KR101395875B1 - Prebiotics Containing Extractions of Persimmon Leaves for Useful Strains in intestine - Google Patents

Abstract

The present invention relates to prebiotics containing extracts of persimmon leaves for promoting probiotic bacteria proliferation and suppressing growth of pathogenic bacteria. According to the present invention, as a water extract of the persimmon leaves is treated to promote the proliferation of probiotic bacteria and an ethanol extract of the persimmon leaves is treated to suppress growth of intestinal pathogenic bacteria, the prebiotics containing the extracts of persimmon leaves can control the growth of both probiotic bacteria and pathogenic bacteria. The extracts of persimmon leaves also can have an antioxidant effect by having a superior ability in electron donation.

Description

{Prebiotics Containing Extracts of Persimmon Leaves for Useful Strains in intestine}

The present invention relates to a prebiotics for promoting the proliferation of beneficial bacteria in the intestines including persimmon leaf extract and for inhibiting the growth of harmful bacteria in the intestines and a method for producing the same.

Diospyrus kaki is one of the fruit trees that grow well in the middle of Korea. It is used as a fruit with a distinctive flavor, and persimmon leaves have long been used in the private sector as a source of persimmon leaves (Park et al., 1995) . Persimmon leaves are known to contain a variety of physiologically active substances, and their clinical pharmacokinetics and efficacy are well documented in various documents such as Dongbokbohang and Boncho Gangmok. Recently, studies on the components and effects of persimmon leaves have been made recently. Domestic and foreign researches on persimmon leaves have been carried out to investigate the composition of persimmon leaf, the preparation method of persimmon leaf tea, the composition of vitamins C according to the cooking method, (Park et al., 1995, Woo et al., 2005, Moon & Park, 2000, An et al., 2003, Bae et al., 2000).

More than 400 kinds of germs in the human intestine are composed of complicated ecosystems which are inhabited by the number of about 100 trillion per gram of contents of the bowel and maintaining a constant balance. Intestinal microflora can be divided into useful bacteria and harmful bacteria, and their health condition is controlled by their balance. In order to maintain health, it is necessary to maintain the intestinal microflora with a lot of beneficial bacteria and few harmful bacteria. Staphylococcus aureus can be changed according to the state of the body such as external food, stress, hormone secretion, etc., and the enzymes and metabolites that they produce are involved in the metabolism of various substances in the body, Lt; / RTI > Therefore, dietary control is useful for promoting intestinal proliferation of beneficial bacteria and inhibiting harmful bacteria. For the study on the proliferation of intestinal beneficial bacteria using extracts of natural materials, there are Enzymugus japonica, Jujube, Omija and Minari fermentation.

Numerous papers and patent documents are referenced and cited throughout this specification. The disclosures of the cited papers and patent documents are incorporated herein by reference in their entirety to better understand the state of the art to which the present invention pertains and the content of the present invention.

The present inventors have tried to develop prebiotics for intestinal health from natural products. As a result, the present inventors confirmed that the persimmon leaf extract has an activity of promoting the proliferation of beneficial bacteria in the intestines and inhibiting the growth of harmful bacteria in the intestines, thereby completing the present invention.

Accordingly, an object of the present invention is to provide a prebiotics comprising a persimmon leaf extract.

Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

According to one aspect of the present invention, there is provided a prebiotics for promoting the proliferation of beneficial bacteria in the intestines including persimmon leaf extract and for inhibiting the growth of harmful bacteria in the intestines.

The present inventors have tried to develop prebiotics for intestinal health from natural products. As a result, the present inventors confirmed that the persimmon leaf extract has an activity of promoting the proliferation of the beneficial bacteria in the intestines and inhibiting the growth of the intestinal pest.

Accordingly, the present invention relates to a prebiotics comprising persimmon leaf extract.

As used herein, the term "Enterobacteriaceae in the intestines" refers to strains that have the same meaning as "probiotics" and have a beneficial effect on the intestinal environment when they reach the intestines. They survive in the stomach acid and the ascites juice and reach the small intestine, And which has a beneficial effect in the intestine, and which does not have toxicity, and which satisfies the non-mutagenic conditions. For example, includes the Lactobacillus genus (Lactobacillus), Lactococcus genus (Lactococcus), Enterococcus genus (Enterococcus), in Streptococcus (Streptococcus) and Bifidobacterium (Bifidobacterium). Preferably, the intestinal beneficial bacteria are Bifidobacterium, Lactobacillus or Streptococcus, more preferably Bifidobacterium or Streptococcus.

More preferably, the Bifidobacterium adolescentis strain of the present invention is Bifidobacterium adolescentis , the Lactobacillus sp. Strain of the present invention is Lactobacillus reuteri , and the Streptococcus sp. strain is Streptococcus Thermo filler's (Streptococcus thermophilus ). Most preferably, the Bifidobacterium sp. Strain is Bifidobacterium adolescentis KCTC 3352, the Lactobacillus sp. Strain is Lactobacillus luteri KCTC 3594, and the Streptococcus sp. Strain is Streptococcus thermophilus KCTC 3658 .

As used herein, the term "harmful bacteria in intestines " means a strain that is harmful to the intestinal environment when it is ingested and reaches the intestines. For example, in industrial Rouge Pseudomonas (Pseudomonas - aeruginosa), Vibrio genus (Vibrio), Staphylococcus genus (staphylococcus), Clostridium genus (Clostridium perfringens , Eubacterium , Bacteroides , and sulfate reducers . Preferably, the intestinal harmful bacteria are Clostridium genus, Ubertium genus or Bacteroid genus.

More preferably from Clostridium sp of the present invention is Clostridium difficile silica (Clostridium difficile or Clostridium perfringens , and the strain of the genus Yubucketium of the present invention is Eubacterium limosum ), and the strain of the genus Bacteroides of the present invention is Bacteroides fragilis ). Most preferably, the clostridial genus strain is Clostridium difficile KCTC 5009 or Clostridium puffin genus KCTC 3269, the strain of the genus Yubucketium is Yubactherium limnem KCTC 3266, the strain of genus Bacteroides Is the bacteroid Fragilex KCTC 3688.

When the persimmon leaves used in the composition of the present invention are obtained by treating the persimmon leaves with the extraction solvent, various extraction solvents may be used. Preferably, a polar solvent or a non-polar solvent can be used. Suitable polar solvents are (i) water, (ii) alcohols (preferably methanol, ethanol, propanol, butanol, n-propanol, iso-propanol, n-butanol, 1-pentanol, Or ethylene glycol), (iii) acetic acid, (iv) dimethyl-formamide (DMFO) and (v) dimethyl sulfoxide (DMSO). Suitable nonpolar solvents are acetone, acetonitrile, ethyl acetate, methyl acetate, fluoroalkane, pentane, hexane, 2,2,4-trimethylpentane, decane, cyclohexane, cyclopentane, diisobutylene, 1- But are not limited to, pentane, 1-chlorobutane, 1-chloropentane, o-xylene, diisopropyl ether, 2- chloropropane, toluene, 1- chloropropane, chlorobenzene, benzene, diethyl ether, diethylsulfide, Methane, 1,2-dichloroethane, aniline, diethylamine, ether, carbon tetrachloride, and THF.

More preferably, the extraction solvent used in the present invention is (a) water, (b) anhydrous or hydrated lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, propanol, butanol, etc.) (E) ethyl acetate, (f) chloroform, (g) butyl acetate, (h) 1,3-butylene glycol, (i) hexane and (j) diethyl ether. . Most preferably, the extract of the present invention is obtained by treating water, ethanol or a combination thereof with persimmon leaves.

As used herein, the term " extract " means that it is used in the art as a crude extract as described above, but broadly includes fractions obtained by further fractionating the extract. That is, the persimmon leaf extract includes not only those obtained by using the above-described extraction solvent but also those obtained by additionally applying a purification process thereto. For example, a fraction obtained by passing the above extract through an ultrafiltration membrane having a constant molecular weight cut-off value, and a separation by various chromatography (manufactured for separation according to size, charge, hydrophobicity or affinity) The fraction obtained through the purification method is also included in the persimmon leaf extract of the present invention.

The persimmon leaf extract used in the present invention may be prepared in powder form by an additional process such as vacuum distillation and freeze-drying or spray drying.

According to a preferred embodiment of the present invention, the persimmon leaf extract of the present invention is a mixture of a water extract and an ethanol extract.

More preferably, the water extract of the present invention promotes the proliferation of beneficial bacteria in the intestines, and most preferably promotes the proliferation for 0.1-12 hours from the start of the culture of the genus Lactobacillus or Bifidobacterium, or cultivation of Streptococcus Promoting proliferation for 8-24 hours from the start.

More preferably, the water extract of the present invention contains 0.1-1.0 g / 100 g, more preferably 0.1-0.7 g / 100 g, still more preferably 0.3-0.7 g / 100 g of free sugar , Most preferably 0.4-0.6 g / 100 g.

More preferably, the water extract of the present invention contains 3-15 mg / g of polyphenol, more preferably 6-12 mg / g, and most preferably 7-11 mg / g.

More preferably, the water extract of the present invention contains 30-60 μg / g, even more preferably 40-60 μg / g, even more preferably 45-55 μg / g of flavonoids , Most preferably 45-50 μg / g.

More preferably, the water extract of the present invention has an antioxidative effect of 30-70%, more preferably 40-60%, and most preferably 45-55%.

 More preferably, the ethanol extract of the present invention inhibits the growth of harmful bacteria in the intestines.

More preferably, the ethanol extract of the present invention has a minimal inhibitory concentration at 1-25 mg / mL, even more preferably at a minimum inhibitory concentration at 3-22 mg / mL, most preferably 3-7 mg / mL or at a minimum inhibitory concentration of 18-22 mg / mL.

More preferably, the ethanol extract of the present invention contains 0.1-1.0 g / 100 g, more preferably 0.1-0.8 g / 100 g, still more preferably 0.4-0.8 g / 100 g of free sugar , Most preferably 0.5-0.5 g / 100 g.

More preferably, the ethanol extract of the present invention contains 3-15 mg / g of polyphenol, more preferably 5-12 mg / g, and most preferably 7-10 mg / g.

More preferably, the ethanol extract of the present invention contains 10-40 μg / g, even more preferably 15-35 μg / g, even more preferably 20-30 μg / g of flavonoids , And most preferably 24-28 μg / g.

More preferably, the ethanol extract of the present invention has an antioxidative effect of 15-55%, more preferably an antioxidative effect of 25-45%, and most preferably an antioxidative effect of 30-40%.

The features and advantages of the present invention are summarized as follows:

(a) The present invention provides a prebiotics comprising persimmon leaf extract.

(b) According to the present invention, the water extract of persimmon leaves is treated to proliferate beneficial bacteria in the intestines, and the ethanol extract of persimmon leaves can be treated to inhibit the growth of harmful bacteria in the intestines. Thus, the prebiotics including persimmon leaves extract There is an advantage that all the growth can be controlled.

(b) The persimmon leaf extract of the present invention has an excellent electron donating ability and also has an advantage of antioxidative action.

FIGS. 1A to 1C show the effects of the persimmon leaf extract on the proliferation of Lactobacillus luteri KCTC 3594 (1a), Streptococcus thermophilus KCTC 3658 (1b) and Bifidobacterium adiales KCTC 3352 (1c).
Fig. 2 shows antioxidant activity of DPPH free radical scavenging ability of persimmon leaf extract.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

Example

Throughout this specification, "%" used to denote the concentration of a particular substance is intended to include solids / solids (wt / wt), solid / liquid (wt / The liquid / liquid is (vol / vol)%.

Experimental Methods and Materials

Samples and strains used

The persimmon leaf powder used in this experiment was purchased from Baekjangsyo Co., Ltd., which was lyophilized from Cheongsong-gun, Gyeongbuk. The strains were distributed from KCTC (Korea Collection for Type cultures, Korea). Enteric bacteria were enteric bacteria such as Clostridium difficile KCTC 5009, Clostridium perfringens KCTC 3269, Yubactherium remnant KCTC 3266, Bacteroides Fragilis KCTC 3688 was used. Bifidobacterium adolescentis KCTC 3352, Lactobacillus luteri KCTC 3594, and Streptococcus thermophilus KCTC 3658 were used as the beneficial bacteria derived from intestines. RCM (Reinforced Clostridial Medium) broth (Difco, USA) was used as a propagation medium for preincubation and main culture to cultivate enteric bacteria in the intestines, and the intestinal noxious bacteria were incubated for 48 hours under anaerobic conditions at 37 ° C. Lactobacilli MRS broth (Dipco, USA) was used as a growth medium for culture of intestinal beneficial bacteria, and the strain was used after being subcultured for 3 or more times before use in the experiment.

Persimmon leaf extract  Way

Water and 80% EtOH were used as extracts of the persimmon leaves. 1,000 g of distilled water was added to 200 g of the persimmon leaf powder and the mixture was treated with hot water at 70 ° C for 10 minutes in a water bath to obtain a third extract. The persimmon leaf extract was filtered with filter paper (Whatman No. 2) and dried in a freeze dryer (EYELA FDU-2100, Japan). In addition, 80% EtOH was used for the persimmon leaf powder and the extract was used by the above method. The dried samples were stored in a -40 ° C freezer.

Persimmon leaves extract Beneficial bacteria  Effect on proliferation

The number of generated colonies was counted, and the average number of colonies was multiplied by a dilution factor to calculate the number of viable cells per mL of the culture. To the 5 mL MRS broth, 1 (v / w)% each of persimmon leaf extract powder (water extract and ethanol extract) was added and sterilized. The test bacteria (Bifidobacterium adolescentis KCTC 3352, Lactobacillus luteri KCTC 3594 and Streptococcus thermophilus KCTC 3658) were added to sterile 5 mL MRS broth medium to a concentration of 660 nm optical density of 0.30 , And then cultured at 37 ° C. The number of bacteria was measured at 0, 4, 8, 12, 16, 20 and 24 hours intervals. Control group was MRS broth without added persimmon leaf extract.

Effect of Persimmon Leaf Extract on the Growth of Intestinal Pests

The minimum inhibitory concentration (MIC) of microorganisms was determined by applying the broth microdissolution method as follows. After culturing the culture medium for 24 hours at 650 nm, the absorbance was adjusted to 0.3, and 160 μL of each culture medium was dispensed into a 96-well plate. The final concentration of the persimmon leaf extract was adjusted to 20, 10, 5 and 1 mg / ml 40 μL was added and then anaerobically cultured for 24 hours. Bacterial growth was measured with an ELISA reader at 650 mn. As a control, 30% DMSO was used instead of the sample, and the experiment was repeated 8 times or more.

Free Sugar Analysis of Persimmon Leaf Extract

The persimmon leaf extract was dissolved in water, added with acetonitrile, filtered through a 0.45 μm filter, and used as an HPLC injection sample. HPLC was performed using a JASCO RI2031 Plus (4.6 mm x 250 mm, 5 μm, Japan) and a carbohydrate column (30 x 300 mm, Waters). The mobile phase was CH ₃ CN: H ₂ O (80:20, v / v) and the flow rate was 1.0 mL / min. A calibration curve was prepared with the standard solution to quantify the individual sugar content of the extract.

Total Polyphenol Content of Persimmon Leaf Extract

The Folin Deinis method (AOAC, Association of official analytical chemists, AOAC) using a phenolic substance reacting with phosphomolybdic acid to give a blue color to determine the total polyphenol compound content of Persimmon leaf extract. Offical methods of analysis . 15th ed., Washington DC. Cd 8-35 (1990)) was partially modified to perform colorimetric determination. 5 mL of distilled water was added to 100 μL of persimmon leaf extract, and 50 μL of Folin-Ciocalteu reagent was mixed. After adding 1.5 mL of saturated Na ₂ CO ₃ and 2.9 mL of distilled water, the absorbance was measured at 765 nm after standing at room temperature for 2 hours. Gallic acid (Sigma, USA) was used as a reference material.

Persimmon leaf extract  Total flavonoid content measurement

And total flavonoid content measurement of the persimmon leaf extract is Moreno (Moreno) (Moreno MIN et al ., J Ethnopharmacol, Comparison of the free radical scavenging activity of propolis from several regions of Argentina . , 71: 109-114 (2000)). 100 μg of persimmon leaf extract was mixed with 2 μl of aluminum nitrate (100 μl), left at room temperature for 10 minutes, and then absorbed at 430 nm. At this time, quercetin (Quercetin, Sigma, USA) was used as a reference material.

Persimmon leaf extract DDPH Radical Scatters  exam

The scavenging activity for DPPH (α, α-diphenyl-β-picrylhydrazyl) radicals was determined by the method of Blios et al. et al., Nature,Antioxidant determination by the use of stable free radicals. 181: 1199 ?? 1200 (1958)). To 2.8 mL of ethanol, add 0.4 mL of persimmon leaf extract and 4 x 10^-4 M DPPH solution was added and mixed for 10 seconds. The mixture was allowed to stand at room temperature for 10 minutes and absorbance was measured at 525 nm. Vitamin C (ascorbic acid, Sigma, USA) was used as a control.

Results and Discussion

Persimmon leaf extract  Intestines Beneficial bacteria  Effect on proliferation

The relationship between the persimmon leaf extract and the intestinal beneficial bacteria proliferation is shown in Fig. In the case of Lactobacillus lutea, the deciduous water extract showed an effect on the mycosis proliferation up to 12 hours, but not significantly after 12 hours. In the case of Streptococcus thermophilus, the effect of persimmon leaf extract on the initial proliferation was not observed, but it was effective in the proliferation of the water extract from 8 hours. In addition, the extract of Persimmon leaves showed an effect on the growth of bacteria in Bifidobacterium adolescentis, but after 12 hours, there was no effect on the growth of bacteria. These results suggest that persimmon leaves stimulate the growth of beneficial bacteria in the intestines and that water extracts are more effective than ethanol extracts. Jung et al., J Korean Soc Food Sci Nutr, Effects of Zizyphus jujuba there is . boeunesis Extracts on the Growth of Intestinal Microflora and Its Antioxidant Activities , 40 (4), 500-508 (2011))) were used to investigate intestinal microbial proliferation. The initial absorbance was high, but the absorbance at the time of reaching the greatest stage had little effect on the control and intestinal bacteria. .

The addition of 10% of the persimmon leaf water extract to the growth of common lactic acid bacteria (Lactobacillus acidophilus MG501, Lactobacillus brevis MG19, Lactobacillus bulgaricus MG515, Lactobacillus lactis MG530, Lactobacillus casei MG311) (Woo et al., Korean J. Food & Nutr, Studies on Antioxidative Effect and Lactic Acid Bacteria Growth of Persimmon Leaf Extracts , 18 (1): 28-38 (2005)). In contrast to the present study in which 1% of persimmon leaves were added, the persimmon leaf extract was added at a high concentration (10%) in the study of Wu et al. In this study, it was found that the addition of persimmon leaves extract helps the growth of beneficial bacteria in the intestines there was.

Effect of Persimmon Leaf Extract on the Growth of Enteric Bacteria in the Intestine

MIC (Minimum Inhibitory Concentration) of persimmon leaves extracts were measured by MIC of persimmon leaf extract and ethanol extract (Table 1). Microbial proliferation was observed at 20 mg / mL in all strains of Clostridium difficile, Clostridium perfringens, Eubacterium limmoth, and Bacteroides fulgillis in the case of the persimmon leaf water extract. In the case of the persimmon leaf ethanol extract, however, Growth was inhibited at a concentration of 5 mg / mL in Clostridium difficile, Clostridium perfringens and Bacteroides furigillis, and at a concentration of 10 g / mL in case of Streptococcus pneumoniae, and the MIC was shown in Table 1 . As a result, it was found that the ethanol extract of persimmon leaves exhibited a higher inhibitory effect on the growth of harmful bacteria than the water extract.

Minimum inhibitory concentration (MIC) Strain Growth by concentration (mg / mL) MIC 1.0 5.0 10.0 20.0 (mg / mL) Clostridium difficile water + ¹⁾ +  + + - EotH + - ²⁾ - - 5.0 Clostridium perfringens water + + + + - EotH + - - - 5.0 Yubat Terium
Limo-breath water + + + + - EotH + + - - 10.0 Bacteroid
Furagillis water + + + + - EotH + - - - 5.0

¹⁾ : not inhibited, ²⁾ : inhibited

Free Sugar Analysis of Persimmon Leaf Extract

The free sugar analysis of the persimmon leaf extract by HPLC is shown in Table 1. The contents of fructose, glucose and sucrose in the 80% ethanol extract of persimmon leaves were 0.30, 0.21 and 0.11 g / 100g, respectively, and 0.25 and 0.19, respectively, 0.06 g / 100 g.

Oh et al., J Korean Soc Food Sci Nutr, Effects of Artemisia capillaris Extracts on Intestinal Microflora In vitro and In vivo, 39 (11), 1587-1594 The higher the content of free sugar, such as (2010)) from fructose, glucose and sucrose has been reported to promote the fermentation by lactic acid bacteria, study information such as (Jeong et al. J Korean Soc Food Sci Nutr, Effects of Zizyphus I have jujuba . boeunesis Extracts on the Growth of Intestinal Microflora and Its Antioxidant Activities . 40 (4), 500-508 (2011)) also reported that the sugars present in the jujube extract act as prebiotics and thus have an enhancing effect. However, the free sugar content of persimmon leaves was relatively low in this study.

Free sugar composition in persimmon leaf extract (g / 100 g) Sample Fructose Glucose Sucrose Water extract 0.25 ¹⁾ 0.19 0.06 80% EtOH extract 0.30 0.21 0.11

¹⁾ : Free sugar content determined according to the HPLC method described in the Experimental Methods and Materials. Each value represents the average of the results from three repeated experiments (SD = 0.002 or less).

Total polyphenol content of persimmon leaf extract

Table 2 shows the total polyphenol contents of the persimmon leaves. When gallic acid was used as a standard substance, the polyphenol content of the persimmon leaf hot water extract was 8.86 mg / g. And 8.32 mg / g for persimmon leaf ethanol extract. Total polyphenol contents were not significantly different according to the extraction solvent. The total polyphenol content of the persimmon leaves extract was found to be 25.30-148.25 mg / 100g in a study of the total polyphenol contents obtained using caffeic acid as a standard material. In contrast, when catechin was used as a standard material, the total polyphenol content of the persimmon leaves was determined by harvesting time (Jung et al., J Korean J. Rep. Of Food Preservation, Chemical Components and Antioxidant Activity of Persimmon (Diospyros Kaki Thunb) Leaves, 11 (2) : 175-181 (2004)) was in the range of the polyphenol content of the extract of persimmon leaves ethanol was wide (105.7-182.2 mg / g), the total polyphenol content of the reference material is evident difference appear.

Total flavonoid content of persimmon leaf extract

When quercetin was used as a standard, the flavonoid content of persimmon leaf hot water extract was 47.59 μg / g. And 25.81 μg / g for ethanol extracts. (R) et al., Journal of Agriculture & Life Science, Effects , Vol. of Cactus Extracts on Human Intestinal Microflora , 43 (3): 45-54 (2009)), the flavonoid content of the cactus extract was 1.04-2.10 mg / g. The flavonoid contents of the extracts were 1.20 mg / 10 g of ethanol extract, 0.12 mg / 10 g of hexane extract, 0.28 mg / 10 g of chloroform extract, 1.91 mg / 10 g of ethyl acetate extract, 0.34 mg / 10 g of butanol extract, 10g, respectively.

Total Polyphenol and Flavonoid Concentration in Persimmon Leaf Extract Sample Total polyphenol (mg / g) ^{1 )} Total flavonoid (μg / g) ^{2 )} Water extract 8.96 ± 0.00 47.59 + - 0.94 80% EtOH extract 8.32 ± 0.00 25.81 ± 0.00

¹⁾ : When gallic acid was used as a standard substance, total polyphenol

²⁾ : When quercetin was used as a standard substance, total flavonoid

DPPH Radical Scatters  exam

The DPPH method is based on the ability of DPPH, a stable free radical, to react with a hydrogen donor (H-donor). DPPH is a relatively stable free radical, which is reduced by antioxidants and discolored, so it is widely used for measuring antioxidant efficacy. As a control, antioxidant activity by DPPH free radical scavenging activity was ascertained using ascorbic acid (95.92%, Sigma) as a natural antioxidant. The antioxidant activity of persimmon leaf extract was high at 49.06% and that of persimmon leaf ethanol was 33.66% when the concentration of control was 100 ppm. The persimmon leaf extract showed excellent electron donating ability. In addition, the higher the concentration of persimmon leaf extract, the better the electron donating ability. Therefore, the persimmon leaf extract not only promotes beneficial bacteria in the intestines but also has antioxidant activity.

summary

This study was carried out to investigate the effects of persimmon leaf extract and intestinal bacteria (Lactobacillus luterium KCTC 3594, Streptococcus thermophilus KCTC 3658, Bifidobacterium adolescentis KCTC 3352) on growth. To determine the growth factors, free sugar content, total polyphenol content, total flavonoid content and DDPH radical scavenging activity were measured. In the case of Streptococcus thermophilus, the extract of the persimmon leaves showed an effect on the proliferation of the beneficial bacteria in the intestinal flora in the intestinal lactic acid bacteria Lactobacillus lutea and Bifidobacterium adolescentis. The free sugar content of ethanol extract of persimmon leaves was higher than that of hot water extract, but the content of free sugar was very low. And total polyphenol contents were not significantly different between hot water extract and ethanol extract. Total polyphenol contents were higher than hot water extract and ethanol extract. The antioxidant activity of DPPH radical scavenging activity was higher than that of the decanter water extract.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the same is by way of illustration and example only and is not to be construed as limiting the scope of the present invention. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

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Claims (15)

Prebiotics for promoting the proliferation of beneficial bacteria in the intestines including persimmon leaf extract and for inhibiting the growth of harmful bacteria in the intestines.
The prebiotics according to claim 1, wherein the intestinal beneficial bacteria are Bifidobacterium, Lactobacillus or Streptococcus.
[Claim 3] The method according to claim 2, wherein the Bifidobacterium sp. Strain is Bifidobacterium adolescentis, the Lactobacillus sp. Strain is Lactobacillus lutea, and the Streptococcus sp. Strain is Streptococcus thermophilus Prebiotics.
The prebiotics according to claim 1, wherein the intestinal harmful bacteria are Clostridium genus, Yubucketium genus or Bacteroid genus.
[Claim 5] The method according to claim 4, wherein the clostridial strain is Clostridium difficile or Clostridium perfringens, and the strain of the genus Yubucketium is Yubactherium limbic, Wherein the prebiotics are glycyrrhizin.
The prebiotics according to claim 1, wherein the persimmon leaf extract is a mixture of water extract and ethanol extract.
The prebiotics according to claim 6, wherein the water extract promotes the proliferation of beneficial bacteria in the intestines.
delete 7. The prebiotics according to claim 6, wherein the water extract contains 3-15 mg / g of polyphenol.
The prebiotics according to claim 6, wherein the water extract contains 30-60 μg / g of flavonoid.
[Claim 7] The prebiotics according to claim 6, wherein the ethanol extract inhibits the growth of harmful bacteria in the intestines.
7. The prebiotics of claim 6, wherein the ethanol extract has a minimal inhibitory concentration at 1-25 mg / mL.
delete 7. The prebiotics according to claim 6, wherein the ethanol extract contains 3-15 mg / g of polyphenol.
7. The prebiotics according to claim 6, wherein the ethanol extract contains 10-40 μg / g of flavonoid.

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