KR20100005552A - Fermented milk using lactobacillus plantarum m23 and preparation method thereof - Google Patents

Abstract

PURPOSE: Fermented milk using Lactobacillus plantarum M23 and a manufacturing method thereof are provided to reduce manufacturing costs of the fermented milk with stable supply of spawn, and to improvemelanin formation inhibition capability. CONSTITUTION: Fermented milk includes Lactobacillus plantarum M23. A manufacturing method of the fermented milk includes a step for inoculating the Lactobacillus plantarum M23 to raw milk, a step for agitating and mixing 70 ~ 90 weight% fermented liquid and 10 ~ 30 weight% of secondary materials, and a step for storing the fermented milk in a wrapping pack.

Description

Fermented milk using Lactobacillus plantarum Μ23 strain and method for preparing the same {Fermented milk Using Lactobacillus plantarum M23 and Preparation method}

The present invention relates to fermented milk using the Lactobacillus plantarum M23 strain and a method for preparing the same, and more specifically, Lactobacillus plantarum M23 strain, a novel lactic acid bacterium capable of inhibiting melanogenesis as a lactic acid bacterium suitable for fermented milk production. It relates to fermented milk and a method for producing the same.

Lactic acid bacteria currently used in fermented milk include Lactobacillus bulgaricus , Lactobacillus casei , Lactobacillus acidophilus , Streptococcus thermophilus and Streptococcus. thermophilus) and Bifidobacterium ronggeom (Bifidobacterium longum ) has been used mainly, but it is dependent on income every year.

Therefore, in addition to the above-mentioned lactic acid bacteria, the need for the development of new lactic acid bacteria that can be smoothly supplied domestically without imports has been constantly emerging.

Accordingly, the present inventors pay attention to the presence of lactic acid bacteria having a variety of properties in the crude milk, and as a result of research efforts to secure lactic acid bacteria with melanin production inhibitory ability suitable for the production of fermented milk, a new lactic acid bacteria Lactobacillus plantarum ( Lactobacillus) from crude oil plantarum ) M23 strain (KACC 91329P) to provide a fermented milk and a method for producing the fermented milk comprising the same.

The present invention is to provide a fermented milk containing a novel lactic acid bacteria and a method for producing the same.

The present invention is to provide a fermented milk comprising a novel lactic acid bacteria isolated from crude milk Lactobacillus plantarum M23 strain (KACC 91329P) and a method for producing the same.

The present invention is a fermented milk containing a novel lactic acid bacterium isolated from crude milk Lactobacillus francarum M23 strain (KACC 91329P) can lower the production cost of fermented milk due to the stable supply of seed.

Fermented milk containing Lactobacillus francarum M23 strain (KACC 91329P) of the present invention not only has excellent palatability in color, taste, texture, etc., but also has superior melanin production inhibition ability than commercially available fermented milk fermented with conventionally found lactic acid fermentation bacteria. Fermented milk may be provided.

The present invention provides a fermented milk comprising a strain of Lactobacillus francarum M23 (KACC 91329P), which is a novel lactic acid bacterium.

The present invention provides a method for producing fermented milk, comprising the step of inoculating raw milk with a novel lactic acid bacterium Lactobacillus francarum M23 strain (KACC 91329P).

Hereinafter will be described in more detail the contents of the present invention.

The present invention provides a fermented milk containing fermented milk, Lactobacillus plantarum M23 strain (KACC 91329P), which is a novel lactic acid bacterium isolated from crude milk.

The Lactobacillus francarum M23 strain (KACC 91329P) has an effect of inhibiting melanin production, and can suppress melanin production when the fermented milk containing the Lactobacillus francarum M23 strain is ingested.

The fermented milk of the present invention can be prepared fermented milk having excellent palatability by adding the fruit fermented sugars such as fructose, oligosaccharide or fruit such as strawberry, blueberry, grapes to fermented milk containing Lactobacillus francarum M23 strain. have. In addition, Lactobacillus plantarum M23 strain is excellent in melanin production inhibitory ability, the fermented milk according to the present invention is superior to the melanin production inhibitory ability than the fermented milk by other strains.

Lactobacillus plantarum M23 strain, a novel lactic acid bacterium of the present invention, was isolated from crude milk, and screened and identified according to the experimental example described below. Identification of the Lactobacillus plantarum M23 strain was made based on its morphological, physiological, biochemical and genetic properties. The novel lactic acid bacteria isolated from crude oil were found to be suitable for fermented milk production because of excellent acid resistance, bile resistance, antibacterial activity and melanin production ability, and completed the present invention.

After culturing in modified MRS medium in crude oil, the inventors separated each colony purely and selected the colonies that turned yellow as potential lactic acid bacteria. The selected strains were plated in electro-modified MRS medium, and then separated by pure aerobic culture. As a result of identifying the purely isolated strains, the strains were Gram-positive bacillus, well grown with or without oxygen, and negative for catalase and motility. In addition, the growth at 15 ℃ but did not grow at 45 ℃, it was confirmed that belongs to Lactobacillus ( Lactobacillus ) did not generate ammonia from gas and alginic acid from glucose (glucose).

On the other hand, 49 kinds of sugar fermentation tests were carried out to confirm the species name, and acid was generated from 25 species such as lactose, and the results obtained by inputting into the ATB identification system and 16S rRNA gene sequence were obtained. Was matched and turned out to be Lactobacillus plantarum.

Therefore, in the present invention, among the skim milk coagulation strains isolated from crude milk, strains excellent in skim milk coagulation rate and melanin inhibition ability were named as Lactobacillus plantarum M23.

As a result of testing the number of viable cells and the growth of lactic acid bacteria, the optimum temperature was 40 ℃ and it took 17 hours to reach the pH condition of pH 4.3-4.4. In addition, by measuring the growth inhibitory concentration (MIC) value through the antibiotic resistance test, compared with other antibiotics, penicillin-G (Penicillin-G), oxacillin, Novobiocin (Novobiocin Antimicrobial susceptibility to) and Chloramphenicol, while resistant to vancomycin. As a result of the enzyme activity test, the enzyme activity is high in leucine arylamidase and beta galactosidase.

In addition, β-glucuronidase, a carcinogenic enzyme that converts benzopyrene to carcinogenic substances, has been shown to have no enzymatic activity, indicating safety. Bile resistance was measured by testing the growth of lactic acid bacteria in MRS medium. The time required to increase the OD value by 0.3 was 4 hours without bile and 5 hours with bile. It appears to be resistant to bile.

In addition, the test of the pH resistance shows that the acid resistance is hardly affected even at pH 2 which is a strong acid compared to the control pH of 6.4. The antibacterial activity test showed a high inhibitory activity of 77.77% against Escherichia coli , 83.76% against Staphylococcus aureus and 86.51% against Salmonella typhimurium . As described above, it can be confirmed that Lactobacillus plantarum M23 of the present invention is a lactic acid bacterium suitable for fermented milk production.

The Lactobacillus plantarum M23 of the present invention was deposited on September 27, 2007 with the accession number KACC 91329P to the Korea Agricultural Microbiological Resources Center (KACC).

The present invention shows a method for preparing fermented milk comprising the novel lactic acid bacteria Lactobacillus francarum M23 strain (KACC 91329P) mentioned above.

The present invention provides a method for producing fermented milk, comprising the step of inoculating crude milk with Lactobacillus plantarum M23 strain (KACC 91329P), which is the novel lactic acid bacterium mentioned above.

The method of producing fermented milk by inoculating Lactobacillus francarum M23 strain of the present invention in crude milk will be described in detail by process.

[Step 1] Preparation of spawn and bulk starter

Lactobacillus plantarum M23 strain (KACC 91329P), isolated from crude oil, was inoculated with 0.5-1% (v / v) of skim milk or reduced skim milk and incubated at 37-42 ° C for 12-24 hours. do. Then, 0.5-1% (v / v) of skim milk or reduced skim milk heat-treated with Lactobacillus plantarum M23 strain (KACC 91329P) was cultured for 30 to 60 minutes at 80-90 ° C. ) And incubated at 37-42 [deg.] C. for 12-24 hours to use this culture as bulk inoculation bacteria.

[Step 2] Preparation of Fermentation Broth

0-100 parts by weight of crude oil, 0-10 parts by weight of skim milk powder and 0-10 parts by weight of yeast extract are added, mixed at 55-65 ° C., completely dissolved, homogenized at a pressure of 180-250 kgf / cm 2 and then at 85-95 ° C. Sterilize for 5-30 minutes.

After cooling the homogenized milk through the above process to 35 ~ 45 ℃ about 100 parts by weight of the homogenized milk Lactobacillus plantarum M23 (KACC 91329P) bulk inoculation bacteria obtained in the first step is Inoculate 0.5 to 1.0 parts by weight of the incubation until the final pH is 4.3 to 4.6 and then stirred with a stirrer to cool the culture to 15 to 20 ℃ containing Ltobacillus plantarum M23 strain (KACC 91329P) Obtain fermentation broth.

The above process is not necessarily limited to one preferred embodiment culture of fermented milk production usable in the present invention.

 [Step 3] Fermented Milk Manufacture

The subsidiary material can be added to the fermentation broth obtained above to improve the functionality.

70-90 weight% of fermentation broth obtained by the said 2nd process, and 10-30 weight% of submaterials are mixed and stirred, and fermented milk is manufactured.

In the above, the subsidiary material may use any one or more selected from the group of syrup, fruit and purified water.

In the above, the syrup may be any one or more selected from the group of oligosaccharides, fructose, sugar, glucose, and dextrin, preferably oligosaccharides and fructose.

The fruit may be any one or more selected from the group of grape concentrate, grape juice, strawberry concentrate, strawberry juice, preferably grape juice and strawberry juice.

As for the subsidiary materials, oligosaccharides 3.0 to 7.0% by weight, liquid fructose 1.0 to 5.0% by weight, grape concentrate or strawberry 쨈 1 to 10% by weight, and purified water 1.0 to 10% by weight.

After the fermented milk is prepared in the above, it can be packed and stored in a container.

The grape concentrate may be used after the grape juice to remove the grape seed and concentrated to have a sugar content of 60 ~ 65 Brix.

Strawberry 에서 in the above can be used strawberry 인 sugar 55-65 Brix, strawberry content 40-50%.

Strawberry 에서 in the above can be used strawberry jam with a sugar content of 61.2Bx, strawberry content 45%.

The storage can be carried out at a temperature of 0 to 10 ° C, preferably 0 to 5 ° C.

Hereinafter, the content of the present invention will be described in more detail by the experimental examples, examples. However, these are only presented to understand the contents of the present invention and should not be construed that the scope of the present invention is limited thereto.

Experimental Example 1 Isolation of Microorganisms

Crude oil used as a sample was collected from ranches in each country in Korea and inoculated in a modified MRS medium containing bromcresol purple and sodium azide (Table 1) by 0.3 ml each in a plan view method 37 Incubated for 48 hours at ℃. Then, each colony was purely separated in electro-modified MRS medium, and the colony that turned yellow was selected as a potential lactic acid bacterium. The selected strains were plated with platinum three times in electro-modified MRS medium and then aerobicly cultured and purified. Pure isolates were inoculated in 10% reduced skim milk (Difco Laboratories, USA) and incubated at 37 ° C. for 24 hours to confirm clotting.

TABLE 1 Composition of Modified MRS Medium Used for Microbial Separation in the Present Invention

ingredient Component ratio (g / ℓ) Proteos peptone 10 Horseradish extract 10 Yeast extract 5 Lactose 20 Tween 80 One Ammonium citrate 2 Sodium acetate 5 Magnesium sulfate 0.1 Manganese Sulfate 0.05 Dipotassium phosphate 2 Sodium azide 0.25 Bromcresol Purple 0.04

Experimental Example 2 Selection of Melanin Inhibitory Active Lactic Acid Strains

The lactic acid bacteria and commercial lactic acid bacteria 1% isolated above were incubated in 10% reduced skim milk until reaching pH 4.4 at 37 ° C., and then the obtained culture solution was centrifuged at 4 ° C. and 15,000 rpm for 10 minutes. The obtained supernatant was filtered with a 0.45 μm PVDF syringe filter (whatman Co.), and 120 μl of 8.0 mM L-Dopa dissolved in 67 mM phosphate buffer (pH 6.8) and 40 μl of the sample were placed in a 96-well microplate. After the measurement, 40 μl of a tyrosinase mushroom (125 unit / mL) solution was added. After reacting for 10 minutes at room temperature, the amount of dopachrome produced was measured by using an ELISA reader (UV-Visible Spectrophotomater). The sample absorbance values before and after the reaction and the absorbance values (C) of the control group were calculated by the following equation, and the inhibition rate of tyrosinase activity (%) was measured. The results are shown in Table 2 below. At this time, when measuring the inhibition rate of melanocyte-derived tyrosinase activity, the tyrosinase produced by Melanoma B16, a mouse-derived melanin-producing cell line, was directly extracted and used.

×100% Inhibition =

× 100

 <Table 2> Non-coagulation of skimmed milk by storage lactic acid strains and isolates from commercial strains, Korea Food Research Institute and inhibition rate of tyrosinase activity after culture

As shown in Table 2, the isolated lactic acid bacterium M23 strain was selected as a strain having excellent melanin production inhibition ability as tyrosinase activity inhibition rate was higher than that of other strains.

Experimental Example 3 Identification of Microorganisms

The taxonomy of microorganisms was performed according to the method of Hammes et al. (The prokaryotes, 1563-1578, 2nd Edition, Springer-Verlag Co. (1992)), the results are shown in Table 3. These strains were both Gram positive and bacillus, well grown with or without oxygen, and negative for catalase and motility. In addition, it grew at 15 ℃ but did not grow at 45 ℃, and did not produce ammonia from gas and alginic acid from glucose (glucose) belonged to the genus Lactobacillus (genus). In order to determine the species, an acid was generated from 25 species including lactose when 49 sugar fermentation tests were performed using the API 50CHL kit (API bioMerieux, France) and input into ATB identification system (API bioMerieux, France). And the results obtained using the 16S rRNA Gene Sequence were found to be Lactobacillus plantarum. Lactobacillus plantarum M23, which has excellent defatting coagulation rate and melanin production ability among skim milk coagulation strains isolated from crude oil, was named Lactobacillus plantarum M23, and was deposited with the Korea Agricultural Microbiological Resources Center (KACC) on September 27, 2007. Deposited as 91329P.

Table 3 Physiological and biochemical properties of the novel microbial L. plantarum M23 of the present invention

 Gram Reactive Cell Form Sporulation Motility Aerobic Growth Anaerobic Growth Catalase Reaction Growing at 15 ° C Growing at 45 ° C Producing Gas from Glucose Generating Ammonia from Arginine + rod--+ +-+--- Party use ability  Glycerol Erititol D-Arabinose L-Arabinose Ribose Glucose Fructose Mannose Sorbose Rhamnose Dulcitol Inositol Mannitol Sorbitol α-Methyl-D-Mannoside α-Methyl-D-Glucoside N Acetyl Glucoseamine Amigalin Arbutin Esculin Salicylic Cellobiose Maltose Lactose Meliviose ----+ + + + +---+ +--+ + + + + + + + +  D-Cyros L-Cyros Adonitol β-Methyl-D-Cyrroside Galactose Sucrose Trehalose Inulin Melezitose Raffinose Starch Glycogen Silitol Genthiobios D-Turanos D-Lysos D-Tagatos D- Fucose L-fucose D-arabitol L-arabitol Gluconate 2-keto-gluconate 5-keto-gluconate ----+ + +--+---+ +----+-+ +-

Experimental Example 4 Growth Test of Microorganisms

The growth of lactic acid bacteria was tested by measuring the number of live bacteria and pH. The number of viable cells was inoculated with 150 ml of 10% skim milk and 1 drop of lactic acid bacteria with a pipette for 1 ml, and then each sample incubated at 34 ° C., 37 ° C. and 40 ° C. for 3 hours at 24 hours was diluted in 0.1% peptone solution. (bromocresol purple) Poured in agar plate (BCP plate count agar) added to the hardened and incubated for 48 hours at 35 ℃, counting the pH change by temperature and time, the optimum temperature is 40 ℃, the optimum of fermented milk It took 17 hours to reach the pH condition pH 4.3-4.4.

Experimental Example 5 Antibiotic Resistance Test

In the antibiotic resistance test, tryptic soy broth (Difco, USA) was used to determine the minimum inhibitory concentration (MIC) value by observing growth by a 2-fold dilution method and the results are shown in Table 4. It was.

TABLE 4 Antibiotic susceptibility of L. plantarum M23

Antibiotic Lowest inhibitory concentration (㎍ / ㎖) Aminoglycosides Amikacin 40 Centamicin 10 Kanamycin 50 Neomycin 25 Streptomycin * 25 β-lactam meter Phenylcillin-G * 0.313 Methicillin 5 Oxacillin 1.875 Ampicillin 5 Gram-positive spectrum Bassitracin * 3.75 Rifampicin 3.75 Novobiocin 0.11 Lincomycin 3.125 Gram-Voice Spectrum Polymycin B * 37.5 Broad spectrum Chloramphenicol 1.25 Vancomycin * 1600

*: units / ml

As can be seen in Table 4, L. plantarum M23 is antibiotic susceptibility to penicillin-G, pentacillin-G, oxacillin, Novobiocin and Chloramphenicol compared to other antibiotics. While high, resistant to vancomycin.

Experimental Example 6 Enzyme Activity Test

Enzyme activity test was prepared by diluting the strain incubated for 18 hours at 37 ℃ in MRS liquid medium with saline to prepare a sample of 10 ⁵ ~ 10 ⁶ cfu / ㎖, using API ZYM kit (API bioMerieux, France) After incubating at 37 ° C. for 5 hours, enzyme reaction was performed. Enzyme activity is shown in Table 5 by comparing the standard color table with a value of 0-5.

Table 5 Comparison of enzymatic activity of L. plantarum M23

enzyme Enzyme activity Alkaline phosphatase esterase (C4) esterase lipase (C8) lipase (C14) leucine arylamidase valine arylamidase cystine arylamidase trypsin chymotrypsin acid phosphatase naphthol-AS-BI-phosphohydrolase α-galactosidase β-galactosidase β-gluronidase α-glucosidase β-glucosidase N-acecyl-β-glucosaminidase α-mannosidase β-fucosidase  1 1 1 0 5 4 2 0 0 3 3 2 5 0 4 4 5 0 0

*: Standard colors are specified in steps from 0 to 5, where 0 is negative and 5 is maximum intensity. 1 is 5 nanomoles (nanomoles), 2 is 10 nanomoles, 3 is 20 nanomoles, 4 is 30 nanomoles, 5 is 40 nanomoles or more enzyme activity.

As can be seen in Table 5, L. plantarum M23 showed high enzymatic activity in leucine arylamidase and β-galactosidase. Β-glucuronidase, a carcinogenic enzyme that converts benzopyrene to carcinogenic substances, was found to have no enzymatic activity, indicating safety.

Experimental Example 7 Bile Resistance Test

Bile tolerance tests were determined according to the method of Gilliland and Walker (Gilliland & Walker, J. Dairy Sci., 73, 905, 1990). At this time, the time required to increase the OD value 0.3 was 4 hours when bile was not added and 5 hours when bile was added.

Experimental Example 8 pH Tolerance Test

The pH resistance test was measured according to Clark et al. (Clark et al., Cultured Dairy Products J., 28 (4), 11, 1993) and the results are shown in Table 6.

<Table 6> L. plantarum M23 resistance after 3 hours in hydrochloric acid solution (unit: CFU per ㎖ (Log number))

Incubation time (hr) pH 2 pH 3 pH 4 pH 6.4 0 7.72 7.78 7.72 7.73 One 7.68 7.75 7.74 7.73 2 7.61 7.64 7.74 7.85 3 7.52 7.61 7.81 7.88

As can be seen in Table 6, it was shown that there was little effect even at pH 2, which is a strong acid, compared to the control pH of 6.4.

Experimental Example 9 Antibacterial Activity Test

The antimicrobial activity was measured according to Gilliland & Speck, J. Food Prot., 40 (12), 820, 1977. The results are shown in Table 7.

TABLE 7 L. plantarum in MRS medium M23 suppression of food poisoning bacteria (*)

Food poisoning bacteria growth % Inhibition Food poisoning bacteria ^a L. plantarum M23 ^a CFU / mL pH CFU / mL pH Esherichia coli 1.15 × 10 ⁸ 6.46 2.55 × 10 ⁷ 5.02 77.77 Salmonella typhimurium 1.25 × 10 ⁸ 5.63 1.68 × 10 ⁷ 4.96 86.51 Saphyloccous aureus 6.59 × 10 ⁸ 5.78 1.07 × 10 ⁸ 4.99 83.76

L. plantarum Initial number of M23: 1.09 × 10 ⁷ CFU / mL

^a measurement after 6 hours of incubation at 37 ° C

As can be seen in Table 7, L. plantarum M23 was 77.77% for Escherichia coli , 83.76% for Staphylococcus aureus , Salmonella typhimurium ) Showed a high inhibitory capacity of 86.51%.

Example 1

96.02 parts by weight of crude oil, 3.85 parts by weight of skim milk powder and 0.13 parts by weight of yeast extract were added, mixed at 65 ° C., completely dissolved, and homogenized at a pressure of 215 ± 35 kgf / cm 2. The homogenate was sterilized at 90 ° C. for 5 minutes, cooled to 37 ° C., and then inoculated with 1.0 part by weight of Lactobacillus plantarum M23 (KACC 91329P) bulk inoculating bacteria with respect to 100 parts by weight of the homogenate. The culture was cooled to 17 ± 2.5 ° C. while slowly incubated with stirrer to 4.4 to obtain a fermentation broth containing Lactobacillus plantarum M23 (KACC 91329P).

84% by weight of the fermentation broth obtained above, 5.0% by weight of oligosaccharide, 4.0% by weight of liquid fructose, 1.0% by weight of grape concentrate, and 6.0% by weight of purified water were added and mixed, followed by stirring and then Lactobacillus plantarum M23 (KACC 91329P). To prepare a fermented milk comprising a.

Table 8 shows the components of the fermented milk.

The Lactobacillus plantarum M23 (KACC 91329P) bulk inoculum is lactobacillus plantarum M23 strain (KACC 91329P), which is a seed isolated from crude milk, at 1% (v / v) in reduced skim milk. Inoculated, incubated at 39.5 ± 2.5 ° C. for 18 ± 6 hours, and then, the culture medium spawn in which Lactobacillus plantarum M23 strain (KACC 91329P) was incubated at 85 ± 5 ° C. for 45 ± 15 minutes It was inoculated at 1% (v / v) to reduced skim milk, and cultured at 39.5 ± 2.5 ℃ for 18 ± 6 hours was used.

Example 2

Fermented milk was prepared in the same manner as in Example 1, except that 3.0% by weight instead of 1.0% by weight of grape concentrate and 4.0% by weight instead of 6.0% by weight of purified water were used.

Example 3

Fermented milk was prepared in the same manner as in Example 1, except that 5.0% by weight instead of 1.0% by weight of grape concentrate and 2% by weight of 6.0% by weight of purified water were used.

Example 4

Fermented milk was prepared in the same manner as in Example 1, except that 2.0% by weight instead of 4.0% by weight of fructose, 5.0% by weight of strawberry 쨈 instead of 1.0% by weight of grape concentrate, and 5.0% by weight of 6.0% by weight of purified water were used. And stored.

Example 5

Fermented milk was prepared in the same manner as in Example 1, except that 2.0% by weight instead of 4.0% by weight of liquid fructose, 6.0% by weight of strawberries and 3.0% by weight of 6.0% by weight of purified water instead of 1.0% by weight of grape concentrate were used. And stored.

Example 6

Fermented milk was prepared in the same manner as in Example 1, except that 2.0 wt% instead of 4.0 wt% of liquid fructose, 1.0 wt% of strawberry 에 instead of 1.0 wt% of grape concentrate, and 1.0 wt% instead of 6.0 wt% of purified water were used. Stored.

Table 8. Fermented milk components of Examples 1 to 6 (unit: weight%)

Item (% by weight) Example 1 Example 2 Example 3 Example 4 Example 5 Example 6 Fermentation solution crude oil 80.66 80.66 80.66 80.66 80.66 80.66 Skim milk powder 3.23 3.23 3.23 3.23 3.23 3.23 Yeast extract 0.11 0.11 0.11 0.11 0.11 0.11 Subsidiary Material oligosaccharide 5.0 5.0 5.0 5.0 5.0 5.0 Liquid fructose 4.0 4.0 4.0 2.0 2.0 2.0 Grape concentrate 1.0 3.0 5.0 - - - Strawberry - - - 4.0 6.0 8.0 Purified water 6.0 4.0 2.0 5.0 3.0 1.0 Sum 100 100 100 100 100 100

Test Example 1 Sensory Test

13 people who worked in dairy related fields for 2 years or more were sensory test agents, and sensory tests such as color, taste, texture, and general preference of the fermented milk prepared in Examples 1 to 6 were conducted by a 9-point grading method. The results are summarized in Table 9 below.

Table 9. Fermented milk sensory test of Examples 1 to 6 and the control group

Item Sensory test * Color flavor Organization Comprehensive Preference Example 1 6.70 ± 1.63 6.82 ± 1.62 6.76 ± 1.83 6.84 ± 1.51 Example 2 6.92 ± 1.11 7.37 ± 1.38 6.86 ± 1.07 7.34 ± 1.43 Example 3 6.84 ± 1.57 7.12 ± 1.26 6.93 ± 0.86 7.10 ± 0.94 Example 4 6.82 ± 1.54 5.64 ± 1.43 6.82 ± 1.08 5.82 ± 1.08 Example 5 7.00 ± 1.34 7.20 ± 0.71 7.00 ± 1.00 7.16 ± 0.92 Example 6 6.55 ± 1.63 7.24 ± 1.17 6.91 ± 1.04 7.12 ± 1.25 Control ** 6.33 ± 1.72 6.42 ± 1.44 6.42 ± 1.38 6.35 ± 1.42

* 1: not very good, 5: normal, 9: very good

** The control is pure milk product of Seoul Milk.

As shown in Table 9, the fermented milk of Examples 1 to 6 of the present invention containing Lactobacillus plantarum M23 strain was able to prepare fermented milk with excellent palatability.

<Test Example 2> Animal test results

In order to determine the whitening effect of fermented milk using Lactobacillus plantarum M23 strain having excellent melanin production inhibitory effects, animal experiments were performed using Brown guinea pigs (females and five) of about 500 g (see Table 10). After removing the hair on the back, and washed, it was fixed and irradiated with 500mJ ultraviolet rays to a predetermined area four times at weekly intervals, the application was started the day after the UV irradiation for 3 weeks. 30 minutes after hair removal and washing, 50 μl of each treatment group was applied to each site (twice a day), applied for 2 to 4 weeks, and the brightness value was measured by a color difference meter.

<Table 10> Whitening effect of fermented milk in Brown guinea pig (n = 5)

Item Week Whitening effect ^{2 )} 0 2 4 Control group 1 50.27 ± 2.04 ¹ 51.65 ± 1.01 52.29 ± 2.73 4.02% Control 2 50.29 ± 2.40 52.06 ± 1.47 52.94 ± 2.45 5.27% S1 9.96 ± 2.20 52.26 ± 1.19 53.77 ± 1.55 7.63% S2 50.01 ± 2.20 52.62 ± 1.22 53.77 ± 2.64 7.52% S3 49.82 ± 1.38 52.88 ± 0.67 53.46 ± 1.75 7.31%

¹⁾ L: Brightness

²⁾ Whitening effect: (brightness value after 4 weeks-initial brightness value) / initial brightness value x 100

Control group 1: Mama Dairy's Domaschinoplane fermented milk

Control group 2: distilled water

S1: grape fermented milk fermented with L. plantarum M23 strain

S2: Plain fermented milk fermented with L. plantarum M23 strain

S3: Whitening Cream Cosmetic

As described above, although described with reference to a preferred embodiment of the present invention, those skilled in the art will be variously modified and modified within the scope of the present invention without departing from the spirit and scope of the invention described in the claims below. It will be appreciated that it can be changed.

Claims (2)

In fermented milk, Fermented milk comprising Lactobacillus francarum M23 strain (KACC 91329P). In the production of fermented milk, A method for producing fermented milk, comprising the step of inoculating Lactobacillus francarum M23 strain (KACC 91329P) in crude milk.