KR20140141785A - Manufacture of stater to fermented fermented milk with lactic of acid bacterial separted traditional fermented foods and development of fermented milk in using this processing - Google Patents

Abstract

The present invention relates to manufacture of a starter for fermented milk by separating lactobacillus from traditional fermented foods and to manufacture of the fermented milk using the same. According to the present invention, the starter for the fermented milk derived from traditional fermented foods is manufactured by the steps of: mixing pulverized kimchi with sterilized physiological saline and separating a kimchi solution; diluting and smearing the kimchi solution, culturing the same, and separating lactobacillus; inoculating the separated lactobacillus into a culture medium and culturing the same; and inoculating the cultured lactobacillus to skim milk and culturing the same.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a starter for fermentation derived from a traditional fermented food, a method for producing the fermented starter, and a method for producing fermented milk using the fermented starter,

The present invention relates to a fermented starter prepared by isolating lactic acid bacteria from a conventional fermented food, and using the fermented milk to produce fermented milk.

 Among microorganisms, lactic acid is produced as a metabolite while growing, and these bacteria are collectively called latic acid bacteria. Lactic acid bacteria have been used unconsciously from ancient civilizations prior to the development of science. They have been used extensively in human life, ranging from fermented dairy products with long history to diverse fermented foods, soy sauce, kimchi, fermented sausages, Has been utilized. Lactobacillus was first known by Pasteur in the process of studying the cause of wine rumble in 1858. There are 12 main lactic acid bacteria and there are 12 lactobacilli, Carnobaterium, Atopobium, Lactococcus, Pediococcus, Tetragenococcus, Leuconostoc, Weisella, Oenococcus, Enterococcus, Streptococcus , Is divided into Vagococcus. Among them, lactic acid bacteria commonly used in fermented milk products are Streptococcus spp. Such as Lactobacillus spp., Lactococcus spp., Streptococcus spp., Leuconostoc spp., And Pediococcus spp. Worldwide producers of fermentation strains in dairy countries include Chr. Hansen, Danisco (formerly Wisby), Marshall, Sodima and Sanofi. Major Korean dairy companies such as Korea Yakult, Pasteur, Namyang Dairy, Maeil Dairy, Seoul Milk, Binggrae, Haitai, It depends on imported seeds.

As a Korean company, Yakult has started cultivating bifidobacteria since 1986 in order to improve the supply and demand structure of domestic bacterium. As a technology that succeeded in cultivating and commercializing bifidobacteria isolated from pure Koreans, Patent No. 10-0142615 Discloses a novel lactic acid bacterium having excellent resistance to acid and cholesterol decomposition which is isolated from the feces of a Korean infant and its use method. However, compared with the market size of the lactic acid fermented milk market in Korea, the level of lactic acid bacteria improvement technology is far behind that of advanced countries, As consumers' preferences for fermented milk products have diversified, the development of new strains with excellent taste and flavor has become important.

Domestic fermented foods use marine biomass and agricultural and livestock biomass as their geographical characteristics. Major fermented foods include vinegar, soy sauce, soybean paste, chonggukjang, kochujang, kimchi, The fermented microorganisms are mostly lactic acid bacteria. The characteristics of fermented foods in Korea are not natural fermentation by starter but natural friendly products in which the yeast germs ultimately influence the flavor, taste and texture of the product in natural symbiotic relationship. The characteristics of these lactic acid bacteria are as follows: And the difference in growth is different. Currently, Chr. Global dairy companies such as Hansen and Dainosco have developed improved lactic acid bacteria by transforming lactic acid bacteria in order to increase the flavor and taste of the final product. Therefore, when lactic acid bacteria derived from domestic fermented foods are used, the characteristics of the fermented milk products can be used as foods and produce highly competitive fermented milk products.

DISCLOSURE Technical Problem The present invention has been conceived to solve the problems as described above, and it is an object of the present invention to prepare a fermentable starter by isolating lactic acid bacteria from kimchi, which is a representative natural fermented food in Korea.

The fermented starter derived from traditional fermented food according to the present invention is characterized in that lactic acid bacteria are isolated and cultured in a conventional fermented food.

In addition, the traditional fermented food is characterized by being a kimchi.

The lactic acid bacteria isolated from kimchi, which is a traditional fermented food, are Lactobacillus plantarum sp., Leuconostoc lctics or Latobacilus acidophilus .

The conventional fermented starter for fermented food derived from traditional fermented food according to the present invention is prepared by mixing the crushed kimchi with sterilized physiological saline and then separating the kimchi liquid, Culturing the isolated lactic acid bacterium inoculated with the separated lactic acid bacteria, and culturing the cultured lactic acid bacteria inoculated with the skimmed milk.

The fermented milk prepared by using the conventional fermented food-derived fermented starter obtained by isolating and culturing the lactic acid bacteria in the conventional fermented food according to the present invention comprises the steps of mixing 5.5 parts by weight of sugar with 100 parts by weight of crude oil to prepare a mixture, Sterilizing the sterilized mixture, fermenting the cooled mixture by adding 2 parts by weight of a fermented starter derived from kimchi to 100 parts by weight of crude oil, and cooling the fermented mixture .

The present invention has the effect of producing a fermentable starter by isolating lactic acid bacteria from typical traditional fermented foods in Korea.

1 is a graph showing the results of culturing characteristics according to a selective medium of lactic acid bacteria isolated from a conventional fermented food according to an embodiment of the present invention
FIG. 2 is a graph showing the activity of Lactobacillus plantarum subsp , Leuconostoc lctics And Latobacilus The appearance of acidophilus
FIG. 3 is a graph showing the activity of Lactobacillus plantarum subsp , Leuconostoc lctics And Latobacilus acidophilus
FIG. 4 is a graph showing the results of a conventional fermented food-derived fermentation starter ( Lactobacillus Production process of fermented milk using acidophilus C)
FIG. 5 is a graph showing the results of a conventional fermented food-derived fermentation starter ( Lactobacillus pH of fermented milk using acidophilus C
FIG. 6 is a graph showing the results of a conventional fermented food-derived starter ( Lactobacillus acidophilus C) was used to measure the titratable acidity of fermented milk.
FIG. 7 is a graph showing the results of a conventional fermented food-derived fermentation starter ( Lactobacillus acidophilus C) was used to measure the number of lactic acid bacteria in fermented milk.
FIG. 8 is a graph showing the results of a conventional fermented food-derived fermentation starter ( Leuconostoc Production process of fermented milk using lactics B)
FIG. 9 is a graph showing the results of a conventional fermented food-derived fermentation starter ( Leuconostoc lactic acid B) was used for pH measurement of fermented milk.
FIG. 10 is a graph showing the results of a conventional fermented food-derived fermentation starter ( Leuconostoc lactic acid B) was used to measure the titratable acidity of fermented milk.
Fig. 11 is a graph showing the results of a conventional fermented food-derived fermentation starter ( Leuconostoc number of lactic acid bacteria fermented milk measurement result graph using lactics B)
FIG. 12 is a graph showing the results of a conventional fermented food-derived fermentation starter ( Lactobacillus Process for the production of fermented milk using plantarum A)
FIG. 13 is a graph showing the results of a conventional fermented food-derived fermentation starter ( Lactobacillus Plantarum A) , a graph of the pH measurement of fermented milk
FIG. 14 is a graph showing the results of a conventional fermented food-derived fermentation starter ( Lactobacillus Plantarum A) was used to measure the titratable acidity of fermented milk.
FIG. 15 is a graph showing the results of a conventional fermented food-derived fermentation starter ( Lactobacillus Plantarum A) , the result of measurement of the number of lactic acid bacteria in fermented milk
FIG. 16 is a graph showing the results of a conventional fermented food-derived fermentation starter ( Lactobacillus acidophilus C, Leuconostoc lactics B and Production process of fermented milk using Lactobacillus plantarum A)
17 is a graph showing the results of a conventional fermented food-derived fermentation starter ( Lactobacillus acidophilus C, Leuconostoc lactics B and PH graph of fermented milk using Lactobacillus plantarum A)
FIG. 18 is a graph showing the results of a conventional fermented food-derived fermentation starter ( Lactobacillus acidophilus C, Leuconostoc lactics B and Lactobacillus plantarum A) was used to measure the titratable acidity of fermented milk.
19 is a graph showing the results of a conventional fermented food-derived fermentation starter ( Lactobacillus acidophilus C, Leuconostoc lactics B and Result of Lactobacillus plantarum A)

The present invention relates to a method for producing fermented starter and a fermented milk using the fermented starter. The present invention relates to a method for producing fermented milk by preparing a fermented starter by separating lactic acid bacteria from a conventional fermented food.

Examples.

Screening of lactic acid bacteria from fermented foods

In order to isolate the functional starter from the fermented food, the fermented food sold in the optimum pH range of fermented milk, 4.0 ~ 4.5, was used as the source of the functional starter. Selected products were Kimchi, .

To separate lactic acid bacteria from selected fermented foods, 25 g of selected samples and 225 ml of sterilized physiological saline were placed in a sterile filter bag and mixed using a stomacher. 1 ml of the separated sample was diluted by deciduous dilution method and 100 μl was taken and plated on MRS (Lactobacilli MRS agar, Difco, USA) plate culture medium and cultured at 37 ° C for 24 hours. A single colony representing a transparent ring for each sample was firstly separated. The primary isolated colonies were randomly separated on a BCP agar medium and cultured at 37 ° C for 24 hours. The colonies showing yellow colonies were separated and the lactic acid bacteria were isolated.

In order to isolate the starter lactic acid bacteria from the selected fermented foods, the pH value of the selective medium was adjusted to 2.5 to separate the lactic acid bacteria from each sample in order to isolate the acidic and brittle lactic acid bacteria which can live from the beginning of experiment to the large intestine. In Kimchi, 5 strains, 1 fermented fish, and 6 strains of lactic acid bacteria were isolated. The identification of isolated strains was commissioned by Macrogen Co., Ltd. The results were as shown in Table 1 below.

Strain
16S RNA sequence Separation source Homology (%) Sientific name One 99 Latobacillus curvatus Kimchi 2 99 Weissella viridescens Kimchi 3 99 Lactobacilus plantarumsubsp Kimchi 4 99 Leuconostoc lctics Kimchi 5 99 Latobacilus acidophilus Kimchi 6 99 Lactobacillus sakei subsp Shrimp 7 99 Lactobacillus sakeisubsp Kochujang 8 99 Pediococcus pentosaceus Kochujang 9 99 Pediococcus sp Cheonggukjang 10 99 Enterococcus faecium Cheonggukjang 11 99 Pediococcus acidiactici Cheonggukjang 12 99 Pediococcus acidiactici Cheonggukjang

Experimental example.

1-1. Application of lactic acid fermented milk

1-1-1. Acid acidity and biliary cholestasis of isolated lactic acid bacteria

MRS liquid medium adjusted to pH 2.5 with 1N HCl was used. The lactic acid bacteria isolated from kimchi, shrimp, kochujang, and chungkukjang shown in Table 1 were cultured in a 37 ° C MRS liquid medium for 24 hours, respectively, and then placed in a 15 ml tube. After centrifugation at 4000 rpm for 15 minutes, the supernatant was discarded and the bacteria were recovered. 1% of the recovered bacteria was inoculated in 10 ml of the MRS liquid medium having adjusted pH, and the cells were incubated at 37 캜 for 2 hours, and the number of viable cells was measured.

In the bile acid test, the concentration of Bacto oxgall (Difco) was added to the MRS liquid medium so as to be 0.3%, and the number of viable cells was measured after culturing at 37 ° C for 4 hours as described above.

1-1-2. Cultivation characteristics according to selective medium

In order to investigate the possibility of lactic acid bacteria isolated from kimchi, shrimp, kochujang, and chungkukjang as shown in Table 1 as a fermentable starter lactic acid bacterium, the growth characteristics of the isolated lactic acid bacteria were examined by differentiating the characteristics of the separated medium. After incubation for 24 hours with casein and lactose adjusted to 5% and 4%, respectively, in the MRS liquid medium, the absorbance was measured.

1-1-3. Antimicrobial activity

The antimicrobial effect was confirmed by disc method. The pathogenic microorganisms (E. coli, Salmonella sp., Listeria monocytogenes) cultured at 37 ° C for 24 hours were plated on a Nutrient agar with a sterilized cotton swab. A paper disc with a diameter of 10 mm was placed on a sterilized plate medium and incubated at 37 ° C for 24 hours at 37 ° C for 24 hours. The cells were incubated at 37 ° C for 24 h. The antimicrobial effect was confirmed by the presence or absence of production.

1-1-4. Experiment result

(1) Acid resistance

Lactic acid bacteria have an increased mortality rate in the culture medium of acidic environment in which lactic acid is produced, and when the human is consumed, the content of hydrochloric acid (HCl) is 0.4 ~ 0.5% (about 0.1N HCl) To about 0.6% of gastric juice. The pH of the gastric juice changes with time, rising from pH 1.8 to pH5 after fasting. Therefore, in order to reach the intestine after ingesting the lactic acid bacteria, the bacterium and acid resistance of the selected three kinds of lactic acid bacteria were analyzed.

The results of the acid tolerance of the selected lactic acid bacteria are shown in Table 2, and the results are shown in Table 3: Lactobacilus plantarumsubsp, Leuconostoc 4 lctics And No. 5 Latobacilus The survival rates of acidophilus were 94%, 99% and 96%, respectively, and the acidity of other isolated lactic acid bacteria was less than 90%.

division One 2 3 4 5 6 7 8 9 10 11 12 Before culture (Log cfu / ml) 7.40 4.72 8.24 6.85 8.45 7.64 8.84 7.05 7.64 7.75 7.70 8.77 After incubation (Log cfu / ml) 3.70 3.51 7.74 6.76 8.10 6.66 5.66 5.82 6.69 7.12 7.35 7.75 Survival rate (%) 50 74 94 99 96 87 64 83 88 92 95 88

1. Latobacillus curvatus, 2. Weissella viridescens, 3. Lactobacilus plantarumusubsp, 4. Leuconostoc lctics, 5. Latobacilus acidophilus, 6. Lactobacillus sakei subsp , 7. Lactobacillus sakeisubsp (hot pepper paste) , 8. Pediococcus pentosaceus , 9. Pediococcus sp , 10. Enterococcus faecium , 11. Pediococcus acidiactici , 12. Pediococcus acidiactici

(2) My bile

As shown in Table 3, the bacteriostasis of the selected lactic acid bacteria showed a survival rate of 90% or more in all the isolated lactic acid bacteria, and the bile bacterium was high. When compared with the lactic acid bacteria having high acid resistance, the Lactobacillus plantarum subsp , # 4 Leuconostoc lctics And No. 5 Latobacilus The acidity of acidophilus was 98%.

division One 2 3 4 5 6 7 8 9 10 11 12 Before culture (Log cfu / ml) 7.40 4.72 8.24 6.85 8.45 7.64 8.84 7.05 7.64 7.75 7.70 8.77 After incubation (Log cfu / ml) 7.24 4.08 8.09 6.73 8.29 7.11 7.95 6.41 7.36 7.09 7.44 8.46 Survival rate (%) 98 86 98 98 98 93 90 91 96 92 97 96

One. Latobacillus curvatus , 2. Weissella viridescens , 3. Lactobacilus plantarumusBSP, 4. Leuconostoc lctics , 5. Latobacilus acidophilus , 6. Lactobacillus sakei subsp , 7. Lactobacillus sakeisubsp (hot pepper paste) , 8. Pediococcus pentosaceus , 9. Pediococcus sp , 10. Enterococcus faecium , 11. Pediococcus acidiactici , 12. Pediococcus acidiactici

(3) Culture characteristics according to the selective medium

As shown in the result graph shown in Fig. 1, the growth characteristics of lactic acid bacteria upon addition of casein and lactose were shown in Lactobacilus plantarum subsp , Leuconostoc lctics And Latobacilus acidophilus was selected as a starter for functional starter, and the applicability of this product as a starter for fermented milk products was analyzed.

(4) Antimicrobial activity

Lactic acid bacteria is an indigenous dominant species present in human and animal fields. It has low pH in the digestive tract, resistance to bile acid and lysozyme, and ability to adsorb to intestinal epithelium, and thus has excellent viability and fixability in digestive tracts In particular, it has been reported that the growth activity at the terminal end of the ileum is high, and that the main activity is stabilization of intestinal microflora, lowering of cholesterol and inhibition of harmful bacteria growth.

FIG. 2 is a graph showing the results of experiments on the acid resistance, biliary properties and growth ability according to the selective medium of the lactic acid bacteria shown in Table 1, and Lactobacilus plantarum subsp , Leuconostoc lctics And Latobacilus acidophilus , and it is expected that the resistance to intestinal microflora and the cholesterol lowering effect, which have been reported in the literature, are high. Especially, accumulation of residual toxins accumulated in the body through stabilization of intestinal microflora And antioxidant activity against pathogenic harmful bacteria in order to measure the efficacy of isolated lactic acid bacteria. As a result, Listeria monocytogenes showed high antimicrobial activity (Figure 3).

2. Application characteristics of isolated lactic acid bacteria to fermented milk

The most important process in the production of fermented milk products is the fermentation of lactic acid by starter, the ingredients of fermentation of lactic acid fermentation, heat treatment, starter bacterium ( Lactobacillus bulgaricus ) and Streptococcus It is important to keep a constant balance between the ratio of the bacteria and the bacillus, and generally use a ratio of 1: 1 or 1: 2 or 1: 3, depending on the ratio of the thermophilus , Reported. When L. bulgaricus and S. thermophilus were cultured together, symbiosis between the two lactic acid bacteria promoted the growth of S. thermophilus . The initial growth rate of S. thermophilus was low (about 45 ° SH) L. bulgaricus, which has a low initial growth rate, produces a large amount of lactic acid (70 ° ~ 80 ° SH) and acid resistance (1.7% ~ 2.7% lactic acid concentration) (over-acidification) and flavor (aroma). L. bulgaricus predominates over 45 ℃ depending on the culture temperature and S. thermophilus predominates over 45 ℃. L. bulgaricus is dominant in yogurt cultured for a long time and S. thermophilus predominates in yogurt cultivated for a short time. Therefore, the fermented milk application characteristics of each bacterium were measured by culturing the selected lactic acid bacteria or by mixing them with a commercial starter.

2-1. Lactobacillus isolated from kimchi, a traditional fermented food acidophilus Possibility of using starter of fermented milk of C and application of fermented milk

In this experiment, we aimed to develop fermented milk products using lactic acid bacteria isolated from kimchi, which is a traditional fermented food, as a starter of fermented milk, which is a fermented milk product. Thus, the quality characteristics of the fermented milk products prepared using the isolates were tested.

2-1-1. Identification of Lactic Acid Bacteria

Seven species were collected from Jeonbuk region and 25g of each sample and 225ml of sterilized physiological saline were put into a sterile filter bag and mixed with a stomacher. 1 ml of the separated sample was diluted by deciduous dilution method and 100 μl was taken and plated on MRS (Lactobacilli MRS agar, Difco, USA) plate culture medium and cultured at 37 ° C for 24 hours. A single colony representing a transparent ring for each sample was firstly separated. The primary isolated colonies were randomly separated on a BCP agar medium and cultured for 24 h at 37 ° C after inoculation with a flat plate method to isolate colonies showing yellow colonies. Lactobacillus acidophilus Similar or identical Lactobacillus acidophilus C Respectively.

2-1-2. Starter manufacturing

Starters for the production of fermented milk include Lactobacillus acidophilus C was inoculated into 10% (w / v) reduced skim milk (Seoul Milk) and cultured at 10 ⁸ CFU / mL or higher. The control group was cultured in the same manner as the commercial strain ABT-5 (Chr. Hansen, Denmark).

2-1-3. Manufacture of fermented milk

5.5 parts by weight per 100 parts by weight of crude oil was mixed and sterilized and then 2 parts by weight of a starter was inoculated with 100 parts by weight of crude oil and cultured at 40 DEG C for 4 hours. (FIG. 4) In the control group, 2 parts by weight of ABT- , Test group 1 was Lactobacillus 2 < / RTI > parts by weight of a single strain of acidophilus C , test group 2 consisted of ABT-5 and Lactobacillus acidophilus C were mixed in a ratio of 1: 1 to prepare a fermented milk.

division Starter for fermented milk production Control group ABT-5 Test group 1 Lactobacillus acidophilus C 2 parts by weight (single strain) Test group 2 ABT-5 and Lactobacillus acidophilus C 2: 1 part by weight (mixed strain)

2-1-4. pH measurement

The fermented milk was measured using a pH meter. During the fermentation, the pH was measured at time intervals of 0, 2, 4 and 6 hours.

2-1-5. Titratable acidity measurement

The titratable acidity is determined by mixing 10 ml of sample with 10 ml of distilled water to make a suspension. Titrate the suspension to pH 8.3 by adding NaOH and titrate the titration of NaOH to titrate the amount of NaOH The conversion factor, 0.9, was multiplied and the weight (g) of the sample was divided to determine the titratable acidity (%). The titratable acidity during fermentation was measured at 0, 2, 4, and 6 hour intervals.

2-1-6. Measure the number of lactic acid bacteria

To measure the changes in the lactic acid bacteria over time during the fermentation of the fermented milk, 1 ml of the sample was sampled at 0, 2, 4 and 6 hours according to Richardson's method, diluted with sterilized physiological saline solution by decidual dilution method, After culturing at 37 ° C for 48 hours, yellow colonies were counted.

2-1-7. Sensory properties of fermented milk

Sensory evaluation of the fermented milk was performed by using the 5 point scaling method.

2-1-8. Research result

(1) pH measurement

As a result of measuring the pH of FIG. 5, the control group did not reach the fermentation end point of 4.7 or lower at the fermentation time of 4 hours, but the fermented milk test group 1 produced by using a single strain reached the fermentation end point at 4.65 at the fermentation time of 4 hours. The fermented milk test group 2, which was prepared by using the combined strain, showed a fermentation end point of 4.75 at the fermentation time of 4 hours, indicating that the activity of the isolate was higher than that of the commercial strain, and thus it could be used as a fermented milk starter do.

(2) Measurement of acidity

As shown in the graph of the acidity measurement result of FIG. 6, the acidity was increased with time in all the test groups including the control group, and a rapid increase was observed after 2 hours of fermentation time. The pH of the fermented milk was higher than that of the control.

(3) Change in the number of lactic acid bacteria

As shown in the graph of the change in the number of lactic acid bacteria in Fig. 7, the number of lactic acid bacteria in the fermented milk was higher in the pre-fermentation time region than in the control group. In the control group, 10 ⁷ CFU / mL after 4 hours and 10 ⁹ CFU / mL or higher in test group 1 were more likely to be used as a starter. Also it is possible the utilization of a composite strain showed that it does not inhibit each other to multiply the production appeared commercially starter and a fermented milk test group 2 at 10 ⁹ CFU / mL or more produced using the composite strain by mixing the strains in combination with commercial strains see.

(5) Sensory properties

The color, flavor, taste and overall acceptability of the prepared fermented milk were evaluated by sensory evaluation using the 5-point scale method, and the results are shown in Table 5.

Test group 2, which was fermented milk prepared by mixing 1: 1 with isolated strains and commercial starters, showed the most sensory results.

division color incense flavor Overall likelihood Control group 4.7 ± 0.5 4.0 ± 0.8 4.1 ± 0.7 4.5 ± 0.7 Test group 1 4.7 ± 0.5 4.2 ± 0.8 4.0 ± 0.7 4.7 ± 0.5 Test group 2 4.8 ± 0.4 4.1 ± 0.7 4.2 ± 0.9 4.8 ± 0.5

Therefore, according to the results of this experiment, microbiological, physicochemical and sensory evaluation of fermented milk prepared from Lactobacillus acidophilus C isolated from traditional fermented food showed superior ability of fermented milk as a starter in comparison with a control strain The possibility of use as a fermented milk starter was confirmed.

2-2. Leuconostoc isolated from traditional fermented food Kimchi lactics Possibility of using starter of fermented milk of B and experiment of application of fermented milk

In this experiment, we aimed to develop fermented milk products using lactic acid bacteria isolated from kimchi, which is a traditional fermented food, as a starter of fermented milk, which is a fermented milk product. The quality characteristics of fermented milk products prepared by using the isolates were studied. The results can be summarized as follows.

2-2-1. Identification of Lactic Acid Bacteria

Seven species were collected from Jeonbuk region and 25g of each sample and 225ml of sterilized physiological saline were put into a sterile filter bag and mixed with a stomacher. 1 ml of the separated sample was diluted by deciduous dilution method and 100 μl was taken and plated on MRS (Lactobacilli MRS agar, Difco, USA) plate culture medium and cultured at 37 ° C for 24 hours. A single colony representing a transparent ring for each sample was firstly separated. The first isolates were randomly divided into two batches of BCP agar medium and cultured at 37 ° C. for 24 hours at 37 ° C. The colonies with yellow colonies were separated and identified. Leuconostoc lactics were found to be similar or identical Leuconostoc lactics B , respectively.

2-2-2. Starter manufacturing

The starter for fermented milk production was prepared by inoculating Kimchi isolate with 10% (w / v) reduced skim milk (Seoul Milk), culturing it at 10 ⁸ CFU / mL or higher, The control group was cultured in the same manner as the commercial strain ABT-5 (Chr. Hansen, Denmark).

2-2-3. Manufacture of fermented milk

5.5 parts by weight per 100 parts by weight of crude oil was mixed and sterilized and then 2 parts by weight of a starter was inoculated with 100 parts by weight of crude oil and cultured at 40 DEG C for 4 hours (FIG. 8). As a control, 2 parts by weight of ABT- , Test group 1 was Leuconostoc 2 parts by weight of a lactic acid B single strain, Test group 2 was prepared by mixing ABT-5 and Leuconostoc lactics B were mixed in a ratio of 1: 1 to prepare a fermented milk.

division Starter for fermented milk production Control group ABT-5 Test group 1 Leuconostoc lactics B 2 parts by weight (single strain) Test group 2 ABT-5 and Leuconostoc lactics B 2: 1 part by weight (mixed strain)

2-2-4. pH measurement

The fermented milk was measured using a pH meter. During the fermentation, the pH was measured at time intervals of 0, 2, 4 and 6 hours.

2-2-5. Titratable acidity measurement

The titratable acidity is determined by mixing 10 ml of sample with 10 ml of distilled water to make a suspension. Titrate the suspension to pH 8.3 by adding NaOH and titrate the titration of NaOH to titrate the amount of NaOH The conversion factor, 0.9, was multiplied and the weight (g) of the sample was divided to determine the titratable acidity (%). The titratable acidity during fermentation was measured at 0, 2, 4, and 6 hour intervals.

2-2-6. Measure the number of lactic acid bacteria

To measure the changes in the lactic acid bacteria over time during the fermentation of the fermented milk, 1 ml of the sample was sampled at 0, 2, 4 and 6 hours according to Richardson's method, diluted with sterilized physiological saline solution by decidual dilution method, After culturing at 37 ° C for 48 hours, yellow colonies were counted.

2-2-7. Sensory properties of fermented milk

Sensory evaluation of the fermented milk was performed by using the 5 point scaling method.

2-2-8 . Research result

(1) pH measurement

As shown in FIG. 9, the pH of the fermented milk was not reached to the fermentation end point of 4.7 or lower in all the test groups at the fermentation time of 4 hours. In the control group, fermented milk was fermented at a fermentation time of 8 hours Test group 2 reached the fermentation end point at the fermentation time of 12 hours. The isolate appears to be slightly less active than the control.

(2) Measurement of acidity

As shown in Fig. 10, the acidity was increased with time in all the test groups including the control group. As in the case of pH, fermented milk produced by commercial starter was higher in acidity than isolate, and test group 1 using single strain was slightly lower, suggesting that the acid producing ability of the isolate is low.

(3) Change in the number of lactic acid bacteria

As shown in Fig. 11, the number of lactic acid bacteria was higher in the control group using the commercial starter after 4 hours of fermentation than in the test groups 1 and 2, which were fermented oils using the isolates. Control group showed 10 ⁷ CFU / mL after 4 hours and test group 1 and 2 showed no significant difference after 6 hours but decreased. In addition, in the fermented milk test group 2 prepared by using the commercial strain and the isolate, 10 ⁷ CFU / mL or more was observed in the fermented milk test group. However, in order to make commercial dairy products with less than 10 ⁸ CFU / mL of lactic acid bacteria in fermented milk, basic conditions are not met and it is necessary to find out how to use them as starters.

(4) Sensory properties

Sensory evaluation was performed on the color, flavor, taste, and overall acceptability of the fermented milk using the 5-point scale method. The results are shown in Table 7.

The control group, which is a fermented milk made using commercial starter, showed higher sensory scores than fermented milk using the isolate. Leuconostoc lactic acid B has a possibility of being used as a starter, but the ability of fermented milk as a starter tends to be somewhat lower than that of a commercial strain, so it is necessary to find a way to utilize it as a starter by increasing its activity.

division color incense flavor Overall likelihood Control group 4.7 ± 0.5 4.0 ± 0.8 4.1 ± 0.7 4.5 ± 0.7 Test group 1 4.2 ± 0.5 4.2 ± 0.5 4.2 ± 0.7 4.4 ± 0.5 Test group 2 4.0 ± 0.4 4.1 ± 0.5 4.0 ± 0.9 4.4 ± 0.5

2-3. Lactobacillus isolated from kimchi, a traditional fermented food Possibility of using starter of fermented milk of plantarum A and application of fermented milk

In this experiment, we aimed to develop fermented milk products using lactic acid bacteria isolated from kimchi, which is a traditional fermented food, as a starter of fermented milk, which is a fermented milk product. The quality characteristics of fermented milk products prepared by using the isolates were studied. The results can be summarized as follows.

2-3-1. Identification of Lactic Acid Bacteria

Seven species were collected from Jeonbuk region and 25g of each sample and 225ml of sterilized physiological saline were put into a sterile filter bag and mixed with a stomacher. 1 ml of the separated sample was diluted by deciduous dilution method and 100 μl was taken and plated on MRS (Lactobacilli MRS agar, Difco, USA) plate culture medium and cultured at 37 ° C for 24 hours. A single colony representing a transparent ring for each sample was firstly separated. The primary isolates were randomly separated on a BCP agar medium and cultured at 37 ° C for 24 hours. The colonies were then isolated and identified. Lactobacillus plantarum was similar or identical to Lactobacillus plantarum plantarum A < / RTI >

2-3-2. Starter manufacturing

The starter for fermented milk production was prepared by inoculating Kimchi isolate with 10% (w / v) reduced skim milk (Seoul Milk), culturing it at 10 ⁸ CFU / mL or higher, The control group was cultured in the same manner as the commercial strain ABT-5 (Chr. Hansen, Denmark).

2-3-3. Manufacture of fermented milk

5.5 parts by weight per 100 parts by weight of crude oil were mixed and sterilized, and 2 parts by weight of a starter was inoculated to 100 parts by weight of crude oil and cultured at 40 DEG C for 4 hours (FIG. 12). As a control, 2 parts by weight of ABT- , Test group 1 was Lactobacillus plantarum A single strain 2 parts by weight, test group 2 ABT-5 and Lactobacillus fermented milk was inoculated with 2 parts by weight of a complex strain obtained by mixing plantarum A at a ratio of 1: 1.

division Starter for fermented milk production Control group ABT-5 Test group 1 Lactobacillus Plantarum A 2 parts by weight (single strain) Test group 2 ABT-5 and Lactobacillus plantarum A 2: 1 part by weight (mixed strain)

2-3-4. pH measurement

The fermented milk was measured using a pH meter. During the fermentation, the pH was measured at time intervals of 0, 2, 4 and 6 hours.

2-3-5. Titratable acidity measurement

The titratable acidity is determined by mixing 10 ml of sample with 10 ml of distilled water to make a suspension. Titrate the suspension to pH 8.3 by adding NaOH and titrate the titration of NaOH to titrate the amount of NaOH The conversion factor, 0.9, was multiplied and the weight (g) of the sample was divided to determine the titratable acidity (%). The titratable acidity during fermentation was measured at 0, 2, 4, and 6 hour intervals.

2-3-6. Measure the number of lactic acid bacteria

To measure the changes in the lactic acid bacteria over time during the fermentation of the fermented milk, 1 ml of the sample was sampled at 0, 2, 4 and 6 hours according to Richardson's method, diluted with sterilized physiological saline solution by decidual dilution method, After culturing at 37 ° C for 48 hours, yellow colonies were counted.

2-3-7. Sensory properties of fermented milk

Sensory evaluation of the fermented milk was performed by using the 5 point scaling method.

2-3-8. Experiment result

(1) pH measurement

As shown in Fig. 13, the pH was measured, and all the test groups reached the fermentation end point with a similar tendency. In all the test groups, the fermentation time was not reached at the end point of 4 hours and the fermentation end point reached 4.7 ~ 4.8 after 8 hours of fermentation. In order to use it as a starter for commercial fermented milk, it is necessary to study how to reach fermentation completion sooner.

(2) Measurement of acidity

As shown in Fig. 14, in all test groups, the acidity increased with time, and all three groups showed similar trends.

(3) Change in the number of lactic acid bacteria

As shown in FIG. 15, in all the test groups including the control group in the pre-fermentation time region, the test group was above 10 ⁷ CFU / mL, indicating that the basic conditions for commercial dairy products having less than 10 ⁸ CFU / mL in the fermented milk were not satisfied see.

(4) Sensory properties

The color, flavor, taste, and overall acceptability of the fermented milk were evaluated by sensory evaluation using the 5-point scale method. Test group 2, which was fermented milk prepared by mixing 1: 1 with isolated strains and commercial starters, showed the most sensory results.

division color incense flavor Overall likelihood Control group 4.7 ± 0.5 4.0 ± 0.8 4.1 ± 0.7 4.5 ± 0.7 Test group 1 4.7 ± 0.5 4.2 ± 0.8 4.0 ± 0.5 4.5 ± 0.5 Test group 2 4.8 ± 0.5 4.4 ± 0.7 4.4 ± 0.9 4.8 ± 0.5

This study was conducted to investigate the effect of Lactobacillus plantarum A As a result of microbiological, physicochemical and sensory evaluation of fermented milk prepared by using the fermented milk, the ability of the fermented milk as a starter was superior to that of the control strain.

2-4. Mixed strains isolated from kimchi, a traditional fermented food ( Lactobacillus acidophilus C, Leuconostoc lactics B, Lactobacillus plantarum A ) fermented milk starter and application of fermented milk

In this experiment, we aimed to develop fermented milk products using lactic acid bacteria isolated from kimchi, which is a traditional fermented food, as a starter of fermented milk, which is a fermented milk product. The quality characteristics of fermented milk products prepared by using the isolates were studied. The results can be summarized as follows.

2-4-1. Identification of Lactic Acid Bacteria

Seven species were collected from Jeonbuk region and 25g of each sample and 225ml of sterilized physiological saline were put into a sterile filter bag and mixed with a stomacher. 1 ml of the separated sample was diluted by deciduous dilution method and 100 μl was taken and plated on MRS (Lactobacilli MRS agar, Difco, USA) plate culture medium and cultured at 37 ° C for 24 hours. A single colony representing a transparent ring for each sample was firstly separated. The primary isolates were randomly separated on a BCP agar medium and cultured at 37 ° C. for 24 hours to isolate colonies showing yellow colonies.

2-4-2. Starter manufacturing

The starter for fermented milk production was prepared by inoculating Kimchi isolate with 10% (w / v) reduced skim milk (Seoul Milk), culturing it at 10 ⁸ CFU / mL or higher, The control group was cultured in the same manner as the commercial strain ABT-5 (Chr. Hansen, Denmark).

2-4-3. Manufacture of fermented milk

5.5 parts by weight per 100 parts by weight of crude oil were mixed and sterilized, and 2 parts by weight of a starter was inoculated on 100 parts by weight of crude oil and cultured at 40 DEG C for 4 hours (FIG. 16).

division Starter for fermented milk production Control group ABT-5 Test group 1 Lactobacillus acidophilus C 2 parts by weight (single strain) Test group 2 Lactobacillus Plantarum A 2 parts by weight (single strain) Test group 3 Leuconostoc lactics B 2 parts by weight (single strain) Test group 4 Lactobacillus acidophilus C and Leuconostoc lactics B and Lactobacillus plantarum A at a ratio of 1: 1: 1 2 parts by weight (complex strain)

2-4-4. pH measurement

The fermented milk was measured using a pH meter. During the fermentation, the pH was measured at time intervals of 0, 2, 4 and 6 hours.

2-4-5. Titratable acidity measurement

The titratable acidity is determined by mixing 10 ml of sample with 10 ml of distilled water to make a suspension. Titrate the suspension to pH 8.3 by adding NaOH and titrate the titration of NaOH to titrate the amount of NaOH The conversion factor, 0.9, was multiplied and the weight (g) of the sample was divided to determine the titratable acidity (%). The titratable acidity during fermentation was measured at 0, 2, 4, and 6 hour intervals.

2-4-6. Measure the number of lactic acid bacteria

To measure the changes in the lactic acid bacteria over time during the fermentation of the fermented milk, 1 ml of the sample was sampled at 0, 2, 4 and 6 hours according to Richardson's method, diluted with sterilized physiological saline solution by decidual dilution method, After culturing at 37 ° C for 48 hours, yellow colonies were counted.

2-4-7. Sensory properties of fermented milk

Sensory evaluation of the fermented milk was performed by using the 5 point scaling method.

2-4-8. Experiment result

(1) pH measurement

As shown in FIG. 17, pH was measured, and as a result, Lactobacillus Test group 4, which was a mixture of test group 1 with acidophilus C single strain and three isolates, reached the end point of fermentation within 4 hours after fermentation end point 4.7. The activity of the test group in which three kinds of the isolates were mixed is higher than that of the control group using the commercial bacteria. Thus, the isolate can be used as a starter of fermented milk. Experimental results showed that the results were similar to those of test group 1 using Lactobacillus acidophilus C monoclonal strain. Lactobacillus acidophilus C was thought to be acting as a bacterium.

(2) Measurement of acidity

As shown in Fig. 18, in all test groups, the fermentation time was rapidly increased after 2 hours, and Lactobacillus acidophilus C single strain and test group 4, which was a mixture of three isolates, showed rapid acidity increase. Leuconostoc lactics C was the slowest increase in test group 3, indicating that acid production was low.

(3) Change in the number of lactic acid bacteria

As shown in Fig. 19, the control group, test group 2 ( Lactobacillus plantarum A ), test group 3 ( Leuconostoc lactics B ) were found to be 10 ⁷ CFU / mL after 4 hours and in test group 2 ( Lactobacillus plantarum A ) and test group 4 ( Lactobacillus acidophilus C , Leuconostoc lactics B and Lactobacillus plantarum A ) showed more than 10 ⁹ CFU / mL after 4 hours of fermentation, indicating that the activity of lactic acid bacteria is very high. In the test group of 3, 3 isolates showed more than 10 ⁹ CFU / mL. Therefore, it is considered that the three bacteria do not inhibit each other and the tendency is similar to that of test group 1. Lactobacillus acidophilus C was thought to be acting as a bacterium. As a result of the experiment, commercial fermented milk products could be produced using a combination of three strains.

(4) Sensory properties

The color, flavor, taste, and overall acceptability of the fermented milk were evaluated by sensory evaluation using the 5-point scale method. The results are shown in Table 11. The most favorable results were obtained from the fermented milk prepared by using the combined strain of three isolates. Next, Lactobacillus acidophilus C fermented milk showed high results.

division color incense flavor Overall likelihood Control group 4.7 ± 0.5 4.0 ± 0.8 4.1 ± 0.7 4.5 ± 0.7 Test group 1 4.7 ± 0.5 4.2 ± 0.8 4.0 ± 0.7 4.7 ± 0.5 Test group 2 4.7 ± 0.5 4.2 ± 0.8 4.0 ± 0.5 4.5 ± 0.5 Test group 3 4.2 ± 0.5 4.2 ± 0.5 4.2 ± 0.7 4.4 ± 0.5 Test group 4 4.8 ± 0.5 4.4 ± 0.5 4.2 ± 0.5 4.8 ± 0.5

This study was conducted to investigate the effect of Lactobacillus acidophilus C, Lactobacillus plantarum , Leuconostoc lactics As a result of microbiological, physicochemical and sensory evaluation of fermented milk prepared by using B , the possibility of its use as a starter of fermented milk was confirmed. In particular, a complex strain containing three microorganisms was able to be used as a starter of fermented milk It is considered that it will be developed as a starter of fermented milk in future.

Claims (5)

A fermented starter derived from a conventional fermented food characterized by culturing the lactic acid bacterium in a conventional fermented food.
The fermentative starter according to claim 1, wherein the traditional fermented food is a kimchi.
3. The fermented food according to claim 1 or 2, wherein the lactic acid bacteria isolated from kimchi, which is a traditional fermented food, is Lactobacillus PlantarumSubsp , Leuconostoc lctics or Latobacilus acidophilus is a fermentable starter derived from a conventional fermented food.
Mixing the crushed kimchi with sterile physiological saline and separating the kimchi liquid;
Smearing the Kimchi liquid, smearing it, and culturing the mixture to separate the lactic acid bacteria;
Culturing the isolated lactic acid bacterium in a medium;
Culturing the cultured lactic acid bacteria in a skim milk;
Wherein the fermented starter is a fermented starter.
Mixing 5.5 parts by weight of sugar with respect to 100 parts by weight of crude oil to prepare a mixture;
Sterilizing the mixture;
Cooling the sterilized mixture;
Adding 2 parts by weight of a kimchi-derived fermentation starter to 100 parts by weight of crude oil to the cooled mixture, and fermenting the mixture;
And cooling the fermented mixture. The method for producing fermented milk using the fermented starter according to claim 1,

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