KR20220045303A - Lactobacillus acidophilus BLA-9 with biological conversion activity of saponin, a method of preparation for fermented red ginseng extracts having increased content of ginsenoside compound K using the same, fermented red ginseng extracts using thereby and health-functional food composition containing the same - Google Patents

Abstract

The present invention is a Lactobacillus acidophilus BLA-9 strain having bioconversion ability for saponin, a method for preparing a fermented red ginseng extract with an increased content of compound K using the same, a fermented red ginseng extract thereby, and a health functional food composition including the same. The Lactobacillus acidophilus BLA-9 strain (accession number: KCTC18848P) having bioconversion ability to saponin has excellent ginsenoside bioconversion ability. When the strain is inoculated into a red ginseng extract and fermented, a fermented red ginseng extract having a high content of ginsenoside F2 and compound K can be obtained. In addition, according to the preparing method of the present invention, the production efficiency of the compound K is remarkably increased, and a fermented red ginseng extract with a further increased content of compound K can be obtained. Since the fermented red ginseng extract of the present invention contains a large amount of the compound K which is easily absorbed by the body, the fermented red ginseng extract is very useful as a healthy functional food or food material.

Description

Lactobacillus acidophilus BLA-9 strain having bioconversion ability for saponin, method for producing fermented red ginseng extract with increased content of compound K using the same, fermented red ginseng extract according thereto, and health functional food composition comprising the same {Lactobacillus acidophilus BLA-9 with biological conversion activity of saponin, a method of preparation for fermented red ginseng extracts having increased content of ginsenoside compound K using the same, fermented red ginseng extracts using thereby and health-functional food composition containing the same}

The present invention relates to a Lactobacillus acidophilus BLA-9 strain having bioconversion ability for saponins, a method for producing a fermented red ginseng extract with an increased content of compound K using the same, a fermented red ginseng extract according to the method, and a health functional food comprising the same to the composition.

Ginseng refers to the root of plants of the genus Araliaceae. It is called Korean ginseng (Korean Peninsula), American ginseng (USA and Canada), Jeonchilsam (China), Bamboo Jeolsam (Japan), etc. depending on the cultivation area. Fresh ginseng, red ginseng, It is divided into white ginseng, sugar ginseng, and balsam ginseng.

Korean ginseng (Panax ginseng C.A. Meyer) has been widely used for health promotion in Korea, China, and Japan since the past. It is known that the active ingredients of ginseng differ significantly depending on the type of ginseng, root part, harvest year, harvest time, and manufacturing method. it is known In addition, it is known that there is a difference in the content of saponin or types of ginsenosides between white ginseng dried fresh ginseng as it is and red ginseng steamed and dried with steam.

'Red ginseng' as referred to in health functional food uses ginseng as a raw material and steams or other methods to steam and dry undried fresh ginseng. It should be powdered or extracted with water or alcohol, and concentrated or fermented to make it suitable for food, and the sum of ginsenosides Rg1 and Rb1 should be controlled to be 0.8 mg/g to 34 mg/g.

A known effective ingredient of ginseng and red ginseng is saponin. Saponins of ginseng and red ginseng are a type of glycoside and have a unique chemical structure different from saponins found in other plants, and their efficacy is also significantly different. Saponins of ginseng and red ginseng are called 'Ginsenoside' in the sense of ginseng glycoside to distinguish them from other plant-based saponins.

Ginsenoside, an active ingredient of ginseng and red ginseng, is a triterpenoid dammarane skeleton with glucose, arabinose, xylose and rhamnose. Currently, more than 100 types have been isolated as glycosides in which one or several sugars are bound (J Ginseng Res 41 (2017), 435-443). Six types of ginsenosides including , Re and Rg1 account for 90% of the total ginsenosides.

Compound K (C-K: compound K, 20-O-β-D-glucopyanosyl 20(S)-protopanasadiol), one of a small amount of low-molecular-weight ginsenosides produced by hydrolysis of high-molecular ginsenosides, is a basic skeleton of aglycone ( aglycon) is one of the PPD-based low-molecular-weight ginsenosides in which one glucose is bound to C-20, and ginsenosides Rb1, Rb2, and Rc are conversion products converted by intestinal microorganisms (Akoa et al., 1998, Akao et al., 1998, Hasegawa et al., 1997; Bae et al., 2000).

The chemical structure of the compound K is shown in Formula 1 below.

[Formula 1]

According to a study by Akao et al. (1998), when ginsenoside Rb1 was administered to rats, it was reported that compound K was detected in plasma for 24 hours after ginsenoside Rb1 administration, and hardly metabolized in the stomach. Karikura et al. (1991) investigated the metabolites of ginsenoside Rb2 in the stomach of rats. Considering that ginsenoside Rb2 is hardly metabolized except for the production of peroxide, ginsenoside is It is judged to be metabolized by intestinal microbes without being metabolized in the digestive system of In addition, according to Professor Dong-hyun Kim's research, the conversion ability of ginsenoside Rb1 was confirmed using the intestinal microbes of 98 Koreans, and as a result, 20% had no or significantly low ginsenoside metabolism ability, and only about 60% of microorganisms capable of converting ginsenoside. It has been reported that many people do not metabolize ginsenosides efficiently. According to these reports, even if ginseng or red ginseng is taken due to the difference in the composition of the intestinal microbes and the enzyme activity of the intestinal microbes for each individual, there are differences in efficacy among individuals, and to reduce this difference, intake of fermented red ginseng is recommended.

In the past, products were manufactured using only the traditional unit processes of red ginseng manufacturing, such as steaming, drying, and shaping. Treatment, enzymatic treatment, and methods using microorganisms are being studied in various ways.

Among them, production by intestinal microorganisms, Bifidobacterium , fungi, or various microorganisms is known as a method for producing compound K. However, the method requires culturing in anaerobic conditions and a long culture time, and the production yield is extremely low. The production of fermented red ginseng containing a large amount of specific low-molecular-weight ginsenoside compound K through fermentation using edible enzymes and lactic acid bacteria because of low yield and edible difficulties due to the production of by-products development is required.

In the process of researching to develop fermented red ginseng with an increased content of compound K, the present inventors selected enzymes and lactic acid bacteria with excellent ginsenoside bioconversion ability, and in fermented red ginseng sequentially fermented using the selected enzymes and strains The present invention was completed by confirming that the content of Compound K was greatly increased as well as that the ginsenoside index was excellent.

KR 10-2010-0054428 A KR 10-1409761 B

An object to be solved by the present invention is to provide a Lactobacillus acidophilus BLA-9 strain having bioconversion ability for saponins.

Another object of the present invention is to provide a method for producing a fermented red ginseng extract having an increased content of compound K and excellent ginsenoside index.

Another object of the present invention is to provide a fermented red ginseng extract prepared according to the above method, the compound K content is increased, and the ginsenoside index is excellent.

Another object of the present invention is to provide a health functional food composition comprising the fermented red ginseng extract as an active ingredient.

In order to achieve the above object, the present invention provides a Lactobacillus acidophilus BLA-9 strain (Accession No.: KCTC18848P) having bioconversion ability for saponins.

In addition, the present invention comprises the steps of (1) preparing a red ginseng extract; (2) adding a complex enzyme of cellulase, β-glucosidase and hemicellulase to the red ginseng extract for enzymatic reaction; and (3) inoculating the Lactobacillus acidophilus BLA-9 strain (Accession No.: KCTC18848P) into the enzyme-treated red ginseng extract and then fermenting it; A method for producing a fermented red ginseng extract is provided.

According to an embodiment of the present invention, the red ginseng extract may be extracted with water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.

According to an embodiment of the present invention, the red ginseng extract may have a solid content of 3 to 50% by weight.

According to an embodiment of the present invention, the complex enzyme of step (2) may be an enzyme derived from Aspergillus niger .

According to an embodiment of the present invention, the complex enzyme of step (2) may be Sumizyme AC.

According to an embodiment of the present invention, the complex enzyme of step (2) may be added in an amount of 0.5 to 50 parts by weight based on 100 parts by weight of the red ginseng extract.

According to an embodiment of the present invention, the enzymatic reaction of step (2) may be performed at 50 to 65 ℃, pH 3.5 to 5.0 for 5 to 60 hours.

According to an embodiment of the present invention, the present invention may not perform the enzyme inactivation step between the steps (2) and (3).

According to an embodiment of the present invention, the fermentation of step (3) may be performed at 30 to 50 ℃ for 5 to 60 hours.

According to an embodiment of the present invention, in the fermented red ginseng extract, the sum of Rb1, Rg1 and Rg3 is at least 7.0 mg/g or more, the content of F2 is at least 5 mg/g or more, and the content of compound K is at least 4.0 mg It can be greater than /g.

According to one embodiment of the present invention, the content of compound K (mg/g) contained in the fermented red ginseng extract after the fermentation is divided by the content (mg/g) of Rb1 contained in the red ginseng extract before the enzyme treatment. The production efficiency may be at least 0.2 or higher.

In addition, the present invention is prepared according to the above method, wherein the sum of Rb1, Rg1 and Rg3 is at least 7.0 mg/g or more, the content of F2 is at least 5 mg/g or more, and the content of compound K is at least 4.0 mg/g It provides a fermented red ginseng extract, characterized in that the above.

In addition, the present invention provides a health functional food composition comprising the fermented red ginseng extract as an active ingredient.

Lactobacillus acidophilus of the present invention ( Lactobacillus acidophilus ) BLA-9 strain (Accession No.: KCTC18848P) has excellent ginsenoside bioconversion ability. When the red ginseng extract is inoculated with the strain and fermented, a fermented red ginseng extract having a high content of ginsenoside F2 and compound K can be obtained.

In addition, according to the production method of the present invention, the production efficiency of compound K is significantly increased, the content of compound K is further increased, and a fermented red ginseng extract having excellent ginsenoside index can be obtained.

The fermented red ginseng extract of the present invention contains a large amount of compound K, which is easily absorbed in the body, and has an excellent ginsenoside index, so it is very useful as a health functional food or food material.

1 is a molecular diagram showing the 16S rRNA gene sequence analysis of the Lactobacillus acidophilus BLA-9 strain of the present invention.
2 is a flowchart illustrating a method for preparing a fermented red ginseng extract according to an embodiment of the present invention.
Figure 3 is a schematic diagram simply showing the process of conversion of PPD-type ginsenoside to compound K (CK) by bioconversion.

Hereinafter, the present invention will be described in detail.

In the present specification, "ginsenoside indicator" refers to ① the content of compound K, ② the sum of Rg1, Rb1 and Rg3 (Rg1+Rb1+Rg3), ③ the content of compound K contained in the fermented red ginseng extract, the red ginseng extract before bioconversion Production efficiency of compound K, which is the value divided by the content of Rb1 contained in (CK/Rb1 before fermentation), ④ the content of compound K in the fermented red ginseng extract after fermentation divided by the content of Rb1 (CK/Rb1), ⑤ fermentation The value obtained by dividing the content of compound K in the post-fermented red ginseng extract by the content of Rg3 (CK/Rg3), ⑥ The value obtained by dividing the content of compound K in the fermented red ginseng extract after fermentation by the sum of Rg1, Rb1 and Rg3 (CK /(Rg1+Rb1+Rg3)), and ⑦ the content of compound K contained in the fermented red ginseng extract after fermentation divided by the sum of Rg1, Rb1, Rg3 and compound K (CK/(Rg1+Rb1+Rg3+CK) ), which may be an indicator of the production efficiency of compound K and/or the quality of the fermented red ginseng extract. The fermented red ginseng extract of the present invention in which the above indicators exceed a certain level has a large sum of Rg1, Rb1 and Rg3 and a high content of compound K, so the quality of the fermented red ginseng extract is very good.

According to one embodiment of the present invention, the present invention provides a Lactobacillus acidophilus BLA-9 strain (Accession No.: KCTC18848P) having bioconversion ability for saponins.

As a result of analyzing the 16S rRNA nucleotide sequence for microbial identification and classification of the BLA-9 strain, which is a kimchi-derived lactic acid bacterium, stored at the headquarters (BTC) research center through the following examples, it was found to have the nucleic acid sequence of SEQ ID NO: 1. . As a result of analyzing the nucleic acid sequence of the strain, it was found to be a strain belonging to Lactobacillus acidophilus , and the nucleic acid sequence of the strain was requested to Macrogen to confirm homology. As a result, Lactobacillus ash It was confirmed that it was 99% identical to the Dophilus HE793099.1 standard strain (FIG. 1). In the nucleic acid sequence of SEQ ID NO: 1, 'n' refers to a base of 'a', 'g', 'c', 't or u', or 'unknown or other'.

Therefore, the microorganism of the present invention having the 16S rRNA nucleotide sequence of SEQ ID NO: 1 was named Lactobacillus acidophilus BLA-9 strain, and as of September 21, 2020, the Korea Research Institute of Bioscience and Biotechnology Biological Resources Center It was deposited (Accession No.: KCTC18848P).

The Lactobacillus acidophilus BLA-9 strain of the present invention has bioconversion ability for saponins, in particular, converts saponins into ginsenosides, and converts polymeric ginsenosides into compound K with increased digestion and absorption in the human body. Therefore, it is very useful for the production of fermented red ginseng.

According to another embodiment of the present invention, the present invention comprises the steps of (1) preparing a red ginseng extract; (2) adding a complex enzyme of cellulase, β-glucosidase and hemicellulase to the red ginseng extract for enzymatic reaction; and (3) inoculating the Lactobacillus acidophilus BLA-9 strain (Accession No.: KCTC18848P) to the enzyme-treated red ginseng extract and then fermenting it; A method for producing a fermented red ginseng extract is provided.

In the production method of the present invention, the "enzyme-treated red ginseng extract" is a product obtained by reacting with the complex enzyme using the "red ginseng extract" of step (2) as a substrate. In addition, the "fermented red ginseng extract" is a product obtained by inoculating and fermenting Lactobacillus acidophilus BLA-9 using the "enzyme-treated red ginseng extract" of step (3) as a substrate.

In this specification, for convenience of description, it is described that the method of the present invention is applied to red ginseng. However, the present invention can be applied not only to red ginseng, but also to all ginseng (eg, fresh ginseng, white ginseng and white ginseng) processed in various forms, and it is clear to those skilled in the art that these are included in the scope of the present invention.

According to an embodiment of the present invention, the red ginseng is manufactured by using ginseng as a raw material, but fresh ginseng that has not been dried is steamed for a long time by steam or other methods and dried to less than 15% moisture, and 4 or 6 year old red ginseng root And/or may be red ginseng, preferably red ginseng root and red rice ginseng in a weight ratio of 50 to 90: 10 to 50, preferably 70 to 80: may be a 6-year-old red ginseng composed of a weight ratio of 20 to 30.

According to one embodiment of the present invention, the red ginseng may be a red ginseng powder obtained by grinding red ginseng, and it is preferable to use the red ginseng powder because the surface area is widened, so that the active substance of the red ginseng can be easily extracted.

The red ginseng is mixed with an extraction solvent in a weight ratio of 1: 1 to 20, preferably 1: 2 to 15, more preferably 1: 3 to 10, at 20 to 100 ° C. for 1 to 15 hours, preferably After extracting single or multiple times with stirring for 2 to 10 hours, the extract can be prepared by concentration under reduced pressure. When the weight ratio of the red ginseng and the extraction solvent is out of the above range, the active ingredient of the red ginseng may be extracted in a small amount in the extract. In addition, the red ginseng extract has a solid content of 0.1 to 70% by weight, preferably 1 to 60% by weight, more preferably 3 to 50% by weight, more preferably 2 to 20% by weight, more preferably 3 to It is preferred to concentrate to 10% by weight, more preferably 3 to 8% by weight.

In the present invention, the red ginseng extract may be extracted with water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. The lower alcohol may include 30 to 80% methanol, ethanol, butanol or propanol.

The extraction solvent is not particularly limited, but the extract extracted with an aqueous solution of 30 to 80%, preferably 40 to 80%, of ethanol preferably acts on the extraction of the active ingredient of red ginseng.

The red ginseng extract of the present invention has a saponin content of 30 to 300 mg/g, preferably 50 to 250 mg/g, more preferably 70 to 200 mg/g, more preferably 100 to 130 mg/g, based on the solid content. g, more preferably 105 to 120 mg/g, more preferably 108 to 110 mg/g.

The term "extract" used while referring to red ginseng in this specification includes not only the crude extract obtained by treating the extraction solvent, but also the processed product of the red ginseng extract. For example, the red ginseng extract may be prepared in a powder state by an additional process such as distillation under reduced pressure and freeze-drying or spray-drying.

High molecular weight ginsenoside, which accounts for more than 90% of ginsenoside, has a very low absorption rate in vivo due to its large size. Therefore, in order to increase the medicinal efficacy of ginsenosides, a process of converting high molecular weight ginsenosides into low molecular weight ginsenosides with relatively good absorption and superior medicinal efficacy is required. That is, the conversion process for removing glucose is required for polymer ginsenosides to effectively exhibit physiological activity in vivo.

To this end, the present inventors selected enzymes and lactic acid bacteria with excellent bioconversion ability for saponins, and by using the selected enzymes and lactic acid bacteria to convert high molecular weight ginsenosides in the red ginseng extract into low molecular weight ginsenosides, the content of compound K was increased. An enhanced fermented red ginseng extract was prepared.

The enzyme for processing the red ginseng extract in step (2) may be a complex enzyme of cellulase, β-glucosidase and hemicellulase. The complex enzyme of step (2) may be an enzyme derived from Aspergillus niger , and specifically may be obtained from a culture of Aspergillus niger.

The complex enzyme of step (2) is Sumizyme AC (manufacturer: ShinNippon, enzyme type: cellulase, β-glucosidase and hemicellulase complex enzyme, enzyme-producing microorganism: Aspergillus niger ) desirable. The Sumizyme AC enzyme has an excellent effect of converting saponins into saponin metabolites, and when used with the Lactobacillus acidophilus BLA-9 strain of the present invention, the ginsenoside bioconversion rate is increased as well as sensory It shows an excellent effect.

The complex enzyme is 0.01 to 70 parts by weight, preferably 0.1 to 60 parts by weight, more preferably 0.5 to 50 parts by weight, more preferably 1 to 40 parts by weight, more preferably based on 100 parts by weight of the red ginseng extract. is preferably added in an amount of 5 to 30 parts by weight, more preferably 10 to 25 parts by weight, and still more preferably 15 to 25 parts by weight.

When the ratio of the complex enzyme to the red ginseng extract is out of the above range, there is a fear that the enzyme is deactivated.

In addition, the enzymatic reaction is 15 to 70 ℃, preferably 30 to 67 ℃, more preferably 40 to 65 ℃, more preferably 50 to 65 ℃, pH 2 to 7, preferably pH 2.5 to 6, More preferably at pH 3.5 to 5 for 5 minutes to 120 hours, preferably 1 to 90 hours, more preferably 5 to 60 hours, more preferably 20 to 48 hours, still more preferably 20 to 30 hours can be performed.

In performing the enzymatic reaction, when the reaction temperature and/or pH is out of the above range, there is a fear that the activity of the enzyme is inhibited.

In addition, in performing the enzymatic reaction, when the reaction time is out of the above range, the conversion rate to saponin metabolites compared to the reaction time may be significantly lowered.

The amount of the enzyme added, reaction temperature and reaction time are optimal conditions for ginsenoside conversion of red ginseng extract.

In the method for producing a fermented red ginseng extract of the present invention, the enzyme inactivation step may not be performed after the enzyme treatment in step (2). The production method of the present invention maximizes the bioconversion of ginsenosides in the red ginseng extract by performing the strain fermentation step of step (3) while maintaining the enzyme activity by not performing the inactivation step after the enzyme treatment of step (2). can do.

The strain for fermenting the red ginseng extract in step (3) may be Lactobacillus acidophilus BLA-9 strain (Accession No.: KCTC18848P).

After inoculating the Lactobacillus acidophilus BLA-9 strain into the red ginseng extract treated with the enzyme in step (2), 10 to 70 ℃, preferably 20 to 60 ℃, more preferably 30 to 50 ℃, 30 to 40°C, more preferably 34 to 39°C, more preferably 36 to 38°C for 30 minutes to 120 hours, preferably 2 to 90 hours, more preferably 5 to 60 hours, still more preferably 12 to 48 hours, more preferably 18 to 32 hours.

The reaction temperature and reaction time of the strain are optimal conditions for conversion of ginsenoside to compound K in the red ginseng extract.

The fermented red ginseng extract is heated at 85 to 95 ℃ for 10 to 30 minutes to inactivate the enzyme and Lactobacillus acidophilus BLA-9 strain.

The reason for inactivating the enzyme and the strain as described above is to prevent conversion of the desired compound K to protopanaxadiol (PPD).

The fermented red ginseng extract of the present invention prepared as described above has a total of Rb1, Rg1 and Rg3 of at least 7.0 mg/g or more, preferably 7.0 to 15.0 mg/g, more preferably 9.0 to 13.0 mg/g, and F2 The content is at least 5.0 mg/g or more, preferably 5.0 to 10.0 mg/g, more preferably 6.0 to 8.0 mg/g, and the content of compound K is at least 4.0 mg/g or more, preferably 4.0 to 7.0 mg /g, more preferably 4.5 to 6.5 mg/g.

The production efficiency of compound K, which is a value obtained by dividing the content (mg/g) of compound K contained in the fermented red ginseng extract after the fermentation by the content (mg/g) of Rb1 contained in the red ginseng extract before the enzyme treatment, is at least 0.2 or more, preferably may be 0.2 to 0.4, more preferably 0.2 to 0.3.

The biggest feature of the present invention is that the red ginseng extract is treated with Sumizyme AC as a mixed enzyme and then fermented with Lactobacillus acidophilus BLA-9 strain, so that the conversion rate of ginsenoside to compound K, that is, the production efficiency of compound K to increase significantly.

Further, according to another embodiment of the present invention, the present invention is prepared according to the above method, so that the sum of Rb1, Rg1 and Rg3 is at least 7.0 mg/g or more, preferably 7.0 to 15.0 mg/g, more preferably is 9.0 to 13.0 mg/g, the content of F2 is at least 5.0 mg/g or more, preferably 5.0 to 10.0 mg/g, more preferably 6.0 to 8.0 mg/g, and the content of compound K is at least 4.0 mg It provides a fermented red ginseng extract, characterized in that /g or more, preferably 4.0 to 7.0 mg/g, more preferably 4.5 to 6.5 mg/g.

Regarding the active ingredient content in the red ginseng extract, if you look at the red ginseng item and index component content part of "Standards and Specifications for Health Functional Foods" (Ministry of Food and Drug Safety Notice No. 2016-143, 2016.12.21.), "Ginsenoside Rg1 , containing 2.5-34 mg/g of Rb1 and Rg3 combined" is described as the reference value. In addition, if you look at the "requirements of the final product" of this document, functionally it meets the requirements of "improves immunity, improves fatigue, improves blood flow/memory through inhibition of platelet aggregation, antioxidant, and can help menopausal women's health" In addition, looking at the detailed items of the above document, in order to help "Immunity enhancement and fatigue improvement", the content of the indicator component (Rg1+Rb1+Rg3) is 3-80 mg/g, "platelet aggregation It is stated that the total amount of the index component should be 2.4-80 mg/g to help with "improving blood flow/memory through inhibition and antioxidant".

As can be seen through the following examples, when the red ginseng extract is subjected to enzyme treatment and/or fermentation, the ginsenoside is converted to compound K and the total of the indicator components is lowered. Although this is at least 4.0 mg/g or more, since the sum of the index components (Rg1, Rb1 and Rg3) is at least 7.0 mg/g or more, it has a very excellent ginsenoside composition.

In addition, according to another embodiment of the present invention, the present invention provides a health functional food composition comprising the fermented red ginseng extract as an active ingredient. The contents of the fermented red ginseng extract in the health functional food composition are the same as described above.

The health functional food composition may be provided in the form of powder, granule, tablet, capsule, syrup or beverage, and the health functional food composition is used together with other foods or food additives in addition to the active ingredient, and is suitable according to a conventional method. can be used The mixed amount of the active ingredient may be suitably determined according to the purpose of its use, for example, prevention, improvement or therapeutic treatment. According to one aspect, the fermented red ginseng extract is 0.001 to 80% by weight, preferably 0.001 to 70% by weight, more preferably 0.01 to 60% by weight, more preferably 0.01 to 30% by weight, more preferably 0.01 It may be included in an amount of from 10% by weight to 10% by weight.

The effective dose of the active ingredient contained in the health food composition is the type and content of the active ingredient and other ingredients contained in the composition, the type of formulation, the age, weight, general health status, sex and diet, administration time, administration It may be adjusted according to various factors including the route, duration of intake, and drugs used at the same time, but is not limited thereto.

Hereinafter, the present invention will be described in detail with reference to examples, but the present invention is not limited by the following examples.

<Example>

Example 1: Isolation and identification of strains

1-1: Isolation and identification of Lactobacillus acidophilus BLA-9 strain

The strain was kimchi-derived lactic acid bacteria, and the kimchi composition was diluted in sterile physiological saline, and the supernatant was diluted in a decimal method and spread on a BL plate medium containing 5% maltese. A single strain was isolated by taking a colony containing a ring unique to Lactobacillus from the BL plate medium, which is a lactic acid bacteria discrimination medium, and this strain was passaged and used in the experiment.

The strain was inoculated in MRS liquid medium, plate medium, and was used by culturing in a constant temperature incubator at 37°C.

After analyzing the beta-glucosidase (β-glucosidase) activity, ginsenoside conversion ability and growth ability in the red ginseng extract, one type of lactic acid bacteria showing the best conversion and growth ability was finally selected. First, each of the 97 types of lactic acid bacteria was inoculated in MRS liquid medium at 0.1% by weight and then cultured with shaking at 37° C. for 24 hours to obtain a lactic acid bacteria culture.

And, in order to select lactic acid bacteria having beta-glucosidase activity, para-nitrophenyl-β-D-glucopyranoside so that the final concentration is 10 mM in 0.2 ml of each of the 97 types of lactic acid bacteria culture (including cells). 0.2 ml of a (pNPG:para-nitrophenyl-β-D-glucopyranoside, SIGMA, USA) solution was added, followed by reaction at 37° C. for 1 hour. Then, 1.6 ml of 0.5 M sodium carbonate (Na ₂ CO ₃ ) was added to terminate the reaction, and absorbance was measured at 420 nm to select 16 strains having an OD ₄₂₀ value of 0.5 or more. And, the amount of para-nitro-phenol (pNP: para-nitro-phenol) generated by substituting it to the standard curve prepared in advance was measured to confirm the relative glucose production. Activity is expressed in U (unit), and 1 U was defined as the amount of enzyme used to decompose pNPG under the above conditions to produce 1 mmol of pNP per minute. The results are shown in Table 1 below.

Lactobacillus β-glucosidase activity (U/mL) BLA-1 1.49 BLA-2 1.47 BLA-3 1.00 BLA-4 0.98 BLA-5 0.81 BLA-6 3.25 BLA-7 3.18 BLA-8 3.51 BLA-9 4.20 BLA-10 3.12 BLA-11 1.33 BLA-12 3.21 BLA-13 3.27 BLA-14 3.25 BLA-15 3.00 BLA-16 3.23

Among the strains in Table 1, 10 strains (BLA-6, BLA-7, BLA-8, BLA-9, BLA-10, BLA-12, BLA-13, BLA-14, BLA-15, BLA-16) were selected, and in order to measure the conversion ability of ginsenoside Rb1 for the selected 10 strains, 1 wt% of an aqueous solution of ginsenoside Rb1 BLA-6, BLA showing ginsenoside conversion ability after adding 1 ml of each of the 16 lactic acid bacteria cultures to 1 ml and reacting for 24 hours at 37° C. while stirring at 200 rpm to drop and develop on TLC -9, BLA-12, BLA-13, BLA-15 strains were selected.

The selected 5 strains were inoculated with 5% by weight of red ginseng extract and then cultured at 37° C. for 48 hours, followed by observation of their growth potential, as shown in Table 2 below.

Lactobacillus Number of viable cells (cfu/mL) BLA-6 5.4 × 10 ⁶ BLA-9 2.1 × 10 ⁸ BLA-12 7.2 × 10 ⁷ BLA-13 1.9 × 10 ⁶ BLA-15 8.3 × 10 ⁷

Looking at Table 2, it can be seen that among the strains, BLA-9 exhibits the best growth ability in the red ginseng extract. Accordingly, the BLA-9 was finally selected.

1-2: Identification of the strain

After separating the chromosomal DNA from the culture medium of each strain in order to investigate the molecular biological characteristics of the BLA-9 strain, the chromosomal DNA was subjected to full sequencing of the 16s rRNA gene to identify microorganisms.

To this end, the DNA was isolated after culturing the strain BLA-9 selected in 1-1 in MRS medium. 16 The rDNA region was amplified using PCR (polymerase chain reaction). The primer sequences for the PCR amplification were 5'-AGAGTTTGATCMTGGCTCAG-3' (forward) and 5'-GGTTACCTTGTTACGACTTC-3' (reverse). For PCR reaction, 50 ng of extracted genomic DNA, 100 μM of each dNTPs, 0.2 μM of each primer, 1×enzyme buffer, and 2 units of Taq polymerase were added, and distilled water was added to make a total volume of 50 μl. The amplification reaction was initially denatured at 94 °C for 90 seconds, then 28 cycles of 94 °C for 30 seconds, 42 °C for 60 seconds, 72 °C for 60 seconds, and further extended at 72 °C for 5 minutes. The PCR product was separated on a 1.5% agarose gel and then eluted. Using the PCR product, the chromosomal DNA of the BLA-9 strain, 16S rRNA sequencing was performed at Cosmogenetech Co., Ltd., and as a result of the nucleotide sequence analysis, the BLA-9 strain of the present invention was Lactobacillus acid It was investigated to be a strain belonging to Pillars, and it was found to have the nucleotide sequence of SEQ ID NO: 1 (see FIG. 1). In addition, as a result of analyzing the base sequence through BLAST Search, it showed about 99% homology with Lactobacillus acidophilus strains previously uploaded to the database (Data Base), SEQ ID NO: 1 It was confirmed that the strain with 100% homology to and did not exist in the past.

Therefore, the present inventors deposited these strains at the Korea Research Institute of Bioscience and Biotechnology Biological Resources Center on September 21, 2020 and were given an accession number KCTC18848P.

Test Example 1: Characteristics of the strain

1-1: type of fungus

The results of observing the morphology of the Lactobacillus acidophilus BLA-9 strain isolated and identified in Examples after anaerobic culture at 37° C. for 48 hours on BL agar plate medium are as follows.

1) Cell shape: various such as club-shaped, Y-shaped

2) Gram staining: positive

1-2: colony type

The results of observing the colony shape after culturing the Lactobacillus acidophilus BLA-9 strain isolated and identified in Examples anaerobically on BL agar plate at 37° C. for 48 hours are as follows.

1) Shape: round

2) Size: 1-3mm

3) Hue: milky white

1-3: physiological properties

Physiological properties of the Lactobacillus acidophilus BLA-9 strain isolated and identified in Examples are as follows.

1) Growth temperature

Growth range: 25~45℃

Optimum temperature: 37~41℃

2) Growth pH

Growth range: pH 2.0~8.0

Optimal pH: pH 6.5~7.0

3) Effect of oxygen: organized anaerobic

4) Catalase: -

5) Ammonia production: -

6) Coagulability: -

7) Litmus milk reduction: -

8) Derived embryo degrading enzyme: -

9) Beta-galactose degrading enzyme: +

10) Alpha-glucose degrading enzyme: +

11) sugar solubility

sugars availability sugars availability xylose - esculin - arabinose - Salicin - ribose - Cellobiose - atonitol - maltose + galactose + lactose - D-glucose + Melibios - D-fructose + saccharose + D-mannose - Trehalose - mannitol - inulin - sorbitol - Melezitose - Alphamethyl-D-mannoside - raffinose - Alphamethyl-D-glucoside - Starch - N-acetylglucoside + Beta-gentiobiose - amygdalin - D-tranose - abutin - D-tagatose -

Example 2: Preparation of fermented red ginseng extract

(1) Preparation of red ginseng extract

Red ginseng powder was prepared by pulverizing 6-year-old red ginseng, which was purchased from Punggi Ginseng Nonghyup, composed of red ginseng root and red rice ginseng in a weight ratio of 75:25.

The red ginseng powder was mixed with a 50 wt% ethanol aqueous solution in a weight ratio of 1:8, extracted while stirring at 80 °C for 8 hours, and filtered, and the filtrate was concentrated under reduced pressure to obtain a solid content of 5 wt% to prepare a red ginseng extract did The red ginseng extract thus prepared had a saponin content of 109.2 mg/g on a solid basis.

(2) Enzyme treatment step

To 100 parts by weight of the red ginseng extract, 20 parts by weight of Sumizyme AC (Sumizyme AC, manufacturer: ShinNippon, enzyme type: cellulase, β-glucosidase and hemicellulase, enzyme-producing microorganism: Aspergillus niger ) are added to 100 parts by weight of the red ginseng extract Then, it was enzymatically treated at 50 °C and pH 4 for 24 hours. Then, the enzyme-treated red ginseng extract was allowed to cool at room temperature for 2 hours.

(3) Fermentation step

First, the Lactobacillus acidophilus ( Lactobacillus acidophilus ) BLA-9 strain of the present invention was inoculated in MRS medium at 0.1% by weight and then cultured with shaking at 37°C for 24 hours to culture the strain (6.2×10 ⁹ cfu/mL). ) was obtained.

The enzyme-treated red ginseng extract was inoculated with the strain culture at 0.5% (v/v) and then fermented at 37° C. for 24 hours. A fermented red ginseng extract was prepared by heating the obtained fermented product at 90° C. for 15 minutes to inactivate enzymes and strains in the fermented product.

Comparative Example 1: Preparation of fermented red ginseng extract - Fermentation omitted

It was carried out in the same manner as in Example 2, except that the fermentation step of step (3) was omitted and a fermented fermented red ginseng extract was prepared.

Comparative Example 2: Preparation of fermented red ginseng extract - enzyme treatment omitted

It was carried out in the same manner as in Example 2, except that the enzyme treatment step of step (2) was omitted and a fermented red ginseng extract was prepared.

Comparative Example 3: Preparation of fermented red ginseng extract - Enzyme treatment after pre-fermentation

It was carried out in the same manner as in Example 2, except that the fermentation step of step (3) was performed first, and then the enzyme treatment of step (2) was performed to prepare a fermented red ginseng extract.

Comparative Example 4: Preparation of fermented red ginseng extract - using Lactobacillus gasseri KCTC3163

Application Examples of Korean Patent Application Laid-Open No. 10-2010-0054428 (Patent Document 1, Korea Food Research Institute, "Extract for improving glucose homeostasis by fermentation of Lactobacillus gasseri KCTC3163, a lactic acid bacterium from ginseng or ginseng extract, and a manufacturing method thereof") With reference to 1-4, a fermented red ginseng extract was prepared.

Specifically, distilled water was added to the red ginseng extract powder and suspended, and then the Lactobacillus gasseri KCTC3163 strain was inoculated and fermented to prepare a fermented red ginseng extract.

Comparative Example 5: Preparation of fermented red ginseng extract - Cytolase PCL5, Sumizyme AC and Rapidase C80Max mixed enzyme and Lactobacillus Sakei HY7802

Korean Patent Registration Publication No. 10-1409761 (Patent Document 2, Korea Yagult Co., Ltd., "Method for producing fermented red ginseng concentrate with enhanced content of compound K using enzyme conversion and lactic acid fermentation, and fermented red ginseng concentrate A fermented red ginseng extract was prepared with reference to Example 6 of "Product containing as an active ingredient").

Specifically, a fermented red ginseng extract was prepared by adding and reacting Cytolase PCL5, Sumizyme AC and Rapidase C80Max mixed enzyme to the red ginseng extract, inoculating and fermenting Lactobacillus Sakei HY7802.

The types of enzymes and strains as raw materials and processing means used in the Examples and Comparative Examples are briefly summarized in Table 4 below.

division Raw material (substrate) means of treatment enzyme strain Example Red Ginseng Extract Sumizyme AC Lactobacillus acidophilus BLA-9 Comparative Example 1 Red Ginseng Extract Sumizyme AC - Comparative Example 2 Red Ginseng Extract - Lactobacillus acidophilus BLA-9 Comparative Example 3 Red Ginseng Extract Sumizyme AC Lactobacillus acidophilus BLA-9 Comparative Example 4 Red Ginseng Extract - Lactobacillus gasseri KCTC3163 Comparative Example 5 Red Ginseng Extract Cytolase PCL5+Sumizyme AC+Rapidase C80Max Lactobacillus Sake HY7802

Test Example 2: Changes in ginsenosides of enzyme-treated red ginseng extract according to enzyme type and enzyme treatment time

In order to confirm the change of ginsenoside according to the type of enzyme and the time of enzyme treatment, 100 parts by weight of the red ginseng extract prepared as in step (1) of the above Example, Sumizyme AC (Example), Rohament CL (Rohament) CL, manufacturer: AB Enzymes) and plantase TCL (Plantase TCL, manufacturer: Bision Biochem) were each added in 20 parts by weight, followed by enzyme treatment time (0.5, 1, 2, 4, 8, 16, 24 and 48 hours), an enzyme-treated red ginseng extract was prepared. Thereafter, the change in ginsenoside (Rg1, Rg3, Rd, F2, CK) content of each enzyme-treated red ginseng extract was measured and shown in Table 5 below. At this time, the control is a red ginseng extract without enzyme treatment and fermentation process.

In order to measure the ginsenoside content, when preparing the test solution, 450 mg of the enzyme-treated red ginseng extract prepared in Example 2 was weighed and placed in a 20 mL volumetric flask, filled with methanol, and sonicated for 30 minutes. After vortexing, it was filtered through a 0.45 μm membrane filter to obtain a test solution.

Specifically, the ginsenoside (Rg1, Rg3, Rd, F2, CK) content was confirmed as follows. HPLC 1260 Series (Agilent) was used as an analytical instrument, and Discovery C18, 5 μm, 250 x 4.0 I.D mm (Supelco) was used as an analytical column. 16 mg of each ginsenoside standard material was weighed, and methanol was added to adjust the volume to 20 mL as a standard stock solution, and the standard stock solution was diluted to an appropriate concentration to prepare a standard solution in a linear section.

division time (h) Ginsenoside content (mg/g) Rg1 Rg3(S) Rb1 Rd F2 CK control 0 5.38 1.99 18.67 2.83 N.D. N.D. Sumizyme 0.5 4.37 1.73 14.93 2.89 N.D. 0.84 One 4.36 1.71 13.96 2.34 0.02 0.78 2 4.33 1.66 11.80 3.34 0.23 0.73 4 4.18 1.58 7.50 5.70 1.02 0.62 8 4.27 1.53 2.75 6.98 4.18 0.55 16 4.09 1.29 0.52 3.45 7.93 0.71 24 3.86 1.00 0.24 2.31 7.37 0.59 48 3.18 0.72 N.D. N.D. 5.78 0.90 Rohament 0.5 5.32 2.05 16.66 3.24 N.D. N.D. One 5.21 1.96 18.01 2.24 N.D. N.D. 2 5.25 2.03 17.54 3.01 N.D. N.D. 4 4.94 1.95 14.26 7.28 N.D. N.D. 8 4.80 2.00 9.61 12.26 N.D. N.D. 16 4.47 2.07 4.81 17.97 N.D. N.D. 24 4.19 1.95 3.06 18.59 N.D. N.D. 48 3.59 2.05 1.08 20.74 N.D. N.D. Plantase 0.5 4.67 1.73 16.17 3.47 N.D. N.D. One 4.67 1.70 16.21 3.16 N.D. N.D. 2 4.68 1.74 16.51 4.07 N.D. N.D. 4 4.57 1.70 15.98 7.94 N.D. N.D. 8 4.41 1.69 14.88 9.73 N.D. N.D. 16 4.28 1.74 13.48 10.30 N.D. N.D. 24 4.17 1.77 12.55 12.50 N.D. N.D. 48 3.53 1.80 8.55 15.08 0.17 0.09

Figure 5 is a schematic diagram simply showing the process of conversion of PPD-type ginsenoside to compound K (CK) by bioconversion.

Looking at Table 5, in the case of the red ginseng extract treated with Lohamant CL and plantase TCL, conversion from Rd to F2 and compound K was not made, whereas in the case of the red ginseng extract treated with sumizyme AC, F2 and compound K It can be seen that the conversion has been performed properly.

Test Example 3: Changes in ginsenosides of enzyme-treated red ginseng extract according to enzyme addition amount and enzyme treatment time

In order to check the change of ginsenoside according to the amount of enzyme added and the enzyme treatment time, the mixed enzyme Sumizyme AC (Example) was added to 100 parts by weight of the red ginseng extract prepared as in step (1) of the above Example, 1.0, 2.0, After adding 4.0 and 5.0 parts by weight, enzyme-treated red ginseng extract was prepared while varying the enzyme treatment time (4, 8, 15, 24, 32 and 48 hours) at 50 °C. Thereafter, the change in ginsenoside (F2, CK) content of each enzyme-treated red ginseng extract was measured and shown in Table 6 below. At this time, the control is a red ginseng extract without enzyme treatment and fermentation process.

Enzyme treatment time (hr) 1 part by weight 2 parts by weight 4 parts by weight 5 parts by weight F2 CK F2 CK F2 CK F2 CK control 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 4 2.45 0.19 5.05 0.00 6.75 0.21 6.55 0.00 8 3.41 0.00 5.55 0.47 5.82 0.75 5.76 1.07 15 4.86 0.91 8.69 1.77 6.71 1.98 3.77 1.37 24 5.04 1.13 8.98 2.12 5.85 1.87 5.12 1.87 32 5.41 1.47 8.84 2.30 5.60 1.90 4.74 1.91 48 5.76 1.68 8.61 3.50 5.11 2.10 5.06 2.06

Looking at Table 6, it can be seen that the content of F2 and compound K of the enzyme-treated red ginseng extract reacted by adding 2 parts by weight of the enzyme to 100 parts by weight of the red ginseng extract was high.

Test Example 4: Changes in ginsenosides of fermented red ginseng extract according to the amount of enzyme added

In order to confirm the change of ginsenoside of the fermented red ginseng extract before and after fermentation according to the amount of enzyme added, in the fermented red ginseng extract of the above example, the amount of enzyme added in step (2) (enzyme 1 part by weight, 2 parts by weight, 5 parts by weight) After measuring the ginsenoside content of each of the enzyme-treated red ginseng extract prepared by using different methods, and the fermented red ginseng extract prepared by fermenting after the enzyme treatment obtained in step (3), it is shown in Table 7 below. At this time, the enzyme in step (2) was treated for 24 hours, and fermentation in step (3) was performed for 8 hours.

division Ginsenoside content (mg/g) Rg1 Rg3(S) Rb1 Rd F2 CK control 8.22 0.82 17.77 1.84 N.D. N.D. 1 part by weight Enzyme treatment 6.36 0.38 0.40 3.35 5.61 1.68 Fermentation after enzyme treatment 5.01 N.D. 0.48 1.33 6.63 1.84 2 parts by weight Enzyme treatment 5.09 0.93 0.86 1.82 10.63 2.90 Fermentation after enzyme treatment 4.38 1.68 3.56 1.52 11.42 3.75 5 parts by weight Enzyme treatment 3.11 0.73 0.74 0.60 5.23 1.74 Fermentation after enzyme treatment 4.49 1.42 3.04 1.43 5.90 2.79

Looking at Table 7, it was confirmed that the ginsenoside content of the fermented red ginseng extract prepared by adding 2 parts by weight of the enzyme, in particular, the content of Rb1, F2, and compound K was the highest.

Test Example 5: Changes in ginsenosides of fermented red ginseng extract according to changes in enzyme treatment time and fermentation time

The purpose of this study was to determine the changes in ginsenosides of the fermented red ginseng extract according to the change in enzyme treatment time and fermentation time.

To this end, in the fermented red ginseng extract of the above example, the enzyme-treated red ginseng extract obtained by varying the enzyme treatment time (20 hours, 24 hours and 48 hours) in the step (2), and the fermentation time (2 hours) in the step (3) , 12 hours, 18 hours, and 24 hours) after measuring the ginsenoside content of each of the fermented red ginseng extracts obtained by fermentation after enzyme treatment, it is shown in Table 8 below.

division Ginsenoside content (mg/g) Rg1 Rg3(S) Rb1 Rd F2 CK Rg1+Rb1+Rg3 control 8.57 0.85 18.97 1.99 0.00 0.00 28.39 Enzyme treatment 20 h 6.06 0.04 1.07 2.41 9.94 1.77 7.17 24 h 5.08 0.74 0.65 1.00 9.76 2.40 6.46 48 h 3.03 0.47 0.08 N.D. 9.25 3.42 3.58 Fermentation after enzyme treatment 2 h 4.45 1.30 2.91 N.D. 9.32 3.86 8.66 12 h 4.57 1.38 3.22 N.D. 9.31 3.98 9.17 18 h 3.51 1.43 3.73 N.D. 6.93 4.13 8.67 24 h 3.77 1.70 4.63 N.D. 7.54 4.41 10.09

Looking at Table 8, it can be seen that the F2 and CK contents of the fermented red ginseng extract fermented after enzyme treatment were increased compared to that of only the enzyme treatment.

On the other hand, the content of Rb1 decreased after the enzyme treatment, but it was confirmed that the content greatly increased when the fermentation was performed after the enzyme treatment. This result is inferred to be because the high glycosides in the red ginseng extract were converted to Rb1 through glucosidase activity by fermentation with the Lactobacillus acidophilus BLA-9 strain of the present invention. Accordingly, it was confirmed that the content of Rb1 also significantly increased as the fermentation time increased.

Test Example 6: Comparison of ginsenoside content

After measuring the ginsenoside content of the fermented red ginseng extract of Examples and Comparative Examples, it is shown in Table 9 below.

division Ginsenoside content (mg/g) Rg1 Rg3(S) Rb1 Rd F2 CK Rg1+Rb1+Rg3 Example jeon 8.57 0.85 18.97 1.99 0.00 0.00 28.39 After enzyme treatment 4.45 1.30 2.91 - 9.32 2.57 8.66 after fermentation 3.40 1.55 4.70 - 6.55 4.50 9.65 Comparative Example 1 jeon 8.57 0.85 18.97 1.99 0.00 0.00 28.39 after 6.06 0.04 1.07 2.41 9.94 1.77 7.17 Comparative Example 2 jeon 5.87 0.59 17.57 - - - 24.04 after 0.18 7.70 1.15 - - - 9.03 Comparative Example 3 jeon 5.87 0.59 17.57 - - - 24.04 after 0.17 8.95 1.61 - - - 10.73 Comparative Example 4 jeon(%) 11.60 - 12.70 - - 0.00 - after(%) 14.60 - 2.40 - - 0.50 - Comparative Example 5 jeon 8.00 - 45.8 - - - 53.8 after - - - - - 1.86 -

Looking at Table 9, the fermented red ginseng extract of Example has a sum of ginsenoside indicators (Rb1, Rg1 and Rg3) of 9.65 mg/g, the content of F2 is 6.55 mg/g, and the content of compound K is 4.50 mg/g g, so the ginsenoside composition is very ideal.

In addition, the ginsenoside content of the fermented red ginseng extracts of Examples and Comparative Examples was calculated as a fermented red ginseng efficacy index and shown in Table 10 below.

division Ginsenoside content (mg/g) Ginsenoside Indicators CK Rg1+Rb1+Rg3 CK/Rb1 before fermentation CK/Rb1 CK/Rg3 CK/(Rg1+Rb1+Rg3) CK/(Rg1+Rb1+Rg3+CK) Example jeon 0.00 28.39 - - - 0.00 0.00 After enzyme treatment 2.57 8.66 0.14 0.88 1.98 0.30 0.23 after fermentation 4.50 9.65 0.24 0.96 2.90 0.47 0.32 Comparative Example 1 jeon 0.00 28.39 - - - 0.00 0.00 After enzyme treatment 1.77 7.17 0.03 0.11 44.25 0.11 0.10 Comparative Example 2 jeon 0.00 24.04 - - - - - after fermentation 0.00 9.03 - - - - - Comparative Example 3 jeon 0.00 24.04 - - - - - Enzyme treatment after fermentation 0.00 6.73 - - - - - Comparative Example 4 jeon(%) 0.00 - - - - - - after(%) 0.50 17.00 0.04 0.21 - 0.03 0.03 Comparative Example 5 jeon - - - - - - - after 1.86 - 0.04 - - - -

Referring to Table 10, the fermented red ginseng extract of the Example is a value obtained by dividing the content of compound K contained in the fermented red ginseng extract after fermentation by the content of Rb1 contained in the red ginseng extract before enzyme treatment (CK/Rb1 before fermentation) ) is 0.237, the content of compound K contained in the fermented red ginseng extract after fermentation divided by the content of Rb1 (CK/Rb1 after fermentation) reaches 0.96, and the content of compound K contained in the fermented red ginseng extract after fermentation is Rg3 Since the value divided by the content of (CK/Rg3) is 2.9, the production efficiency of compound K is very good.

On the other hand, in the fermented red ginseng extract of Comparative Example 4 with reference to Patent Document 1, the production efficiency of compound K, which is a value obtained by dividing the content of compound K contained in the fermented red ginseng extract after fermentation by the content of Rb1 contained in the red ginseng extract before enzyme treatment, was 0.04 , and the value obtained by dividing the content of compound K contained in the fermented red ginseng extract by the content of Rb1 after fermentation was only 0.21, confirming that the production efficiency of compound K was significantly lower than in Examples.

In addition, in the fermented red ginseng extract of Comparative Example 5 with reference to Patent Document 2, the production efficiency of compound K, which is a value obtained by dividing the content of compound K contained in the fermented red ginseng extract after fermentation by the content of Rb1 contained in the red ginseng extract before the enzyme treatment, was 0.04 It was confirmed that the production efficiency of compound K was significantly lower than that of Example.

From the above results, according to the method for producing a fermented red ginseng extract using the Lactobacillus acidophilus BLA-9 strain having bioconversion ability of the present invention, the production efficiency of Compound K is significantly higher than that of the fermented red ginseng extract prepared by the conventional method. It was excellent, and it was specifically confirmed that the ginsenoside index was excellent.

Although the present invention has been described as the above-mentioned preferred embodiment, it is possible to make various modifications and variations without departing from the spirit and scope of the invention. It is also intended that the appended claims cover such modifications and variations as fall within the scope of the present invention.

<110> Bionic Trading Corporation <120> Lactobacillus acidophilus BLA-9 with biological conversion activity of saponin, a method of preparation for fermented red ginseng extracts having increased content of ginsenoside compound K using the same, fermented red ginseng extracts using thereby and health-functional food composition containing the same <130> HPC-9245 <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 1550 <212> DNA <213> Lactobacillus acidophilus <400> 1 ccagtggtgg tggtggggga aagaatggca agtcgaacgc gttggcccaa ttgattgatg 60 gtgcttgcac ctgattgatt ttggtcgcca acgagtggcg gacgggtgag taacacgtag 120 gtaacctgcc cagaagcggg ggacaacatt tggaaacaga tgctaatacc gcataacagc 180 gttgttcgca tgaacaacgc ttaaaagatg gcttctcgct atcacttctg gatggacctg 240 cggtgcatta gcttgttggt ggggtaacgg cctaccaagg cgatgatgca tagccgagtt 300 gagagactga tcggccacaa tgggactgag acacggccca tactcctacg ggaggcagca 360 gtagggaatc ttccacaatg ggcgcaagcc tgatggagca acaccgcgtg agtgaagaag 420 ggtttcggct cgtaaagctc tgttgttaaa gaagaacacg tatgagagta actgttcata 480 cgttgacggt atttaaccag aaagtcacgg ctaactacgt gccagcagcc gcggtaatac 540 gtaggtggca agcgttatcc ggatttattg ggcgtaaaga gagtgcaggc ggttttctaa 600 gtctgatgtg aaagcyttcg gcttaaccgg agaagtgcat cggaaactgg ataacttgag 660 tgcagaagag ggtagtggaa ctccatgtgt agcggtggaa tgcgtagata tatggaagaa 720 caccagtggc gaaggcggct acctggtctg caactgacgc tgagactcga aagcatgggt 780 agcgaacagg attagatacc ctggtagtcc atgccgtaaa cgatgagtgc taggtgttgg 840 agggtttccg cccttcagtg ccggagctaa cgcattaagc actccgcctg gggagtacga 900 ccgcaaggtt gaaactcaaa ggaattgacg ggggcccgca caagcggtgg agcatgtggt 960 ttaattcgaa gctacgcgaa gaaccttacc aggtcttgac atcttgcgcc aaccctagag 1020 atagggcgtt tccttcggga acgcaatgac aggtggtgca tggtcgtcgt cagctcgtgt 1080 cgtgagatgt tgggttaagt cccgcaacga gcgcaaccct tgttactagt tgccagcatt 1140 aagttgggca ctctagtgag actgccggtg acaaaccgga ggaaggtggg gacgacgtca 1200 gatcatcatg ccccttatga cctgggctac acacgtgcta caatggacgg tacaacgagt 1260 cgcgaactcg cgagggcaag caaatctctt aaaaccgttc tcagttcgga ctgcaggctg 1320 caactcgcct gcacgaagtc ggaatcgcta gtaatcgcgg atcagcatgc cgcggtgaat 1380 acgttcccgg gccttgtaca caccgcccgt cacaccatga gagtttgtaa tgcccaaagc 1440 cggtggggta accttttaga aggagccgtc ctaaggcagg gcagatgacn nnnnnnnnnn 1500 ngtaacaagn nnnnnnnnnn ngaacctgnn nnnngatcac ctcctttcta 1550

Claims (14)

Lactobacillus acidophilus ( Lactobacillus acidophilus ) BLA-9 strain (Accession No.: KCTC18848P) having bioconversion ability for saponins. (1) preparing a red ginseng extract;
(2) adding a complex enzyme of cellulase, β-glucosidase and hemicellulase to the red ginseng extract for enzymatic reaction; and
(3) The enzyme-treated red ginseng extract is inoculated with Lactobacillus acidophilus BLA-9 strain (Accession No.: KCTC18848P) and then fermented; A method for preparing a red ginseng extract. 3. The method of claim 2,
The red ginseng extract of step (1) is a method for producing a fermented red ginseng extract with an increased content of compound K, characterized in that it is extracted with water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. 3. The method of claim 2,
The red ginseng extract prepared in step (1) is a method for producing a fermented red ginseng extract with an increased content of compound K, characterized in that the solid content is 3 to 50% by weight. 3. The method of claim 2,
The complex enzyme of step (2) is a method for producing a fermented red ginseng extract with an increased content of compound K, characterized in that it is an enzyme derived from Aspergillus niger . 3. The method of claim 2,
(2) A method for producing a fermented red ginseng extract with an increased content of compound K, characterized in that the complex enzyme of step (2) is Sumizyme AC. 3. The method of claim 2,
The complex enzyme of step (2) is a method for producing a fermented red ginseng extract with an increased content of compound K, characterized in that it is added in an amount of 0.5 to 50 parts by weight based on 100 parts by weight of the red ginseng extract. 3. The method of claim 2,
The enzymatic reaction of step (2) is a method for producing a fermented red ginseng extract with an increased content of compound K, characterized in that it is carried out at 50 to 65 ℃, pH 3.5 to 5.0 for 5 to 60 hours. 3. The method of claim 2,
(2) A method for producing a fermented red ginseng extract with an increased content of compound K, characterized in that not performing an enzyme inactivation step between steps (2) and (3). 3. The method of claim 2,
The fermentation of step (3) is a method for producing a fermented red ginseng extract with an increased content of compound K, characterized in that it is carried out at 30 to 50 ° C. for 5 to 60 hours. 3. The method of claim 2,
The fermented red ginseng extract has a total sum of Rb1, Rg1 and Rg3 of at least 7.0 mg/g or more, an F2 content of at least 5.0 mg/g or more, and a compound K content of at least 4.0 mg/g or more. A method for producing a fermented red ginseng extract with an increased content of 3. The method of claim 2,
The production efficiency of compound K, which is a value obtained by dividing the content (mg/g) of compound K contained in the fermented red ginseng extract after the fermentation by the content (mg/g) of Rb1 contained in the red ginseng extract before the enzyme treatment, is at least 0.2, characterized in that A method for producing a fermented red ginseng extract with an increased content of compound K. 13. Prepared according to the method of any one of claims 2 to 12, wherein the sum of Rb1, Rg1 and Rg3 is at least 7.0 mg/g or more, the content of F2 is at least 5.0 mg/g or more, and the content of compound K is Fermented red ginseng extract, characterized in that at least 4.0 mg/g or more. A health functional food composition comprising the fermented red ginseng extract of claim 13 as an active ingredient.

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