Patent Publication Number: US-2019183943-A1

Title: Composition, for preventing hair loss or promoting hair growth, comprising strains showing lipolysis effect

Description:
TECHNICAL FIELD 
     The present invention relates to a composition for preventing hair loss or promoting hair growth including a strain having lipolysis ability. 
     BACKGROUND ART 
     With the development of modern society, the number of alopecia patients has increased rapidly. Particularly, the age of alopecia patients has dropped gradually, 60% of people who require hair care service are in their twenties and thirties. The number of patients suffering from alopecia is estimated to reach almost 10 million, causing serious socioeconomic problems. 
     Reports say that people with severe hair loss are more likely to experience psychological depression and anxiety, perceive high levels of stress, are faced with difficulty in their relationships with other people and the opposite sex, and fail to maintain good family relationships. Thus, hair loss has become a matter of concern in health science. 
     Although the exact cause of hair loss is clearly unknown, reports say that hair loss is possibly caused by genetic, hormonological and immunological abnormalities. 
     Minoxidil for transdermal application and Finasteride for oral administration approved by the U.S. Food and Drug Administration (FDA) are being currently widely used as therapeutic agents for alopecia. However, these therapeutic agents are limited in their use because of their attendant side effects such as cardiovascular disorders, skin irritation, decreased sexual function, and severe teratogenesis. 
     Previous studies reported that galenic preparations can ameliorate alopecia, but the results are still unsatisfactory. In recent years, some studies and patents reported that the use of fermentation products of plant extracts and hot water-extracted herbal materials such as fleeceflower root by lactic acid bacteria belonging to the genus  Lactobacillus  can promote hair growth. However, most of the results were obtained from experimental rats and no research has been conducted on people who had suffered or were suffering hair loss. 
     DISCLOSURE 
     Technical Problem 
     The present invention is aimed at providing a composition that is effective in preventing hair loss or promoting hair growth or a novel strain that is isolated from kimchi and is effective in decomposing fat. 
     The present invention is aimed at providing a composition for preventing hair loss or promoting hair growth that can be used safely without causing side effects. 
     Technical Solution 
     One embodiment of the present invention provides a composition for preventing hair loss or promoting hair growth including a strain having lipolysis ability. 
     The strain having lipolysis ability may be a lactic acid bacterial strain isolated from kimchi. Specifically, the strain may be one belonging to the genus  Leuconostoc  and  Lactobacillus . More specifically, the strain may be  Leuconostoc holzapfelii, Leuconostoc mesenteroides  or  Lactobacillus sakei . Even more specifically, the strain may be  Leuconostoc holzapfelii  Ceb-kc-003 (KCCM11830P).  Leuconostoc mesenteroides  (KCCM11827P) or  Lactobacillus sakei  (KCCM11841P). 
       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. The strain is effective in preventing hair loss, promoting hair growth, improving sexual function, lowering cholesterol levels, and/or decomposing fat. 
       Leuconostoc mesenteroides  (KCCM11827P) was deposited with the Korean Culture Center of Microorganisms (120-861, Hongje-2ga-Gil, Seodaemun-Ku. Seoul. Korea) on Apr. 6, 2016 and assigned accession number KCCM11827P. The strain is effective in preventing hair loss, promoting hair growth, improving sexual function, lowering cholesterol levels, and/or decomposing fat. 
       Lactobacillus sakei  (KCCM1841P) was deposited with the Korean Culture Center of Microorganisms (120-861. Hongje-2ga-Gil, Seodaemun-Ku, Seoul, Korea) on Jun. 1, 2016 and assigned accession number KCCM11841P. The strain is effective in preventing hair loss, promoting hair growth, improving sexual function, lowering cholesterol levels, and/or decomposing fat. 
     The strain having lipolysis ability may be used per se or in the form of a culture, concentrate or dried product thereof. Accordingly, the composition may include the strain having lipolysis ability or a culture, concentrate or dried product thereof. 
     The strain having lipolysis ability can be cultured by any suitable method known in the art. A natural or synthetic medium can be used to culture the strain. Examples of carbon sources of the medium include, but are not limited to, glucose, sucrose, dextrin, glycerol, and starch. Examples of nitrogen sources of the medium include, but are not limited to, peptone, meat extract, yeast extract, dried yeast, soybean, ammonium salt, nitrate, and other organic and inorganic nitrogenous compounds. One or more inorganic salts may be added to the medium. Examples of such inorganic salts include, but are not limited to, magnesium, manganese, calcium, iron, and potassium salts. The medium may further include one or more compounds selected from amino acids, vitamins, nucleic acids, and compounds related thereto. The strain may be cultured at a temperature of 20 to 40° C. for 12 hours to 4 days. As an example, one or more galenic preparations, an extract thereof or a mixture thereof may be added to and cultured in the culture medium. Examples of the galenic preparations include green kernel black bean.  Cynanchum wilfordii  root, cooked Rehmannia, licorice root, Szechuan lovage root, Eucommia bark, Chinese cinnamon bark, Chinese angelica root, sweetflag rhizome, fleeceflower root, leafy twig of arorvitae, ginger, arbor-vitae seed, fruit of Szechuan pepper, bugbane rhizome, and vitex fruit. The use of the culture further improves the effect of the strain to prevent hair loss or promote hair growth. The addition of green kernel black bean and/or  Cynanchum wilfordii  root to the culture medium is particularly preferred. The green kernel black bean and/or  Cynanchum wilfordii  root may be added in an amount of 0.01% by weight (wt %) to 10 wt %, specifically 0.01 wt % to 5 wt %. 
     The culture of the strain having lipolysis ability may be a culture broth containing the bacterial cells. The culture may also be obtained by removal of the supernatant from the culture broth or concentration of the culture broth. The composition of the culture may further include not only one or more typical ingredients necessary for the culture of strains having lipolysis ability but also one or more ingredients exerting a synergistic effect on the growth of the strains. The composition of the culture can be readily determined by those skilled in the art. 
     The strain may be in a liquid or dry state. The strain can be dried by any suitable technique known in the art. Examples of such drying techniques include, but are not limited to, air drying, natural drying, spray drying, and freeze drying. 
     The composition is effective in preventing hair loss or promoting hair growth. The composition is also effective in improving sexual function or decomposing fat. The composition may be used as a pharmaceutical composition. Alternatively, the composition may be used as a food, health food, health supplement food or health functional food composition. One embodiment of the pharmaceutical composition may be a quasi-drug or a pharmaceutical preparation. 
     As an example, the composition may further include one or more galenic ingredients that are effective in preventing hair loss or promoting hair growth or one or more pharmaceutical ingredients. Examples of the galenic ingredients include galenic preparations such as green kernel black bean.  Cynanchum wilfordii  root, cooked Rehmannia, licorice root. Szechuan lovage root, Eucommia bark, Chinese cinnamon bark, Chinese angelica root, sweetflag rhizome, red fleeceflower root, leafy twig of arorvitae,  Acanthopanax senticosus , pine leaf and flower, buckwheat, Anemarrhena rhizome, ginger, arbor-vitae seed, fruit of Szechuan pepper, bugbane rhizome, and vitex fruit, extracts thereof, and mixtures thereof. Examples of the pharmaceutical ingredients include Minoxidil or Finasteride. 
     A further embodiment of the present invention relates to a composition for preventing hair loss or promoting hair growth including a strain having lipolysis ability and a strain capable of lowering cholesterol levels. The strain capable of lowering cholesterol levels may be derived from cheonggukjang. Specifically, the strain capable of lowering cholesterol levels may be, for example, one belonging to the genus  Brevibacillus, Lactobacillus, Lactococcus, Propionibacterium, Enterococcus  or  Bifidobacterium . More specifically, the strain capable of lowering cholesterol levels may be, for example,  Lactobacillus fermentum, Brevibacillus reuszeri  or  Enterococcus faecium . In one embodiment,  Brevibacillus reuszeri  Ceb-ch-003 (KCCM11911P),  Enterococcus faecium  Ceb-ch-001 (KCCM11909P) and/or  Lactobacillus fermentum  Ceb-ch-002 (KCCM11910P) may be used in combination with the strain having lipolysis ability described herein. 
       Brevibacillus reuszeri  Ceb-ch-003 was isolated from cheonggukjang and was deposited with the Korean Culture Center of Microorganisms (120-861, Hongje-2ga-Gil. Seodaemun-Ku. Seoul. Korea) on Oct. 7, 2016 and assigned accession number KCCM11911P. The strain is effective in improving sexual function or lowering cholesterol levels. 
       Enterococcus faecium  Ceb-ch-001 was isolated from cheonggukjang and was deposited with the Korean Culture Center of Microorganisms (120-861, Hongje-2ga-Gil, Seodaemun-Ku, Seoul, Korea) on Oct. 7, 2016 and assigned accession number KCCM11909P. The strain is effective in improving sexual function or lowering cholesterol levels. 
       Lactobacillus fermentum  Ceb-ch-002 was isolated from cheonggukjang was deposited with the Korean Culture Center of Microorganisms (120-861, Hongje-2ga-Gil. Seodaemun-Ku, Seoul, Korea) on Oct. 7, 2016 and assigned accession number KCCM11910P. The strain is effective in improving sexual function or lowering cholesterol levels. 
     The composition further includes the strain capable of lowering cholesterol levels at a concentration of 2.5×10 4  to 2.5×10 10  CFU/ml, preferably 5×10 5  to 5-10 8  CFU/ml. The strain capable of lowering cholesterol levels may be present in an amount of 0.05 wt % to 50 wt %, based on the weight of the composition. 
     The composition may further include collagen. 
     The composition includes the strain having lipolysis ability at a concentration of 5×10 4  to 5×10 10  CFU/ml, preferably 1×10 6  to 1×10 9  CFU/ml. 
     The composition of the present invention may further include a pharmaceutically or sitologically acceptable carrier. In this case, the composition may be formulated with the carrier to provide a food or pharmaceutical drug. 
     As used herein, the term “pharmaceutically or sitologically acceptable carrier” refers to a carrier or diluent that causes no irritation to target organisms and does not deteriorate the biological activity and properties of the strain. 
     The composition may be formulated into various preparations for oral or parenteral administration. Preferably, the composition is formulated into a liquid oral solution. 
     When formulated into a liquid solution, the pharmaceutically or sitologically acceptable carrier may be selected from saline solution, sterilized water, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, and mixtures thereof that are sterile and biocompatible. If necessary, one or more general additives, such as antioxidants, buffer solutions, and bacteriostatic agents may be added to the composition. The composition of the present invention may be formulated into liquid preparations (such as aqueous solutions, suspensions or emulsions), pills, capsules, granules, and tablets. In this case, the composition of the present invention may further include one or more additives selected from diluents, dispersants, surfactants, binders, and lubricants. A binding agent, an emulsifier or preservative may be further added to the composition to prevent the quality of the composition from deteriorating. The composition of the present invention may further include one or more additives selected from amino acids, vitamins, enzymes, flavoring agents, non-protein nitrogen compounds, silicates, buffers, extractants, and oligosaccharides. The strain having lipolysis ability may be administered in the form of a liquid. In this case, the strain having lipolysis ability at a concentration of 5×10 4  to 5×10 8  CFU/ml, preferably 1×10 6  to 1×10 8  CFU/ml, may be administered 1 to 4 times (30 ml to 100 ml each time) daily. As an example, the strain having lipolysis ability at a concentration of 1×10 6  to 1×10 8  CFU/ml may be administered 2 to 4 times (50 ml each time) daily. More specifically, the strain having lipolysis ability at a concentration of 1×10 6  to 1×10 8  CFU/ml may be administered once or twice (50 ml to 100 ml each time) daily. Even more specifically, the strain having lipolysis ability at a concentration of 1×10 6  to 1×10 8  CFU/ml may be administered in an amount of 50 ml to 100 ml once 30 minutes before breakfast and in an amount of 50 ml to 100 ml once 30 minutes before dinner or before bed. The strain capable of lowering cholesterol levels at a concentration of 2.5×10 4  to 2.5×10 8  CFU/ml, preferably 5×10 5  to 5×10 7  CFU/ml, may be administered 1 to 4 times (15 ml to 50 ml each time) daily. As an example, the strain capable of lowering cholesterol levels at a concentration of 5×10 5  to 5×10 7  CFU/ml may be administered 2 to 4 times (25 ml to 50 ml each time) daily. More specifically, the strain capable of lowering cholesterol levels at a concentration of 5×10 5  to 5×10 7  CFU/ml may be administered once or twice (25 ml to 100 ml each time) daily. Even more specifically, the strain capable of lowering cholesterol levels at a concentration of 5×10 5  to 5×10 7  CFU/ml may be administered in an amount of 20 ml to 100 ml once 30 minutes before breakfast and in an amount of 20 ml to 100 ml once 30 minutes before dinner or before bed. The strain capable of lowering cholesterol levels may be administered simultaneously with the strain having lipolysis ability. Alternatively, the two strains may be administered sequentially. The administration cycle and dose of the strain capable of lowering cholesterol levels may be different from those of the strain having lipolysis ability. 
     Examples of oral preparations including the strain having lipolysis ability include tablets, troches, lozenges, water-soluble or oily suspensions, powders, granules, emulsions, hard or soft capsules, syrups, and elixirs. The composition of the present invention may be formulated into desired preparations such as tablets and capsules. In this case, the composition may include: a binder such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose or gelatin; an excipient such as dicalcium phosphate; a disintegrant such as corn starch or sweet potato starch; or a lubricant such as magnesium stearate, calcium stearate, sodium stearyl fumarate or polyethylene glycol wax. For capsule preparations, the composition may further include a liquid carrier such as a fatty oil. 
     The daily dose of the composition, the strain having lipolysis ability or the strain capable of lowering cholesterol levels may vary depending on the weight, age, sex, health condition, main symptoms to be treated, prevented or ameliorated, time and mode of administration, and severity of disease and may be in the range of about 0.0001 mg/kg to about 10 g/kg, specifically about 0.01 mg/kg to about 500 mg/kg. The composition, the strain having lipolysis ability or the strain capable of lowering cholesterol levels may be administered or applied 1 to 6 times, for example, 1 to 4 times, a day. 
     The strain having lipolysis ability may be present in a liquid or dry state, preferably a liquid state, in the composition. The strain can be dried by any suitable technique known in the art. Examples of such drying techniques include, but are not limited to, air drying, natural drying, spray drying, and freeze drying. The strain having lipolysis ability may be used in the form of a powder. In this case, the strain may be added in an amount of 0.05 to 80 wt %, for example, 0.1 to 70 wt %, 0.1 to 50 wt % or 1 to 30 wt %, based on the weight of the composition. 
     A further embodiment of the present invention relates to a method for preventing hair loss or promoting hair growth including administering a composition including the strain having lipolysis ability to a subject. The composition including the strain having lipolysis ability is the same as that described in the previous embodiment and a detailed description thereof is thus omitted to avoid duplication. 
     Another embodiment of the present invention relates to a  Leuconostoc mesenteroides  strain (KCCM11827P) or a  Lactobacillus sakei  strain (KCCM11841P), a culture thereof, a concentrate thereof or a dried product thereof. The strains are lactic acid bacterial strains isolated from kimchi and have the ability to decompose fat. The strains are effective in preventing hair loss, promoting hair growth or decomposing fat. 
     Another embodiment of the present invention relates to a method for preparing a composition for preventing hair loss or promoting hair growth, including preparing a strain having lipolysis ability, a culture thereof, a concentrate thereof or a dried product thereof and applying a pharmaceutically or sitologically acceptable carrier to the strain or the culture, concentrate or dried product thereof. The strain having lipolysis ability and the culture, concentrate or dried product thereof are the same as those described in the previous embodiment and a detailed description thereof is thus omitted to avoid duplication. 
     One embodiment of the present invention relates to a composition for ameliorating sexual dysfunction including a strain having lipolysis ability, a culture thereof, a concentrate thereof or a dried product thereof. The sexual dysfunction may be erectile dysfunction. A further embodiment of the present invention relates to a method for preventing or treating sexual dysfunction including administering to a subject in need of such prevention or treatment a therapeutically or prophylactically effective amount of the strain having lipolysis ability or the culture, concentrate or dried product thereof. The therapeutically or prophylactically effective amount is the same as that described above. 
     Advantageous Effects 
     The strain having lipolysis ability and the composition including the strain are effective in preventing hair loss, promoting hair growth or improving sexual function. 
     In addition, the strain having lipolysis ability and the composition including the strain can be used safely without causing side effects to prevent hair loss, promote hair growth, lower cholesterol levels or improve sexual function. 
    
    
     
       DESCRIPTION OF DRAWINGS 
         FIG. 1  is a histogram comparing the effects of  Leuconostoc holzapfelii  Ceb-kc-003 (KCCM11830P) and another lactic acid bacterial strain on fat decomposition. 
         FIG. 2  is a histogram comparing the effects of  Leuconostoc holzapfelii  Ceb-kc-003 (KCCM11830P) and another lactic acid bacterial strain on NO production. 
         FIG. 3  is a histogram comparing the effects of  Lactobacillus sakei  (KCCM11841P) and another lactic acid bacterial strain on fat decomposition. 
         FIG. 4  is a histogram comparing the effects of  Leuconostoc mesenteroides  KCCM11827P and another lactic acid bacterial strain on fat decomposition. 
         FIG. 5  shows photographs comparing scalp hair before and ˜3.5 months after taking  Leuconostoc holzapfelii  Ceb-kc-003 (KCCM11830P). 
         FIG. 6  shows photographs comparing scalp hair before and ˜2.5 and ˜3.5 months after taking  Leuconostoc holzapfelii  Ceb-kc-003 (KCCM11830P). 
         FIG. 7  shows photographs comparing scalp hair before and ˜1.7 months after taking  Leuconostoc holzapfelii  Ceb-kc-003 (KCCM11830P). 
         FIG. 8  shows photographs comparing scalp hair before and ˜3 months after taking  Leuconostoc holzapfelii  Ceb-kc-003 (KCCM11830P). 
         FIG. 9  shows photographs showing changes in the gloss and thickness of hairs and the growth of new bangs after taking  Leuconostoc mesenteroides  (KCCM11827P) for ˜3 months. 
         FIG. 10  shows photographs comparing scalp hair before and ˜45 days after taking  Leuconostoc mesenteroides  (KCCM11827P). 
         FIG. 11  shows photographs comparing hair thicknesses, strengths, and colors before and ˜1 month after taking  Leuconostoc mesenteroides  (KCCM1827P). 
         FIG. 12  shows photographs comparing scalp hair before and ˜27 days after taking  Lactobacillus sakei  (KCCM11841P). 
         FIG. 13  shows photographs comparing scalp hair before and ˜2 months after taking  Lactobacillus sakei  (KCCM11841P). 
         FIG. 14  shows photographs comparing scalp hair before and ˜1.5 months after taking  Lactobacillus sakei  (KCCM11841P). 
         FIG. 15  shows photographs comparing scalp hair before and ˜2 months after taking a combination of  Leuconostoc mesenteroides  (KCCM11827P) and  Lactobacillus fermentum  Ceb-ch-002 (KCCM11910P). 
         FIG. 16  shows photographs comparing scalp hair before and ˜1.5 months after taking a combination of  Leuconostoc mesenteroides  (KCCM11827P) and  Lactobacillus fermentum  Ceb-ch-002 (KCCM11910P). 
         FIG. 17  shows photographs comparing scalp hair before and ˜1.5 months after taking a combination of  Lactobacillus sakei  (KCCM11841P) and  Lactobacillus fermentum  Ceb-ch-002 (KCCM11910P). 
         FIG. 18  shows photographs comparing scalp hair before and ˜3 months after taking a combination of  Lactobacillus sakei  (KCCM11841P) and  Lactobacillus fermentum  Ceb-ch-002 (KCCM11910P). 
         FIG. 19  is a histogram comparing the ability of  Lactobacillus fermentum  (KCCM11910P) isolated from cheonggukjang. Korean fermented soybean, to remove cholesterol with that of another lactic acid bacterial strain. 
     
    
    
     MODE FOR INVENTION 
     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 1-3: Isolation of  Leuconostoc holzapfelii  Strain Ceb-Kc-003 (KCCM11830P),  Leuconostoc mesenteroides  Strain (KCCM11827P) and  Lactobacillus sakei  Strain (KCCM11841P) from Kimchi and Identification of the Strains 
     (1) Sampling and Strain Isolation 
     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 &lt;−70° C. 
     (2) Investigation of Morphological and Taxological Properties 
     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 a homology of 99% with  Leuconostoc holzapfelii.    
     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. 
     A dominant strain was isolated from another kimchi sample after culture under the conditions described above. As a result of morphological and biochemical investigation, the strain was found to have a homology of 99% with  Leuconostoc mesenteroides.    
     The newly isolated strain  Leuconostoc mesenteroides  was deposited with the Korean Culture Center of Microorganisms (120-861, Hongje-2ga-Gil. Seodaemun-Ku, Seoul, Korea) on Apr. 6, 2016 and assigned accession number KCCM11827P. 
     A dominant strain was isolated from another kimchi sample after culture under the conditions described above. As a result of morphological and biochemical investigation, the strain was found to have a homology of 99% with  Lactobacillus sakei.    
     The newly isolated strain  Lactobacillus sakei  was deposited with the Korean Culture Center of Microorganisms (120-861. Hongje-2ga-Gil, Seodaemun-Ku. Seoul, Korea) on Jun. 1, 2016 and assigned accession number KCCM11841P. 
     The strains were freeze-dried and powdered. 
     (3) Preparation of Culture Solutions of the Strains 
     A culture solution of  Leuconostoc holzapfelii  Ceb-kc-003 was prepared by the following procedure (Example 1): 
     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  Leuconostoc holzapfelii  Ceb-kc-003, followed by culture at around 35° C. for 2-3 days. 
     Culture solutions of  Leuconostoc mesenteroides  (KCCM11827P) and  Lactobacillus sakei  (KCCM1841P) were prepared in the same manner as described above for the culture solution of  Leuconostoc holzapfelii  Ceb-kc-003 (Examples 2-3, respectively). 
     Example 4: Effect of the  Leuconostoc holzapfelii  Strain Ceb-Kc-003 (KCCM11830P) on Fat Decomposition 
     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. 
     The results are shown in  FIG. 1 . 
     From these results, it can be concluded that the powder of the  Leuconostoc holzapfelii  strain Ceb-kc-003 (KCCM11830P) 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  Leuconastoc holzapfelii  Strain Ceb-Kc-003 (KCCM11830P) on Improving Sexual Function and Preventing Sexual Dysfunction 
     The powders of the  Leuconostoc holzapfelii  strain Ceb-kc-003 (KCCM11830P) at concentrations of 1×10 6  to 1×10 8  CFU/ml were administered orally to a total of 10 male smokers or non-smokers with or without sexual dysfunction, 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 I-month intervals after administration of the strain. 
     4 out of the 10 men answered questionnaires for 1-3 months after taking. The results are shown in Table 1. 
     
       
         
           
               
             
               
                 TABLE 1 
               
             
            
               
                   
               
               
                 Data on effects of the strain on improving in sexual function 
               
            
           
           
               
               
               
               
               
            
               
                 Subject 
                 Male non-smoker at 
                 Male non-smoker at 
                 Male smoker at age 
                 Male smoker at 
               
               
                 Time 
                 age 45 
                 age 54 
                 63 
                 age 57 
               
               
                   
               
               
                 Before taking 
                 Good erectility 
                 Good erectility 
                 Impotence 
                 Poor erectility 
               
               
                 1 month after 
                 Increased frequency 
                 Good erectility 
                 Impotence 
                 Poor erectility 
               
               
                 taking 
                 of erections in the 
               
               
                   
                 early morning 
               
               
                 2 months after 
                 Increased frequency 
                 Increased duration of 
                 Erection in the early 
                 Increased frequency 
               
               
                 taking 
                 of erections in the 
                 erection 
                 morning 
                 of erections in the 
               
               
                   
                 early morning 
                   
                   
                 early morning 
               
               
                 3 months after 
                 Increased duration of 
                 Increased frequency of 
                 Increased frequency 
                 Increased duration 
               
               
                 taking 
                 erection 
                 erections 
                 of erections in the 
                 of erection 
               
               
                   
                   
                   
                 early morning 
               
               
                   
               
            
           
         
       
     
     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. 
     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  FIG. 2 . 
     Example 6: Effect of the  Lactobacillus sakei  Strain (KCCM111841P) on Fat Decomposition 
     In this example, the effect of the  Lactobacillus sakei  strain (KCCM11841P) 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. 
     The results are shown in Table 2 and  FIG. 3 . 
     
       
         
           
               
               
               
             
               
                   
                 TABLE 2 
               
               
                   
                   
               
               
                   
                   Lactobacillus sakei  (KCCM11841P) 
                 
                   Lactobacillus plantarum 
                 
               
               
                   
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                 20 ml 
                 7% 
                 3% 
               
               
                 40 ml 
                 13% 
                 7% 
               
               
                 60 ml 
                 25% 
                 11% 
               
               
                 80 ml 
                 32% 
                 15% 
               
               
                 100 ml  
                 41% 
                 18% 
               
               
                   
               
            
           
         
       
     
     From these results, it can be concluded that the powder of the  Lactobacillus sakei  strain (KCCM11841P) is significantly effective in fat decomposition in proportion to the strain concentration compared to the other lactic acid bacterial species  Lactobacillus plantarum.    
     Example 7: Effect of the  Leuconostoc mesenteroides  Strain (KCCM1827P) on Fat Decomposition 
     In this example, the effect of the  Leuconostoc mesenteroides  strain (KCCM11827P) 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. 
     The results are shown in Table 3 and  FIG. 4 . 
     
       
         
           
               
               
               
             
               
                   
                 TABLE 3 
               
               
                   
                   
               
               
                   
                 
                   Leuconostoc mesenteroides 
                 
                   
               
               
                   
                 (KCCM11827P) 
                 
                   Lactobacillus plantarum 
                 
               
               
                   
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                 20 ml 
                 8% 
                 3% 
               
               
                 40 ml 
                 13% 
                 5% 
               
               
                 60 ml 
                 27% 
                 9% 
               
               
                 80 ml 
                 31% 
                 11% 
               
               
                 100 ml  
                 37% 
                 15% 
               
               
                   
               
            
           
         
       
     
     From these results, it can be concluded that the powder of the  Leuconostoc mesenteroides  strain (KCCM11827P) is significantly effective in fat decomposition in proportion to the strain concentration compared to the other lactic acid bacterial species  Lactobacillus plantarum.    
     Example 8: Preventive Effect of the  Leuconostoc holzapfelii  Strain Ceb-Kc-003 (KCCM11830P) on Hair Loss 
     The culture solution of the  Leuconostoc holzapfelii  strain Ceb-kc-003 (KCCM11830P) prepared in Example 1 at a concentration of 1×10 6  to 1×10 8  CFU/ml was administered orally 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. 
     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. 
     The experimental results are shown in Table 4. 
     
       
         
           
               
             
               
                 TABLE 4 
               
             
            
               
                   
               
               
                 Numbers of hairs lost before and after taking 
               
            
           
           
               
               
               
               
               
            
               
                 Subject 
                 Man at 
                 Man at 
                 Woman 
                 Men at 
               
               
                 Time 
                 age 45 
                 age 52 
                 at age 38 
                 age 57 
               
               
                   
               
               
                 Before taking 
                 60 hairs lost 
                 43 hairs lost 
                 74 hairs lost 
                 23 hairs lost 
               
               
                 10 days after 
                 40 hairs lost 
                 32 hairs lost 
                 55 hairs lost 
                 10 hairs lost 
               
               
                 taking 
               
               
                 20 days after 
                 22 hairs lost 
                 15 hairs lost 
                 23 hairs lost 
                  4 hairs lost 
               
               
                 taking 
               
               
                 30 days after 
                 12 hairs lost 
                  5 hairs lost 
                  8 hairs lost 
                  2 hairs lost 
               
               
                 taking 
               
               
                   
               
            
           
         
       
     
     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  FIGS. 5-8 . 
     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 concluded that the  Leuconostoc holzapfelii  strain Ceb-kc-003 (KCCM11830P) is effective in treating hair loss. 
     Example 9: Preventive Effect of the  Leuconostoc mesenteroides  Strain (KCCM11827P) on Hair Loss 
     The culture solution of the  Leuconostoc mesenteroides  strain (KCCM11827P) prepared in Example 2 at a concentration of 1×10 6  to 1×10 8  CFU/ml was administered orally to a woman aged 43 years, a man aged 60 years and a woman aged 49 years who suffered from alopecia twice daily (50-100 ml each time) over 1-3 months. 
     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. 
     The experimental results are shown in Table 5. 
     
       
         
           
               
             
               
                 TABLE 5 
               
             
            
               
                   
               
               
                 Numbers of hairs lost before and after taking 
               
            
           
           
               
               
               
               
            
               
                 Subject 
                   
                 Man 
                   
               
               
                 Time 
                 Woman at age 43 
                 at age 60 
                 Woman at age 49 
               
               
                   
               
               
                 Before taking 
                 39 hairs lost 
                 43 hairs lost 
                 25 hairs lost 
               
               
                 10 days after taking 
                 35 hairs lost 
                 32 hairs lost 
                 21 hairs lost 
               
               
                 20 days after taking 
                 29 hairs lost 
                 28 hairs lost 
                 20 hairs lost 
               
               
                 30 days after taking 
                 21 hairs lost 
                 16 hairs lost 
                 12 hairs lost 
               
               
                   
               
            
           
         
       
     
     The hairs of the 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  FIGS. 9-11 . 
     These results indicate that the  Leuconostoc mesenteroides  strain (KCCM11827P) is effective in preventing hair loss and markedly improving the thickness, number, and gloss of hairs. Therefore, it can be concluded that the  Leuconostoc holzapfelii  strain Ceb-kc-003 (KCCM11830P) is effective in treating hair loss. 
     Example 10: Preventive Effect of the  Lactobacillus sakei  Strain (KCCM11841P) on Hair Loss 
     The culture solution of the  Lactobacillus sakei  strain (KCCM1841P) prepared in Example 3 at a concentration of 1×10 6  to 1×10 8  CFU/ml was administered orally to a man aged 52 years, a woman aged 42 years and a man aged 60 years who suffered from alopecia twice daily (50-100 ml each time) over 0.8-3 months. 
     The numbers of hairs lost were counted before and 10, 20, and 27 days after taking the strain. The patients were instructed to wash their hair before hair counting. 
     The experimental results are shown in Table 6. 
     
       
         
           
               
             
               
                 TABLE 6 
               
             
            
               
                   
               
               
                 Numbers of hairs lost before and after taking 
               
            
           
           
               
               
               
               
            
               
                 Subject 
                   
                   
                   
               
               
                 Time 
                 Man at age 52 
                 Woman at age 42 
                 Man at age 60 
               
               
                   
               
               
                 Before taking 
                 47 hairs lost 
                 40 hairs lost 
                 47 hairs lost 
               
               
                 10 days after taking 
                 40 hairs lost 
                 32 hairs lost 
                 37 hairs lost 
               
               
                 20 days after taking 
                 35 hairs lost 
                 25 hairs lost 
                 26 hairs lost 
               
               
                 27 days after taking 
                 30 hairs lost 
                 24 hairs lost 
                 16 hairs lost 
               
               
                   
               
            
           
         
       
     
     The hairs of the patients before and at the predetermined time points after administration of the strain were photographed to determine whether the patients&#39; scalps were improved and to measure the hair thicknesses, numbers, and glosses. Some of the results are shown in  FIGS. 12-14 . 
     These results indicate that the  Lactobacillus sakei  strain (KCCM11841P) is effective in preventing hair loss, improving the condition of scalp, and markedly improving the thickness, number, and gloss of hair. Therefore, it can be concluded that the  Lactobacillus sakei  strain (KCCM11841P) is effective in treating hair loss. 
     Example 11: Isolation and Identification of  Lactobacillus fermentum  Strain Ceb-Ch-002 (KCCM11910P) 
     (1) Sampling and Strain Isolation 
     A sample was taken from cheonggukjang, a Korean fermented soybean, 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 &lt;−70° C. 
     (2) Investigation of Morphological and Taxological Properties 
     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 a homology of 99% with  Lactobacillus fermentum.    
     The newly isolated strain  Leuconostoc fermentum  Ceb-ch-002 was deposited with the Korean Culture Center of Microorganisms (120-861, Hongje-2ga-Gil, Seodaemun-Ku, Seoul, Korea) on Oct. 7, 2016 and assigned accession number KCCM11910P. 
     The strain was freeze-dried and powdered. 
     (3) Preparation of Culture Solution of the Strain 
     A culture solution of  Lactobacillus fermentum  (KCCM11910P) was prepared by the following procedure: 
     2 kg of dextrose, 1.5 kg of whole milk powder, and 0.05 kg of an enzyme 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  Lactobacillus fermentum  Ceb-ch-002, followed by culture at around 35° C. for 2-3 days. 
     Example 12: Effect of the  Lactobacillus fermentum  Strain Ceb-Ch-002 (KCCM11910P) on Reducing Cholesterol Levels 
     The effect of the powder of the  Lactobacillus fermentum  strain Ceb-ch-002 (KCCM11910P) on reducing cholesterol levels was examined. To this end, solutions of the freeze-dried powder of the  Lactobacillus fermentum  strain Ceb-ch-002 (KCCM11910P) (1-5 mg/ml) and solutions of a powder of  Lactobacillus plantarum  (1-5 mg/ml) as controls were used. 
     &lt;Ability to Remove Cholesterol&gt; 
     The ability of the strain 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 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 a 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. 
     The results are shown in  FIG. 19  and Table 7. This experiment was conducted in triplicate and the data were averaged. 
     
       
         
           
               
               
               
             
               
                   
                 TABLE 7 
               
               
                   
                   
               
               
                   
                   Lactobacillus fermentum  Ceb-ch-002 
                   
               
               
                   
                 (KCCM11910P) 
                 
                   Lactobacillus plantarum 
                 
               
               
                   
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                 1 mg/ml 
                 21% 
                 11% 
               
               
                 2 mg/ml 
                 27% 
                 16% 
               
               
                 3 mg/ml 
                 39% 
                 22% 
               
               
                 4 mg/ml 
                 51% 
                 31% 
               
               
                 5 mg/ml 
                 62% 
                 35% 
               
               
                   
               
            
           
         
       
     
     Example 13: Effect of Combination of the  Leuconostoc mesenteroides  Strain (KCCM11827P) and the  Lactobacillus fermentum  Strain Ceb-Ch-002 (KCCM11910P) on Hair Loss 
     The culture solution of the  Leuconostoc mesenteroides  strain (KCCM11827P) prepared in Example 2 at a concentration of 0.5×10 6  to 0.5×10 8  CFU/ml was combined with the culture solution of the  Lactobacillus fermentum  strain (KCCM11910P) prepared in Example 3 at a concentration of 0.5×10 6  to 0.5×10 8  CFU/ml. The combination of the two strain culture solutions was administered orally to a man aged 56 and a man aged 55 who suffered from alopecia twice (50-100 ml each time) daily over 1-3 months. 
     The numbers of hairs lost were counted before and 10, 20, and 40 days after taking the strains. The patients were instructed to wash their hair before hair counting. 
     The experimental results are shown in Table 8. 
     
       
         
           
               
             
               
                 TABLE 8 
               
             
            
               
                   
               
               
                 Numbers of hairs lost before and after taking 
               
            
           
           
               
               
               
               
            
               
                   
                 Subject 
                   
                   
               
               
                   
                 Time 
                 Man at age 56 
                 Man at age 55 
               
               
                   
                   
               
               
                   
                 Before taking 
                 40 hairs lost 
                 50 hairs lost 
               
               
                   
                 10 days after taking 
                 34 hairs lost 
                 42 hairs lost 
               
               
                   
                 20 days after taking 
                 26 hairs lost 
                 28 hairs lost 
               
               
                   
                 40 days after taking 
                 14 hairs lost 
                 11 hairs lost 
               
               
                   
                   
               
            
           
         
       
     
     The hairs of the patients before and at the predetermined time points after administration of the strains were photographed to measure their thicknesses, numbers, and glosses. Some of the results are shown in  FIGS. 15-16 . 
     These results indicate that the combination of the  Leuconostoc mesenteroides  strain (KCCM11827P) and the  Lactobacillus fermentum  strain (KCCM11910P) is particularly effective in preventing hair loss and markedly improving the thickness, number, and gloss of hairs. Therefore, it can be concluded that the combined administration of the two strains is effective in treating hair loss compared to single administration of the  Leuconostoc mesenteroides  strain (KCCM11827P). 
     Example 14: Preventive Effect of Combination of the  Lactobacillus sakei  Strain (KCCM11841P) and the  Lactobacillus fermentum  Strain Ceb-Ch-002 (KCCM11910P) on Hair Loss 
     The culture solution of the  Lactobacillus sakei  strain (KCCM11841P) prepared in Example 3 at a concentration of 0.5×10 6  to 0.5, 10 8  CFU/ml was combined with the culture solution of the  Lactobacillus fermentum  strain (KCCM11910P) prepared in Example 9 at a concentration of 0.5×10 6  to 0.5×10 8  CFU/ml. The combination of the two strain culture solutions was administered orally to a man aged 28 and a man aged 54 who suffered from alopecia twice (50-100 ml each time) daily over 1-3 months. 
     The numbers of hairs lost were counted before and 10, 20, and 40 days after taking the strains. The patients were instructed to wash their hair before hair counting. 
     The experimental results are shown in Table 9. 
     
       
         
           
               
             
               
                 TABLE 9 
               
             
            
               
                   
               
               
                 Numbers of hairs lost before and after taking 
               
            
           
           
               
               
               
               
            
               
                   
                 Subject 
                   
                   
               
               
                   
                 Time 
                 Man at age 28 
                 Man at age 54 
               
               
                   
                   
               
               
                   
                 Before taking 
                 43 hairs lost 
                 51 hairs lost 
               
               
                   
                 10 days after taking 
                 24 hairs lost 
                 41 hairs lost 
               
               
                   
                 20 days after taking 
                 14 hairs lost 
                 30 hairs lost 
               
               
                   
                 40 days after taking 
                  9 hairs lost 
                 14 hairs lost 
               
               
                   
                   
               
            
           
         
       
     
     The hairs of the patients before and at the predetermined time points after administration of the strains were photographed to measure their thicknesses, numbers, and glosses. Some of the results are shown in  FIGS. 17-18 .