LEUCONOSTOC HOLZAPFELII STRAIN FOR PREVENTING DEPILATION IMPROVING HAIR GROWTH OR IMPROVING SEXUAL DISFUNTION AND COMPOSITION COMPRISING THE SAME
20230167402 · 2023-06-01
Assignee
Inventors
Cpc classification
C12R2001/01
CHEMISTRY; METALLURGY
A61K35/744
HUMAN NECESSITIES
A23L33/135
HUMAN NECESSITIES
International classification
A23L33/135
HUMAN NECESSITIES
A61K35/744
HUMAN NECESSITIES
Abstract
The present invention relates to a novel Leuconostoc holzapfelii Ceb-kc-003 (deposited as Korea Accession Number accession KCCM11830P) strain and a composition comprising the same. The novel strain has abilities of preventing hair loss, promoting hair growth, or improving sexual function.
Claims
1. A Leuconostoc holzapfelii strain Ceb-kc-003 (KCCM11830P), a culture solution, concentrate or dried product thereof.
2. The strain, the culture solution, concentrate or dried product thereof according to claim 1, wherein the strain is effective in preventing hair loss, promoting hair growth, ameliorating erectile dysfunction, lowering cholesterol levels or decomposing fat.
3. A composition comprising a Leuconostoc holzapfelii strain Ceb-kc-003 (KCCM11830P) or a culture solution, concentrate or dried product thereof.
4. The composition according to claim 3, wherein the composition is a food composition or a pharmaceutical composition.
5. The composition according to claim 4, wherein the composition is effective in preventing hair loss or promoting hair growth.
6. The composition according to claim 3, wherein the daily dose of the Leuconostoc holzapfelii Ceb-kc-003 (KCCM11830P) strain is from 1.5×10.sup.5 to 5×10.sup.10 CFU.
7. The composition according to claim 3, wherein the Leuconostoc holzapfelii Ceb-kc-003 (KCCM11830P) strain at a concentration of 5×10.sup.4 to 5×10.sup.8 CFU/ml is administered 1 to 4 times (30 ml to 100 ml each time) daily.
8. The composition according to claim 5, wherein the composition is further effective in ameliorating erectile dysfunction.
9. The composition according to claim 3, further comprising a strain belonging to the genus Lactobacillus, Lactococcus, Propionibacterium, Enterocuccus or Bifidobacterium.
10. The composition according to claim 9, wherein the strain is a Leuconostoc mesenteroides strain (KCCM11827P), a Lactobacillus sakei strain (KCCM11841P), a culture solution thereof, a concentrate thereof, a dried product thereof or a mixture thereof.
11. A method of preventing hair loss or promoting hair growth, wherein the method comprises administering a Leuconostoc holzapfelii strain Ceb-kc-003 (KCCM11830P), a culture solution, concentrate or dried product thereof to a subject in need.
12. The method according to claim 11, wherein the method comprises that the Leuconostoc holzapfelii Ceb-kc-003 (KCCM11830P) strain at a concentration of 5×10.sup.4 to 5×10.sup.8 CFU/ml is administered 1 to 4 times (30 ml to 100 ml each time) daily.
13. The method according to claim 11, wherein the method further ameliorates erectile dysfunction, lowers cholesterol levels or decomposes fat.
14. The method according to claim 11, wherein the method further comprising administering a strain belonging to the genus Lactobacillus, Lactococcus, Propionibacterium, Enterocuccus or Bifidobacterium to the subject.
15-18. (canceled)
Description
DESCRIPTION OF DRAWINGS
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MODE FOR INVENTION
[0042] The present invention will be described in more detail with reference to the following examples. However, these examples are provided for illustrative purposes only and the present invention is not limited thereto.
Example 1: Isolation and Identification of Leuconostoc holzapfelii Ceb-Kc-003 (KCCM11830P)
[0043] (1) Sampling and Strain Isolation
[0044] A sample was taken from kimchi, a traditional Korean fermented food, diluted stepwise, plated on a BHI solid medium (Difco, USA) supplemented with 3% sodium chloride, and cultured at 37° C. for 24 h. A dominant strain was isolated from the sample. Colonies were selected and passaged three times in fresh media. The pure cultured bacterial strain was placed in a medium supplemented with 20% glycerol and stored at ≤−70° C.
[0045] (2) Investigation of Morphological and Taxological Properties
[0046] The isolated strain was identified. To this end, the strain was primarily morphologically and biochemically investigated. The strain was found to be morphologically Gram-positive by Gram staining. The taxological properties of the strain were analyzed by 16s rRNA partial sequencing. As a result, the isolated strain was found to have the sequence set forth in SEQ ID NO: 1 and a homology of 99% with Leuconostoc holzapfelii.
[0047] The newly isolated strain Leuconostoc holzapfelii Ceb-kc-003 was deposited with the Korean Culture Center of Microorganisms (120-861, Hongje-2ga-Gil, Seodaemun-Ku, Seoul, Korea) on Apr. 11, 2016 and assigned accession number KCCM11830P.
[0048] The strain was freeze-dried and powdered.
[0049] (3) Preparation of Culture Solution of the Strain
[0050] A culture solution of the Leuconostoc holzapfelii strain Ceb-kc-003 was prepared by the following procedure:
[0051] 2 kg of dextrose, 1.5 kg of whole milk powder, and 0.05 kg of an yeast extract were added to 100 liters of purified water. The mixture was sterilized in an autoclave at 121° C. for 15-30 min and cooled to around 35° C. Thereafter, the sterile mixture was inoculated with 0.2-0.4 liters of the Leuconostoc holzapfelii strain Ceb-kc-003, followed by culture at around 35° C. for 2-3 days.
Example 2: Preventive Effect of the Leuconostoc holzapfelii Strain Ceb-Kc-003 (KCCM11830P) on Hair Loss
[0052] The culture solution of the Leuconostoc holzapfelii strain Ceb-kc-003 (KCCM11830P) at a concentration of 1×10.sup.6 to 1×10.sup.8 CFU/ml was administered to a total of 5 men aged 40-85 years and one woman who suffered from alopecia twice daily (50-100 ml each time) over 1-4 months.
[0053] The numbers of hairs lost were counted before and 10, 20, and 30 days after taking the strain. The patients were instructed to wash their hair before hair counting.
[0054] The experimental results are shown in Table 1.
TABLE-US-00002 TABLE 1 Subject Man at Man at Woman at Men at Time age 45 age 52 age 38 age 57 Before taking 60 hairs lost 43 hairs lost 74 hairs lost 23 hairs lost 10 days 40 hairs lost 32 hairs lost 55 hairs lost 10 hairs lost after taking 20 days 22 hairs lost 15 hairs lost 23 hairs lost 4 hairs lost after taking 30 days 12 hairs lost 5 hairs lost 8 hairs lost 2 hairs lost after taking
[0055] The hairs of the male patients before and at the predetermined time points after administration of the strain were photographed to measure their thicknesses, numbers, and glosses. Some of the results are shown in
[0056] These results indicate that the Leuconostoc holzapfelii strain Ceb-kc-003 (KCCM11830P) is effective in preventing hair loss and markedly improving the thickness, number, and gloss of hairs. Therefore, it can be conclude that the Leuconostoc holzapfelii strain Ceb-kc-003 (KCCM11830P) is effective in treating hair loss.
Example 3: Effect of the Leuconostoc holzapfelii Strain Ceb-Kc-003 (KCCM11830P) on Reducing Cholesterol Levels
[0057] The effect of the Leuconostoc holzapfelii strain Ceb-kc-003 (KCCM11830P) on reducing cholesterol levels was examined. To this end, solutions of the powder of the Leuconostoc holzapfelii strain Ceb-kc-003 (KCCM11830P) (1-5 mg/ml) and solutions of a powder of Lactobacillus plantarum (1-5 mg/ml) as controls were used.
[0058] <Ability to Remove Cholesterol>
[0059] The ability of the strains to remove cholesterol was specifically measured by the following procedure. 30 ml of an MRS culture ground was added to a 50 ml Falcon tube, inoculated with the test strain (glycerol stock) at a density of 1×10.sup.7 counts/ml, and cultured at 30° C. for 24 h. 300 μl of the culture solution was added to 30 ml of an MRS culture ground containing 350 mg/L cholesterol (Wako Pure Chemical) and 0.2% bile acid (w/v). After culture at 37° C. for 24 h, the culture solution was subjected to centrifugal separation at 8,000 rpm and 4° C. for 10 min. The cholesterol level of the supernatant was measured using Determiner FC (Kyowa Medex Co., Ltd.). The level (%) of cholesterol removed from the culture ground was calculated by subtracting the remaining cholesterol level from the initial cholesterol level of the cholesterol culture ground.
[0060] The results are shown in
[0061] From these results, it can be concluded that the powder of the Leuconostoc holzapfelii strain Ceb-kc-003 (KCCM11830P) is significantly effective in reducing cholesterol levels in proportion to the strain concentration compared to the other lactic acid bacterial species Lactobacillus plantarum.
Example 4: Effect of the Leuconostoc holzapfelii Strain Ceb-Kc-003 (KCCM11830P) on Fat Decomposition
[0062] In this example, the effect of the Leuconostoc holzapfelii strain Ceb-kc-003 (KCCM11830P) on fat decomposition was examined. To this end, 1000 cc of pork fat was treated with 20-100 ml of culture solutions of the strain, followed by shaking at 37° C. for 24 h. This procedure was repeated for Lactobacillus plantarum as a control.
[0063] The results are shown in
[0064] From these results, it can be concluded that the powder of the Leuconostoc holzapfelii strain Ceb-kc-003 is significantly effective in fat decomposition in proportion to the strain concentration compared to the other lactic acid bacterial species Lactobacillus plantarum.
Example 5: Effects of the Leuconostoc holzapfelii Strain Ceb-Kc-003 (KCCM11830P) on Improving Sexual Function and Preventing Sexual Dysfunction
[0065] The powders of the Leuconostoc holzapfelii strain Ceb-kc-003 (KCCM11830P) at concentrations of 1×10.sup.6 to 1×10.sup.8 CFU/ml were administered orally to a total of 10 male smokers or non-smokers, aged 45-70 years, once or twice (50 ml to 100 ml each time) daily over a total of 3 months. The frequency of erections in the early morning, erectility, and duration of erection were self-evaluated before and at 1-month intervals after administration of the strain.
[0066] 4 out of the 10 men answered questionnaires for 1-3 months after taking. The results are shown in Table 2.
TABLE-US-00003 TABLE 2 Data on effects of the strain on improving in sexual function Male Male Male Male Subject non-smoker non-smoker smoker smoker Time at age 45 at age 54 at age 63 at age 57 Before Good Good Impotence Poor taking erectility erectility erectility 1 month Increased Good Impotence Poor after taking frequency of erectility erectility erections in the early morning 2 months Increased Increased Erection Increased after taking frequency of duration of in the early frequency of erections erection morning erections in the early in the early morning morning 3 months Increased Increased Increased Increased after taking duration of frequency of frequency duration of erection erections of erections erection in the early morning
[0067] The above results indicate that continuous taking of the powder of the Leuconostoc holzapfelii strain Ceb-kc-003 is effective in preventing and treating sexual dysfunction.
[0068] In addition, the concentrations of NO released from Hacat cells for 24 h were evaluated. NO is helpful in improving erectility due to its ability to increase blood circulation and dilate capillaries. The concentrations of NO produced after taking the novel strain of the present invention were compared with those after taking Lactobacillus plantarum. This experiment was conducted in triplicate and the data were averaged. The results are shown in
[0069] The effects of culture solutions of Leuconostoc mesenteroides (KCCM11827P) and Lactobacillus sakei (KCCM11841P) strains together with the Leuconostoc holzapfelii strain Ceb-kc-003 (KCCM11830P) on the prevention and the treatment of hair loss and the improvement of sexual function were evaluated in the same manner as described above. As a result, better results were obtained compared to the single use of the Leuconostoc holzapfelii strain Ceb-kc-003.