KR102466825B1 - 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 relates to a Lactobacillus acidophilus BLA-9 strain having bioconversion ability to 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 according to the method, and a health functional food containing the same It's about the composition.
The Lactobacillus acidophilus BLA-9 strain (accession number: KCTC18848P) having bioconversion ability to saponin of the present invention has excellent ginsenoside bioconversion ability. When the strain is inoculated into the 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 production method of the present invention, the production efficiency of Compound K is remarkably increased, so that 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 compound K that is easily absorbed by the body, it is very useful as a health functional food or food material.

Description

Lactobacillus acidophilus BLA-9 strain having bioconversion ability to saponin, method for producing fermented red ginseng extract with increased content of compound K using the same, fermented red ginseng extract according to the method, and health functional food composition containing 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 to 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 according to the method, and a health functional food containing the same It's about the composition.

Ginseng refers to the root of a plant in the Araliaceae family. Depending on the place of cultivation, it is called by names such as Korean ginseng (Korean Peninsula), American ginseng (USA and Canada), Jeonchil ginseng (China), and Jukjeol ginseng (Japan). It is classified into white ginseng, tang ginseng and bongmil ginseng.

Korean ginseng (Panax ginseng C.A. Meyer) has been widely used in Korea, China, and Japan since the past for the purpose of improving health. It is known that the active ingredients of ginseng differ significantly depending on the type of ginseng, the root part, the year of harvest, the harvest time, and the manufacturing method. By lotus root of ginseng, in general, younger roots have a higher sugar content and older ones have a high saponin content. It is known. In addition, it is known that white ginseng, in which fresh ginseng is dried as it is, and red ginseng, which is steamed and dried, show differences in the content of irradiated ginseng or the types of ginsenosides.

'Red ginseng' referred to in health functional food uses ginseng as a raw material, and undried fresh ginseng is steamed, cooked, and dried by steam or other methods. It should be powdered or extracted with water or alcohol, concentrated or fermented to make it edible, and managed so that the sum of ginsenosides Rg1 and Rb1 is 0.8 mg/g to 34 mg/g.

Saponin is known as an effective component of ginseng and red ginseng. Saponin of ginseng and red ginseng is a type of glycoside and has a unique chemical structure different from saponin found in other plants, and its efficacy is also greatly different. Saponins of ginseng and red ginseng are called 'ginsenosides' in the sense of ginseng glycosides to distinguish them from saponins of other plant origin.

Ginsenoside, an active ingredient of ginseng and red ginseng, contains glucose, arabinose, xylose and rhamnose in the dammarane skeleton of triterpenoids. Ginsenosides Rb1, Rb2, Rc, Rd, which are high-molecular ginsenosides, are currently isolated over 100 species as glycosides in which one or several sugars are linked (J Ginseng Res 41 (2017), 435-443). Six ginsenosides, including , Re and Rg1, account for 90% of the total ginsenosides.

Compound K (C-K: compound K, 20-O-β-D-glucopyranosyl 20(S)-protopanasadiol), which is one of a small amount of low-molecular-weight ginsenosides produced by hydrolysis of high-molecular ginsenosides, has an aglycone ( It is one of the PPD-based low-molecular-weight ginsenosides in which one glucose is linked to C-20 of the aglycon), and is a conversion product of ginsenosides Rb1, Rb2, and Rc 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, compound K was detected in plasma 24 hours after administration of ginsenoside Rb1, and it was reported that it was hardly metabolized in the stomach. According to Karikura et al. (1991) examining the metabolites of ginsenoside Rb2 in the stomach of rats, ginsenoside Rb2 is hardly metabolized except for the production of peroxide. Almost no metabolism occurs in the digestive system of , and it is judged that it is metabolized by intestinal microorganisms. In addition, according to Professor Kim Dong-hyun's study, the conversion ability of ginsenoside Rb1 was confirmed using the intestinal microorganisms of 98 Koreans, and as a result, 20% had no or significantly low ginsenoside metabolism ability, and only about 60% of the microorganisms could convert ginsenoside. It has been reported that many people do not metabolize ginsenosides efficiently. These reports show that even if ginseng or red ginseng is taken, there is a difference in efficacy depending on the individual due to differences in the composition of intestinal microorganisms and enzyme activity of intestinal microorganisms, and it is recommended to consume fermented red ginseng to reduce these differences.

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

Among them, as a method for producing compound K, production by intestinal microorganisms such as Bifidobacterium , fungi, or various microorganisms is known, but the method requires incubation under anaerobic conditions and a long incubation time, and the production yield is extremely low. Research on the manufacturing method of fermented red ginseng containing a large amount of compound K, a specific low-molecular-weight ginsenoside, through fermentation using edible enzymes and lactic acid bacteria, due to the low yield in terms of productivity and difficulty in eating due to the formation of by-products. development is required

In the course of research to develop fermented red ginseng with increased compound K content, the present inventors selected enzymes and lactic acid bacteria with excellent ginsenoside bioconversion ability, and 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 and the ginsenoside index was excellent.

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

The problem to be solved by the present invention is to provide a Lactobacillus acidophilus BLA-9 strain having bioconversion ability for saponin.

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

Another object of the present invention is to provide a fermented red ginseng extract prepared according to the above method, in which 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 containing 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 number: KCTC18848P) having a bioconversion ability for saponin.

In addition, the present invention comprises the steps of (1) preparing a red ginseng extract; (2) enzymatic reaction by adding complex enzymes of cellulase, β-glucosidase and hemicellulase to the red ginseng extract; and (3) inoculating the enzyme-treated red ginseng extract with Lactobacillus acidophilus BLA-9 strain (accession number: KCTC18848P) and then fermenting it; It provides a method for preparing fermented red ginseng extract.

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

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

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

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

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

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

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

According to one embodiment of the present invention, the fermented red ginseng extract has a total of Rb1, Rg1 and Rg3 of at least 7.0 mg/g, an F2 content of at least 5 mg/g, and a compound K content of at least 4.0 mg. /g or more.

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

In addition, 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, 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 containing the fermented red ginseng extract as an active ingredient.

The Lactobacillus acidophilus BLA-9 strain (accession number: KCTC18848P) of the present invention has excellent ginsenoside bioconversion ability. When the strain is inoculated into the 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 production method of the present invention, the production efficiency of Compound K is remarkably increased, so that the content of Compound K is further increased and a fermented red ginseng extract having an excellent ginsenoside index can be obtained.

The fermented red ginseng extract of the present invention contains a large amount of compound K that is easily absorbed into 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 schematic diagram showing the analysis of the 16S rRNA gene sequence of the Lactobacillus acidophilus BLA-9 strain of the present invention.
2 is a flowchart showing a method for preparing a fermented red ginseng extract according to an embodiment of the present invention.
3 is a schematic diagram showing a simple process in which PPD-type ginsenoside is converted into compound K (CK) by bioconversion.

Hereinafter, the present invention will be described in detail.

In the present specification, "ginsenoside index" 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, 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 value obtained by dividing the content of compound K contained in the fermented red ginseng extract by the content of Rb1 after fermentation (CK / Rb1), ⑤ fermentation The content of Compound K contained in the fermented red ginseng extract after fermentation divided by the content of Rg3 (CK/Rg3), ⑥ The content of Compound K contained in the fermented red ginseng extract after fermentation divided by the sum of Rg1, Rb1 and Rg3 (CK/Rg3) /(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 can be an indicator of the production efficiency of Compound K and/or the quality of fermented red ginseng extract. The fermented red ginseng extract of the present invention in which the above indexes exceed a certain level has a high sum of Rg1, Rb1 and Rg3 and a high content of compound K, so the quality of the fermented red ginseng extract is very excellent.

According to one embodiment of the present invention, the present invention provides a Lactobacillus acidophilus BLA-9 strain (accession number: KCTC18848P) having bioconversion ability to saponin.

As a result of analyzing the 16S rRNA sequence for microbial identification and classification of the BLA-9 strain, a lactic acid bacteria derived from kimchi stored in the headquarters (BTC Co., Ltd.) laboratory through the following example, 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 as a result of confirming homology by requesting the nucleic acid sequence of the strain to Macrogen, 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' means 'a', 'g', 'c', 't or u', or 'unknown or other' base.

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 the Korea Research Institute of Bioscience and Biotechnology, as of September 21, 2020, It was deposited (accession number: KCTC18848P).

The Lactobacillus acidophilus BLA-9 strain of the present invention has a bioconversion ability to saponin, in particular converts saponin into ginsenoside, and converts the high-molecular ginsenoside into compound K with increased digestion and absorption in the human body. Therefore, it is very useful in the production of fermented red ginseng.

According to another embodiment of the present invention, the present invention comprises (1) preparing a red ginseng extract; (2) enzymatic reaction by adding complex enzymes of cellulase, β-glucosidase and hemicellulase to the red ginseng extract; and (3) inoculating the enzyme-treated red ginseng extract with Lactobacillus acidophilus BLA-9 strain (accession number: KCTC18848P) and then fermenting; It provides a method for preparing fermented red ginseng extract.

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

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

According to one embodiment of the present invention, the red ginseng is prepared by using ginseng as a raw material, but steaming undried fresh ginseng with steam or other methods for a long time and drying it to a moisture content of 15% or less and processing it, 4 or 6 year old red ginseng root And / or it may be red ginseng, preferably 6-year-old red ginseng composed of red ginseng roots and red ginseng roots in a weight ratio of 50-90: 10-50, preferably 70-80: 20-30.

According to an embodiment of the present invention, the red ginseng may be red ginseng powder obtained by pulverizing red ginseng, and it is preferable to use red ginseng powder because it is easy to extract effective substances of red ginseng due to a large surface area.

The red ginseng is mixed with an extraction solvent at a weight ratio of 1: 1 to 20, preferably 1: 2 to 15, and more preferably 1: 3 to 10, and maintained at 20 to 100 ° C for 1 to 15 hours, preferably The extract may be prepared by performing single or multiple extractions while stirring for 2 to 10 hours, followed by concentration under reduced pressure. When the weight ratio of the red ginseng to the extraction solvent is out of the above range, a small amount of active ingredients of red ginseng may be extracted 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, still more preferably 3 to 60% by weight. It is preferable 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, lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. Examples of the lower alcohol include 30 to 80% methanol, ethanol, butanol or propanol.

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

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

The term "extract" used herein while referring to red ginseng includes not only a crude extract obtained by treating an extraction solvent, but also a processed product of 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.

The major 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 efficacy of ginsenosides, a process of converting high-molecular ginsenosides into low-molecular ginsenosides that are relatively easily absorbed and have more excellent efficacy is required. That is, high-molecular ginsenosides require a conversion process to remove glucose in order to effectively exhibit physiological activity in vivo.

To this end, the present inventors selected enzymes and lactic acid bacteria with excellent bioconversion ability for saponin, and converted high-molecular ginsenosides in the red ginseng extract to low-molecular ginsenosides using the selected enzymes and lactic acid bacteria, thereby increasing the content of compound K. An increased fermented red ginseng extract was prepared.

The enzyme for treating 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 in step (2) is Sumizyme AC (manufacturer: ShinNippon, type of enzyme: complex enzyme of cellulase, β-glucosidase and hemicellulase, enzyme-producing microorganism: Aspergillus niger ) desirable. The Sumizyme AC enzyme has an excellent effect of converting saponin into saponin metabolites, and when used together with the Lactobacillus acidophilus BLA-9 strain of the present invention, not only the ginsenoside bioconversion rate increases, but also sensory shows excellent effect.

The complex enzyme is present in an amount of 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 1 to 40 parts by weight, based on 100 parts by weight of the red ginseng extract. It is preferable to add 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 concern that the enzyme may be inactivated.

In addition, the enzyme reaction is carried out at 15 to 70 ° C, preferably 30 to 67 ° C, more preferably 40 to 65 ° C, still more preferably 50 to 65 ° C, 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, still more preferably 5 to 60 hours, still 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 are out of the above range, there is a concern that the activity of the enzyme may be 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 addition amount of the enzyme, reaction temperature and reaction time are optimal conditions for converting red ginseng extract to ginsenoside.

In the method for preparing the fermented red ginseng extract of the present invention, an enzyme inactivation step may not be performed after the enzyme treatment in step (2). The manufacturing method of the present invention maximizes the bioconversion of ginsenosides in red ginseng extract by performing the strain fermentation step of step (3) in a state where the enzyme activity is maintained without 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 ° C, preferably 20 to 60 ° C, more preferably 30 to 50 ° C, 30 to 40°C, more preferably 34 to 39°C, still 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 ° C. for 10 to 30 minutes to inactivate the enzyme and the Lactobacillus acidophilus BLA-9 strain.

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

In the fermented red ginseng extract of the present invention prepared as described above, 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 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 the value obtained by dividing the amount (mg/g) of Compound K contained in the fermented red ginseng extract after fermentation by the content (mg/g) of Rb1 contained in the red ginseng extract before 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 conversion rate of ginsenoside to compound K, that is, the production efficiency of compound K, is improved by treating red ginseng extract with Sumizyme AC as a mixed enzyme and then fermenting it with Lactobacillus acidophilus BLA-9 strain. 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 /g or more, preferably from 4.0 to 7.0 mg/g, more preferably from 4.5 to 6.5 mg/g.

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

As can be confirmed through the following examples, when the red ginseng extract is treated with enzymes and/or fermented, the total of the index components is lowered as ginsenosides are converted into compound K. The content of compound K in the fermented red ginseng extract of the present invention is reduced. Since the total of the index components (Rg1, Rb1, and Rg3) is at least 7.0 mg/g or more while at least 4.0 mg/g, 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 containing the fermented red ginseng extract as an active ingredient. The contents of the fermented red ginseng extract in the health functional food composition are as described above.

The health functional food composition may be provided in the form of a powder, granule, tablet, capsule, syrup or beverage. can be used The mixing amount of the active ingredient may be appropriately determined depending on the purpose of use thereof, 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, still more preferably 0.01 to 10% by weight.

The effective dose of the active ingredient contained in the health food composition is determined by the type and amount of the active ingredient and other ingredients contained in the composition, the type of formulation, the age, weight, general health condition, gender and diet of the ingested person, administration time, administration It may be adjusted according to various factors including, but not limited to, the route, duration of intake, and concurrently used drugs.

Hereinafter, the present invention will be described in detail by 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 is a lactic acid bacteria derived from kimchi, and the kimchi composition was diluted in sterilized physiological saline, and the supernatant was diluted in a decimal method and spread on a BL plate medium containing 5% malfi. A single strain was isolated by taking a colony containing a ring specific to Lactobacillus from the BL plate medium, which is a medium for distinguishing lactic acid bacteria, and this strain was subcultured and used in experiments.

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

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

In addition, in order to select lactic acid bacteria having beta-glucosidase activity, para-nitrophenyl-β-D-glucopyranoside was added to each of the 97 lactic acid bacteria cultures (including cells) to a final concentration of 10 mM in 0.2 ml each. (pNPG: para-nitrophenyl-β-D-glucopyranoside, SIGMA, USA) After adding 0.2 ml of the solution, the mixture was reacted at 37° C. for 1 hour. Thereafter, 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. In addition, the relative glucose production was confirmed by measuring the amount of para-nitro-phenol (pNP) produced by substituting it into a standard curve prepared in advance. Activity was expressed as U (unit), and 1 U was defined as the amount of enzyme used to degrade 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 having a β-glucosidase activity (U/mL) of 3.0 or more (BLA-6, BLA-7, BLA-8, BLA-9, BLA-10, BLA-12, BLA-13, BLA-14, BLA-15, BLA-16) were selected, and 1% by weight of ginsenoside Rb1 aqueous solution was used to measure the conversion ability of ginsenoside Rb1 for the selected 10 strains. After adding 1 ml of each of the 16 types of lactic acid bacteria cultures to 1 ml, reacting at 37 ° C. for 24 hours while stirring at 200 rpm, dropping and developing on TLC, BLA-6 and BLA showing ginsenoside conversion ability -9, BLA-12, BLA-13, and BLA-15 strains were selected.

The five strains selected above were inoculated into 5% by weight of red ginseng extract, incubated at 37 ° C. for 48 hours, and their growth ability was observed and shown in Table 2 below.

Lactobacillus Viable cell count (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 highest growth potential in red ginseng extract. Accordingly, the BLA-9 was finally selected.

1-2: Identification of strains

After isolating chromosomal DNA from each strain culture medium to investigate the molecular biological characteristics of the BLA-9 strain, the chromosomal DNA was intended to be analyzed for the entire base sequence of the 16s rRNA gene to identify microorganisms.

To this end, after culturing the selected strain BLA-9 of 1-1 in MRS medium, DNA was isolated. 16 rDNA regions were amplified using PCR (polymerase chain reaction). The primer sequences for PCR amplification were 5'-AGAGTTTGATCMTGGCTCAG-3' (forward) and 5'-GGTTACCTTGTTACGACTTC-3' (reverse). For the PCR reaction, 50 ng of extracted genomic DNA, 100 μM of each dNTP, 0.2 μM of each primer, 1×enzyme buffer, and 2 units of Taq polymerase were added and distilled water was added to a total volume of 50 μl. The amplification reaction was initially denatured at 94 °C for 90 seconds, followed by 28 cycles of 94 °C for 30 seconds, 42 °C for 60 seconds, and 72 °C for 60 seconds, followed by extension at 72 °C for 5 minutes. PCR products were separated on a 1.5% agarose gel and then eluted. Using the chromosomal DNA of the PCR product, the BLA-9 strain, 16S rRNA sequencing was performed by requesting Cosmogenetech Co., Ltd. As a result of the sequencing, the BLA-9 strain of the present invention is Lactobacillus acid It was investigated to be a strain belonging to Philus, and 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 the Lactobacillus acidophilus strains previously uploaded to the database, and SEQ ID NO: 1 It was confirmed that strains with 100% homology do not previously exist.

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 accession number KCTC18848P.

Test Example 1: Characteristics of strain

1-1: Morphology of bacteria

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

1) Shape of cell: club-shaped, Y-shaped, etc. various

2) Gram stain: positive

1-2: Colony type

After culturing the Lactobacillus acidophilus BLA-9 strain isolated and identified in Example anaerobically on BL agar plate medium at 37° C. for 48 hours, the colony morphology was observed and the results are as follows.

1) Shape: Round

2) Size: 1~3mm

3) Color: 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

Optimum pH: pH 6.5~7.0

3) Effect of Oxygen: Organized anaerobic

4) Catalase: -

5) Ammonia production:-

6) Coagulability: -

7) Litmus Milk Reduction: -

8) Degenerative enzyme: -

9) beta-galactose lyase: +

10) alpha-glucose lysing enzyme: +

11) Glycoavailability

sugars usability sugars usability xylose - esculin - arabinose - Salicin - ribose - cellobiose - atonitol - maltose + galactose + lactose - D-glucose + melibios - D-fructose + saccharose + D-mannose - trehalose - mannitol - inulin - Sorbitol - melegitose - 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

Purchased from Punggi Ginseng Agricultural Cooperative Federation, 6-year-old red ginseng consisting of red ginseng root and red ginseng in a weight ratio of 75: 25 was ground to prepare red ginseng powder.

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

(2) enzyme treatment step

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

(3) fermentation step

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

The strain culture was inoculated with 0.5% (v/v) of the enzyme-treated red ginseng extract and then fermented at 37° C. for 24 hours. 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, but 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 - Omission of enzyme treatment

It was carried out in the same manner as in Example 2, but 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 - enzymatic treatment after pre-fermentation

It was carried out in the same manner as in Example 2, but 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 Example of Korea Patent Publication 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 its preparation method") Fermented red ginseng extract was prepared by referring to 1-4.

Specifically, a fermented red ginseng extract was prepared by adding and suspending the red ginseng extract powder in distilled water, inoculating and fermenting the Lactobacillus gasseri KCTC3163 strain.

Comparative Example 5: Preparation of Fermented Red Ginseng Extract - Using Cytolase PCL5, Sumizyme AC and Rapidase C80Max mixed enzyme and Lactobacillus Sakei HY7802

Korea Patent Registration Publication No. 10-1409761 (Patent Document 2, Korea Yagurt Co., Ltd., "Method for manufacturing fermented red ginseng concentrate with enhanced content of compound K using enzyme conversion and lactic acid bacteria fermentation and fermented red ginseng concentrate prepared by the manufacturing method) Fermented red ginseng extract was prepared with reference to Example 6 of "Products Containing as an Active Ingredient").

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

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

division raw material (substrate) processing means 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+Smizyme AC+Rapidase C80Max Lactobacillus Sakei HY7802

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

In order to confirm the change in ginsenoside according to the enzyme type and enzyme treatment time, the mixed enzymes Sumizyme AC (Example) and Rohament CL (Rohament After adding 20 parts by weight of CL, manufacturer: AB Enzymes) and Plantase TCL (Plantase TCL, manufacturer: Bision Biochem), respectively, the enzyme treatment time at 50 ° C. (0.5, 1, 2, 4, 8, 16, 24 and 48 hours), enzyme-treated red ginseng extracts were prepared. Thereafter, changes in ginsenosides (Rg1, Rg3, Rd, F2, CK) contents of each enzyme-treated red ginseng extract were measured and are shown in Table 5 below. At this time, the control group is a red ginseng extract without enzyme treatment and fermentation.

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 put into a 20 mL volumetric flask, filled with methanol, and sonicated for 30 minutes. Then, after vortexing, the test solution was filtered through a 0.45 μm membrane filter.

Specifically, the content of ginsenosides (Rg1, Rg3, Rd, F2, CK) was confirmed as follows. An HPLC 1260 Series (Agilent) was used as an analytical instrument, and a 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 make 20 mL of the standard stock solution. The standard stock solution was diluted to an appropriate concentration to prepare a standard solution in the linear section.

division time (h) Ginsenoside content (mg/g) Rg1 Rg3(S) Rb1 Rd F2 C.K. control group 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

5 is a schematic diagram showing a simple process in which PPD-type ginsenoside is converted into compound K (CK) by bioconversion.

Referring to Table 5, in the case of the red ginseng extract treated with Lohament CL and Plantase TCL, conversion from Rd to F2 and compound K was not achieved, whereas in the case of the red ginseng extract treated with Sumizyme AC, F2 and compound K You can check that the conversion was done properly.

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

In order to confirm the change in ginsenoside according to the amount of enzyme added and the enzyme treatment time, 1.0, 2.0, 2.0, After adding 4.0 and 5.0 parts by weight, enzyme-treated red ginseng extracts were prepared while varying the enzyme treatment time (4, 8, 15, 24, 32 and 48 hours) at 50 ° C. Then, after measuring the change in ginsenoside (F2, CK) content of each enzyme-treated red ginseng extract, it is shown in Table 6 below. At this time, the control group is a red ginseng extract without enzyme treatment and fermentation.

Enzyme treatment time (hr) 1 part by weight 2 parts by weight 4 parts by weight 5 parts by weight F2 C.K. F2 C.K. F2 C.K. F2 C.K. control group 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

Referring to Table 6, it can be confirmed that the enzyme-treated red ginseng extract, which was reacted by adding 2 parts by weight of enzyme with respect to 100 parts by weight of red ginseng extract, had high contents of F2 and compound K.

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

In order to confirm the change in ginsenosides 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 different methods and the fermented red ginseng extract prepared by fermenting after enzyme treatment obtained in step (3), the contents are shown in Table 7 below. At this time, the enzyme in step (2) was treated for 24 hours, and the fermentation in step (3) was performed for 8 hours.

division Ginsenoside content (mg/g) Rg1 Rg3(S) Rb1 Rd F2 C.K. control group 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 fermented red ginseng extract prepared by adding 2 parts by weight of the enzyme had the highest content of ginsenosides, particularly Rb1, F2 and compound K.

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

The changes in ginsenosides in the fermented red ginseng extract according to the enzyme treatment time and fermentation time were investigated.

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

division Ginsenoside content (mg/g) Rg1 Rg3(S) Rb1 Rd F2 C.K. Rg1+Rb1+Rg3 control group 8.57 0.85 18.97 1.99 0.00 0.00 28.39 enzyme treatment 20h 6.06 0.04 1.07 2.41 9.94 1.77 7.17 24h 5.08 0.74 0.65 1.00 9.76 2.40 6.46 48h 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 12h 4.57 1.38 3.22 N.D. 9.31 3.98 9.17 18h 3.51 1.43 3.73 N.D. 6.93 4.13 8.67 24h 3.77 1.70 4.63 N.D. 7.54 4.41 10.09

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

On the other hand, although the content of Rb1 decreased after enzyme treatment, it was confirmed that the content greatly increased when fermentation was performed after the enzyme treatment. These results are 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 significantly increased as the fermentation time increased.

Test Example 6: Comparison of ginsenoside content

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

division Ginsenoside content (mg/g) Rg1 Rg3(S) Rb1 Rd F2 C.K. Rg1+Rb1+Rg3 Example I'm 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 I'm 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 I'm 5.87 0.59 17.57 - - - 24.04 after 0.18 7.70 1.15 - - - 9.03 Comparative Example 3 I'm 5.87 0.59 17.57 - - - 24.04 after 0.17 8.95 1.61 - - - 10.73 Comparative Example 4 I'm(%) 11.60 - 12.70 - - 0.00 - after(%) 14.60 - 2.40 - - 0.50 - Comparative Example 5 I'm 8.00 - 45.8 - - - 53.8 after - - - - - 1.86 -

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

In addition, the ginsenoside contents of the fermented red ginseng extracts of Examples and Comparative Examples were calculated as fermented red ginseng efficacy indicators and are shown in Table 10 below.

division Ginsenoside content (mg/g) Ginsenoside Indicator C.K. Rg1+Rb1+Rg3 CK/pre-fermentation Rb1 CK/Rb1 CK/Rg3 CK/(Rg1+Rb1+Rg3) CK/(Rg1+Rb1+Rg3+CK) Example I'm 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 I'm 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 I'm 0.00 24.04 - - - - - after fermentation 0.00 9.03 - - - - - Comparative Example 3 I'm 0.00 24.04 - - - - - Enzyme treatment after fermentation 0.00 6.73 - - - - - Comparative Example 4 I'm(%) 0.00 - - - - - - after(%) 0.50 17.00 0.04 0.21 - 0.03 0.03 Comparative Example 5 I'm - - - - - - - after 1.86 - 0.04 - - - -

Looking at Table 10, the fermented red ginseng extract of Example is the production efficiency of Compound K, which is the 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 value of dividing the content of Compound K contained in the fermented red ginseng extract by the content of Rb1 after fermentation (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 excellent.

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 the 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 that of the Example.

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 the 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 Only, it was confirmed that the production efficiency of compound K was significantly lower than that of Examples.

From the above results, according to the production method of the 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. Excellent, and it was specifically confirmed that the ginsenoside index was excellent.

Although the present invention has been described in terms of the preferred embodiments mentioned above, various modifications and variations are possible without departing from the spirit and scope of the invention. The appended claims also cover such modifications and variations as fall within the subject matter of this 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 660 720 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 having bioconversion ability for saponin ( Lactobacillus acidophilus ) BLA-9 strain (accession number: KCTC18848P). (1) preparing a red ginseng extract;
(2) enzymatic reaction by adding complex enzymes of cellulase, β-glucosidase and hemicellulase to the red ginseng extract; and
(3) inoculating the enzyme-treated red ginseng extract with Lactobacillus acidophilus BLA-9 strain (accession number: KCTC18848P) and then fermenting; fermentation with increased content of compound K, including Manufacturing method of red ginseng extract. According to claim 2,
Method for producing a fermented red ginseng extract with increased content of compound K, characterized in that the red ginseng extract in step (1) is extracted with water, lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof. According to claim 2,
Method for producing a fermented red ginseng extract with increased content of compound K, characterized in that the red ginseng extract prepared in step (1) has a solid content of 3 to 50% by weight. According to claim 2,
The complex enzyme in step (2) is a method for producing a fermented red ginseng extract with increased content of compound K, characterized in that the enzyme is derived from Aspergillus niger . According to claim 2,
Method for producing a fermented red ginseng extract with increased content of compound K, characterized in that the complex enzyme in step (2) is Sumizyme AC. According to claim 2,
The 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 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 claim 2,
The method for producing a fermented red ginseng extract with increased content of compound K, characterized in that the enzyme reaction of step (2) is performed at 50 to 65 ° C., pH 3.5 to 5.0 for 5 to 60 hours. According to claim 2,
Method for producing a fermented red ginseng extract with increased content of compound K, characterized in that the enzyme inactivation step is not performed between steps (2) and (3). According to claim 2,
Method for producing a fermented red ginseng extract with increased content of compound K, characterized in that the fermentation in step (3) is performed at 30 to 50 ° C. for 5 to 60 hours. According to claim 2,
The fermented red ginseng extract has a sum of Rb1, Rg1 and Rg3 of at least 7.0 mg/g, an F2 content of at least 5.0 mg/g, and a compound K content of at least 4.0 mg/g. Method for producing fermented red ginseng extract with increased content of According to claim 2,
Characterized in that the production efficiency of compound K, which is the value obtained by dividing the content (mg / g) of compound K contained in the fermented red ginseng extract after fermentation by the content (mg / g) of Rb1 contained in the red ginseng extract before enzyme treatment, is at least 0.2 or more Method for producing a fermented red ginseng extract with increased content of compound K. It is prepared according to the method of any one of claims 2 to 12, so that 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.

Images