.63±0.51 pH 1.01±0.08 1.09±0.11 1.23±0.06 1.60±0.23 1.38±0.09 Titratable acidity (μEq/ml) 157.5±0.4 117.3±3.2 108.5±1.8 69.2±3.2 97.0±3.6 Total acidity
(μEq/6h) 1599±23 991±40 723±17 332±22 558±26

As a result, as shown in Table 17 and FIGS. 21 to 24, the gastric acid volume of the control group was measured the most, and the amount of gastric juice decreased as the concentration increased in the FGKP experimental group. In the positive control, Sucralfate, it was confirmed that the volume of gastric acid was reduced compared to the control group. In addition, when looking at the pH, the lowest was measured in the control group, and the pH tended to increase as the concentration increased. The pH of the positive control group also increased compared to that of the control group. Similarly, acidity and free acidity were found to be the highest in the control group, and in the positive control group as well as in the FGKP experimental group, lower measured values were obtained as the concentration increased.

Experimental Example 12: Confirmation of pepsin activity

1) Animal model

Experimental animals were divided into 5 groups with 8 animals per group. After acclimatization, the experimental animals were fasted for 24 hours, and then each test substance was orally administered. Distilled water was orally administered to the control group (CON), and the experimental group was orally administered with the fermented kiwifruit powder (FGKP) prepared in Example at concentrations of 50 mg/kg, 125 mg/kg, and 250 mg/kg. To the positive control group (Sucralfate), 50 mg/kg of Sucralfate was orally administered. After 30 minutes of sample administration, each group was anesthetized with isoflurane, and then the abdominal cavity was opened about 2 cm, and the pyloric region connecting the stomach and duodenum was ligated. The incised abdominal cavity was sutured, and then moved to a cage after staying in a recovery chamber.

2) Experiment method

After 6 hours of pyloric ligation, they were anesthetized and euthanized, and then the stomach was removed and gastric juice was collected and used for the experiment. Put 500 μl of 2% hemoglobin into each tube, heat it at 37°C for 3 to 4 minutes, and then add 100 μl of gastric juice to each tube. Heat is then applied at 37° C. for 10 minutes. Then, 1 ml of TCA 16% was added to each tube and centrifuged at 6000xg for 30 minutes. After taking the supernatant from each tube from there and reacting at 20 ℃ for a certain time, the absorbance was measured at 280 nm. Pepsin activity was calculated according to 6 below, and the results are shown in Table 18 and FIG. 25.

[Equation 6]

CON FGKP 50 FGKP 125 FGKP 250 Sucralfate 50 Pepsin activity
(per ml/h) 5.02±0.22 4.51±0.10 4.26±0.30 2.26±0.05 2.89±0.04

As a result, as shown in Table 18 and FIG. 25, the highest activity was confirmed in the control group, and pepsin activity decreased as the concentration increased in the FGKP experimental group and positive control group.

<110> VITECH Co.,Ltd. <120> COMPOSITION FOR IMPROVING STOMACH FUNCTION COMPRISING FERMENTED KIWI <130> P21-0256 <160> 2 <170> KoPatentIn 3.0 <210> 1 <211> 1414 <212> DNA <213> unknown <220> <223> Lactococcus Lactis VI-01 KCTC14351BP <400> 1 tgcaagttga gcgctgaagg ttggtacttg taccgactgg atgagcagcg aacgggtgag 60 taacgcgtgg ggaatctgcc tttgagcggg ggacaacatt tggaaacgaa tgctaatacc 120 gcataaaaac tttaaacaca agttttaagt ttgaaagatg caattgcatc actcaaagat 180 gatcccgcgt tgtatagct agttggtgag gtaaaggctc accaaggcga tgatacatag 240 ccgacctgag agggtgatcg gccacattgg gactgagaca cggcccaaac tcctacggga 300 ggcagcagta gggaatcttc ggcaatggac gaaagtctga ccgagcaacg ccgcgtgagt 360 gaagaaggtt ttcggatcgt aaaactctgt tggtagagaa gaacgttggt gagagtgggaa 420 agctcatcaa gtgacggtaa ctacccagaa agggacggct aactacgtgc cagcagccgc 480 ggtaatacgt aggtcccgag cgttgtccgg atttattggg cgtaaagcga gcgcaggtgg 540 tttattaagt ctggtgtaaa aggcagtggc tcaaccattg tatgcattgg aaactggtag 600 acttgagtgc aggagaggag agtggaattc catgtgtagc ggtgaaatgc gtagatatat 660 ggaggaacac cggtggcgaa agcggctctc tggcctgtaa ctgacactga ggctcgaaag 720 cgtggggagc aaacaggatt agataccctg gtagtccacg ccgtaaacga tgagtgctag 780 atgtagggag ctataagttc tctgtatcgc agctaacgca ataagcactc cgcctgggga 840 gtacgaccgc aaggttgaaa ctcaaaggaa ttgacggggg cccgcacaag cggtggagca 900 tgtggtttaa ttcgaagcaa cgcgaagaac cttaccaggt cttgacatac tcgtgctatt 960 cctagagata ggaagttcct tcgggacacg ggatacaggt ggtgcatggt tgtcgtcagc 1020 tcgtgtcgtg agatgttggg ttaagtcccg caacgagcgc aacccctatt gttagttgcc 1080 atcattaagt tgggcactct aacgagactg ccggtgataa accggaggaa ggtggggatg 1140 acgtcaaatc atcatgcccc ttatgacctg ggctacacac gtgctacaat ggatggtaca 1200 acgagtcgcg agacagtgat gtttagctaa tctcttaaaa ccattctcag ttcggattgt 1260 aggctgcaac tcgcctacat gaagtcggaa tcgctagtaa tcgcggatca gcacgccgcg 1320 gtgaatacgt tcccgggcct tgtacacacc gcccgtcaca ccacgggagt tgggagtacc 1380 cgaagtaggt tgcctaaccg caaggagggc gctc 1414 <210> 2 <211> 1441 <212> DNA <213> unknown <220> <223> Lactobacillus paracasei VI-02 KCTC14352BP <400> 2 tgcagtcgaa cgagttctcg ttgatgatcg gtgcttgcac cgagattcaa catggaacga 60 gtggcggacg ggtgagtaac acgtgggtaa cctgccctta agtgggggat aacatttgga 120 aacagatgct aataccgcat agatccaaga accgcatggt tcttggctga aagatggcgt 180 aagctatcgc ttttggatgg acccgcggcg tattagctag ttggtgaggt aatggctcac 240 caaggcgatg atacgtagcc gaactgagag gttgatcggc cacattggga ctgagacacg 300 gcccaaactc ctacgggagg cagcagtagg gaatcttcca caatggacgc aagtctgatg 360 gagcaacgcc gcgtgagtga agaaggcttt cgggtcgtaa aactctgttg ttggagaaga 420 atggtcggca gagtaactgt tgtcggcgtg acggtatcca accagaaagc cacggctaac 480 tacgtgccag cagccgcggt aatacgtagg tggcaagcgt tatccggatt tattgggcgt 540 aaagcgagcg caggcggttt tttaagtctg atgtgaaagc cctcggctta accgaggaag 600 cgcatcggaa actgggaaac ttgagtgcag aagaggacag tggaactcca tggttagcgg 660 tgaaatgcgt agatatatgg aagaacacca gtggcgaagg cggctgtctg gtctgtaact 720 gacgctgagg ctcgaaagca tgggtagcga acaggattag ataccctggt agtccatgcc 780 gtaaacgatg aatgctaggt gttggagggt ttccgccctt cagtgccgca gctaacgcat 840 taagcattcc gcctggggag tacgaccgca aggttgaaac tcaaaggaat tgacgggggc 900 ccgcacaagc ggtggagcat gtggtttaat tcgaagcaac gcgaagaacc ttaccaggtc 960 ttgacatctt ttgatcacct gagagatcag gtttcccctt cgggggcaaa atgacaggtg 1020 gtgcatggtt gtcgtcagct cgtgtcgtga gatgttgggt taagtcccgc aacgagcgca 1080 acccttatga ctagttgcca gcatttagtt gggcactcta gtaagactgc cggtgacaaa 1140 ccggaggaag gtggggatga cgtcaaatca tcatgcccct tatgacctgg gctacacacg 1200 tgctacaatg gatggtacaa cgagttgcga gaccgcgagg tcaagctaat ctcttaaagc 1260 cattctcagt tcggactgta ggctgcaact cgcctacacg aagtcggaat cgctagtaat 1320 cgcggatcag cacgccgcgg tgaatacgtt cccgggcctt gtacacaccg cccgtcacac 1380 catgagagtt tgtaacaccc gaagccggtg gcgtaaccct tttagggagc gagccgtcta 1440 a 1441

Claims (12)

A composition for improving gastric function comprising fermented kiwifruit fermented with lactic acid bacteria as an active ingredient,
The lactic acid bacteria are Lactococcus lactis VI-01 KCTC 14351BP, Lactobacillus paracasei VI-02 KCTC 14352BP, Lactobacillus acidophilus, Lactobacillus casei ) And Lactobacillus Helveticus ( Lactobacillus Helveticus ) A composition for improving gastric function, characterized in that.
delete delete delete According to claim 1,
The kiwi fermented product is a composition for improving gastric function, characterized in that it is included in 30 to 80% by weight relative to the total weight of the composition.
According to claim 1,
The composition is a composition for improving gastric function, characterized in that the health functional food composition.
According to claim 6,
The health functional food composition is a composition for improving gastric function, characterized in that at least one dosage form selected from the group consisting of capsules, tablets, powders, granules, liquids, pills, flakes, pastes, syrups, gels, drinks, jellies and bars .
According to claim 1,
The composition is a composition for improving gastric function, characterized in that the food composition.
According to claim 8,
The food composition is a composition for improving gastric function, characterized in that at least one formulation selected from the group consisting of dairy products, beverages, sauces, jams and confectionery.
A composition for improving digestive function comprising fermented kiwifruit fermented with lactic acid bacteria as an active ingredient,
The lactic acid bacteria are Lactococcus lactis VI-01 KCTC 14351BP, Lactobacillus paracasei VI-02 KCTC 14352BP, Lactobacillus acidophilus, Lactobacillus casei ) And Lactobacillus Helveticus ( Lactobacillus Helveticus ) A composition for improving digestive function, characterized in that.
A composition for improving gastric mucosal damage comprising fermented kiwifruit fermented with lactic acid bacteria as an active ingredient,
The lactic acid bacteria are Lactococcus lactis VI-01 KCTC 14351BP, Lactobacillus paracasei VI-02 KCTC 14352BP, Lactobacillus acidophilus, Lactobacillus casei ) And Lactobacillus Helveticus ( Lactobacillus Helveticus ) A composition for improving gastric mucosal damage, characterized in that.
A composition for inhibiting gastric acid secretion comprising fermented kiwifruit fermented with lactic acid bacteria as an active ingredient,
The lactic acid bacteria are Lactococcus lactis VI-01 KCTC 14351BP, Lactobacillus paracasei VI-02 KCTC 14352BP, Lactobacillus acidophilus, Lactobacillus casei ) And Lactobacillus Helveticus ( Lactobacillus Helveticus ) A composition for inhibiting gastric acid secretion, characterized in that.

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