KR20180104518A - Prebiotics Containing Grape seed flour - Google Patents

Abstract

The present invention relates to prebiotics for suppressing growth of harmful bacteria in the intestines and promoting beneficial bacteria in the intestines, containing grape seed powder. According to the present invention, the prebiotics containing grape seed powder can selectively affect the growth of beneficial bacteria and harmful bacteria by suppressing the growth of harmful bacteria while promoting the growth of beneficial bacteria, thereby promoting intestinal health.

Description

Prebiotics Containing Grape seed flour < RTI ID = 0.0 >

The present invention relates to a prebiotics for promoting the proliferation of beneficial bacteria in the intestines containing grape seed powder and for inhibiting the growth of harmful bacteria in the intestines.

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 >

Probiotics refers to microorganisms that are beneficial to the human body, which means living bacteria that produce lactic acid by reaching the intestine, which can not grow harmful bacteria, and convert them to acid so that beneficial bacteria can increase.

Prebiotics are substances that help to improve the intestinal environment by becoming a nutrient source of probiotics, an indigestible ingredient that helps the growth of beneficial bacteria in the intestines. In order for any food ingredient to be utilized as a prebiotic to be used by the beneficial microorganism in the intestine to promote the growth or activity of the microorganism and thereby exhibit a beneficial effect on host health, it must not be digested or absorbed from the upper part of the gastrointestinal tract, It is necessary to selectively activate the bacterium and to inhibit harmful bacteria such as pathogens.

Polyphenol, which is known to be contained in tea, is known to promote the growth of lactic acid bacteria . It has also been reported that grape seeds rich in catechin can promote the growth of certain Lactobacillus strains. However, these components have problems in that they do not simultaneously work to promote beneficial bacteria and inhibit phe- nomena.

Accordingly, the inventors of the present invention invented a prebiotics including grape seed powder by confirming that the grape seed powder promotes the growth of beneficial bacteria in the intestines and can inhibit the growth of specific harmful bacteria.

It is an object of the present invention to provide a prebiotics for promoting the proliferation of beneficial bacteria in the intestines and for inhibiting the growth of noxious bacteria in the intestines.

Another object of the present invention is to provide a health functional food and a pharmaceutical composition for promoting the proliferation of beneficial bacteria in the intestines and for inhibiting the growth of noxious bacteria in the intestines.

In order to achieve the above object, the present invention provides a prebiotics for promoting the proliferation of beneficial bacteria in the intestines including grape seed powder and for inhibiting the growth of harmful bacteria in the intestines.

Preferably, the grape seed powder may have both a function of promoting the proliferation of the beneficial bacteria in the intestines and a function of inhibiting the growth of the intestinal noxious bacteria.

Preferably, the grape seed powder may be produced from by-products generated during the manufacturing process of the wine.

The present invention also provides a health functional food for promoting the proliferation of beneficial bacteria in the intestines containing the prebiotics and inhibiting the growth of harmful bacteria in the intestines.

The present invention also provides a pharmaceutical composition for promoting the proliferation and inhibiting the growth of harmful bacteria in the intestines comprising the prebiotics.

The prebiotics including the grape seed powder according to the present invention can selectively affect the growth of beneficial bacteria and harmful bacteria, thereby enhancing the growth of beneficial bacteria and inhibiting the growth of harmful bacteria. .

In addition, the prebiotics including the grape seed powder according to the present invention can be utilized as various health functional foods, and can be utilized as a pharmaceutical composition for promoting the proliferation of enteric bacteria in the intestines and for inhibiting the growth of harmful bacteria in the intestines.

1 is in the grape seed powder, control, grape seed extract, respectively, yuikgyun of Leuconostoc mesenterorides were cultured for 48 hours.
FIG. 2 is a graph showing the growth activity of Lactobacillus kefiri DH5 strain, which is a useful bacterium, after culturing for 96 hours in grape seed powder, control group and grape seed extract, respectively.
FIG. 3 is a graph showing growth activity of Clostridium perfringens , a harmful bacterium, in the grape seed powder, control group and grape seed extract after 12 hours of culture.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In general, the nomenclature used herein is well known and commonly used in the art.

In one aspect, the present invention relates to a prebiotics for promoting the proliferation of beneficial bacteria in the intestines including grape seed powder and for inhibiting the growth of harmful bacteria in the intestines.

The present inventors confirmed that the "grape seed powder" has a function of promoting the proliferation of beneficial bacteria in the intestinal tract and inhibiting the growth of harmful bacteria in the intestines, and has a prebiotic effect Respectively.

The grape seed powder according to the present invention has both a function of promoting the proliferation of the beneficial bacteria in the intestines and a function of inhibiting the growth of the intestinal harmful bacteria. The other ingredients known as prebiotics in the past can only function in promoting the proliferation of beneficial bacteria in the intestines and the inhibition of growth of harmful bacteria in the intestines while the grape seed powder according to the present invention has the function of promoting the proliferation of intestinal benefit bacteria, It can be used as a very useful prebiotics.

&Quot; Prebiotics " means an indigestible food ingredient that is used by useful microorganisms in the large intestine to promote the growth or activity of microorganisms, thereby exhibiting a healthful effect.

There are no particular restrictions on the type, area or harvest date of the grapes used in the present invention. In one embodiment of the present invention, Chardonnay grapes were used, but the present invention is not limited thereto.

The grape seed powder of the present invention can be produced from beverages made from grapes or from by-products generated in the course of producing liquors. Preferably, the grape seed powder may be produced from by-products generated during the manufacturing of wine or wine.

Here, " by-product " is generically referred to as wine or wine distillate produced in a distillation process after crushing and fermentation processes using grapes as raw materials.

Typically, by-products that occur during the manufacture of wine or wine are being discarded. However, these wastes amount to as much as 40% of raw grapes and are economically disastrous. According to the present invention, byproducts generated in the production of wines can be utilized as prebiotics, there is a great advantage in terms of environment and economy.

 Grape seed flour (GSF) according to one embodiment of the present invention separates and dries grape seeds from the remaining byproducts that produce wine, and then extracts grape seed from the grape seed using a "cold press" Extracting the oil, and then pulverizing it into a powder.

The method of separating, drying, compressing and pulverizing the grape seed powder can be carried out in various forms by known techniques.

Grain size of the grape seed powder can be varied according to the grinding environment, and sieving operation can be added to produce a powder with smaller particles. The particle size of the grape seed powder can be adjusted to suit the application purpose and purpose such as food, medicines.

The present inventors not only discovered the selective prebiotic effect of the grape seed powder on the growth of beneficial bacteria and harmful bacteria, but also found that the "grape seed powder" is more effective than the "grape seed extract" in promoting the growth of beneficial bacteria in the intestines, And the inhibitory effect was more excellent.

Herein, " grape seed extract " is obtained by treating grape seed with various extraction solvents, and water, alcohol (preferably, methanol, ethanol) and the like can be used as an extraction solvent.

In one embodiment of the present invention, i) grape seed powder was extracted with water after ultrasonication, and ii) the grape seed powder was sonicated and extracted with 80% methanol.

As a result of the experiment, it was confirmed that the growth of the lactic acid bacteria, which is a useful bacterium, was further promoted compared with the case that the grape seed powder was cultured in the control group (grape seedless group) and the grape seed extract.

On the other hand, it was confirmed that the growth inhibitory activity of the grape seed powder was excellent, while the control group (grape seedless group) and the grape seed extract showed little growth inhibitory activity.

Therefore, it was confirmed that the effect of the grape seed powder was superior to that of the grape seed extract in addition to the prebiotic effect which selectively affects the growth of beneficial bacteria and harmful bacteria.

In the present invention, the term " Enterobacteriaceae in the intestines " means useful bacteria living in the intestines of the host, and strains having an effect on the intestinal environment when ingested and reached the intestines. These are bacteria that survive in stomach acid and succulent acid to reach the small intestine, proliferate and settle in the intestines, exhibit useful effects in the intestinal tract, and are free of toxicity and non-pathogenic conditions. That is, it can be understood as having the same meaning as probiotics.

"Intestinal yuikgyun" is a lactic acid (Lactic acid bacteria), Lactobacillus genus (Lactobacillus), flow Pocono stock in (Leuconostoc), Lactococcus genus (Lactococcus), can include a Bifidobacterium (Bifidobacterium), Enterobacter Enterococcus , Streptococcus , and the like.

Preferably, the enteric yuikgyun the Lactobacillus genus (Lactobacillus), flow Pocono stock in (Leuconostoc), Lactococcus genus (Lactococcus), Bifidobacterium (Bifidobacterium) in the strain, and the Pocono more preferably acids stock ( Leuconostoc), the genus Lactobacillus (Lactobacillus) strain.

Preferably, the strain belonging to the genus Leuconostoc is Leuconostoc mesenteroides), and strains belonging to the genus Lactobacillus (Lactobacillus) may be a Lactobacillus khepri (Lactobacillus kefiri).

In one embodiment of the invention, grape seed powder flow Pocono stock mesen teroyi des (Leuconostoc mesenteroides) was confirmed that that the strain and Lactobacillus Kane promote the growth of free (Lactobacillus kefiri) DH5 strain, as compared to grape seed non-addition group, and grape seed extract It was confirmed that the growth promoting effect was better.

The term " harmful bacteria in the intestines " means harmful bacteria living in the intestines of the host, and strains which are harmful to the intestinal environment when they are ingested and reached the intestines.

"Intestinal harmful bacteria," the luge Pseudomonas labor in (Pseudomonas - aeruginosa), Vibrio genus (Vibrio), Staphylococcus genus (staphylococcus), Clostridium genus (Clostridium), oil cake Te Solarium in (Eubacterium), night steroid in ( Bacteroides strains and sulfate reducers .

Preferably, the enteric harmful bacteria is Clostridium genus, oil cake Te Solarium in steroid or night and in strain, and more preferably, Clostridium sp of the present invention, Clostridium perfringens (Clostridium perfringens).

In one embodiment of the present invention, it was confirmed that the grape seed powder inhibited the growth of Clostridium perfringens strain, and the group without grape seed and the grape seed extract showed no growth inhibitory effect.

Therefore, it has been proved that the grape seed powder has selective prebiotic effect not only promoting the proliferation of the beneficial bacteria in the intestines but also inhibiting the growth of the intestinal harmful bacteria.

The grape seed powder can be used as a very useful prebiotic because it can promote the proliferation of beneficial bacteria and the growth inhibition of harmful bacteria at the same time as compared with the conventional components which are capable of promoting the proliferation of beneficial bacteria or the inhibition of the growth of harmful bacteria .

In another aspect, the present invention relates to a health functional food for promoting the proliferation of beneficial bacteria in the intestines containing grape seed powder and for inhibiting the growth of harmful bacteria in the intestines.

The health functional food of the present invention may be a beverage, a gum, a tea, a vitamin complex, a health supplement food, and more specifically, a dairy product, a confectionery, a seasoning, a drink and a drink, a snack, a candy, a jelly, Dairy products, dairy drinks and milk drinks, tea, soy milk and soybean milk products, such as pasta, soups, sauces and dressings, confectionery products, oil and fat products, chocolate products, processed foods, but are not limited to, soy dairy-like products, frozen foods, cooked and alternate foods, meat products, cheese, yogurt, bread and buns, yeast products, cakes, cookies and crackers Do not.

The health functional food may be prepared in the form of capsules, tablets, powders, liquid suspensions, pills and granules containing grape seed powder.

In addition, the health functional food may further include not only the grape seed powder as an active ingredient, but also a component that is ordinarily added at the time of food production.

The health functional food of the present invention can be used as a nutritive, a vitamin, a mineral (electrolyte), a flavoring agent such as a synthetic flavor agent and a natural flavor agent, a coloring agent and an aging agent (cheese, chocolate etc.), a pectic acid and its salt, Salt, organic acid, protective colloid thickener, pH adjusting agent, stabilizer, preservative, glycerin, alcohol, carbonating agent used in carbonated beverage, and may contain flesh for producing fruit juice or vegetable beverage.

The health functional food according to the present invention can enhance the growth of beneficial microorganisms and inhibit the growth of harmful microorganisms, thereby improving the health by changing intestinal flora.

In another aspect, the present invention relates to a pharmaceutical composition for promoting the proliferation of beneficial bacteria in the gut containing grapeseed powder and for inhibiting the growth of harmful bacteria in the intestines.

The pharmaceutical composition according to the present invention promotes the proliferation of beneficial bacteria in the intestines and inhibits the growth of harmful bacteria in the intestines, thereby preventing intestinal diseases or improving intestinal health.

The pharmaceutical composition comprising the grape seed powder of the present invention contains grape seed powder as an active ingredient and may further comprise an appropriate carrier, excipient or diluent according to a conventional method.

Examples of the carrier, excipient and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

In addition, the pharmaceutical composition of the present invention may be formulated into various oral or parenteral administration forms.

Examples of formulations for oral administration include tablets, pills, hard, soft capsules, liquids, suspensions, emulsions, syrups, granules and the like. These formulations may contain, in addition to the active ingredient, a diluent such as lactose, dextrose, (For example, silica, talc, stearic acid and magnesium or calcium salts thereof and / or polyethylene glycol), in addition to the active ingredient (s). The tablets may also contain binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and / or polyvinylpyrrolidine, optionally mixed with starch, agar, alginic acid Or a disintegrating or boiling mixture such as its sodium salt and / or an absorbent, a colorant, a flavoring agent and a sweetening agent. The formulations may be prepared by conventional mixing, granulating or coating methods.

Representative examples of formulations for parenteral administration include injectable preparations, and water, Ringer's solution, isotonic saline or suspensions may be mentioned as a solvent for the injectable preparation. The sterile, fixed oils of the injectable preparations may be used as a solvent or suspending medium, and any non-irritating fixed oils, including mono-, di-glycerides, may be used for this purpose. The injectable preparation may be a fatty acid such as oleic acid.

The dosage of the pharmaceutical composition of the present invention may vary depending on the age, sex, weight, and severity of the disease of the patient. The amount and the administration frequency of the pharmaceutical composition of the present invention to be actually administered is appropriately determined by various factors such as the kind of symptom to be treated, route of administration, sex, health condition, diet, age and weight of the individual, .

Particularly, the grape seed powder according to the present invention inhibits the growth of Clostridium perfringens , which is a major component of older grains. Therefore, by improving the intestinal health when the pharmaceutical composition of the present invention is administered to a higher-grade layer, Or treatment.

Example

Hereinafter, the present invention will be described in more detail with reference to Examples. It will be apparent to those skilled in the art that these embodiments are for illustrative purposes only and that the scope of the present invention is not construed as being limited by these examples.

Manufacture of grape seed powder and grape seed extract

Chardonnay grapes were used to make wine, and the remaining by-products were cold-pressed to remove grease from the grape seeds. Then, the remaining product was hot-air dried and the skin was removed by air blow. After the hot air drying, the remaining product was filtered using a sieve having a size of 100 mesh (150 micron) to prepare fine grained seed flour (GSF).

Grape seed extracts (GSE) were prepared by i) sonicated grape seed water extracts (SGWE) and ii) grape seed powder was extracted with 80% methanol after ultrasonication Sonicated grape seed MeOH 80% extracts, SGME).

Standardization of samples

In order to standardize the content of catechin contained in grape seed extract and grape seed powder used in the experiment, the content of catechin contained in grape seed powder and grape seed extract was analyzed by HPLC (high performance liquid chromatography). Using the catechin content obtained here, grape seed extracts with the same catechin content in 5 mg or 10 mg of grape seed powder per ml were used.

<Yield of extract> Sample code Extraction solvent Yield ( % ) SGWE water 8.22% SGME 80% MeOH 5.69%

<Catechin Content in Grape Seed Extract and Grape Seed Powder> Sample code Contents of phenolic compound (mg / g) (+) - Catechin GSF 7.01 SGWE 10.06 ± 0.18 SGME 8.70 ± 0.25

(GSF: Grape Seed Powder, SGWE: Grape Seed Powder Ultrasonically Extracted with Water, SGME: Grape Seed Powder Extract Extracted with 80% Methanol after Ultrasonic Processing)

Experimental Example  One: For profit  Promoting proliferation

(1) Ryukono Stock Messeneroides ( Leuconostoc mesenterorides )  Germ

A yuikgyun, was used as the current stock Pocono mesen teroyi des (Leuconostoc mesenteroides) strain belonging to lactic acid bacteria (lactic acid bacteria). The strain was isolated from Kefir grain using a selection medium, and the method of separation is as follows.

0.002% BPB-MRS medium or Rogosa medium was used, and anaerobic culture was performed for 72 hours using an anaerobic jar at 30 (this allows excluded competitive compartment of yeast or bacteria). Then, colonies of various lactic acid bacteria were selected based on the color and shape of the colonies, and 16S rRNA seqeuncing was performed to identify them.

Two days before, the turbidity of the pre-awakened fungus was adjusted to 2.0, injected into a tube containing 3 mL of MRS broth, diluted to 10 ¹ and 10 ⁰ CFU / mL, and spreading was performed to measure the initial number of bacteria. In the other vial containing 3 mL of MRS broth, the graft seed powder (GSF), the sonicated grape seed extract (SGWE), and the sonicated grape seed powder were sonicated and extracted with 80% methanol After 5 minutes of sonication (50 sec operation, 10 sec interval, 5 times repeated) with 5 mg and 10 mg of MeOH 80% extracts and SGME, respectively, 0.1% DMSO was injected (in this case, Corrected value).

Then, the turbidity of the bacteria which had been pre-awakened two days before was adjusted to 2.0, and cultured in an incubator at 30 ° C for 48 hours and 96 hours after col- lation in the solid medium.

As a result, as shown in Fig. 1, when the Leuconostoc mesenteroides were cultured for 48 hours and cultured in the 5 mg and 10 mg grape seeds (GSF), the control and grape seed extracts (SGWE, SGME) The growth was significantly increased. Specifically, the growth of 5 mg of grape seed powder (GSF) increased 27.82% compared to 5 mg of grape seed extract and 14.98% of SGME 5 mg, and that of 10 mg of grape seed powder (GSF) 32.3% and 15.9%, respectively, compared to SGME 10mg.

(2) Lactobacillus Kepri ( Lactobacillus kefiri )  DH5

Another yuikgyun, were tested using the Lactobacillus khepri (Lactobacillus kefiri) DH5 strain belonging to lactic acid bacteria (lactic acid bacteria).

Lactobacillus khepri (Lactobacillus kefiri) DH5 strain was identified isolated from Lactobacillus kefir (Lactobacillus kefir),.

The method of isolating the strain is as follows. 0.002% BPB-MRS medium or Rogosa medium was used, and anaerobic culture was performed for 72 hours using an anaerobic jar at 30. (Which enabled the elimination of competitive colonies of yeast and bacteria, etc.), various colonies of lactic acid bacteria were selected based on the color and shape of the colonies, and 16S rRNA seqeuncing was carried out to identify them.

In the same manner as above, the number of colonies was measured after culturing in grape seed powder (GSF), control group and grape seed extract (SGWE, SGME).

As a result, as shown in FIG. 2, Lactobacillus khepri (Lactobacillus kefiri) DH5 after a 96 hours culture, it is incubated in the grape seed powder (GSF) of 5mg, growth as compared to the control group, and grape seed flour extract (SGWE, SGME) Respectively. Specifically, the growth of 5 mg of grape seed powder (GSF) increased 3.2% compared to SGWE 5 mg and 1% of SGME 5 mg.

Experimental Example  2: Suppression of growth against harmful bacteria  effect

Clostridium perfringens strain (KCTC 5009) belonging to Clostridium was used as a harmful bacteria.

In the same manner as above, the number of colonies was measured after culturing in grape seed powder (GSF), control group and grape seed extract (SGWE, SGME).

As a result, as shown in Fig. 3, the cultivation of Clostridium perfringens bacteria in 10 mg of grape seed powder (GSF) after culturing for 12 hours was superior to the control and grape seed powder extracts (SGWE, SGME) Growth was greatly reduced. Specifically, the growth of 10 mg of grape seed (GSF) decreased 33.8% compared to 10 mg of grape seed extract and 33.6% of SGME 10 mg.

That is, the 10 mg / mL grape seed powder (GSF) significantly inhibited the growth of Clostridium perfringens, but the grape seed extracts containing the same amount of total polyphenols (catechins) as SGWE , SGME) had no effect on the growth of harmful bacteria.

Thus, the grape seed extract does not inhibit the growth of harmful microorganisms, while the grape seed powder can inhibit the growth of harmful microorganisms. Therefore, the grape seed extract can be used as a better prebiotic than the grape seed extract.

In conclusion, the grape seed powder promotes the proliferation of beneficial bacteria in the intestines and inhibits the growth of harmful bacteria in the intestines. Thus, it can be said that it is a very useful prebiotics which selectively affects the growth of beneficial bacteria and harmful bacteria.

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will appreciate that such specific embodiments are merely preferred embodiments and that the scope of the invention is not limited thereby. It will be obvious. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (10)

Prebiotics for promoting the proliferation of beneficial bacteria in the intestines and inhibiting the growth of harmful bacteria in the intestines, including grape seed powder.
The method according to claim 1,
Wherein the grape seed powder has a function of promoting the proliferation of beneficial bacteria in the intestines and a function of inhibiting the growth of harmful bacteria in the intestines.
The method according to claim 1,
Wherein the grape seed powder is produced from by-products generated during the manufacturing process of the wine.
The method according to claim 1,
Wherein the beneficial bacteria in the intestines are lactic acid bacteria .
5. The method of claim 4,
The lactic acid bacterium belongs to the genus Leuconostoc , Lactobacillus, Lt; RTI ID = 0.0 &gt; prebiotics. &Lt; / RTI &gt;
6. The method of claim 5,
Strains belonging to the stock flow in Kono (Leuconostoc) is stock flow Pocono mesen teroyi des (Leuconostoc mesenteroides), and strains belonging to the genus Lactobacillus (Lactobacillus) is Lactobacillus khepri (Lactobacillus kefiri) vertices landscape bio, characterized in that.
The method according to claim 1,
Wherein the intestinal harmful bacteria are strains belonging to the genus Clostridium .
8. The method of claim 7,
The Claus tree strains belonging to the genus Stadium (Clostridium) is Clostridium ticks landscape bio, characterized in that the puff Lin presence (Clostridium perfringens).
, A health functional food for promoting the proliferation and inhibiting the growth of harmful bacteria in the intestines comprising the prebiotics according to claim 1.
A pharmaceutical composition for promoting the proliferation and inhibiting the growth of harmful bacteria in the intestines comprising the prebiotics according to claim 1.

Images