KR101482873B1 - Fermentation metabolite of Dendropanax morbiferus produced by liquid-state fermentation and manufacturaring process for the same - Google Patents

Abstract

The present invention relates to a Dendropanax morbiferus fermented metabolite produced by liquid fermentation and a method for producing the same. The method according to an aspect of the present invention comprises: a Dendropanax morbiferus raw ingredient pretreating step of pulverizing and cutting Dendropanax morbiferus leaves and stems; a fermentation substrate producing step of producing a hydrothermal extract liquid of the pretreated Dendropanax morbiferus raw ingredients; a microorganism pre-culturing step of culturing microorganisms, wherein the microorganism pre-culturing step is simultaneously performed with the Dendropanax morbiferus raw ingredient pretreating step or is selectively performed before or after the Dendropanax morbiferus raw ingredient pretreating step; a liquid fermenting step of inoculating and culturing the microorganisms in the fermentation substrate; and obtaining a Dendropanax morbiferus fermented metabolite by sterilizing, filtering, and concentrating the fermented liquid obtained from the liquid fermenting step.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a fermentation fermentation metabolite of liquid fermentation,

FIELD OF THE INVENTION The present invention relates to a sulfuric acid fermentation metabolite by liquid fermentation and a preparation method thereof, and more particularly to a sulfuric acid fermentation metabolite by liquid fermentation with improved bioavailability and physiological activity, and a preparation method thereof.

In the modern society, chemical synthesis technology has been used for industrial development. However, these chemical synthesis technologies have caused serious environmental problems by using high temperature, high pressure, highly toxic reagents and solutions. .

Biotransformation refers to the production of a desired product from a precursor using the physiological function of a biological organism. The term biotransformation refers to bioprocessing, bioconversion, biosythesis, biocatalysis, etc. As well. Bio-conversion technology is used in many fields because it has less energy consumption and environment-friendly than chemical synthesis technology. As a result, food, medicine, and cosmetics are used in many fields. 50% of the total.

Among the biotransformation technologies, researches have been conducted on methods of increasing biologic activity by converting glycosides into aglycons through microorganism-based methods and enzyme conversion.

Fermentation using microorganisms can be expected to increase the absorption of the active substance as a result of the low molecular weight of the active ingredient, and to exhibit the same effect normally to a person who does not have a specific intestinal bacteria.

Research through biotransformation techniques has been conducted in many places, mainly through the saponins of ginseng. Among them, ginseng contains saponin called gisenoside. Research has been conducted to develop a method for increasing the yield of compound K, which is a strong physiologically active ingredient. Studies on the possibility of developing functional food materials using bioconversion technology for flavonoids, which are known to have excellent antioxidant activity, and studies on inhibition of melanin synthesis by using biotransformation technology have been conducted. And so on.

Research on the development of food materials is progressing gradually through bioconversion technology.

On the other hand, Hwangchil is an evergreen tree of the dicotyledonous plant, and its international name is Dendropanax morbifera, which means panacea. It is also called a golden tree. It is a Korean native species native to the southern coast of Korea and Jeju Island. Its height is about 15m. The flower is bloomed at the end of June branch. The fruit is elliptical nucleus with a length of 7 ~ 19㎜. It ripens in color. The constituents and effects of this woody wood are mainly sesquiterpene, which contains water, gum, alcohol, ester and the like. Hwangchil, which is a sap extracted from Hwangchilgi bark, is said to have been used only by the royal family because it has a remarkable pharmacological effect and a delicate environment.

It has been widely accepted that nutritional value, pharmacological value, and therapeutic value have been studied in modern scientific studies and it has been studied in various directions and forms in order to utilize useful materials of Huangchil. Korean Patent Laid-Open Publication No. 2004-0107852 discloses a method for preparing a Huangchil extract having a skin whitening effect and a Huangchil extract according to the method, Korean Patent Registration No. 10-0318019 discloses a Huangchil extract having anticancer activity, 0663284 discloses a seedling extract of Hwangchil, which is excellent in physiological activity. Korean Patent Registration No. 10-0988072 discloses a fermented product of Hwangchulnam and a pharmaceutical composition containing the same, Korean Patent Publication No. 2003-0005098 discloses a fermented product And Korean Unexamined Patent Application Publication No. 2005-0036093 discloses a technique of an ultraviolet cut cosmetic composition containing an extract of Hwangjak tree as an active ingredient.

It is an object of one embodiment according to the present invention to provide a fermented yellowish fermented metabolite by liquid fermentation with improved bioavailability and physiological activity and a method for producing the same.

One embodiment of the present invention relates to a method for the preparation of a method for the preparation of a papermaking plant, Preparing a fermentation substrate to produce a hot water extract of the pretreated greasy raw material; A microorganism pre-culture step in which the microorganism is selectively cultivated simultaneously with or before or after the pretreatment step of the above-described greenhouse material; A liquid fermentation step of inoculating and culturing the microorganism in the fermentation substrate; And a step of sterilizing, filtering and concentrating the fermentation broth obtained in the liquid fermentation step to obtain a yellowish fermented metabolite, and a process for producing the yellowish fermentation metabolism by liquid fermentation.

The solid content of the hot-water extract in the fermentation substrate may be 1 to 10 Brix.

The microorganism is an edible mold Aspergillus oryzae , Saccharomyces cerevisiae which is a brewer's yeast, Phellinus mushroom strain baumii and Ganoderma lucidum , Bacillus subtilis , or lactobacillus, Lactobacillus plantarum .

The microorganism pre-culturing step may be carried out at 25 to 37 DEG C for 2 to 10 days.

The microorganism is Bacillus subtilis , The solid content of the Welsh Hot Water Extract is 5 to 10 Brix or the microorganism is Lactobacillus plantarum , and the content of the solid content of the Welsh hot-water extract may be 1 to 5 Brix.

In the liquid fermentation step, the juice may be further mixed with the fermentation substrate.

The solid content of the juice may be 1 to 10 Brix.

The liquid culture can be carried out at 25 ° C to 40 ° C for 5 days to 20 days after 10% of the microorganism culture liquid is inoculated.

The sulfuric acid fermented metabolite may be processed in the form of a concentrate or powder.

Another embodiment of the present invention provides a yellowish fermented metabolite produced by the above method.

The sulfuric acid fermented metabolite may be used in a cosmetic composition, a pharmaceutical composition or a food composition.

The fermented metabolism of sulfuric acid by liquid fermentation according to the present invention can be understood as a state in which a useful substance of woody wood is low-molecularized by microorganisms and a secondary metabolite resulting from fermentation into microorganisms. According to one embodiment of the present invention, the specific form of the fermented fermentation broth can be determined according to the processing method of the liquid fermentation broth completed by liquid fermentation by the above-described production method, and the liquid fermentation broth can be in the form of a concentrated liquid, May be in the form of a solid powder that has undergone a process.

The fermented fermented metabolite according to one embodiment of the present invention not only has high yield by liquid fermentation, but also has excellent antioxidant ability, bioavailability and physiological activity.

The fermentation metabolic body according to one embodiment of the present invention uses fermentation methods such as liquid fermentation and pre-culture of microorganisms, thereby shortening the fermentation time, improving the yield and facilitating the production, May be prepared in the form of a powder. Accordingly, it can be changed into various types of formulations and can be used for various purposes. For example, the above sulfuric acid fermentation metabolite can be used as a cosmetic composition, a pharmaceutical composition, a food composition, and the like.

1 is a flow chart schematically showing a step of manufacturing a method for manufacturing a yellowish fermented metabolite by liquid fermentation according to an embodiment of the present invention.

Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art can easily carry out the present invention. Embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art. Therefore, the embodiments of the present invention can be modified into various other forms, and the scope of the present invention is not limited to the following embodiments.

Throughout the description of the present invention, when a component is referred to as " comprising ", it means that it can include other components as well, without excluding other components unless specifically stated otherwise.

In the specification of the present invention, it is to be understood that when a step is located "on" or "before" another step, this is not only the case where a step is in a direct time series relationship with another step, And may have the same rights as in the case of an indirect temporal relationship in which the temporal order of the two phases can be changed.

The terms " about "," substantially ", etc. used to the extent that they are used throughout the specification of the present invention are used in their numerical values or in close proximity to their numerical values when the manufacturing and material tolerances inherent in the meanings mentioned are presented, Is used to prevent unauthorized exploitation by an unscrupulous infringer of precise or absolute disclosures in order to aid in the understanding of the disclosure. The term " step " or " step of ~ " used throughout the specification does not mean " step for.

The present invention relates to a fermented fermented lean body by liquid fermentation and a method of producing the fermented fermented metabolite by liquid fermentation. A fermentation substrate preparation step; Microorganism pre-culture step; Liquid fermentation step; And a step of obtaining a yellowish fermented metabolite, wherein the yellowish fermented metabolite by liquid fermentation according to an embodiment of the present invention is prepared by the above method.

The fermented metabolism of sulfuric acid by liquid fermentation according to the present invention can be understood as a state in which a useful substance of woody wood is low-molecularized by microorganisms and a secondary metabolite resulting from fermentation into microorganisms. According to one embodiment of the present invention, the specific form of the yellowish fermentation metabolite can be determined according to the processing method of the fermented liquid fermented by the liquid fermentation according to the above-described production method, and the form of the concentrated yellowish lye fermentation broth can be determined, May be in the form of a solid powder that has undergone a process.

The fermented fermented metabolite according to one embodiment of the present invention not only has high yield by liquid fermentation, but also has excellent antioxidant ability, bioavailability and physiological activity.

The major component of Hwangchil is Sesquiterpene, which contains water, gum, alcohol and esters. It is known that Hwangchil, which is a sap extracted from Hwangchilbok tree, has a remarkable pharmacological effect and a delicate environment.

In recent years, nutritional value, pharmacological value, and therapeutic value have been recognized by the scientific research of Hwangchu. It has been reported that yellowtail is effective for many diseases such as improving blood circulation (cholesterol, blood pressure), antioxidant (anti-aging), liver function improvement (hangover, fatigue recovery, detoxification), immunity enhancement and regeneration.

However, the active ingredient of Hwigae-gil is slow in absorbing human body and has limited use in human body. There have been attempts to utilize the active ingredient of Hwangchujang in the human body. However, there has been a problem that it is difficult to process and mass-produce Hwangchilchu and by-products in large quantities due to difficulty in changing formulations in various forms.

However, since the fermentation metabolic body according to one embodiment of the present invention uses a method such as liquid fermentation and pre-culture of microorganisms, the fermentation time is shortened, the yield thereof is improved and the production is easy, and the fermented fermented metabolite is a concentrate Or in the form of a dry powder. Accordingly, it can be changed into various types of formulations and can be used for various purposes. For example, the above sulfuric acid fermentation metabolite can be used as a cosmetic composition, a pharmaceutical composition, a food composition, and the like.

When the sulfuric acid fermentation metabolite is used in a cosmetic composition, it is preferable to use a commonly used ingredient such as purified water, a moisturizer, an oil, a surfactant, a high alcohol, a thickener, a chelating agent, a colorant, a fatty acid, an antioxidant, Such as solutions, suspensions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations, and the like, And spray or the like.

In addition, when used in a pharmaceutical composition, it can be formulated using a pharmaceutically acceptable carrier and / or excipient, and can be mixed with a lubricant, wetting agent, sweetening agent, flavoring agent, emulsifying agent, suspending agent, preservative and the like.

In addition, when used as a food composition, it can be mixed with components such as protein, carbohydrate, fat, nutrients, seasoning and flavor, which are conventionally added in food production, and can be formulated into various forms.

Hereinafter, each step of the method for producing a yellowish fermented metabolite by liquid fermentation according to an embodiment of the present invention will be described in detail, and the yellowish fermented metabolite by liquid fermentation according to one embodiment of the present invention will be described more concretely And can be understood.

The method for preparing a fermented yellowish fermented metabolite by liquid fermentation according to an embodiment of the present invention comprises the steps of: pretreatment of a fermented raw material for crushing and cutting crudaceae leaves and stems; Preparing a fermentation substrate to produce a hot water extract of the pretreated greasy raw material; A microorganism pre-culture step which is carried out simultaneously with or before and after the pre-treatment step of the above-described greasy raw material and in which a fermenting microorganism is cultured; A liquid fermentation step of inoculating and cultivating the microorganism in the yellowish hot water extract solution; And sterilizing, filtering and concentrating the liquid fermentation broth to obtain a fermented fermented broth.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a flow chart schematically showing a step of manufacturing a fermented yellowish fermented metabolite by liquid fermentation according to an embodiment of the present invention; FIG.

First, the pretreatment step of the crudeness raw material for washing, crushing and cutting crudiferous leaves and stems can be carried out (S ₁ ).

According to one embodiment of the present invention, the pretreatment step of the raw material for yellowing is performed as a pre-stage for performing the yellowing extract, and the washing, crushing and cutting of the yellowing raw material is performed to facilitate the extraction.

More specifically, after washing and drying the yellowish leaves and stems, the stems are crushed and cut to 1 to 2 mm, and the leaves can be crushed and cut to a size of 2 to 3 cm. According to an embodiment of the present invention, the water content may be about 10% in washing and drying the leaves and stems.

Next, a step of preparing a fermentation substrate for producing a hot-water extract of the pretreated roughage material can be performed (S ₂ ).

According to one embodiment of the present invention, the litter leaf and the stem can be extracted by a hot water extraction method. More specifically, the pre-treated Yellowish leaves and stems are added 10 times with water and reflux-extracted at 90 to 95 캜 for 8 to 10 hours. The extract can be filtered and then concentrated. But is not limited to, it can be concentrated at 60 ° C or less in a reduced pressure concentrator, and the solids content can be 1 to 10 Brix. The concentrated extract can be sterilized at 110 to 125 DEG C for 10 to 20 minutes.

The extract of Huangchil has antimicrobial activity and it may be difficult to grow microorganisms. Therefore, it is necessary to control the solid content of the extract to provide an environment in which the fermenting microorganism can grow. According to one embodiment of the present invention, the solid content of the extract can be 1 to 10 bricks. However, the content of the solid content may be slightly different depending on the kind of the fermenting microorganism. More specifically, Aspergillus oryzae , Saccharomyces cerevisiae , Phellinus When using baumii , Ganoderma lucidum , the solid content may be 1 to 10 Bricks, and Bacillus In the case of using subtilis , the content of the solid may be 5 to 10 Bricks, and Lactobacillus If plantarum is used, it may be 1 to 5 bricks.

In addition, a microorganism pre-culture step may be carried out (S ₃ ) in which a fermenting microorganism is selectively cultured at the same time as or before or after the pretreatment step of the above-described yellowing raw material.

In one embodiment of the present invention, microorganisms may be cultured and then used for the fermentation of the Phellinus linteus extract. The strains used for liquid fermentation were edible fungi Aspergillus oryzae , the brewer's yeast Saccharomyces cerevisiae , mushroom strain Phellinus baumii and Ganoderma lucidum , Bacillus subtilis , lactic acid bacteria Lactobacillus plantarum .

All of the above strains can be pre-cultured prior to the main culture and can be cultured after pure culture. The microbial pre-cultivation can shorten the liquid fermentation time.

But are not limited to, edible fungus Aspergillus The oryzae are cultured in PDA medium at 30 ° C for 5 days, and the spores formed can be harvested with 0.1% physiological saline, then seeded again on the PDB medium, and cultured for 5 days at 30 ° C and 120 rpm.

The brewer's yeast Saccharomyces The cerevisiae can be inoculated on a YM broth medium with a single colony formed by solid culturing in an agar medium at 30 占 폚 for 3 days and cultured at 30 占 폚 for 2 days with shaking at 120 rpm.

Mushroom myceliumPhellinus baumiiWowGanoderma lucidum(1 × 1 cm, 3 pieces) formed by culturing in a PDB medium at 25 ° C. for 20 days can be used as a liquid seed bacterium by inoculating the PDB medium and culturing at 25 ° C. at 130 rpm for 10 days.

Bacillus Subtilis can be cultured in NB medium at 37 ° C for 2 days at 37 ° C and 120 rpm in a single colony cultured in NA medium for 2 days.

Lactobacillus Plantarum can be used as a liquid seed after colonies formed by first culturing in MRS solid medium are cultured again in MRS broth at 37 ° C for 24 hours.

Next, a liquid fermentation step in which the pre-cultured microorganism is inoculated to the fermentation substrate and cultured can be performed (S ₄ ).

According to an embodiment of the present invention, the fermentation substrate may be a yellowish hot water extract, and the liquid fermentation microorganism may be inoculated after sterilizing the yellowish hot water extract. The solid content of the Welsh Hot Water Extract may be 1 to 10 Bricks, and the microorganism culture liquid may be inoculated into the hot water extract. More specifically, the concentration of the microorganism culture solution may be 10%, and may be shake cultured or stationary culture at 25 ° C to 40 ° C for 5 days to 20 days depending on the kind of the microorganism. But are not limited to, for example, Aspergillus with oryzae Saccharomyces S. cerevisiae was cultured at 30 ° C for 5 days with shaking at 120 rpm, Phellinus baumiI and Ganoderma luidum was incubated for 20 days at 25 ° C and 130 rpm for 20 days, Bacillus subtilis was shake cultured at 40 ° C and 120 rpm for 5 days, Lactobacillus Plantarum can be cultured for 5 days at 37 ° C.

According to one embodiment of the present invention, the fermentation microorganism can be pre-cultured before inoculation in the pyrochlore extract to reduce the time of liquid fermentation. In addition, the yield can be greatly improved by being used as a fermentation substrate of a hot-water extract solution.

In addition, the yellowish fermented metabolite according to one embodiment of the present invention may contain various secondary metabolites depending on the kind of the microorganism, as well as the useful substance of the sulfuric acid. According to one embodiment of the present invention, Bacillus Subtilis and Lactobacillus plantarum, which are lactic acid bacteria, may have better antioxidant capacity.

Further, according to one embodiment of the present invention, a juice may be prepared, and a juice may be further added to the fermentation substrate (S ₅ ).

The juice may be provided as a source of sugar in the liquid fermentation step. When the fermentation substrate contains a juice, the fermentation metabolite may be different from that of the fermentation substrate.

According to one embodiment of the present invention, the juice may be prepared by adding water twice to a boat and refluxing the mixture at 90 to 100 ° C for 5 hours, followed by filtration and concentration under reduced pressure (60 ° C or less). The solids content of the juice may be controlled to be between 1 and 10 Brix and may be sterilized at 121 占 폚 for 15 minutes prior to inoculation with the microorganism.

The juice may be mixed with the same amount of the same amount of solid matter as that of the yellowish hot-water extract. Even when the juice is further contained in the fermentation substrate, 10% of the pure fermentation microorganism solution can be inoculated as described above, and the fermentation can proceed by shaking culture or stationary culture. The specific incubation temperature and period are as described above, and a description thereof will be omitted.

Next, the liquid fermentation broth can be sterilized, filtered and concentrated to obtain a fermented fermented broth (S ₆ ).

A fermented fermentation broth can be obtained from the fermented fermented fermentation broth.

More specifically, the above-described fermentation broth can be sterilized at 100 to 121 ° C for 15 to 20 minutes, and then filtered and concentrated under reduced pressure (60 ° C or less) to be used as a concentrate or as a solid powder. The concentrate may be used as a concentrate of 10 to 60 Brix, and the concentrate may be prepared as a solid powder by lyophilization and spray drying.

The sulfuric acid fermented metabolite may be used in a cosmetic composition, a pharmaceutical composition, a food composition, etc., and may be processed into a concentrate or a solid powder according to the intended use and formulation.

The fermented hydrogen peroxide metabolism is a metabolite resulting from the fermentation of microorganisms, and its ingredients and structure can not be clearly specified. However, the useful substance of sulfurylcholine can be reduced in molecular weight to increase the absorption rate in the body, Especially, it has been proved that antioxidant effect is excellent.

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in more detail with reference to the following examples, but they should not be construed as limiting the scope of the present invention.

[Example]

[Preparation of sulfuric acid fermentation metabolite]

Burrows  Pretreatment of raw materials

The leaves and stems were washed and dried to a moisture content of about 10%. The stems were broken into 1 ~ 2mm and the leaves were broken into 2 ~ 3㎝.

Burrows Heat number  Preparation of Extracts

The water content was adjusted to 1, 5, and 10Brix by filtration and concentration under reduced pressure (60 ° C or lower) at 90 ° C to 95 ° C for 10 hours. And sterilized for 15 minutes.

Juice  Produce

Water was added to the pellet twice, and the mixture was refluxed at 90 to 100 ° C for 5 hours, filtered and concentrated under reduced pressure (60 ° C or less). The solid content was adjusted to 1, 5, and 10 Brix and sterilized at 121 ° C for 15 minutes.

Pure culture of liquid fermentation microorganisms

Aspergillus oryzae were cultured for 5 days at 30 ° C and 120 rpm in a PDB medium inoculated with spores obtained by pure culture in a PDA medium. Saccharomyces cerevisiae was two days incubation a single colony isolated from a YM agar YM liquid culture medium was inoculated in 30 ℃, 120rpm, Phellinus baumii and Ganoderma lucidum was cultured for 10 days at 25 ° C and 130 rpm by inoculating a single spore into PDB medium. Bacillus Subtilis was cultured in NB medium at 37 ℃ and 120 rpm for 2 days in single colony isolated from NA medium. Lactobacillus plantarum was cultured in MRS liquid medium at 37 ℃ for 1 day in a single colony formed in MRS solid medium.

Microbial inoculation and liquid culture

10% of the liquid fermented microbial broth was inoculated in the same amount of the same amount of 10grix of Huangchun hot water extract (10Brix), Huangchu hot water extract (10Brix) and juice (10Brix), and then Aspergillus oryzae and Saccharomyces cerevisiae were cultured at 30 ° C for 5 days with shaking at 120 rpm, Phellinus baumii and Ganoderma lucidum is 25 ℃, 20 days with shaking culture at 130rpm, Bacillus Subtilis was incubated at 40 ℃ for 5 days with shaking at 120 rpm and Lactobacillus plantarum was incubated at 37 ℃ for 5 days.

Fermentation Metabolite  purchase

The incubated fermented broth was sterilized at 100 to 121 ° C for 15 to 20 minutes, filtered and concentrated under reduced pressure (60 ° C or less) to obtain a concentrate having a concentration of 10 to 60 Brix.

[evaluation]

One. Burrows  Depending on the solids concentration of the extract, Proliferative ability

The growth of six strains was investigated according to the solids content of Hwangchil hot - water extract because of its antimicrobial activity. After 1% inoculation of pure culture broth of each strain into the broth extract of 1, 5 and 10 Brix, which was prepared as above, the number of colony formed in the solid medium after incubation under the same condition as the pure culture of each strain was compared with the control The results are shown in the following Table 1 as "- (non-viability), + (poor growth), ++ (normal growth) and +++ (excellent growth).

Strain Concentration / Brix One 5 10 Aspergillus oryzae +++ +++ +++ Saccharomyces cerevisiae +++ +++ +++ Phellinus baumii +++ +++ +++ Ganoderma lucidum +++ +++ +++ Bacillus subtilis ++ +++ +++ Lactobacillus plantarum + +++ -

As shown in Table 1, Lactobacillus plantarum was not only the growth in 10Brix concentration, five strains the growth was good at 10Brix concentration. Bacillus subtilis showed better growth at 5 and 10Brix than 1Brix. In other words, all six strains selected in this experiment were judged to be usable for fermentation metabolism of Huangchil extract.

In addition, the same amount of juice (1, 5, 10 Brix) of 1, 5, and 10 Brix of Huangchun hot-water extract was mixed with the same amount, and the degree of growth of the six strains was examined. The results are shown in Table 2 below , And the result almost coincided with the growth pattern in the yellowish hot water extract.

Strain Concentration / Brix One 5 10 Aspergillus oryzae +++ +++ +++ Saccharomyces cerevisiae +++ +++ +++ Phellinus baumii +++ +++ +++ Ganoderma lucidum +++ +++ +++ Bacillus subtilis ++ +++ +++ Lactobacillus plantarum + +++ -

2. Yellowish fermentation Metabolite  Antioxidant experiment

To the mixture of the same amount of Hwangchil hot water extract (10Brix), Huangchil hot water extract (10Brix) and juice (10Brix), Aspergillus oryzae (Hwang Kuk Kyun), Phellinus baumii , Ganoderma lucidun , Saccharomyces, cerevisiae (brewer's yeast), Bacillus subtilis (bacillus subtilis ), Lactobacillus plantarum (lactic acid bacterium), 10% pure culture broth of 10 strains of plantarum (lactobacillus) were inoculated, respectively, for 30 days at 30 ℃ for 5 days, shaking culture at 25 ℃ for 3 days, shaking culture at 30 ℃ for 5 days, After incubation for 5 days at 37 ° C, lactic acid bacteria were cultured at 37 ° C for 5 days, filtered, concentrated, and lyophilized.

Free radical scavenging activity and measured by the reducing power of the sample on the electron donating effect on 1,1-diphenyl-2-picrylhydrazyl (DPPH, WaKo) radicals. To 1 ml of each concentration sample, 1 ml of 500 μM DPPH solution was added and stirred, followed by reaction at 37 ° C for 30 minutes in a dark place. The absorbance of the reaction solution was measured at 517 nm, and the radical scavenging activity was calculated in the same manner as described below.

Free radical scavenging activity (%) = (1-A / B) x 100

A: sample addition absorbance, B: sample no addition absorbance

Fermentation
temperament Fermentation metabolism Sieve concentration
(占 퐂 / mg) DPPH Scatters (%) Control group A.
oryzae S.
cerevisiae P.
baumii G.
lucidun B.
subtilis L.
plantarum Burrows  Heat Water extract (10 Brix ) 0 0.21 0.21 0.21 0.21 0.21 0.21 0.21 50 7.56 11.27 13.46 7.58 7.85 11.52 17.30 100 14.92 22.74 26.31 13.33 15.52 34.92 31.69 250 39.86 55.72 58.97 28.13 33.96 65.73 67.41 500 64.36 84.81 86.14 47.19 64.86 97.18 99.75 750 84.30 86.81 88.84 66.39 79.58 99.158 100.98 1000 86.28 90.15 91.25 72.16 86.11 99.19 100.73 Burrows  Heat Water extraction Water and ship Juice blend Solution 10 Brix ) 0 0.21 0.21 0.21 0.21 0.21 0.21 0.21 50 5.29 8.97 7.96 16.51 11.37 6.66 13.27 100 9.13 15.85 15.72 28.22 21.83 16.88 20.69 250 22.45 40.82 33.17 63.5 46.41 34.44 38.64 500 43.12 59.12 64.11 85.10 84.04 63.44 66.52 750 60.32 83.98 76.43 85.43 85.50 87.90 77.97 1000 76.42 85.92 84.21 85.63 86.05 94.94 92.64

The DPPH scavenging activity was higher in fermented metabolites prepared from fermented hot water extract than fermented hot water extract. In general, the DPPH scavenging activity was higher in the fermented broth of lactic acid bacteria and the fermented broth of Chungkookjang than that of mushroom, hwanggukyun and yeast.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, but many variations and modifications may be made without departing from the scope of the present invention. It is evident that many variations are possible by those skilled in the art. It will be apparent to those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. something to do.

Claims (11)

Pretreatment of raw materials for crudeness to crush and cut crustacean leaves and stems;
Preparing a fermentation substrate to produce a hot water extract of the pretreated greasy raw material;
A microorganism pre-culture step in which the microorganism is selectively cultivated simultaneously with or before or after the pretreatment step of the above-described greenhouse material;
A liquid fermentation step of inoculating and culturing the microorganism in the fermentation substrate; And
Sterilizing, filtering and concentrating the fermentation broth obtained in the liquid fermentation step to obtain a fermented fermented broth;
≪ / RTI > wherein the fermentation broth is a fermented fermented broth.
The method according to claim 1,
Wherein the solid content of the hydrothermal extract in the fermentation step is 1 to 10 Brix.
The method according to claim 1,
The microorganism is an edible mold Aspergillus oryzae , Saccharomyces cerevisiae which is a brewer's yeast, Phellinus mushroom strain baumii and Ganoderma lucidum , Bacillus subtilis , or Lactobacillus plantarum , which is a lactic acid bacterium .
The method according to claim 1,
Wherein the microorganism pre-culturing step is performed at 25 to 37 DEG C for 2 to 10 days.
The method according to claim 1,
The microorganism is Bacillus subtilis , The solid content of the Welsh Hot Water Extract is 5 to 10 Brix or the microorganism is Lactobacillus plantarum , and the content of solid content of the extract is 1 to 5 Brix.
The method according to claim 1,
Wherein the liquid fermentation step further comprises adding a juice to the fermentation substrate.
The method according to claim 6,
Wherein the solid content of the juice is 1 to 10 Brix. ≪ RTI ID = 0.0 > 8. < / RTI >
The method according to claim 1,
Wherein the liquid culture is carried out at 25 占 폚 to 40 占 폚 for 5 to 20 days inoculated with 10% of the microorganism culture solution.
The method according to claim 1,
Wherein the sulfuric acid fermented metabolite is processed in the form of a concentrate or powder.
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