KR100989638B1 - Fermented milk Using Lactobacillus zeae RMK354 and Preparation method thereof - Google Patents

Abstract

The present invention relates to a fermented milk comprising a Lactobacillus Zia RMK354 strain and a method for preparing the same, and more specifically, to a Lactic acid bacterium Lactobacillus zeae ( Lactobacillus zeae ) RMK354 strain having a lactic acid and ACE inhibitory activity ability suitable for producing fermented milk. It relates to a fermented milk comprising and a preparation method thereof.

Fermented milk containing Lactobacillus Jia RMK354 strain (KACC 91314P) of the present invention is not only excellent palatability in color, taste, texture, etc., but also excellent in ACE inhibitory activity than commercially available fermented milk fermented with conventionally found lactic acid fermentation bacteria. Can be provided.

ACE inhibitory activity, fermented milk, Lactobacillus

Description

Fermented milk containing Lactobacillus giae RMＫ354 strain and its preparation method {Fermented milk Using Lactobacillus zeae RMK354 and Preparation method}

The present invention relates to a fermented milk comprising a Lactobacillus Zia RMK354 strain and a method for preparing the same, and more specifically, to a Lactic acid bacterium Lactobacillus zeae ( Lactobacillus zeae ) RMK354 strain having a lactic acid and ACE inhibitory activity ability suitable for producing fermented milk. It relates to a fermented milk comprising and a preparation method thereof.

Lactic acid bacteria currently used in fermented milk include Lactobacillus bulgaricus , Lactobacillus casei , Lactobacillus acidophilus , Streptococcus thermophilus and Bifidobacte. Leeum long gum ( Bifidobacterium longum ) and the like are mainly used, the lactic acid bacteria used in the fermented milk is a situation that depends on the import every year.

Therefore, the necessity for the development of new lactic acid bacteria that can be supplied smoothly in the country without importing from abroad other than the above-mentioned lactic acid bacteria has emerged constantly.

Accordingly, the present inventors pay attention to the presence of lactic acid bacteria having a variety of properties in the crude milk, researched to ensure the lactic acid bacteria that are suitable for the production of fermented milk, and ACE inhibitory activity, as a result, a novel lactic acid bacteria Lactobacillus jiae ( Lactobacillus zeae ) RMK354 strain (KACC 91314P) to isolate and provide a method for producing fermented milk and 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 Lactobacillus Jia RMK354 strain (KACC 91314P) isolated from crude milk and a method for producing the same.

The present invention provides a fermented milk comprising Lactobacillus Jia RMK354 strain (KACC 91314P), a lactic acid fermentation bacterium isolated from crude milk, thereby lowering the production cost of fermented milk due to the stable supply of seed.

Fermented milk containing Lactobacillus Jia RMK354 strain (KACC 91314P) of the present invention is not only excellent palatability in color, taste, texture, etc., but also excellent in ACE inhibitory activity than commercially available fermented milk fermented with conventionally found lactic acid fermentation bacteria. Can be provided.

The present invention provides fermented milk, comprising the Lactobacillus strain RMK354 strain (KACC 91314P).

The present invention provides a method for producing fermented milk, comprising the step of inoculating a crude milk with Lactobacillus strain RMK354 strain.

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

Hereinafter, the configuration of the present invention in detail.

The present invention provides a fermented milk comprising the seed Lactobacillus Gi RMK354 (KACC 91314P) isolated from the crude oil. The Lactobacillus strain RMK354 (KACC 91314P) of the present invention can increase the ACE inhibitory activity, the fermented milk containing the Lactobacillus strain RMK354 (KACC 91314P) can increase the ACE inhibitory activity.

The fermented milk of the present invention can be prepared fermented milk having excellent palatability by adding a food grade acceptable sugar such as fructose, oligosaccharide and / or fruit using strawberry, blueberry, grape, etc. have. In addition, Lactobacillus Jia RMK354 strain has excellent ACE inhibitory activity, the fermented milk according to the present invention is superior to the ACE inhibitory activity than the fermented milk by other strains.

Lactobacillus Jia RMK354 strain, a novel microorganism of the present invention, was isolated from crude oil, and screened and identified according to the method of the experimental example described below. Identification of microorganisms was made based on their morphological, physiological and biochemical properties. The novel lactic acid bacteria isolated from crude oil was found to be suitable for fermented milk production with excellent lactic acid and ACE inhibitory ability, thus completing the present invention.

After culturing in modified MRS medium in crude milk, 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, it was grown at 15 ℃ and 45 ℃, it did not generate ammonia from gas and alginic acid from glucose (glucose) was confirmed to belong to Lactobacillus ( Lactobacillus ).

On the other hand, 49 kinds of sugar fermentation tests were carried out to confirm the species name, acid was generated from 20 species such as lactose, inputted into the ATB identification system, and the results obtained by using the 16S rRNA gene sequence. Was matched and turned out to be Lactobacillus.

Therefore, the present invention is Lactobacillus zeae strains excellent in skim milk coagulation rate and ACE inhibitory ability among the skim milk coagulation strains isolated from crude milk Named RMK354.

As a result of testing the growth of lactic acid bacteria by measuring the number of viable cells and pH, the optimum temperature is 40 ℃, it takes 10 hours to reach the pH value of the fermented milk, pH 4.3 ~ 4.4. In addition, by measuring the growth inhibitory concentration (MIC) value through the antibiotic resistance test, the resistance to polymycin B and vancomycin compared to other antibiotics, while penicillin-G, bacitracin and novobio High sensitivity to God. As a result of enzyme activity test, enzyme activity was high in esterase and leucine arylamidase.

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 7.3 hours without bile and 8 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. In the antimicrobial activity test, there is no inhibitory activity against Escherichia coli and Staphylococcus aureus , but it shows 60.0% inhibition against Salmonella typhimurium . As described above, the Lactobacillus geranium of the present invention. It can be seen that RMK354 is a suitable lactic acid bacterium for the production of fermented milk.

Lactobacillus geranium of the present invention RMK354 was deposited on May 4, 2007 with the accession number KACC 91314P to the Korea Institute of Agricultural Biotechnology (KACC).

Hereinafter, in the present invention, the Lactobacillus jiae RMK354 is referred to as L. zeae RMK354 for convenience.

The present invention shows a method for producing fermented milk comprising the novel lactic acid bacteria Lactobacillus Jia RMK354 strain (KACC 91314P) mentioned above.

The present invention provides a method for producing fermented milk, comprising the step of inoculating crude milk with lactobacillus Jia RMK354 strain (KACC 91314P), the novel lactic acid bacteria mentioned above.

When the Lactobacillus Jia RMK354 strain of the present invention is inoculated in crude milk to explain in detail the process for producing fermented milk as follows.

[Step 1] Preparation of spawn and bulk starter

Lactobacillus zeae RMK354 strain (KACC 91314P), isolated from crude oil, is inoculated in skim milk or reduced skim milk at 0.5-1% (v / v) and incubated at 37-42 ° C for 12-24 hours. . Then, 0.5-1% (v / v) of skim milk or reduced skim milk heat-treated with Lactobacillus zeae RMK354 strain (KACC 91314P) was heat-treated at 80-90 ° C. for 30 minutes to 60 minutes. Inoculated at, 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 and 0-10 parts by weight of skim milk powder are added, mixed at 55-65 ° C., completely dissolved, homogenized at a pressure of 180-250 kgf / cm 2 and sterilized at 85-95 ° C. for 5-30 minutes.

Lactobacillus zeae RMK354 strain (KACC 91314P) bulk inoculating bacteria obtained in the first step with respect to 100 parts by weight of the cooled homogenized milk after cooling the homogenized crude milk through the above process 0.5 Incubate at ~ 1.0 parts by weight to incubate until the final pH is 4.3 ~ 4.6, and the culture medium is cooled to 15 ~ 20 ℃ while stirring with a stirrer to obtain a fermentation broth containing Lactobacillus zeae RMK354 strain (KACC 91314P) .

[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 used strawberry 쨈 or blueberry 쨈.

As for the subsidiary materials, oligosaccharides 3.0 to 7.0% by weight, liquid fructose 1.0 to 5.0% by weight, strawberries or blueberries 4 to 10% by weight and purified water 1.0 to 10% by weight can be used.

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

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%.

Blueberry 쨈 can be used in the blueberry 당 sugar of 55 to 65 Brix, blueberry content of 40 to 50%.

Blueberry 에서 in the above can be used blueberry jam with a sugar content of 62Brix, blueberry content 40%.

In the above storage may be carried out at a temperature of 0 ~ 10 ℃, preferably 0 ~ 5 ℃.

Hereinafter, the content of the present invention will be described in more detail with reference to Examples. However, these examples are only presented to understand the content of the present invention, and the scope of the present invention should not be construed as being limited to these embodiments.

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 for pure separation. 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 Isolation

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 ACE Inhibitory Active Lactic Acid Strains

The lactic acid bacteria and commercial lactic acid bacteria 1% isolated above were incubated in 10% reduced skimmed milk until pH 4.4 was reached at 37 ° C., and the results of measuring the ACE inhibition rate with the whey obtained from each are shown in Table 2 below.

Table 2. The results of coagulation of skimmed milk and the results of ACE inhibition after cultivation in commercial strains, Korea Food Research Institute

Note) * Commercial Strains

As shown in Table 2, the isolated lactic acid bacteria RMK354 strain was selected as a strain having excellent ACE inhibitory activity as the ACE inhibition rate was similar or higher than that of other strains.

Experimental Example 3 Identification of Microorganisms

The taxonomy of microorganisms is described by Hammes et al. The prokaryotes, 1563-1578, 2nd Edition, Springer-Verlag Co. (1992), and the results are shown in Table 3. These strains were both Gram-positive and bacillus, well grown with or without oxygen, and catalase and motility were negative. In addition, it was grown at 15 ℃ and 45 ℃ belonged to the genus Lactobacillus (genus) because it does not produce gas from glucose (glucose), and ammonia from arginine (arginine). In order to determine the species, when 50 kinds of sugar fermentation tests were conducted using API 50CHL kit (API bioMerieux, France), acid was generated from 20 species such as lactose and inputted into the ATB identification system. The results obtained using the gene sequence were matched and found to be Lactobacillus zeae . Lactobacillus zeae strains from the crude milk coagulation strains that were isolated from crude milk were found to have excellent skim milk coagulation rate and ACE inhibitory activity. It was named RMK354 and deposited on May 4, 2007 with the accession number KACC 91314P to the Korea Agricultural Microbiological Resources Center (KACC).

Table 3. Physiological and Biochemical Properties of New Microorganism L. zeae RMK354

Gram reaction
Cell morphology
Spore formation
motility
Aerobic growth
Anaerobic growth
Catalase reaction
Grow at 15 ℃
Grow at 45 ℃
Gas generation from glucose
Production of Ammonia from Arginine +
rod
-
-
+
+
-
+
+
-
- Party use ability Glycerol
Erythritol
D-Arabinose
L-Arabinose
Ribose
Glucose
Fructose
Mannose
Sorbos
Rhamnos
Dulcitol
Inositol
Mannitol
Sorbitol
α-methyl-D-mannoside
α-methyl-D-glucoside
N Acetyl Glucamine
Amigdalin
Arbutin
Esculin
Salinity
Cellobiose
Maltose
Lactose
Melibiose -
-
-
+
+
+
+
+
-
+
-
-
+
-
-
-
+
+
+
+
+
+
+
+
- D-cyros
L-cyros
Adonitol
β-methyl-D-cyroside
Galactose
Sucrose
Trehalose
Inulin
Melezitos
Raffinos
Starch
Glycogen
Silitol
Genthiobiose
D-Turanos
D-Lithus
D-tagatose
D-after course
L-hukos
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, 37, and 40 ° C for 3 hours at 3 hours intervals was diluted in 0.1% peptone solution. Poured from the agar plate and hardened, incubated for 48 hours at 35 ℃, counting the pH change by temperature and time, the optimum temperature is 40 ℃, it took 10 hours to reach the pH pH 4.3 ~ 4.4 of the fermented milk .

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. zeae RMK354

Antibiotic Minimum inhibitory concentration (㎍ / ㎖) Aminoglycosides Amikacin 1.25 Centamicin 1.25 Kanamycin 100 Neomycin 12.5 Streptomycin * 50 β-lactam meter Phenylcillin-G * 0.156 Methicillin 160 Oxacillin 7.5 Ampicillin 20 Gram-positive spectrum Bassitracin * 0.312 Rifampicin 30 Novobiocin 0.312 Lincomycin 20 Gram-Voice Spectrum Polymycin B * 1200 Broad spectrum Chloramphenicol 10 Vancomycin * 1600

 *: units / ml

As can be seen in Table 4, L. zeae RMK354 was more resistant to polymycin B and vancomycin than other antibiotics, while it was highly susceptible to penicillin-G, bacitracin and novobiocin.

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 Enzyme Activities of L. zeae RMK354

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-glucronidase
α-glucosidase
β-glucosidase
N-acetyl-β-glucosaminenidase
α-mannosidase
β-fucosidase 0
3
2
0
3
0
0
0
0
0
One
0
One
0
0
0
0
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. zeae RMK354 showed high enzyme activity in esterase and leucine arylamidase. 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 of 0.3 was 7.3 hours without bile and 8 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. Comparison of L. zeae RMK354 resistance after 3 hours in hydrochloric acid solution

                                          Unit: CFU / mL (Log number)

Incubation time (hr) pH 2 pH 3 pH 4 pH 6.4 0 7.79 7.82 7.83 7.83 One 7.70 7.72 7.79 7.86 2 7.64 7.71 7.78 7.88 3 7.51 7.67 7.74 7.89

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. zeae in MRS medium RMK354 Suppression of Food Poisoning Bacteria (*)

Food poisoning bacteria growth % Inhibition Food poisoning bacteria ^a L. zeae RMK354 ^a CFU / mL pH CFU / mL pH Esherichia coli 9.2 × 10 ⁶ 6.56 2.3 × 10 ⁸ 5.64 - Salmonella
typhimurium 1.45 × 10 ⁷ 6.55 5.8 × 10 ⁶ 5.67 60.0 Saphyloccous aureus 3.2 × 10 ⁴ 6.55 7.7 × 10 ⁸ 5.76 -

* L. zeae Initial number of RMK354: 3.8 × 10 ⁶ CFU / mL

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

As can be seen in Table 7, L. zeae RMK354 is Escherichia but for coli) and Stephen Pyrococcus fluorescein-house (Staphylococcus aureus) was a deterrent, Salmonella Thai Phi hair Stadium (Salmonella typhimurium ) showed a suppression of 60.0%.

Experimental Example 10 Setting Optimal Culture Conditions for ACE Inhibition Activity

end. Effect of Reduced Skim Milk and Inoculation on ACE Inhibitory Activity

After culturing for 15 hours at 37 ℃ for each reduced skim milk content and inoculum inoculation, the results of measuring the ACE inhibitory activity is shown in Table 8. When the amount of reduced skim milk was 10%, the addition amount of the bacterium showed the highest ACE inhibitory activity when 0.1% was added.

Table 8. Effect of reduced skim milk content and bacterial inoculum on ACE inhibitory activity

Reduced skim milk
density(%) % Inhibition of ACE according to inoculation amount * 0.1 parts by weight 0.5 parts by weight 1 part by weight 8 62.0 ± 5.7 76.4 ± 2.1 80.9 ± 7.2 10 89.2 ± 3.3 88.9 ± 6.7 88.5 ± 2.3 12 88.9 ± 8.3 75.5 ± 3.1 80.9 ± 7.2

* The amount of inoculation is strain ( L. zeae) for 100 parts by weight of reduced skimmed milk according to each reduced skimmed milk concentration. RMK354).

I. Effect of Incubation Temperature and Incubation Time on ACE Inhibitory Activity

The strain ( L. zeae) was reduced to 10% of the reduced skim milk concentration. RMK354) 0.1% of the inoculation amount was added, and after culturing at 24 hours for 3 hours at each culture temperature, the results of measuring the ACE inhibition rate are shown in Table 9 below. As shown in Table 9, the ACE inhibitory activity was optimum culture condition as 89.2% at 37 ° C for 15 hours and 89.4% at 40 ° C for 15 hours.

 Table 9. Effect of incubation temperature and incubation time on ACE inhibitory activity

Incubation time (hr) ACE inhibition rate according to culture temperature (%) 34 ℃ 37 ℃ 40 ℃ 0 33.9 55.1 51.4 3 34.8 69.1 52.7 6 36.9 73.2 53.7 9 45.6 76.4 73.2 12 54.8 88.2 77.5 15 56.6 89.2 89.4 18 48.0 83.5 53.0 24 39.0 73.8 38.0

Example 1

96.15 parts by weight of crude oil and 3.85 parts by weight of skim milk powder were added and blended at 65 ° C. to completely dissolve and homogenized at a pressure of 215 ± 35 kgf / cm 2. Sterilized at 90 ° C. for 5 minutes, cooled to 40 ° C., and then inoculated with 1.0 parts by weight of Lactobacillus zeae RMK354 (KACC 91314P) bulk inoculating bacteria to 100 parts by weight of the cooled homogenate to reduce the final pH to 4.4. The culture was cooled to 17 ± 2.5 ° C. while slowly stirring with a stirrer to obtain a fermentation broth including Lactobacillus zeae RMK354 strain (KACC 91314P).

To 84% by weight of the fermentation broth obtained above, 5.0% by weight of oligosaccharide, 2.0% by weight of liquid fructose, 4.0% by weight of strawberry ,, 5.0% by weight of purified water were added and mixed, followed by stirring to prepare Lactobacillus zeae RMK354 strain (KACC 91314P). Fermented milk containing was obtained.

Table 10 shows the components of the fermented milk.

The Lactobacillus zeae RMK354 (KACC 91314P) bulk inoculum was inoculated with Lactobacillus zeae RMK354 strain (KACC 91314P), which was isolated from crude oil, at 1% (v / v) in reducing skim milk. After incubation at 39.5 ± 2.5 ° C. for 18 ± 6 hours, the cultured spawn of the Lactobacillus zeae RMK354 strain (KACC 91314P) was incubated at 85 ± 5 ° C. for 45 ± 15 minutes for reduced skim milk. Was inoculated at 1% (v / v) and incubated at 39.5 ± 2.5 ° C. for 18 ± 6 hours.

[Example 2]

Fermented milk containing Lactobacillus zeae RMK354 strain (KACC 91314P) in the same manner as in Example 1, except that 6.0% by weight instead of 4.0% by weight of strawberry and 3.0% by weight of 5.0% by weight of purified water were used. Was prepared.

Example 3

Fermented milk containing Lactobacillus zeae RMK354 strain (KACC 91314P) in the same manner as in Example 1 except for using 8.0% by weight instead of 4.0% by weight of strawberry and 1% by weight instead of 5.0% by weight of purified water. Was prepared.

Example 4

Fermented milk containing Lactobacillus zeae RMK354 strain (KACC 91314P) was prepared in the same manner as in Example 1, except that 4.0 wt% of blueberries were used instead of 4.0 wt% of strawberries.

Example 5

Lactobacillus zeae RMK354 strain (KACC 91314P) in the same manner as in Example 1, except that 6.0 wt% of strawberries and 3.0 wt% instead of 5.0 wt% of purified water were used instead of 4.0 wt% of strawberries. To prepare a fermented milk comprising a.

 Example 6

Lactobacillus zeae RMK354 strain (KACC 91314P) in the same manner as in Example 1, except that 8.0% by weight of strawberries and 1.0% by weight of 5.0% by weight of purified water were used instead of 4.0% by weight of strawberries. To prepare a fermented milk comprising a.

Table 10. 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 crude oil 80.77 80.77 80.77 80.77 80.77 80.77 Skim milk powder 3.23 3.23 3.23 3.23 3.23 3.23 oligosaccharide 5.0 5.0 5.0 5.0 5.0 5.0 Liquid fructose 2.0 2.0 2.0 2.0 2.0 2.0 Strawberry 쨈 * 4.0 6.0 8.0 - - - Blueberries 쨈 ** - - - 4.0 6.0 8.0 Purified water 5.0 3.0 1.0 5.0 3.0 1.0

* Strawberry 당 with sugar content of 61.2Bx, strawberry content 45%.

** Blueberry 62 with sugar 62Brix, blueberry content 40%.

Test Example 1 Sensory Test

Sensory tests such as color, taste, texture and overall acceptability of the fermented milk and the control fermented milk prepared by the method of Examples 1 to 6 were carried out using 13 persons who worked for 2 years or more in the field of fermented milk. Performed by the point scoring method, the results are summarized in Table 11 below.

Table 11. Sensory tests of fermented milk and control fermented milk of Examples 1 to 6

Item Sensory evaluation Color flavor Organization Comprehensive likelihood Example 1 7.07 ± 1.60 6.46 ± 1.80 6.61 ± 1.26 6.61 ± 1.50 Example 2 7.00 ± 1.29 7.15 ± 1.14 6.69 ± 1.25 7.23 ± 1.01 Example 3 6.84 ± 1.57 7.53 ± 1.26 7.07 ± 0.86 7.30 ± 0.94 Example 4 6.46 ± 1.33 6.53 ± 1.45 6.61 ± 1.80 6.84 ± 1.34 Example 5 7.15 ± 1.34 6.69 ± 1.60 6.92 ± 1.32 6.84 ± 1.40 Example 6 7.53 ± 1.50 7.07 ± 1.55 7.15 ± 1.28 7.53 ± 1.12 Control ** 6.15 ± 1.51 6.38 ± 0.96 6.57 ± 1.80 6.46 ± 1.26

* 1: Not very good 5: Fair 9: Very good

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

As shown in Table 11, the fermented milk containing Lactobacillus Jia RMK354 strain of the present invention was added to the fermented milk and fruit 당시 could produce a fermented milk with excellent palatability.

<Test Example 2>: Animal test result

In order to investigate the antihypertensive effect of fermented milk using ACE inhibitory strains, the fermented milk input group (A) prepared in Example 6 containing the Lactobacilus zeae RMK354 strain in 30 male SHR rats was marketed and commercially available. Each group (B) with fermented milk (Project 102-80, Namyang Dairy Products) was set up, and the oral administration method was selected. week) and the results are shown in Table 12-1 and Table 12-2 below.

As a control group, commercial fermented milk (Pureore Co., Ltd.) of Seoul Milk Company was placed.

Sample doses were determined on a daily basis with two doses of 150 ml of commercial fermented milk / 60 kg of adult body weight.

Table 12-1. Changes in hypertension of SHR rats (Unit: mmHg)

Item
Week 0 One 2 3 control ¹ 172.92 ±
20.53 ^a 171.56 ±
14.41 ^a 168.45 ±
10.86 ^a 68.86 ±
11.60 ^a A ²⁾ 176.20 ±
11.81 ^a 181.59 ±
7.83 ^a 174.52 ±
4.68 ^a 170.40 ±
3.40 ^a B ³⁾ 177.88 ±
21.35 ^a 172.50 ±
22.24 ^a 164.14 ±
11.44 ^a 160.86 ±
13.02 ^a

Mean ± Std (n = 10)

¹⁾ Feed commercial fermented milk (Seoul Milk Company's Puree product) as a control

²⁾ Lactobacillus Feed fermented milk fermented with zeae RMK354.

³⁾ Feed 120-80 fermented milk from Namyang Dairy.

^The same letter in the same criteria means a statistically insignificant level at the 5% level of multiple comparisons.

Table 12-2. Changes in hypertension of SHR rats (Unit: mmHg)

Item
Week 4 5 6 7 control ¹ 168.86 ±
11.60 ^a 167.86 ±
7.31 ^a 172.57 ±
4.58 ^a 170.43 ±
5.16 ^a A ²⁾ 167.60 ±
5.31 ^a 161.00 ±
8.62 ^a 160.40 ±
4.92 ^b 160.40 ±
5.96 ^b B ³⁾ 159.14 ±
7.52 ^a 158.57 ±
4.23 ^a 158.57 ±
5.86 ^b 159.43 ±
7.57 ^b

Mean ± Std (n = 10)

¹⁾ Feed commercial fermented milk (Seoul Milk Company's Puree product) as a control

²⁾ Feed fermented milk fermented with Lactobacillus zeae RMK354.

³⁾ Feed 120-80 fermented milk from Namyang Dairy.

^The same letter in the same criteria means that there is no statistical significance at the 5% level of the SNK multiple comparison.

Compared with the commercial fermented milk as a control group, the A and B product groups showed significance after 6 weeks, and the fermented milk containing the Lactobacilus zeae RMK354 strain of the present invention was found to have an effect of lowering blood pressure.

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 strain RMK354 strain (KACC 91314P). In the production of fermented milk, A method for producing fermented milk, comprising the step of inoculating Lactobacillus RMK354 strain (KACC 91314P) in crude milk.