KR101388918B1 - Lactobacillus rhamnosus HK-9 derived from human body, and fermented material and cosmetic composition using it - Google Patents

Abstract

The present invention relates to a lactic acid bacterium Lactobacillus rhamnosus strain HK-9 derived from a human body and a utilization method thereof, and more particularly to a lactic acid bacterium strain Lactobacillus rhamnosus strain HK-9 having a physiological activity enhancing activity through natural product fermentation, Lactobacillus lambosus HK-9 having an antioxidant effect and a fermented product and cosmetic product using the same.
The lactic acid bacterium according to the present invention is a strain derived from a human body, not a general food, and thus is very effective for activity such as whitening as well as food fermentation. As a result, it is possible to effectively dissolve the effective substance by overcoming the problem that it is not eluted from the existing natural product or the elution of a very small amount and the fermentation efficiency is inferior, and the effect as a raw material for cosmetics can be obtained.

Description

Lactobacillus rhamnosus HK-9 derived from human body Lactobacillus lambosus HK-9 and fermented product using the same and a cosmetic composition {Lactobacillus rhamnosus HK-9 derived from human body, and fermented material and cosmetic composition using it}

The present invention relates to a lactic acid bacterium Lactobacillus rhamnosus strain HK-9 derived from a human body and a utilization method thereof, and more particularly to a lactic acid bacterium strain Lactobacillus rhamnosus strain HK-9 having a physiological activity enhancing activity through natural product fermentation, Lactobacillus Lambosus HK-9 having an antioxidative effect and a fermented product and a cosmetic product using the same.

Since the lactic acid bacterium according to the present invention is a strain derived from a human body, not a general food, it is very effective for activity such as whitening as well as food fermentation. As a result, it is possible to effectively dissolve the effective substance by overcoming the problem that it is not eluted from the existing natural product or the elution of a very small amount and the fermentation efficiency is inferior, and the effect as a raw material for cosmetics can be obtained.

Lactobacillus ( Lactobacillus ) is an important microorganism widely used worldwide in the production of various fermented foods including fermented dairy products such as cheese, fermented milk and lactic acid bacteria drink.

Lactic acid bacteria have been developed and used for a long time in order to effect the dressing effect of treatment and prevention of fermentation and intestinal diseases in the food industry. There are several reports on the use of lactic acid bacteria. As an example, 70% of patients suffering from diarrhea have been treated since they reported that they were treated with lactic acid bacteria for infantile diarrhea (RO Clock, JAMA Vol. 61, P.164, 1913) (LM Gornpertz, et al., JAMA Vol. 80, P. 90, 1923), inhibiting the growth of intestinal noxious bacteria and providing favorable conditions for the growth of beneficial bacteria to prevent food poisoning, prevent intestinal decline, prevent constipation, , And that it has various physiological functions such as inhibition of carcinogenesis.

Fermented products using these lactic acid bacteria are produced in the form of yogurt or cheese, or have a great influence on the food industry such as kimchi and chonggukjang.

On the other hand, when a product is fermented using a common lactic acid bacterium, there is a limit to the production of various physiologically active substances produced during the fermentation and fermentation of the fermented product. In addition, depending on the type of lactic acid bacteria, the yield of fermentation can be changed when fermenting natural products. Some of them are used for foods, and the fermentation activity is used for cosmetics. It is true. However, most of the lactic acid bacteria used for producing the fermented product having such functionality are imported from foreign countries, and the source of lactic acid bacteria is a strain isolated from fermented dairy products such as cheese.

Therefore, securing new lactic acid bacteria having excellent fermentation ability can have a very large economic effect on economics. Therefore, the present inventors have focused on the fact that the food is the taste of man's hand and that lactic acid bacteria are present in the hands of women [ In fact, there are reports that lactic acid bacterium is present in many female hands (PNAS 105 (46) 17994-17999). The bacteria that are present in many female hands are identified and their physiological activity Lactobacillus strain Lactobacillus strain HK-9, which is a novel fermented lactic acid bacterium, was isolated.

Lactobacillus Lactobacillus Lymphosus HK-9 isolated from the present invention is a lactic acid bacterium isolated from a woman's hand in view of the fact that the female hand is more exquisite than that of the male hand. In addition to fermentation ability, which is a function of lactic acid bacteria, the strain has activity to whitening and so on.

The usual lactic acid bacteria are mostly used for food fermentation since most of the strains obtained from food are used, but the strains identified by the present inventor can be applied to foods and cosmetics.

Although it has been fully illustrated by the above description, the object of the present invention is to provide a novel microorganism which can be used both for fermentation of natural materials and for cosmetics.

The present inventors isolated the strains corresponding to the above purposes and confirmed the increase of the effective physiological activity and the improvement of the yield of fermentation through the fermentation of natural materials and verified the whitening effect.

The present invention as described above is characterized in that the lactic acid bacteria Lactobacillus lambosus HK-9 (accession number: KCCM11254P) isolated from a human body.

The present invention is also characterized in that it is a fermented product fermented with the above-mentioned lactic acid bacteria.

And a liquid (extract of the lactic acid bacteria itself) extracted from the lactic acid bacteria. At this time, the extract of the lactic acid bacteria itself was obtained by centrifuging the Lactobacillus lambosus strain HK-9 (accession number: KCCM11254P) cultured in the MRS medium to obtain a strain, subjecting the obtained strain to ultrasonic treatment, And the like.

Further, the present invention is characterized in that it is a cosmetic composition or food containing the above-mentioned fermented product.

The present invention relates to Lactobacillus lambosus strain HK-9 isolated from a woman's hand. The present invention relates to an anti-oxidant effect , It can be supplied as a raw material for food and cosmetic materials and can have a large economic impact.

FIG. 1 shows the 16S rDNA sequence of the strain of Lactobacillus lambosus HK-9, which was examined by the Korean Microorganism Preservation Center.
Fig. 2 is a diagram showing phylogenetic locations of Lactobacillus lambatus HK-9.
Fig. 3 is a diagram showing the morphological appearance of lactic acid bacteria by SEM photograph of Lactobacillus lambosus HK-9.
FIG. 4 is a graph showing the cell concentration of Lactobacillus laminocus HK-9 using the MRS medium over time. FIG.
5 is a graph showing ginsenoside content of ginseng following fermentation of Lactobacillus lambosus strain HK-9 and other common strains.
FIG. 6 is a graph showing the result of the production of a new substance in the broth fermented using Lactobacillus lambosus HK-9. FIG.
7 is a graph showing the results of DPPH activity before and after fermentation of the above fermented broccoli.
8 and 9 show results of the whitening-related efficacy experiments of Lactobacillus lambosus strain HK-9 itself, FIG. 8 shows the melanin synthesis inhibition activity experiment, FIG. 9 shows the tyrosinase inhibition experiment Fig.

Hereinafter, the present invention will be described in more detail by way of examples.

Example 1. Method for isolating lactic acid bacteria

In order to isolate the new lactic acid bacteria, 40 people in their 20s and 40s were asked to participate in the experiment about 7 days before the experiment. Experiments were carried out on the unspecified day .

The initial microorganisms were inoculated into the previously prepared MRS Agar medium by the palms of the experimental subjects. The inoculated initial samples were cultured at 30 ° C for 24 hours, and the obtained microorganism colonies were inoculated again onto the MRS agar medium to obtain strains. The most active microorganisms were selected as a result of fermentation by each colony obtained from the second inoculation.

The microorganisms obtained by the above method were referred to the Korean Microorganism Preservation Center and lactic acid bacteria were identified using PCR. The results are shown in Fig. 1 (base sequence of 16S rDNA, sequence is attached to Sequence Listing 1) and Fig. 2 Position).

As a result of the Korean Microorganism Conservation Center, it was confirmed that the strain obtained in the present invention is a strain belonging to the genus Lactobacillus . The inventors named the above strain Lactobacillus rhamnosus strain HK-9 and deposited this strain with the deposit number KCCM11254P on Feb. 21, 2012 at the Korean Culture Center of Microorganism.

The morphology of the lactic acid bacteria confirmed by the SEM photograph of the new strain Lactobacillus lambosus HK-9 is shown in Fig.

Example 2. Culture method of lactic acid bacteria

The lactic acid bacteria isolated in Example 1 were cultured in MRS medium and the optimal culture time was determined according to the characteristics of the strain. The culture medium was prepared by adding 55% of MRS medium and 0.5% of L-cysteine hydrochloride to the culture medium. The incubation temperature was 36 ° C. and the experiment was carried out under anaerobic conditions in the form of a sealed fermentation strain Therefore, the culture was carried out at a minimum stirring speed (50 to 60 rpm), and the volume of the flask was 400 ml in a 1 L flask for 72 hours.

FIG. 4 shows the cell concentration of Lactobacillus lambosus strain HK-9 cultured by the above method. According to the results, when the cells were cultured up to 12 hours, the cells were present at an amount of about 644 x 10 ⁶ , but when they were cultured for 12 hours to 24 hours, the maximum concentration was increased to 3255 x 10 ⁶ , The cell concentration was maintained, and after 48 hours, the cells were gradually reduced.

Therefore, the optimum incubation time of the Lactobacillus lambosus strain HK-9 of the present invention was determined to be 48 hours.

Example 3. Validation of Enhancement of Effective Physiological Activity through Fermentation of Natural Products

1) Analysis of ginsenoside content of fermented pancake

The thus-isolated Lactobacillus lambosus strain HK-9 was used to ferment the panting. As a fermentation method, 100 g of pansam was added in a powder form to a 2 L flask, and then 1 L of a culture medium was prepared. Then, Lactobacillus lambosus strain HK-9 was inoculated 5% and fermented for 48 hours.

In addition, Lactobacillus rhamnosus GG, Lactobacillus paracasei, Lactobacillus casei and Lactobacillus acidophilus Plantarum + Bifidobacterium lactis Longum strains were used as a control group together with Lactobacillus lambosus strain HK-9 in the same manner as Lactobacillus lambosus strain HK-9 .

The total saponin content (Rg3 + Rg2 + Rh2 + CK + Rg1 + Rb1 + Rb2 + Rc + Re + Rd) was measured by using the butanol extraction method after making the fermented product into powder form for ginsenoside analysis.

As can be seen from the results of the content of ginsenosides in the fermentation of the pancake according to the lactic acid bacteria of FIG. 5, it was confirmed that the highest amount of ginsenoside was detected in the Lactobacillus lambosus strain HK-9, which is the lactic acid bacterium of the present invention, Of lactic acid bacteria up to 200%.

2) Antioxidant activity test of golden fermented product

The fermented product of gold was made by the same method as the fermentation of pansam. In a 2 L flask, 100 g of gold was ground and added to a 1 L culture medium, followed by 5% inoculation with Lactobacillus lambosus strain HK-9 and fermentation for 48 hours.

In addition, Lactobacillus rhamnosus GG, Lactobacillus paracasei, and Lactobacillus casei strains were used as a control in combination with Lactobacillus lambosus strain HK-9 and cultured in the same manner as the Lactobacillus lambosus strain HK-9.

For the experiment of fermentation activity, the fermentation broth was prepared by lyophilization and powdered. The DPPH scavenging activity, reducing power and total phenol content were measured. The results are shown in Table 1.

The antioxidant activity of Lactobacillus lambosus strain HK-9, which is a lactic acid bacterium of the present invention, was confirmed to be superior in antioxidant activity of gold in Table 1 above.

Especially, the results of total phenol content of antioxidant activity showed 2 ~ 4 times higher activity than gold fermented by other lactic acid bacteria. Based on these results, it can be confirmed that the lactic acid bacteria of the present invention, Lactobacillus lambosus strain HK-9, has a fungal activity (antioxidative activity related to anti-aging), which is also related to the separation from the female hand Seems to be.

From these results, it can be seen that fermentation through Lactobacillus lambosus strain HK-9 facilitates dissolution of useful physiological activity and fermentation activity by fermentation compared to other lactic acid bacteria.

3) Generation of new substances by fermentation

The inventors measured the change of physiologically active substance before and after fermentation of the dodec using the strain according to the present invention.

Before fermentation, Doduck was dried in a hot air drier at 40 ℃ for 24 hours, and then pulverized into 2 ~ 3mm using a ball mill and extracted with 70% ethanol.

In addition, fermented Doduck was fermented in the same manner as the above-mentioned fermentation of the above-mentioned phage, and 100 g of dried Doduck was ground into a powder form and added to a 2 L flask. Then, 1 L of a culture medium was prepared and then Lactobacillus lambosus strain HK-9 was inoculated 5% And fermented for 48 hours.

The results of HPLC analysis of dodec prepared by the above method are shown in FIG. From the results of FIG. 6, only one peak at 2.7 minutes was detected in the broth before fermentation, but a new peak was detected at 3.1 minutes along with the peak detected at 2.7 minutes after fermentation.

As shown in FIG. 7 showing the DPPH activity of fermentation broth and fermented broodstock after fermentation, these novel peaks exhibit a new physiological activity. As a result, it can be confirmed that DPPH activity of broodstock increased by fermentation, It can be confirmed that a novel substance produced through the fermentation of Lymphocus strain HK-9 has an effect on DPPH activity.

4) Measurement of the activity of the fungus activity according to the Lactobacillus Lymphosus strain HK-9 itself

Melanin synthesis inhibition activity and tyrosinase inhibition experiments were carried out to determine the efficacy of the fragrant active substances from the Lactobacillus Lymphosus HK-9 strain itself.

First, in order to disrupt Lactobacillus lambosus strain HK-9, Lactobacillus lambosus strain HK-9 cultivated in a conventional MRS medium was centrifuged to obtain a total of 100 ml of strain. The strain was then sonicated for 20 to 50 minutes to disrupt the strain. The shattered strain and its solution were subjected to hot - water extraction at 60 ℃ to obtain a strain extract.

Figure 8 shows the results of the melanin synthesis inhibition activity. In order to observe the effects on the melanin biosynthesis and cell viability after administration of the sample using the melanocyte Clon-M3 cells, the inhibition rate of melanin formation was measured after 24 hours of treatment. As a result, it was confirmed that the inhibition rate was increased from 12.5 mg / ml to 100 mg / ml, and finally, the inhibition rate reached about 40%. These results show that the component of the strain of Lactobacillus lambosus strain HK-9 has an effect of reducing the production of melanin.

9 is a result of measurement of tyrosinase inhibition. Tyrosinase is an enzyme that promotes the production of melanin pigment in the skin, and by measuring the inhibitory activity of tyrosinase, a whitening effect can be recognized. As a result of FIG. 9, it was confirmed that the inhibition rate was finally about 22%.

This result is a result of a definite experiment that contains a component having a fumaric activity in the strain itself, which is lower than the rate of inhibition of melanin using a natural product. Thus, it is possible to produce relatively easy fermentation By using the strain, it is expected to be able to produce cosmetics-related raw materials and products more competitively.

Korea Microorganism Conservation Center (overseas) KCCM11254P 20120217

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Claims (6)

delete Lactic acid bacteria derived from human body Lactobacillus Laminocus HK-9 (Accession No .: KCCM11254P) fermented with fermented or golden fermented product. delete delete A cosmetic composition containing the phaetam or golden fermented product of claim 2. A food containing the phasmatical or golden fermented product of paragraph 2.

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