KR101423433B1 - Compositions for anti-inflammatory or prevention or treatment of food poisoning comprising extracts of fermented Dendropanax morbifera - Google Patents

Abstract

The present invention relates to a fermented Dendropanax morbifera extract having increased content of salicylic acid, theaflavin, and theaflavin-3-gallate. The fermented Dendropanax morbifera extract has effects of anti-inflammation or inhibition of food poisoning bacteria growth. The fermented Dendropanax morbifera extract can be safely applied to a pharmaceutical or a functional food composition without side effects since the extract is derived from natural products.

Description

TECHNICAL FIELD The present invention relates to a composition for preventing or treating anti-inflammatory or food poisoning,

The present invention relates to a composition for preventing or treating anti-inflammatory or food poisoning comprising an extract of Fusarium oxysporum fungi having an increased content of components of salicylic acid, tephalin, and teffrafin-3-gallate.

Along with an increase in average life expectancy, food and food consumption has been moving in the direction of safety and health since 2000. As a result, organic agricultural products, non-polluting foods, functional foods, and natural foods are continuously gaining popularity. An example of such a fermented food is a fermented food prepared by adding a sugar or a microorganism. Fermented foods are fermented by decomposition of polymeric materials into low molecular weight substances to increase the digestion and absorption rate, to increase the palatability due to the generation of new flavor, and to enhance the components having physiological activity. In addition to drugs for maintenance and treatment of health, Research on the effect of physiological activity on ingestible substances has been extensively conducted, and functional foods and ancillary medicines have been actively developed (Heo SI et al. , 2010).

Dendropanax morbifera is a plant belonging to Araliaceae. It is a medicinal plant with excellent pharmacological efficacy. It has a scientific name Dendro-panax morbifera Nakai, meaning a panacea. In the past, it was used as a natural coating material because it is composed of 66.7% of nonvolatile matter, 10.8% of directional component, 8.1% of moisture, and 14.4% of solid content. However, recent studies have shown that diabetic, hypertensive, , Jeong IM et al., Pharmazie, 2009; Park, BY et al., Ethnopharmacol 2004. Jeong, IM et al., Phytother Res., 2009).

These yellowtail trees can be ingested as foods, such as yellowtail, berries, leaves, branches and roots, or added to liquor oil, flavored oil, liquor, tea, food additives, etc. However, in order to take a pharmacological effect, There is a limit to the intake in the form. Methods for extracting useful components and taking them for a long time have been studied. The extract has a weak pharmacological activity per unit weight, and the fruit contains a large amount of fat, which is an oil component, so that absorption into the body is slow. There is a problem that it is difficult to change formulations in various forms such as powders and granular extracts of by-products.

Korean Patent No. 10-1054041 discloses a process and a composition for mass-processing a fermented product and a fermented extract using Hwangchujang, and Korean Patent No. 10-0988072 discloses a fermented product And an antioxidant pharmaceutical composition containing the same. Korean Patent Laid-Open Publication No. 2011-0082992 discloses a composition for treating atopic and acne skin including extracts of Hwangjak tree sap and extract of Hwangchujigae bark. In Korean Patent No. 10-1194947, A pharmaceutical composition for the treatment of liver diseases is disclosed. However, it has not yet been disclosed in the present invention on the extracts of fermented Yellow-throated tree which have increased content of components of salicylic acid, teaplavin and tephavalin-3-gallate.

Accordingly, the present inventors have found that the fermentation process using microorganisms can increase the content of salicylic acid, teaplavin, and tephalin-3- As a result, the present invention has been completed.

It is an object of the present invention to provide a pharmaceutical composition for preventing or treating anti-inflammatory or food poisoning, comprising a fermented extract of Fusarium oxysporum species with an increased content of salicylic acid, teaplavin and tefflavin-3-galate component.

It is another object of the present invention to provide an anti-inflammatory food composition comprising a fermented extract of Fusarium oxysporum sp. Having an increased content of salicylic acid, tephalin and tefflavin-3-galate component.

In one aspect, the present invention relates to a pharmaceutical composition for the prevention or treatment of anti-inflammatory or food poisoning, comprising a fermented extract of Fusarium oxysporum species with an increased content of salicylic acid, teaflavin and tefflavin-3-galate component.

In the present invention, the salicylic acid is 2-hydroxybenzoic acid having a structure represented by the following formula (1), and is one of the phenolic compounds of the woody spruce tree.

Salicylic acid is used as a composition of analgesics, antipyretics, and dermatological treatment drugs because of its effects on antipyretic, analgesic, antibacterial and anti-inflammatory effects.

In the present invention, theaflavin (TF-1) is one of the phenolic compounds produced by oxidation and polymerization of Cetechin species with an oxidizing enzyme, and has the following structural formula (2).

In the present invention, theaflavin-3-gallate (TF-2) is one of the phenolic compounds produced by oxidation and polymerization of catechins by oxidative enzymes, 3 < / RTI >

The above-mentioned teffavirin and teffavirin-3-gallate have antioxidative, anti-bacterial, antitumor, anti-inflammatory and cardiovascular disease-preventing effects which eliminate free-radicals.

In the present invention, the content of salicylic acid, tephalavin and tefflavin-3-gallate components is increased by 2 to 3 times the content of salicylic acid, teaplavin and tefflavin- 60-fold, preferably 8 to 10-fold increase in salicylic acid, 1.8- to 2.5-fold increase in the amount of teaflavin, and 2 to 50-fold increase in tephalin-3-gallate.

In one specific embodiment, the content of polyphenolic compounds in the extracts of fermented leaves and fermented branches and the extracts of the leaves and leaves of the leaves of the leaves of the leaves and leaves of the leaves, which did not undergo fermentation, were measured. As a result, the contents of salicylic acid, The amount of tephavalin - 3 - gallate was increased about 9 times, 2 times and 57 times as compared with the extracts of. In addition, it was confirmed that the contents of teabrabin and teafflabin - 3 - galitrate, which are the extracts of fermented yellowtail millet, were increased about 1.1 times and 2 times, respectively.

Thus, the content of salicylic acid, teaplavin and tefflavin-3-galate component is remarkably increased through the extract of Fusarium oxysporum species, and thus it has an excellent effect on the efficacy of these components, in particular, anti-inflammation and prevention or treatment of food poisoning.

In one specific implementation, the causative organisms of food poisoning include Escherichia coli , Salmonella enterica , Pseudomonas aeruginosa , Bacillus cereus , and Staphylococcus aureus The effects of Staphylococcus aureus on the inhibition of growth of food poisoning bacteria by the concentration of the extracts of Hwangchujangguli and Kwangju leaves, And extracts from the leaves of the Hwangchu - mackerel which have not undergone the fermentation process inhibit the growth of Salmonella enterica and Pseudomonas aeruginosa , and have an excellent effect of inhibiting the growth of food poisoning bacteria as the concentration of the extract increases .

In the present invention, the wood hwangchil fermentation extract hwanggukgyun (Aspergillus oryzae), back Aspergillus (Aspergillus kawauchi), black Aspergillus (Aspergillus niger) in hwangchil wood dry powder, Bacillus subtilis (Bacillus refers to an extract obtained by inoculating one or more microorganisms selected from the group consisting of subtilis or hiochi bacteria , fermenting them, and extracting with an extraction solvent.

In the present invention, the dry powder may be any one or more selected from the group consisting of leaves, flowers, branches, fruits and roots, preferably leaves, branches and roots Preferably one or more members selected from the group consisting of leaves and branches is cut, dried and pulverized by a conventional method, and powdered. The drying is not limited as long as it is a method for removing water from the perennial tree. For example, natural drying, freeze drying, vacuum drying and the like can be used.

In the present invention, it is preferable that the fermentation is carried out at a temperature of 30 to 45 ° C and a relative humidity of 88 to 92%. When the fermentation is carried out under these conditions, the maximum increase effect of the content of salicylic acid, teaflavin and tefflavin-3-gallate, which is one of the objects of the present invention, can be obtained.

In the present invention, the Fusarium oxysulfurum fermentation extract can be obtained by using an extracting method and an extraction solvent known in the art, preferably a hot water, an anhydrous or hydro-lower alcohol having 1 to 4 carbon atoms , Ethanol, propanol, butanol, etc.), acetone, ethyl acetate, chloroform, and 1,3-butylene glycol may be used singly or in combination of two or more. More preferred are anhydrides or lower alcohols having 1 to 4 carbon atoms And still more preferably methanol or ethanol. The extraction method is not particularly limited, and examples thereof include cold extraction, ultrasonic extraction, and reflux cooling extraction.

In addition, the above-mentioned fermentation extract of Huangchu tree can be obtained through a conventional purification process in addition to the extraction method using the above-mentioned extraction solvent. For example, separation using an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatographies (made for separation by size, charge, hydrophobicity or affinity) The obtained fractions can also be obtained.

In the present invention, the Fusarium oxysporum fermentation extract includes all of the extract and the purified product obtained in each step of extraction, fractionation or purification, their diluted solution, concentrate or dried product.

The pharmaceutical composition of the present invention may contain components commonly used in pharmaceutical compositions in addition to the above-mentioned fermented extract of Huaxia wood. For example, lubricants, wetting agents, sweeteners, flavoring agents, emulsifying agents, suspending agents, preservatives and carriers.

The pharmaceutical composition according to the present invention may additionally contain components commonly used in pharmaceutical compositions in addition to the fermented extract of P. japonicus. For example, lubricants, wetting agents, sweeteners, flavoring agents, emulsifying agents, suspending agents, preservatives and carriers.

Suitable carriers for such pharmaceutical compositions include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, , Syrups, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, but are not limited thereto. Carriers and preparations which are acceptable as suitable medicaments are described in detail, for example, in Remington's Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical composition may be administered to mammals such as rats, mice, livestock, humans, and the like by various routes such as parenteral, oral, and the like, and all manner of administration may be expected, for example, oral, rectal or intravenous , Muscle, subcutaneous, intrauterine or intracerebroventricular injection.

The pharmaceutical composition is administered in a pharmaceutically effective amount. In the present invention, a pharmaceutically effective amount means an amount sufficient to treat or prevent a disease at a reasonable benefit / risk ratio applicable to medical treatment or prevention. The effective dose level will depend on the type of disease and its severity, Including the age, weight, health and sex of the patient, sensitivity of the patient to the drug, time of administration of the particular extract used, route of administration and rate of release, duration of treatment, Can be determined according to factors well known in the art. Generally, a dose of 0.001 to 100 mg / kg, preferably 10 to 100 mg / kg on an adult basis can be administered once to several times per day. When the composition is an external preparation, it is preferable to apply the composition in an amount of 1.0 to 3.0 ml on an adult basis once to five times a day and continue for 1 month or more. However, the scope of the present invention is not limited by the dose.

The pharmaceutical composition may be prepared in a unit dosage form by formulating it with a pharmaceutically acceptable carrier and / or excipient according to a method which can be easily carried out by a person having ordinary skill in the art to which the present invention belongs. Into a capacity container. The formulations may be formulated into tablets, capsules, powders, granules, suspensions, emulsions, syrups, emulsions, alerts, ointments, sprayers, oils A tablet, a tincture, a bath, a liniment, a lotion, a patch, a pad, a cream, and the like. In addition, it can be preferably used as an external preparation for skin intended for topical administration for application directly to the affected area. In this case, a form such as an ointment, a lotion, a spray or a gel is preferable. These external preparations for skin may also be included in a support base or matrix or the like, which can be applied directly to the treatment site, and examples of the support substrate include gauze or bandages.

The active ingredient of the Fusarium oxysporum fermentation extract is 0.01 to 5% by weight, preferably 0.01 to 3% by weight, based on the whole composition. When the weight of the Fusarium oxysporum fermentation extract is less than 0.01% by weight, the effect of salicylic acid, teaplavin and tephavalin-3-gallate is low, and if it is more than 5% by weight, The content and effect of salicylic acid, teaplavin, and tephavalin-3-gallate components are not increased compared to those used, resulting in uneconomical problems.

In another aspect, the present invention relates to a food composition for preventing inflammation, which comprises fermented extract of Fusarium oxysporum species having increased contents of salicylic acid, tephalavin and tefflavin-3-gallate component.

The extracting solvent, extraction method, and efficacy of the fermentation extract of Fusarium oxysporum are as described above.

In the present invention, the food composition may include components that are conventionally added in the manufacture of food, in addition to the above fermented extract of Huangchu tree. For example, proteins, carbohydrates, fats, nutrients, flavoring agents, and flavoring agents. The above-mentioned carbohydrates are polysaccharides such as disaccharides such as monosaccharide, glucose, sucrose and oligosaccharides of glucose and fructose sugars, dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. The flavoring agent may be a natural flavoring agent (tau martin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.)) and synthetic flavorings (saccharin, aspartame, etc.).

There is no particular limitation on the kind of the food. Examples of the food to which the Huangchuang fermentation extract can be added include dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, ice cream, , A drink, an alcoholic beverage, and a vitamin complex, all of which include health foods in a conventional sense.

When the food of the present invention is a beverage, it may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. The natural carbohydrates may be monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, natural sweeteners such as dextrin and cyclodextrin, synthetic sweeteners such as saccharin and aspartame, and the like .

The food may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, A carbonating agent used in a carbonated beverage, and the like. In addition, the health food of the present invention may contain flesh for the production of natural fruit juice, fruit juice beverage and vegetable beverage. These components may be used independently or in combination.

The active ingredient of the Fusarium oxysporum fermentation extract is 0.01 to 5% by weight, preferably 0.01 to 3% by weight, based on the whole composition. When the weight of the Fusarium oxysporum fermentation extract is less than 0.01% by weight, the effect of salicylic acid, teaplavin and tephavalin-3-gallate is low, and if it is more than 5% by weight, The content and effect of salicylic acid, teaplavin, and tephavalin-3-gallate components are not increased compared to those used, which is inefficient.

The fermented extract of Huangchu tree of the present invention is safely applied to pharmaceutical and functional food compositions as described above because it is derived from natural materials and has no toxicity and side effects and has been used for a long time and can be used safely even in long-term use .

The extract of Fusarium oxysporum species of the present invention having an increased content of salicylic acid, teaplavin and tephavalin-3-gallate has an anti-inflammatory activity or an effect of inhibiting the growth of food poisoning bacteria. In addition, it can be safely applied to pharmaceutical and functional food compositions since it is derived from natural products and has no side effects.

1 is a graph showing the growth inhibitory activity of Salmonella Enterica and Pseudomonas aeruginosa , which are food poisoning bacteria, according to the concentration of the extract of Fermented Green Mulberry tree of the present invention.

Hereinafter, the present invention will be described in more detail with reference to Production Examples and Examples. These preparations and examples are only for illustrating the present invention more specifically, and the scope of the present invention is not limited by these preparations and examples according to the gist of the present invention.

Production Example 1: Fermentation microorganism culture

After the inoculation of 10 μg of Hwang Guk-gyun into TSB liquid medium (TSB broth: tryptone 17 g, Soytone 3 g / L, Dextrose 2.5 g / L, Sodium chloride 5 g / L, Dipotassium phosphate 2.5 g / L, pH 7.3-7.5; BBL / Difoc) The cells were cultured at 40 ° C for 24 hours to obtain a cell count of about 1 × 10 ⁹ cfu / ml.

Production Example 2: Preparation of Fermented Extracts of Wooden Millet

2-1. Pickling and washing

Leaves and branches of Korean wild grasses collected in Jeollanam - do in August were thoroughly cleaned to remove foreign matter and impurities, and moisture of the surface was removed.

2-2. cut

The leaves and branches of the washed Hwigulak tree of Production Example 2-1 were each cut into lengths of 0.5 to 2 cm using a cutting machine.

2-3. Freeze-dried and crushed

The cut leaves and branches of Preparation Example 2-2 were lyophilized using a freeze drier and then pulverized to a particle size of 1 mm or less.

2-4. Inoculation of Hwang Kook Kyun

2 kg of Hwanggukjun of Preparation Example 1 was inoculated to 1 kg of the leaves and branches of the perennials of Preparation Example 2-3.

2-5. Fermentation

The leaves and branches of Hwangchu-kaku inoculated with Hwang-guk-gum of Preparation Example 2-4 were placed in plastic baskets of 50, 40 and 30 cm in length, height and height, respectively, and the surface of the barrel was covered with a polyethylene film of 0.01 mm. , 30 ° C, 40 ° C and 50 ° C and 90% relative humidity for 48 hours.

2-6. Extraction and Filtration

95% ethanol was added at 10 times the weight of the leaves and branches of the prepared fermented yellowtail of Preparation Example 2-5, and the leaves and leaves of the perennials were grown for 48 hours using an ASE accelerated solvent extractor (Dimnes, USA) The extracts were firstly filtered using gauze, and then subjected to secondary filtration using a filter paper (Advante No. 2).

2-7. concentration

The filtered leaves and branches of the above-mentioned Preparative Example 2-6 were frozen in a deep freezer at -45 ° C for 12 hours, dried using a freeze dryer, and stored at 4 ° C in a refrigerator.

COMPARATIVE EXAMPLE 1: Preparation of Wortwood extract

Other than the fermentation processes of Production Examples 2-4 and 2-5, the same method as in Production Example 2 was used in the production of Hokutogi extract.

Example 1: Determination of the total polyphenol content of fermented Extracts

The total polyphenol contents of the fermented extracts of the leaves and branches of Wuchulia crassifolia prepared in Preparation Example 2 and the leaves and the branch extracts of the Wuchuchunae prepared in Comparative Example 1 were measured. Specifically, 1 g of each of the extracts of Preparation Example 2 and Comparative Example 1 was added to 2 ml of sodium carbonate, and then 100 μl of a 50% Folin-Ciocalteu reagent was added. After that, the absorbance was measured at 720 nm using a spectrophotometer and the total polyphenol content of each extract was measured based on the calibration curve of gallic acid. The significance test was performed using Duncan's multiple test (SPSS Inc, Chicago, IL, USA) software package program, version 12 SPSS (Statistical Package of Social Sciences, multiple range test). The results are shown in Table 1 below.

Comparative Example 1 Production Example 2 20 ℃ 30 ℃ 40 ℃ 50 ℃ Total polyphenol
(mg GAE / gDM) leaf 429 ± 2.6 398 ± 4.2 411 ± 3.1 402 ± 3.7 399 ± 2.4 Branch 342 ± 8.9 334 ± 6.8 328 ± 5.4 337 ± 7.2 341 ± 7.7

As shown in Table 1, there was no significant difference in the total polyphenol contents of the leaves of Leucumber leaf and the leaves of Leucigenia sp.

Example 2: Determination of total flavonoid content of Fermented Extracts

In order to compare the total flavonoid contents of the leaves and the fermented extracts of Leucaceae tree prepared in Preparation Example 2 and the leaf and branch extracts of the Leucigeniae prepared in Comparative Example 1, 0.15 ml of 5% sodium nitrite, And 1 g of the extract of Preparation Example 2 and Comparative Example 1, respectively, and the mixture was reacted at 25 DEG C for 6 minutes. Next, 0.3 ml of 10% aluminum chloride was added, followed by reaction at 25 ° C for 5 minutes. Then, 1 ml of 1N sodium hydroxide was added to each sample to mix well. Then, the absorbance at 510 nm was measured using a spectrophotometer, and the total flavonoid content of each extract was measured based on the calibration curve of rutin hydrate. The significance test was performed using Duncan's multiple test (SPSS Inc, Chicago, IL, USA) software package program, version 12 SPSS (Statistical Package of Social Sciences, multiple range test). The results are shown in Table 2 below.

Comparative Example 1 Production Example 2 20 ℃ 30 ℃ 40 ℃ 50 ℃ Total flavonoid
(mg RU / gDM) leaf 75 ± 1.6 72 ± 2.1 69 ± 4.2 62.9 ± 3.7 61.4 ± 2.8 Branch 32 ± 5.7 29 ± 4.8 31 ± 6.1 55.3 ± 7.2 42 ± 5.6

As shown in Table 2, there was no significant difference in the total flavonoid content of the leaf of Leucumber leaf and the leaf extract of the Leucigenia sp.

Example 3: Antioxidant activity measurement of Fermented Extracts of Wollaeng tree

The ABTS radical scavenging activity of the leaf and root fermented extract prepared in Preparation Example 2 and the leaf and branch extracts prepared in Comparative Example 1 were measured and compared. Specifically, 7.4 mM ABTS (2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt) and 2.6 mM potassium persulfate were mixed and allowed to stand at room temperature and in a dark place for 24 hours, The ABTS solution was diluted with phosphate buffered saline (PBS, pH 7.4) at 732 nm so that the absorbance was 0.700 ± 0.030. 50 μl of the extract was added to 950 μl of the diluted solution, reacted in the dark for 10 minutes, and the absorbance was measured at 732 nm. BHA (Butylated hydroxyanisole), a synthetic antioxidant, was used as a control, and ACTS radical scavenging activity was measured using Equation 1. The significance test was performed using Duncan's multiple test (SPSS Inc, Chicago, IL, USA) software package program, version 12 SPSS (Statistical Package of Social Sciences, multiple range test). The results are shown in Table 3.

[Experimental Equation 1]

ABTS radical scavenging ability (%) = 100 - (OD value of experimental group / OD value of control group) x 100

BHA
(0.1%) Comparative Example 1 Production Example 2 20 ℃ 30 ℃ 40 ℃ 50 ℃ ABTS radical scavenging ability (%) 90 ± 2.4 leaf 78.5 ± 2.4 71 ± 1.8 72 ± 4.3 74 ± 8.6 73 ± 7.7 Branch 84.3 ± 3.8 75 ± 2.9 77 ± 2.4 79.3 ± 3.6 76 ± 4.1

As shown in Table 3, the ABTS radical scavenging activity of non-fermented Hwangchuki leaves and eggplant extracts was different. The extracts of Hwangchuki (leafy leaves) and Kwangwol (fermented) extracts were 74 ± 8.6% and 79.3 ± 3.6% Respectively. Antioxidant contents of non - fermented eggplant extracts were relatively high.

Example 4: Determination of nitrite scavenging ability of the fermented extract of Wuchulia chinense

The nitrite scavenging ability of the leaf and root fermented extracts prepared in Preparation Example 2 and the leaf and branch extracts prepared in Comparative Example 1 were measured and compared. Specifically, 1 ml of fermented extract was added to 1 ml of 1 mM NaNO ₂ solution, and the mixture was adjusted to pH 1.2, pH 3.0 and pH 4.0 with 0.1 N HCl and 0.2 M citrate buffer (pH 2.5). The total volume of the reaction solution was then adjusted to 10 ml Respectively. Subsequently, 1 ml of the mixture was reacted at 37 ° C for 1 hour, and then 3 ml of a 2% acetic acid solution and 0.4 ml of a grease reagent (1% sulfanilic acid: 1% naphthylamine = 1: 1) dissolved in a 30% , And then left at room temperature for 15 minutes. Absorbance was measured at 520 nm. As a control, distilled water was used instead of the grease reagent. The nitrite scavenging activity was calculated using Equation 2. The significance test was performed using Duncan's multiple test (SPSS Inc, Chicago, IL, USA) software package program, version 12 SPSS (Statistical Package of Social Sciences, multiple range test). The results are shown in Table 4 below.

[Experimental Equation 2]

Nitrite scavenging ability (%) = 100- (OD value of experimental group / OD value of control group) x 100

Fermentation temperature pH Extract concentration 0.1% 0.25% 0.5% EC ₅₀ (%) Production Example 2 20 ℃ leaf 1.2 18.4 67.2 92.7 0.18 3.0 39.8 46.2 61.4 0.31 4.0 8.1 11.2 20.9 > 0.5 Branch 1.2 28.6 48.9 61.7 0.25 3.0 20.4 31.7 38.9 > 0.5 4.0 10.2 13.4 19.9 > 0.5 30 ℃ leaf 1.2 20.4 70.4 97.4 0.16 3.0 42.7 51.4 68.2 0.23 4.0 9.8 10.4 21.1 > 0.5 Branch 1.2 34.4 51.2 58.9 0.23 3.0 23.5 32.1 55.8 0.45 4.0 8.9 40.6 24.7 > 0.5 40 ℃ leaf 1.2 27.1 77.4 99.8 0.14 3.0 46.6 55.5 70.9 0.21 4.0 9.1 11.9 21.8 > 0.5 Branch 1.2 41.0 54.4 70.8 0.20 3.0 25.9 33.3 48.0 0.81 4.0 12.5 17.1 30.4 > 0.5 50 ℃ leaf 1.2 26.4 75.4 98.7 0.16 3.0 45.1 54.8 68.4 0.24 4.0 8.8 11.4 21.1 > 0.5 Branch 1.2 40.7 54.1 68.8 0.21 3.0 25.1 32.1 47.4 0.84 4.0 11.5 16.8 29.6 > 0.5 Comparative Example 1 - leaf 1.2 50.7 78.9 96.0 0.09 3.0 38.7 43.7 60.6 0.37 4.0 8.4 11.4 21.2 > 0.5 Branch 1.2 24.4 58.7 82.6 0.20 3.0 31.0 37.8 41.8 > 0.5 4.0 9.1 12.7 17.8 > 0.5

As shown in Table 4, in the sample concentration according to pH, the leaf and leafy fermentation extracts of the Huchilki leaves have superior nitrite scavenging ability compared to the non-fermented Huchilchia leaves and branching extracts, The fermented extract showed the best nitrite scavenging ability.

All extracts showed better nitrite scavenging ability as pH was lowered, and nitrite scavenging ability of leaf extract was better than eggplant extract.

Example 5: Measurement of the inhibitory effect of fermented extract of Wuchulia chinense on growth inhibition of food poisoning bacteria

first. Escherichia coli , Salmonella enterica , Pseudomonas aeruginosa , Bacillus cereus and Staphylococcus aureus strains were cultivated in TSB liquid medium (Tryptic soy broth, pH 7.0, BBL / Difco) at 25 ° C. These cultures were spot inoculated in TSA solid medium and then cultured in a 25 ° C incubator for 24 hours.

Then, the filter paper (Advantes, No. 2) was placed on the culture medium on which the strain was cultured, and the leaves and the branched fermented extracts prepared by fermenting the same at a temperature of 40 ° C in the above Preparation Example 2, Wheatgrass leaves and eggplant extracts were treated with 0.1, 1, 5 and 10 mg / dish to confirm the formation of inhibition zone. As a control, penicillin was used instead of Hwangchujang leaf and eggplant fermentation extracts or Hwangchujanggwa leaf and eggplant extracts. The results are shown in Table 5 and Fig.

Strain Concentration (mg / dish) 0.1 One 5 10 Production Example 2
(Fermentation temperature: 40 DEG C) leaf Escherichia coli
( Escherichia coli) - - - - Salmonella enterica
( Salmonella enterica) - - + +++ Pseudomonas aeruginosa
(Pseudomonas) - - + ++ Bacillus cereus
(Bacillus cereus) - - + + Staphylococcus aureus
( Staphylococcus aureus ) - - + + Branch - - - - Escherichia coli
( Escherichia coli ) Salmonella enterica
( Salmonella enterica ) - - - - Pseudomonas aeruginosa
( Pseudomonas ) - - - - Bacillus cereus
( Bacilllus cereus ) - - - - Staphylococcus aureus
( Staphylococcus aureus ) - - - - Comparative Example 1 leaf Escherichia coli
( Escherichia coli ) - - - - Salmonella enterica
( Salmonella enterica ) - + +++ +++ Pseudomonas aeruginosa
( Pseudomonas ) - - ++ +++ Bacillus cereus
( Bacilllus cereus ) - - + + Staphylococcus aureus
( Staphylococcus aureus ) - - + + Branch Escherichia coli
( Escherichia coli ) - - - - Salmonella enterica
( Salmonella enterica ) - - + - Pseudomonas aeruginosa
( Pseudomonas ) - - - - Bacillus cereus
( Bacilllus cereus ) - - - - Staphylococcus aureus
( Staphylococcus aureus ) - - - - Control group Escherichia coli
( Escherichia coli ) + ++ ++ +++ Salmonella enterica
( Salmonella enterica ) + ++ ++ +++ Pseudomonas aeruginosa
( Pseudomonas ) + ++ ++ +++ Bacillus cereus
( Bacilllus cereus ) + ++ ++ +++ +++ ++ ++ + Staphylococcus aureus
( Staphylococcus aureus ) +++: strongest growth inhibition, zone diameter,> 20 mm; ++: strong growth inhibition, ring diameter, 16 to 20 mm; +: Growth inhibition, ring diameter, 10 to 15 mm; -: No growth inhibition, <10mm

As shown in Table 5 and FIG. 1, it was confirmed that the extracts of Fermented Leaf Leaf Leaf Leaf Leaf Leaf Leaf Leaf Leaf Leaf Leaf Leaf Leaf Leaf Leaf Leaf Leaf Leaf Leaf Extract and Salmonella Enterica Leaf and Pseudomonas Alegus Leaf Leaf Suppress Growth Dependently. However, the fermented extracts of P. japonicus and P. japonicus showed no inhibitory effect on the pathogenic bacteria.

Example 6: Analysis of Polyphenolic Compounds Contents of Fermented Extracts from Woodenwood

The leaves and the fermented extracts of P. ganoderma lucidum prepared by fermenting at a temperature of 40 ° C. of Preparation Example 2 and the extracts of P. ganoderma leaves and eggplant prepared in Comparative Example 1 were analyzed by HPLC (High pressure liquid chromatography, 1200 series, agilent technologies, USA ) Was used to analyze the phenolic compound content. Qualitative and quantitative analysis of polyphenolic compounds was performed by diluting a standard substance of 18 kinds of phenolic compounds at a certain ratio, obtaining peak areas through HPLC, and measuring the concentration using a rare-curved equation. The HPLC column was a shiseido (Tokyo, Japan) Capcell Pak C18 column (5 탆, 250 mm ㅧ 4.6 mm) and the flow rate of the mobile phase was 1.0 mL / min. The regression curve of the reference material is shown in Table 6. The results are shown in Table 7.

Standard material Regression curve equation Resveratrol y = 40.34x + 320.6 Gentisic acid y = 0.841x-14.32 4-Hydroxybenzoic acid (4-hydroxybenzoic acid) y = 18.09x + 117.07 Chlorogenic acid y = 7.77x-663.78 Syringic acid y = 30.38x-33.57 Para-coumaric acid (ρ-Coumaric acid) y = 53.95x + 282.75 Cathechin y = 3.32x + 29.20 Myricetin y = 12.54x + 305.49 Caffeic acid y = 34.86x + 172.86 Ferulic acid y = 25.97x + 172.4 Salicylic acid y = 7.07x-41.7 Trans- Shinan acid (trans -Cinnamic acid) y = 83.82x + 743.78 Naringin y = 16.29x + 113.33 Quercetin y = 16.32x-43.8 Hesperidin y = 0.562x + 1.99 Gallic acid y = 29.28x + 1205.65 Theaflavin y = 24.647x-189.01 Theaflavin-3-gallate &lt; / RTI &gt; y = 6.560x-35.929

Unit: ug / g Production Example 2 (fermentation temperature: 40 DEG C) Comparative Example 1 Standard material leaf Branch leaf Branch Gallic acid 24.7 0.00 11.1 17.4 2,5-Dihydroxybenzoic acid
(2,5-Dihydroxybenzoic acid) 337 24.1 420.4 259.1 Catechine 413.07 13.1 520.2 359.1 4-hydroxybenzoic acid
(4-Hydroxybenzoic acid) 1.1 19.7 232.9 70.6 Chlorogine acid 38.0 16.0 51.5 16.1 Caffeic acid 12.5 5.2 19.8 32 Syringic acid 25.8 0.6 64.1 42.1 Para-cumaric acid
(ρ-Coumaric acid) 21.2 0.04 26.6 2.1 Trans-ferulic acid
(trans-Ferulic acid) 113 13.3 147.5 34.9 Salicylic acid 403 6.4 45.8 248.9 Hesperidin 1631 1769 1365.3 1576.7 Naringin 49.1 30.5 67.8 53.2 Resveratrol 1.3 32 13.18 66.2 Myricetin 65.5 79.8 4.0 97.4 Quercetin 40.0 3.6 47.9 5.4 Trans-synnamic acid
( trans- Cinnamic acid) 0.6 0.2 0.03 0.6 Theaflavin 5.4 34 3.3 31 Tefflaven-3-gallate
(Theaflavin-3-gallate) 56.7 89 1.0 40 Sum 3,239.1 2,136.5 3,042.4 2,952.6

As shown in Table 7 above, the fermented extract of Hwangbukki leaf showed higher contents of salicylic acid, Myricetin, trnas-cinnamic acid, Theaflavin, Theaflavin-3-gallate content was increased by about 9, 16, 20, 2 and 57 times, respectively.

In addition, the content of theaflavin and theaflavin-3-gallate increased in the fermented extract of P. japonica compared to the extract of P. japonicus, about 1.1 times and 2.2 times Respectively.

One ; Penicillin treatment
2 ; Horticultura leaf extract treatment
3; Treating leafy fermented leek extract
4 ; Hwangchil tree branch egg extract treatment
5; Processed with Fermented Extract of Wooden Branches

Claims (10)

Wherein the fermented extract of Hwangchujang leaves is obtained by inoculating Aspergillus oryzae into the crushed leaf of Hwangcholgwa leaf and fermenting at a temperature of 30 to 45 DEG C and a relative humidity of 88 to 92% Wherein the content of salicylic acid, teaplavin, and teffrafin-3-gallate components is increased by 2 to 60 times as compared to the extract of Hokutogi tree leaf.
delete delete delete delete Wherein the fermented extract of Hwangchujang leaves is obtained by inoculating Aspergillus oryzae into the crushed leaf of Hwangcholgwa leaf and fermenting at a temperature of 30 to 45 DEG C and a relative humidity of 88 to 92% Wherein the content of salicylic acid, teaplavin and tefflavin-3-gallate components is increased by 2 to 60 times as compared to the extract of Hokutogama leaf. delete delete delete delete

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