KR20100113717A - Korean intestine-derived microorganisms having saponin-biotransforming activity and processes for preparing fermented ginseng using the same - Google Patents

Abstract

PURPOSE: A novel Lactobacillus sp. strain with saponin bioconversion ability and a method for producing ginseng fermentation using the same are provided to obtain ginseng fermentation with high ginsenoside Rg3 and Δ20-ginsenoside Rg3 content. CONSTITUTION: A Lactobacillus plantarum KFCC11434P has saponin bioconversion ability. A Lactobacillus sakei KFCC11435P has saponin bioconversion ability. The Lactobacillus plantarum KFCC11434P or Lactobacillus sakei KFCC11435P is used for producing ginseng fermentation by inoculating to ginseng and culturing. The ginseng is raw ginseng, white ginseng or red ginseng.

Description

Korean intestine-derived microorganisms having saponin-biotransforming activity and processes for preparing fermented ginseng using the same}

The present invention relates to a novel Lactobacillus strain having a saponin bioconversion ability derived from Korean intestine and a method for preparing ginseng fermented product using the same.

Ginseng has approximately 11 species known on Earth as a perennial belonging to the genus sukgeuncho haksang ohgagwa ginseng plant classification, typical species include Korean ginseng (Panax ginseng CA Meyer), American ginseng (Panax quinquefolium L.), jeonchilsam (Panax notoginseng FH Chen), Panax japonicus CA Meyer. Korean ginseng ( Panax ginseng CA Meyer) is native to the Far East of Asia (33-48 ° N, Korea, Northern Manchuria, and parts of Russia), and has excellent medicinal effects. American ginseng ( Panax quinquefolium L.) grows and grows in the United States and Canada. Panax notoginseng FH Chen is grown or grown in southwestern Guangxi from southeastern Yunnan Province. Also, Panax japonicus CA Meyer is distributed in Japan, southwest China and Nepal.

As research on the major ingredients and pharmacological effects of ginseng has been published, it has been spotlighted as a natural health food, its demand is gradually increasing, and it has been accelerated by economic growth and improvement of living standard, and widely used as a popular natural health food. It is becoming. In addition, ginseng has been used for various diseases such as psychiatry, nervous system diseases and diabetes in the form of herbal medicine mainly in Asian countries such as Korea, and saponins, the main components of the ginseng, are tonic, tonic, soothing, hematopoietic and antihypertensive It is reported to have an effect.

In general, ginseng is used in the form of red ginseng, which is produced by washing ginseng as it is grown and harvested, dried white ginseng or fresh ginseng at room temperature, and then steamed with steam at about 98 to 100 ° C. Of these, red ginseng is known to be much stronger than white ginseng.

Recently, the active ingredients of ginseng are easily absorbed into the human body, and as a method of reinforcing the components present in ginseng in trace amounts, the method of fermenting ginseng using intestinal microorganisms or lactic acid bacteria or treating enzymes is used.

Republic of Korea Patent Application No. 1980-4291 enzymatically decomposes ginseng residues from the intrinsic fragrance of ginseng to produce organic pH when the organic nitrogen concentration is 0.2-0.8% and the glucose content is 3% or more suitable for lactic acid bacteria fermentation. Disclosed is a method for preparing a lactic acid bacteria ginseng beverage, comprising the step of adjusting the concentration of 3.8 to 4.8 to remove insoluble polymer protein and fiber, culturing the lactic acid bacteria in the eluate, and then adding a flavor component unique to ginseng.

Korean Patent Publication No. 1997-061909 discloses a compound K as an intestinal bacterial metabolite of ginseng saponin and a metabolite of ginsenoside Rb1, ginsenoside Rb2, ginsenoside Rc, and ginsenoside Rd, which are protofaanaxadiol-based saponins. , [20-0- [aL-arabinofuranosyl (16) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol] designated as compound Y and ginsenoside Mc; It has been disclosed that 20 (S) -protopanaxatriol was isolated and identified as a metabolite of ginsenoside Rg1 and ginsenoside-Re, which are protoanax triol saponins.

In addition, Korean Patent Registration Nos. 479,803, 497,895, 517,899, 618,171, etc., ferment the ginseng or ginseng extract with lactic acid bacteria or enteric bacteria, 20-0-β-D-glucopyranosyl-20 (S Fermented ginseng (i.e. ginseng extract) with high content of) -protopanaxadiol (Compound K), ginsenoside F, 20 (S) -protopanaxadiol, 20 (S) -protopanaxatriol, and the like, and The preparation method thereof has been disclosed.

Meanwhile, Korean Patent Registration No. 228,510 heats ginseng at a temperature of 110-180 ° C. for 0.5-20 hours, and processes the obtained ginseng with alcohol, hexane, ether, dichloromethane, chloroform, ethyl acetate, and mixed solvents thereof. After the extraction with an organic solvent selected from the group consisting of chromatographed the extract and ginsenosides Rg3 and / or Rg5 comprising a step of separating and obtaining a method for preparing ginsenosides Rg3 and / or Rg5. According to Korean Patent Registration No. 228,510, the ginsenoside Rg3 and / or Rg5 is a metabolite of saponin having an excellent vasorelaxant effect.

The ginsenoside Rg3 and Δ20-ginsenoside Rg3 are known to have excellent anti-tumor and anticancer effects as well as excellent vasodilating effect (Kim et al., Eur. J. Pharmacol. 1999, 367 (1): 51 -57; Kim et al., Arch. Pharm. Res. 2004, 27 (4): 429-435; Li X. et al., Sichuan Da Xue Xue Bao Yi Xue Ban. 2005, 36 (2): 217- 220).

However, as disclosed in Korean Patent Registration No. 228,510, when preparing ginseng extracts having a high content of ginsenoside Rg3 and 20-ginsenoside Rg3, generation of carcinogens such as HMF (Hydroxylmethylfurfual) generated during heat treatment Inevitably, it is difficult to use directly as a food because it must inevitably undergo a neutralization process according to a heat and / or acid treatment process, and an extraction and purification process must be performed separately.

In addition, when ginseng extract is prepared by fermenting ginseng with lactic acid bacteria or enteric bacteria as disclosed in Korean Patent Registration Nos. 479,803, 497,895, 517,899, 618,171, Ginsenoside Rg3 and △ The content of 20-ginsenoside Rg3 is very low, making it difficult to expect vasodilation, antitumor, and anticancer effects.

In addition, the Republic of Korea Patent No. 789 678, 1 - 856 790 and No. 866 504 discloses a novel microorganism that is, Phedi OKO Gus in (Pediococcus Sp.) Strains and Lu Pocono stock in (Leuconostoc Sp.) Strain, Lactobacillus Ali menta Julius (Lactobacillus alimentarius ) M-2 strain (KCTC 11054 BP), Leuconostoc mensenteroides M-3 strain (KCTC 11055 BP), Lactobacillus brevis M2 strain (KCTC11390BP) and Lactobacillus plantarta A method of preparing fermented ginseng with high content of ginsenoside Rg3 has been disclosed by fermenting ginseng with rum ( Lactobacillus plantarum ) P2 strain (KCTC11391BP). However, even if the ginseng is fermented according to the conventional manufacturing method, because the content of ginsenoside Rg3 is as low as up to about 10%, it is still not satisfactory.

The present inventors have extensively searched for microorganisms capable of producing ginseng fermentation products having high content of ginsenoside Rg3 and Δ20-ginsenoide Rg3, which are known to be excellent in vasorelaxation, antitumor, and anticancer effects. As a result, dozens of microorganisms having ginseng fermentation activity were isolated from feces and kimchi liquid collected from all over the country. Among them, two strains belonging to the genus Lactobacillus have a significantly higher saponin bioconversion capacity. It has been found that ginseng fermentation with high content of ginsenoside Rg3 and Δ20-ginsenoide Rg3 can be prepared.

Therefore, an object of the present invention is to provide a novel Lactobacillus genus microorganism having a saponin bioconversion ability.

In addition, an object of the present invention is to provide a method for producing a ginseng fermented product using the microorganism.

According to one aspect of the present invention, Lactobacillus plantarum KFCC11434P or Lactobacillus sakei KFCC11435P having saponin bioconversion ability is provided.

According to another aspect of the present invention, ginseng comprising inoculating and culturing ginseng with at least one strain selected from the group consisting of Lactobacillus plantarum KFCC11434P and Lactobacillus sakei KFCC11435P. A method for producing a fermentation product is provided.

In the production method of the ginseng fermentation, the ginseng may be ginseng, white ginseng, or red ginseng. In addition, the method of producing the ginseng fermentation may further include the step of aging the culture solution obtained after the culture at 80 to 95 ℃ for 12 to 48 hours ("aging step"). In addition, the manufacturing method of the ginseng fermentation may further comprise the step of concentrating or lyophilizing the aging liquid obtained by performing the aging step or the supernatant taken by centrifuging the aging liquid.

Lactobacillus plantarum KFCC11434P and Lactobacillus sakei KFCC11435P newly isolated by the present invention have excellent saponin bioconversion ability. In particular, when the microorganism is inoculated with ginseng such as ginseng, white ginseng, red ginseng, and cultured, ginseng fermentation products having a high content of ginsenoside Rg3 and Δ20-ginsenoide Rg3 can be obtained.

"Ginseng fermented product" as used herein is inoculated with microorganisms, especially Lactobacillus plantarum KFCC11434P and / or Lactobacillus sakei KFCC11435P to ginseng, such as ginseng, white ginseng, red ginseng, and cultured It refers to a product obtained by, and may also be referred to as 'fermented ginseng' and the like.

The present invention is a microorganism having a saponin bioconversion ability, a novel microorganism having excellent ginsenoside Rg3 and Δ20-ginsenoide Rg3 generating ability, that is, Lactobacillus plantarum KFCC11434P or Lactobacillus Sakii ( Lactobacillus) sakei ) to provide KFCC11435P.

The present inventors collected fecal and kimchi solution from all over the country, and isolated strains forming a single colony using lactic acid bacteria separation medium (MRS-BCP medium). The obtained strains were added to the red ginseng concentrate to measure metabolic activity, and dozens of microorganisms having fermentation activity were isolated. From the obtained microorganisms, two kinds of microorganisms having particularly high saponin bioconversion ability, particularly ginsenoside Rg3 and Δ20-ginsenoide Rg3 generating ability, were selected. As a result of analyzing 16S rRNA gene sequences of two isolated microorganisms, it was found that they had nucleotide sequences of SEQ ID NOs: 1 and 2, respectively, and compared the homology using BLAST, and analyzed the characteristics of the strains. It was found to be a new strain belonging to Lactobacillus plantarum and Lactobacillus sakei , respectively. The two strains, Lactobacillus plantarum CKDHC 0801 and Lactobacillus sakei CKDHC 0802, were deposited to the Korea Microorganism Conservation Center on Dec. 18, 2008, and deposited with KFCC 11434P and KFCC 11435P was granted.

The present invention also provides a method for producing ginseng fermentation comprising inoculating ginseng and incubating at least one strain selected from the group consisting of Lactobacillus plantarum KFCC11434P and Lactobacillus sakei KFCC11435P. To provide. As described above, Lactobacillus plantarum KFCC11434P and Lactobacillus sakei KFCC11435P newly isolated by the present invention have excellent saponin bioconversion ability, in particular, ginseng using When fermented, ginseng fermented with high content of ginsenoside Rg3 and Δ20-ginsenoide Rg3 can be obtained.

The ginseng used as a substrate of the strain in the production method of the present invention is not particularly limited, and both ginseng itself and ginseng processed materials can be used, for example, ginseng, red ginseng, red rice ginseng, white ginseng, misam, ginseng fruit, ginseng Leaves and the like, or ginseng extract and ginseng powder may be used. The origin of the ginseng is not limited, and for example, Korean ginseng, samchil ginseng, American ginseng, bamboo shoot ginseng, Himalayan ginseng, Vietnamese ginseng and the like can be used without limitation. In addition, the ginseng may be used by sterilization in a conventional manner as needed.

In addition, ginseng in the form of a dry powder, acid treated ginseng, high temperature treated ginseng, and pressurized ginseng disclosed in Korean Patent Registration No. 497,895 may also be used. However, when the acid treatment is lowered the pH of the culture medium may lower the activity of the strain, and when treated at a high temperature may cause carcinogens such as HMF during heat treatment, it is preferable to use ginseng or ginseng powder as it is. . The ginseng powder may have a particle size of at least 100 mesh or more to increase the surface area of ginseng to increase the speed of fermentation of microorganisms.

The method of fermenting ginseng, that is, inoculation and culture of the strain may be performed according to conventional methods (Korean Patent Registration Nos. 479,803, 497,895, 517,899, 618,171, etc.). For example, ginseng powder or ginseng extract (for example, alcohol or alcohol solution extract, hot water extract, etc.) is suspended in water, and the above strain is added thereto, and at 20 to 50 ° C., preferably at 30 to 37 ° C. , 12 hours to 15 days, preferably 24 hours to 5 days.

By going through the fermentation process as described above, ginsenosides Rb1, Rb2, Rc, and Rd contained in the ginseng raw material are ginsenosides Rg3, Δ20-ginsenoside Rg3, compound K (Compound K), ginsenoside Rh2 Bioconverted to the back.

The production method of the present invention may further comprise the step of aging the culture solution obtained after the culture at 80 to 95 ℃ for 12 to 48 hours ("aging step"). When the aging step is performed under the above conditions, sterilization is also performed at the same time as aging, and further, ginsenoside Rg3 and Δ20-ginsenoside Rg3 may be further increased.

The ginseng fermented product obtained according to the production method of the present invention can be used directly as a food without separate purification. That is, the ginseng fermented product obtained according to the production method of the present invention may be concentrated or lyophilized and powdered according to a conventional method and applied to food as it is. That is, the concentrated or lyophilized aging liquid obtained through the aging process may be applied to the food as it is, or the supernatant taken by centrifugation of the aging liquid may be applied to the food as it is. .

Hereinafter, the present invention will be described in more detail with reference to examples. However, these examples are for illustrating the present invention, and the scope of the present invention is not limited to these examples.

Example 1. Isolation and Identification of Strains

(1) Strain Isolation

Fecal and kimchi collected from all over the country were refrigerated at 4 ℃. The collected fecal and kimchi solution was stirred uniformly, and then diluted stepwise using 0.9% sterile saline solution (10 ^-2 , 10 ^-4 , 10 ^-6 and 10 ^-8 ), and MRS (lactic acid bacteria separation medium) ( deMan Rogosa Sharpe, Difco ™, Becton Dickinson and Company, USA) -BCP (Bromocresol Purple) flat plates. The smeared medium observed colony formation at 30 ° C. and incubated for 24-48 hours until significant levels of colony were formed. Strain colonies were selected in which the index purple of the original medium formed yellow colonies and borders through the lactic acid formation of the strains in the media at diluent concentrations that were distinguishable as single colonies. Selected colonies were subcultured on MRS-BCP plate medium again to confirm that they were single colonies, and metabolic activity was measured for the strains identified as single colonies.

The activity was performed by adding the obtained strains to the red ginseng concentrate to check whether they had metabolic activity. Specifically, 1 ml of red ginseng concentrate (Deokwon ginseng herbal farming combination of Geumsan, Chungnam) was appropriately diluted with 0.9% saline. MRS agar (Difco Laboratories, dextrose 20.0 g / l; meat peptone 10.0 g / l; beep extract 10.0 g / l; yeast extract 5.0 g / l; sodium acetate 5.0 g / l; 2.0 g / l disodium phosphate; 2.0 g / l ammonium citrate; 1.0 g / l tween 80; 0.1 g / l magnesium sulfate; 0.05 g / l manganese sulfate; 15 g / l agar) After incubation at 30 ℃ and 37 ℃ was incubated for 72 hours and 48 hours, respectively.

As a result of measuring the metabolic activity as described above, about 10 kinds of microorganisms were isolated. 90 g of water was added to 10 g of red ginseng extract using each microorganism isolated, sterilized at 85 ° C., and then inoculated with the microorganism at a concentration of about 5%, followed by incubation at 37 ° C. for 2 days. The obtained culture was aged and sterilized at 85 ° C. for 24 hours, and then analyzed by ginsenoside by HPLC. As a result, two microorganisms (CKDHC 0801 and CKDHC) having particularly high ability to produce ginsenoside Rg3 and Δ20-gingenide Rg3 were detected. 0802). Of the two microorganisms, CKDHC 0801 is a strain isolated in human intestine, CKDHC 0802 is a strain isolated from kimchi liquid.

(2) Identification of isolated strain by 16S rRNA gene sequencing

16S rRNA gene sequencing of two isolated microorganisms was performed. 16S rRNA gene sequencing, see Lane, D. J. (1991), 16S / 23S sequencing. In: Nucleic Acids Techniques in Bacterial Systematics, Eds .: Stackebrandt, E. and Goodfellow, M. pp. 115-175, John Wiley and Sons, New York and Harju, S., et al., 2004, Rapid isolation of yeast genomic DNA: Bust n 'Grab. BMC Biotech. It was carried out according to the method disclosed in 21: 4-8.

The two microorganisms were inoculated in 10 ml of MRS liquid medium and incubated in a rotary incubator at 80 rpm and 30 ° C. for 2 days. 1 ml of the culture was placed in a 1.5 ml centrifuge tube and centrifuged at 15,000 x g for 5 minutes. The supernatant was removed to extract genomic DNA and then the tube was placed in Lysis buffer (2% (v / v) Triton X-100, 1% (w / v) SDS, 100 mM NaCl, 10 mM Tris-Cl and 1 mM). EDTA; pH 8.0) 200 μl was added to make it homogeneous, and the tube was frozen at −80 ° C. for 4 minutes and then bathed again in boiling water for 2 minutes. This freezing and melting process was performed twice, followed by vortexing for 30 seconds. Then, 200 μl of chloroform was added and vortexed again for 2 minutes, and then centrifuged at 15,000 × g for 5 minutes at 4 ° C. Only the supernatant was separated and treated with 2 μl of RNaseI solution (mg / ml). Genomic DNA was isolated using ethanol precipitation and dissolved in 200 μl of TE solution, which was used to amplify the 16S rRNA gene.

16S rRNA gene from the chromosomal DNA of the microorganism was PCR amplified using primers 8F (AGAGTTTGATCCTGGCTCAG, SEQ ID NO: 3) and 1541R (AAGGAGGTGATCCAGCC, SEQ ID NO: 4). PCR program for amplification was performed as follows. After reacting at 94 ° C. for 5 minutes, the cycle consisting of 30 seconds at 94 ° C., 40 seconds at 55 ° C., and 1 minute 30 seconds at 72 ° C. was repeated 35 times and reacted at 72 ° C. for 5 minutes as the final elongation step. PCR products amplified from genomic DNA were purified using Accu Prep gel purification kit (Accu. Chem. Sci. Corp. Westbury, NY). Each PCR amplification product thus obtained was introduced into a pPCR2.1-TOPO vector (Invitrogen, USA) and transformed with E. coli DH5a (Real Biotech, Inc., Taiwan) using this vector. After valid E. coli DH5a strains were identified, the plasmids were isolated and purified. The sequencing of the 16S rRNAs in pM-2 and pM-3 which were thus obtained plasmids was performed. As a result of sequencing, the 16S rRNA sequences of the two microorganisms, CKDHC 0801 and CKDHC 0802, were identified as SEQ ID NOs: 1 and 2, respectively (1474 bp and 1455 bp, respectively). As a result of analyzing the homology of the gene sequence using BLAST, the CKDHC 0801 strain belongs to Lactobacillus plantarum , and the CKDHC 0802 strain turns out to be a new strain belonging to Lactobacillus sakei . It became.

(3) Analysis of sugar fermentation ability (biochemical characterization)

For two microorganisms obtained through sequencing of the 16S rRNA gene, namely, Lactobacillus plantarum CKDHC 0801 and Lactobacillus sakei CKDHC 0802 strains, the API 50 CHL kit (BioMerieux, France) The fermentation test was carried out for 49 sugars using), the results are shown in Table 1a and 1b.

Biochemical Properties of Isolated Strains No. Metric Lb. plantarum
CKDHC 0801 Lb. sakei
CKDHC 0802 One Glycerol - - 2 Erythritol - - 3 D-Arabinose - - 4 L-Arabinose + + 5 D-Ribose + + 6 D-Xylose - - 7 L-Xylose - - 8 D-Adonitol - - 9 Methyl-β-D-Xylopyranoside - - 10 D-galactose + + 11 D-Glucose + + 12 D-Fructose + + 13 D-Mannose + + 14 L-Sorbose - - 15 L-Rhamnose - - 16 Dulcitol - - 17 Inositol - - 18 D-Mannitol + - 19 D-Sorbitol + - 20 Methyl-a-D-Mannopyranoside + - 21 Methyl-a-D-Glucopyranoside - - 22 N-acetylglucosamine + + 23 Amygdalin + - 24 Arbutin + - 25 Esculin ferric citrate - -

Biochemical Properties of Isolated Strains No. Metric Lb. plantarum
CKDHC 0801 Lb. sakei
CKDHC 0802 26 Salincin + - 27 D-Cellobiose + - 28 D-Maltose + + 29 D-Lactose + - 30 D-Melibiose + + 31 D-Saccharose + + 32 D-trehalose + + 33 Inulin + - 34 D-Melezitose + - 35 D-Raffinose + - 36 Amidon (starch) - - 37 Glycogen - - 38 Xylitol - - 39 Gentiobiose + + 40 D-Turanose - - 41 D-Lyxose - - 42 D-Tagatose - - 43 D-Fucose - - 44 L-Fucose - - 45 D-Arabitol - - 46 L-Arabitol - - 47 Potassium Gluconate - - 48 Potassium 2-Ketogluconate - - 49 Potassium 5-Ketogluconate - -

In Tables 1a and 1b, "+" indicates fermentation using sugars and "-" indicates no fermentation using the corresponding sugars.

(4) Strain Deposit

The two strains isolated above, namely Lactobacillus plantarum CKDHC 0801 and Lactobacillus sakei CKDHC 0802, were deposited at the Korea Microorganism Conservation Center on Dec. 18, 2008, and deposited no. KFCC 11434P and KFCC 11435P were given.

Example 2.

250 g of water was added to 50 g of Korean ginseng powder, sterilized at 85 ° C., and then inoculated with Lactobacillus plantarum CKDHC 0801 (microbial accession number KFCC11434P) at a concentration of about 5%, followed by 2 days at 37 ° C. Incubate with stirring. At this time, the culture was carried out under reduced pressure to easily diffuse the culture solution into the particle pores of ginseng powder. The resulting culture was aged and sterilized at 85 ° C. for 24 hours. The obtained aging solution was concentrated in vacuo at 75 ° C. to prepare ginseng fermented product (ie, fermented ginseng).

Example 3.

American ginseng was used instead of Korean ginseng, and fermented ginseng was prepared in the same manner as in Example 2 using Lactobacillus sakei CKDHC 0802 (Microbial Accession No. KFCC11435P) as a strain.

Example 4.

Bamboo ginseng was used instead of Korean ginseng, and fermented ginseng was prepared in the same manner as in Example 2 using Lactobacillus sakei CKDHC 0802 (Microbial Accession No. KFCC11435P) as a strain.

Example 5.

Fermented ginseng was prepared in the same manner as in Example 2, using Himalayan ginseng instead of Korean ginseng.

Example 6.

Using ginseng instead of Korean ginseng, fermented ginseng was prepared in the same manner as in Example 2.

Test Example 1.

As a comparative example, fermented ginseng was prepared as disclosed in Korean Patent Registration No. 479,803 (Comparative Example 1). The changes in the components of the fermented ginseng obtained in the above Examples and Comparative Examples were measured as follows. 5 g of the ginseng fermentation product obtained is precisely weighed and taken into a 100 ml concentrated flask. After concentration under reduced pressure, 50 ml of saturated butanol is added, a reflux condenser is added and the mixture is heated and extracted at 70 to 80 ° C. in a water bath for about 1 hour, and then filtered and the residue The same operation was repeated twice for. The filtrate was washed with saturated butanol 10 and the combined filtrate and washings were placed in a 250 ml separatory funnel and shaken well with 20 ml of water and washed with water. Transfer the entire saturated butanol extract to a concentrated flask with a constant amount, remove it under reduced pressure in a water bath, remove 50 mL of butanol, add 50 ml of ether to the residue, attach it to a reflux condenser, heat it for 30 minutes in a water bath, and degrease the ether. Removed. 10 times methanol was added to the obtained residue to completely dissolve it, and then filtered using a filter (0.45 μm), followed by quantitative analysis of ginsenoside components by high performance liquid chromatography (HPLC). The results are shown in Table 2 below. The HPLC conditions are as follows.

Organization: Shimadzu Prominence

Column: Gromsil ODS 4.6 (i, d) × 250 mm

Mobile phase A: acetonitrile: distilled water = 15: 85

Mobile phase B: acetonitrile: distilled water = 80: 20

Flow rate: 1.0 ml / min

Detector: ELSD (Alltech)

division Ginsenoside (%) Example 2 Example 3 Example 4 Comparative Example 1 Ginsenoside Rb1 0.6 0.5 0.5 2.7 Ginsenoside Rb2 0.3 0.3 0.3 2.2 Ginsenoside Rc 0.2 0.2 0.2 1.9 Ginsenoside Rd 0.1 0.2 0.2 1.3 Ginsenoside Rg3 27.8 28.2 28.3 9.00 Δ20-ginsenoside Rg3 30.2 30.4 31.2 10.52 Ginsenoside Rh2 0.8 1.4 1.2 0.4 Compound K 0.2 0.1 0.2 1.5

As can be seen from the results of Table 2, in the case of fermented ginseng prepared through conventional acid treatment and lactic acid bacteria fermentation, ginsenoside Rg3 and Δ20-ginsenoside Rg3 content was only about 10%, The ginseng fermentation product prepared using the newly isolated strain according to the present invention showed high levels of ginsenoside Rg3 and Δ20-ginsenoside Rg3 of about 30% or more.

<110> CHONGKUNDANGHEALTHCARE CO. <120> Korean intestine-derived microorganisms having          saponin-biotransforming activity and processes for preparing          fermented ginseng using the same <130> PN0240 <160> 4 <170> KopatentIn 1.71 <210> 1 <211> 1474 <212> DNA <213> Lactobacillus plantarum <400> 1 gcggttacgg cgtgctatac atgcaagtcg aacgaactct ggtattgatt ggtgcttgca 60 tcatgattta catttgagtg agtggcgaac tggtgagtaa cacgtgggaa acctgcccag 120 aagcggggga taacacctgg aaacagatgc taataccgca taacaacttg gaccgcatgg 180 tccgagcttg aaagatggct tcggctatca cttttggatg gtcccgcggc gtattagcta 240 gatggtgggg taacggctca ccatggcaat gatacgtagc cgacctgaga gggtaatcgg 300 ccacattggg actgagacac ggcccaaact cctacgggag gcagcagtag ggaatcttcc 360 acaatggacg aaagtctgat ggagcaacgc cgcgtgagtg aagaagggtt tcggctcgta 420 aaactctgtt gttaaagaag aacatatctg agagtaactg ttcaggtatt gacggtattt 480 aaccagaaag ccacggctaa ctacgtgcca gcagccgcgg taatacgtag gtggcaagcg 540 ttgtccggat ttattgggcg taaagcgagc gcaggcggtt ttttaagtct gatgtgaaag 600 ccttcggctc aaccgaagaa gtgcatcgga aactgggaaa cttgagtgca gaagaggaca 660 gtggaactcc atgtgtagcg gtgaaatgcg tagatatatg gaagaacacc agtggcgaag 720 gcggctgtct ggtctgtaac tgacgctgag gctcgaaagt atgggtagca aacaggatta 780 gataccctgg tagtccatac cgtaaacgat gaatgctaag tgttggaggg tttccgccct 840 tcagtgctgc agctaacgca ttaagcattc cgcctgggga gtacggccgc aaggctgaaa 900 ctcaaaggaa ttgacggggg cccgcacaag cggtggagca tgtggtttaa ttcgaagcta 960 cgcgaagaac cttaccaggt cttgacatac tatgcaaatc taagagatta gacgttccct 1020 tcggggacat ggatacaggt ggtgcatggt tgtcgtcagc tcgtgtcgtg agatgttggg 1080 ttaagtcccg caacgagcgc aacccttatt atcagttgcc agcattaagt tgggcactct 1140 ggtgagactg ccggtgacaa accggaggaa ggtggggatg acgtcaaatc atcatgcccc 1200 ttatgacctg ggctacacac gtgctacaat ggatggtaca acgagttgcg aactcgcgag 1260 agtaagctaa tctcttaaag ccattctcag ttcggattgt aggctgcaac tcgcctacat 1320 gaagtcggaa tcgctagtaa tcgcggatca gcatgccgcg gtgaatacgt tcccgggcct 1380 tgtacacacc gcccgtcaca ccatgagagt ttgtaacacc caaagtcggt ggggtaacct 1440 tttaggaacc agccgcctaa ggtgacaatg atgg 1474 <210> 2 <211> 1455 <212> DNA <213> Lactobacillus sakei <400> 2 agtcgaacgc actctcgttt agattgaagg agcttgctcc tgattgataa acatttgagt 60 gagtggcgga cgggtgagta acacgtgggt aacctgccct aaagtggggg ataacatttg 120 gaaacagatg ctaataccgc ataaaaccta acaccgcatg gtgtagggtt gaaagatggt 180 ttcggctatc actttaggat ggacccgcgg tgcattagtt agttggtgag gtaaaggctc 240 accaagaccg tgatgcatag ccgacctgag agggtaatcg gccacactgg gactgagaca 300 cggcccagac tcctacggga ggcagcagta gggaatcttc cacaatggac gaaagtctga 360 tggagcaacg ccgcgtgagt gaagaaggtt ttcggatcgt aaaactctgt tgttggagaa 420 gaatgtatct gatagtaact gatcaggtag tgacggtatc caaccagaaa gccacggcta 480 actacgtgcc agcagccgcg gtaatacgta ggtggcaagc gttgtccgga tttattgggc 540 gtaaagcgag cgcaggcggt ttcttaagtc tgatgtgaaa gccttcggct caaccgaaga 600 agtgcatcgg aaactgggaa acttgagtgc agaagaggac agtggaactc catgtgtagc 660 ggtgaaatgc gtagatatat ggaagaacac cagtggcgaa ggcggctgtc tggtctgtaa 720 ctgacgctga ggctcgaaag catgggtagc aaacaggatt agataccctg gtagtccatg 780 ccgtaaacga tgagtgctag gtgttggagg gtttccgccc ttcagtgccg cagctaacgc 840 attaagcact ccgcctgggg agtacgaccg caaggttgaa actcaaagga attgacgggg 900 gcccgcacaa gcggtggagc atgtggttta attcgaagca acgcgaagaa ccttaccagg 960 tcttgacatc ctttgaccac tctagagata gagctttccc ttcggggaca aagtgacagg 1020 tggtgcatgg ttgtcgtcag ctcgtgtcgt gagatgttgg gttaagtccc gcaacgagcg 1080 caacccttat tactagttgc cagcatttag ttgggcactc tagtgagact gccggtgaca 1140 aaccggagga aggtggggac gacgtcaaat catcatgccc cttatgacct gggctacaca 1200 cgtgctacaa tggatggtac aacgagttgc gagaccgcga ggtttagcta atctcttaaa 1260 accattctca gttcggattg taggctgcaa ctcgcctaca tgaagccgga atcgctagta 1320 atcgcggatc agcatgccgc ggtgaatacg ttcccgggcc ttgtacacac cgcccgtcac 1380 accatgagag tttgtaacac ccaaagccgg tgaggtaacc cttcggggag ccagccgtct 1440 aagtgaacag atgca 1455 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer <400> 3 agagtttgat cctggctcag 20 <210> 4 <211> 17 <212> DNA <213> Artificial Sequence <220> <223> Primer <400> 4 aaggaggtga tccagcc 17  

Claims (6)

Lactobacillus plantarum KFCC11434P with saponin bioconversion ability. Lactobacillus sakei KFCC11435P with saponin bioconversion ability. Lactobacillus plantarum KFCC11434P and Lactobacillus sakei KFCC11435P at least one strain selected from the group consisting of ginseng fermentation method comprising the step of inoculating and culturing. The method of claim 3, wherein the ginseng is ginseng, white ginseng, or red ginseng. The method according to claim 3 or 4, further comprising the step of aging the culture solution obtained after the culture at 80 to 95 ° C. for 12 to 48 hours (“aging step”). The method according to claim 5, further comprising the step of concentrating or lyophilizing the aging solution obtained by performing the aging step or the supernatant taken by centrifuging the aging solution.