KR101313120B1 - Manufacturing method of rice Kefir by using Lactobacillus plantarum and Saccharomyces cerevisiae - Google Patents

Abstract

The present invention relates to the production of a kefir using rice, and selected strains suitable for preparing rice kefir using lactic acid bacteria and yeasts known for their functional activity, and established conditions for producing rice kepers using these selection strains. Seven strains of lactic acid bacteria and two yeast strains were selected for the fermentation of rice, and experiments were carried out to determine the selection, inoculation and fermentation temperature of rice fermentation medium. A dry product was produced. The present invention enables the production of a new rice fermented product which is estimated to have a functional activity from the established rice kefer manufacturing process and can contribute to the expansion of rice consumption by the production of rice kefer fermented products.

Description

Manufacturing method of rice kefer characterized in that the fermentation using lactic acid bacteria Lactobacillus plantarum strain and yeast Saccharomyces cerevisiae strain {Manufacturing method of rice Kefir by using Lactobacillus plantarum and Saccharomyces cerevisiae}

The present invention relates to a rice kefir (Kefir), and more specifically, Lactobacillus plantarum ( Lactobacillus) plantarum ) MG207 strain and yeast Saccharomyces cerevisiae ( Saccharomyces cerevisiae ) MG111 strain using a technique for producing a fermented rice kefer.

In the present invention relates to the production of a kefir using rice was selected strains suitable for producing rice kefir using lactic acid bacteria and yeast bacteria known functional activity. In the selection process of these strains, the production of polysaccharides, fast acid production, and bacterial growth of known functional activity were considered. Lactic acid bacteria selection experiment suitable for fermentation of rice was carried out through fermentation suitability experiment using 7 kinds of lactic acid bacteria and 2 kinds of yeast, and after experiment to determine the selection of rice fermentation medium, inoculation amount and fermentation temperature A manufacturing experiment and a rice kefir lyophilized product production experiment were conducted. Selection of strains for the production of rice kepers of the present invention, establishment of rice kefer fermentation processes, and production of rice keffer products have the meaning of production of rice kepers, which are functional new products using rice.

Kefir is an effervescent fermented milk originally consumed in the mountainous region of Caucasus, and refers to fermented milk made by adding kefir grain to milk as a starter. It contains about 1% lactic acid and about 0.8% alcohol, sour and slightly alcoholic. Kefer's origin comes from the Turkish kef, meaning that milk is added to the yeast and lactobacillus-containing yeasts that ferment lactose to produce alcohol, carbon dioxide, and lactic acid, and have a distinctive flavor. Similar to making it from goat's milk, sheep's milk or milk, there are Eastern Europe's Urda, Chile's Skuta, and horse milk Kumiss has a strong alcohol content. To make it, put the cooled and heated milk in Kepper spawn in hot water, leave it for 1 week at 20 ℃, make it under the drunk and add about 5 times of heating coolant and mix it well. have

The kefer product is a fermentation product made from milk and using lactic acid bacteria and yeast strains. For the production of kefer, in principle, the fermentation starter is used as a fermentation starter, which is a lactic acid bacterium and yeast cell mass. However, since many kefer grains are not sterilized, many kefir producers use purely isolated lactic acid bacteria and yeast strains. It is known to make a kefir using.

Kefer, a fermented milk product, is known to have not only unique palatability but also many functional activities. Some studies have shown that in some countries with a high intake of meat, some countries have significantly lower incidence of colorectal cancer. In case of high intake of yogurt fermented milk, it is reported that the incidence of colorectal cancer is significantly lowered. Kefer products are known to have the benefits of fermented milk and at the same time have additional functional activity due to the production of polysaccharides.

In Korea, kimchi, a representative lactic acid bacterium fermented food, contains lactic acid bacteria such as Leuconostoc and Lactobacillus , and studies have shown that they are effective in anti-cancer, cancer prevention, and immunity. In addition, by reducing the pH of the intestine has a formal effect of inhibiting the growth of harmful microorganisms and pathogenic microorganisms, has been reported to inhibit the activity of enzymes that convert carcinogenic precursors to carcinogenic substances.

Although there are many studies and patents on the production of rice fermented products using lactic acid bacteria, there is no research on the rice fermented products using lactic acid bacteria and yeast, and there is no patent.

The present invention is to establish a manufacturing process of rice kefer which is a rice fermented food using lactic acid bacteria and yeast strains having functional activity. The technical objects to be achieved by the present invention are not limited to the above-mentioned technical problems, and other technical subjects which are not mentioned can be clearly understood by those skilled in the art from the description of the present invention .

The present invention is Lactobacillus plantarum Lactobacillus plantarum ) MG207 strain and yeast Saccharomyces Saccharomyces cerevisiae ) Provides a method for preparing rice kefer (Kefir) fermented using MG111 strain. Preferably, the lactic acid bacteria Lactobacillus plantarum MG207 strain is inoculated 1 to 10wt%, the yeast Saccharomyces cerevisiae MG111 strain may be characterized by inoculating 1 ~ 10wt%. Preferably, the fermentation may be characterized in that the first fermentation for 4 hours to 10 hours at 25 ℃ ~ 30 ℃, the second fermentation for 21 hours to 27 hours at 15 ℃ ~ 25 ℃. Preferably, the fermentation may be characterized by using a medium consisting of rice flour 1 ~ 10wt%, peptone 0.1 ~ 3wt% and the balance of purified water.

In another aspect, the present invention provides a rice kepper prepared by any one of the above method. Preferably, the rice hopper may be characterized in that it is lyophilized using sucrose (sucrose) as a cryoprotectant. More preferably, the rice hopper may be characterized in that the freeze-dried using 5 ~ 30wt% sucrose as a cryoprotectant.

The present invention investigated the fermentation method for the production of rice kefir using lactic acid bacteria and yeast having a functional activity, to provide a liquid product and a freeze-dried product of rice kefer after the fermentation temperature, inoculation amount determination experiment to establish a rice kefer manufacturing process It was.

Meanwhile, the method for obtaining strains required for the fermentation of the present invention will be described in detail. The following picture is the website of 'Medogen'.

After selecting 'Product' on the website, and selecting 'health functional food raw material', Lactobacillus plantarum ( Lactobacillus) through the product information column as shown below plantarum ) MG207 strain information can be confirmed and purchased.

On the other hand, if you select 'Product' and then 'General Food Ingredients' on the website, yeast saccharomyces cerevisiae ( Saccharomyces) through the product information column as shown below. cerevisiae ) MG111 strain information can be confirmed and purchased.

Through the present invention, it is possible to select a strain suitable for the production of rice kefir from functionally active lactic acid bacteria and yeast, and to establish a rice kefer production process through determination of rice fermentation broth components, inoculation amount and fermentation temperature determination experiment.

Furthermore, through the present invention, it is possible to produce a new rice fermented product which is estimated to have functional activity from the established rice kefer manufacturing process, and contribute to the expansion of rice consumption by the production of the rice kefer fermented product.

1 is a lactic acid bacteria ( L. brevis) in agar medium and MRS agar medium added with 5% saccharification solution of rice MG18, L. casei MG311, L. plantarum MG207, L. rhamnosus MG316, Leu . The result of polysaccharide production was inoculated (5 μl drop) of mesenteroides MG865, L. kefiranofaciens KCTC5075 and L. bulgaricus MG515) and incubated at 37 ° C. for 48 hours.
FIG. 2 shows lactic acid bacteria ( L. brevis) in a medium containing only 5% rice and 1% peptone in rice 5% saccharified solution. MG18, L. casei MG311, L. plantarum MG207, L. rhamnosus MG316, Leu . mesenteroides MG865 and L. bulgaricus MG515) were inoculated at 1% and fermented at 37 ° C.
3 is lactic acid bacteria ( L. brevis) in the medium of only 5% of rice and the medium of peptone 1% in rice 5% saccharified solution MG18, L. casei MG311, L. plantarum MG207, L. rhamnosus MG316, Leu . mesenteroides MG865 and L. bulgaricus MG515) were inoculated at 1% and fermented at 37 ° C.
4 is lactic acid bacteria ( L. brevis) in the medium of only 5% rice and 1% peptone in rice 5% saccharified solution MG18, L. casei MG311, L. plantarum MG207, L. rhamnosus MG316, Leu . mesenteroides MG865 and L. bulgaricus MG515) were inoculated at 1% and fermented at 37 ° C.
5 is inoculated with 1% each of two kinds of lactic acid bacteria ( L. plantarum MG207, L. bulgaricus MG515) in a medium having a peptone added amount of 0.1%, 0.5%, and 1%, respectively, and then fermented at 37 ° C. The experimental results of the pH change are shown.
Figure 6 is Lactobacillus 1% of plantarum MG207) and yeast strains ( Saccharomyces exiguus KFRI138 and S. cerevisiae MG111) were inoculated at 1% for 1st fermentation at 30 ℃ for 24 hours, and the number of viable cells was measured during 2 days fermentation at 15 ℃ and 25 ℃ for 2 days. One experimental result is shown.
Figure 7 is Lactobacillus plantarum MG207) and yeast ( Saccharomyces) Exiguus KFRI138 and S. cerevisiae MG111) were inoculated at 1% each for 1st fermentation at 30 ° C for 24 hours and 2 days of fermentation at 15 ° C and 25 ° C for 2 days.
8 is a lactobacillus ( Lactobacillus plantarum ) and yeast ( Saccharomyces ) in established rice liquid medium (rice 5% saccharification, peptone 0.1%) cerevisiae MG111) was inoculated in 1% increments. And after the primary fermentation for 8 hours at 30 ℃, 25 ℃, 20 ℃, while the second fermentation at 15 ℃ for 2 days shows the result of measuring the number of viable cells, pH and acidity.
9 is inoculated 1%, 3%, 5% of lactic acid bacteria and yeast in the established rice liquid medium (rice 5% saccharification, 0.1% peptone), respectively, after primary fermentation for 8 hours at 30 ℃, 2 at 15 ℃ It shows the result of measuring the number of viable cells and pH while fermenting the second day.

The present invention is Lactobacillus plantarum Lactobacillus plantarum ) MG207 strain and yeast Saccharomyces Saccharomyces cerevisiae ) relates to a method for preparing rice kefer fermented with MG111 strain (Kefir).

Preferably, the lactic acid bacteria Lactobacillus plantarum MG207 strain is inoculated 1 to 10wt%, the yeast Saccharomyces cerevisiae MG111 strain may be characterized by inoculating 1 ~ 10wt%.

Preferably, the fermentation may be characterized in that the first fermentation for 4 hours to 10 hours at 25 ℃ ~ 30 ℃, the second fermentation for 21 hours to 27 hours at 15 ℃ ~ 25 ℃.

Preferably, the fermentation may be characterized by using a medium consisting of rice flour 1 ~ 10wt%, peptone 0.1 ~ 3wt% and the balance of purified water.

The present invention also relates to a rice kefir prepared by any one of the above methods.

Preferably, the rice hopper may be characterized in that it is lyophilized using sucrose (sucrose) as a cryoprotectant. More preferably, the rice hopper may be characterized in that the freeze-dried using 5 ~ 30wt% sucrose as a cryoprotectant.

Hereinafter, the present invention will be described in detail with reference to the accompanying drawings.

L. brevis MG18, L. casei MG311, L. plantarum MG207, L. rhamnosus MG316, Leu. The results of investigating the polysaccharide production ability using seven kinds of lactic acid bacteria such as mesenteroides MG865, L. kefiranofaciens KCTC5075, and L. bulgaricus MG515 are shown in FIG.

In general, all strains are poor in growth and difficult to confirm the production of slime, but L. plantarum, L. casei, L. rhamnosus and Leu. More milky products can be observed in mesenteroides than in other strains (Figs. 1-A and 1-B). Even in MRS agar medium, these strains can identify distinct products, especially Leu. mesenteroides can be seen to produce a large amount of viscous material (Fig. 1-C, Fig. 1-D).

In order to examine the growth of lactic acid bacteria in rice medium, only distilled water was added so that the rice powder became 5% (w / v), and 1% peptone was added to the saccharified solution treated with α-amylase and glucoamylase enzymes. 6% of lactic acid bacteria ( L. brevis MG18, L. casei MG311, L. plantarum MG207, L. rhamnosus MG316, Leu. Mesenteroides MG865, L. bulgaricus MG515) were inoculated into the culture medium at pH, Changes in acidity and viable cell numbers are as shown in FIGS. 2, 3, and 4.

To make rice saccharification liquid, distilled water was added to make the rice powder 5% (w / v), followed by preheating at 60 ° C. for 40 minutes and warming at 100 ° C. for 40 minutes to increase rice saccharification efficiency. Α-amylase 0.13 unit / g rice powder was added to the gelatinized solution for 40 minutes at 80 ℃, glucoamylase 3.5 units / g rice powder was added for 40 minutes at 75 ℃ to prepare a saccharified solution.

As shown in Figures 2, 3 and 4, the growth and acid production of lactic acid bacteria were not achieved at all in the medium of rice alone, and L. plantarum among the six lactic acid bacteria used in the rice saccharification medium. The MG207 strain showed the lowest pH and the most acid production and the highest growth in the viable cell number.

In order to determine the amount of peptone added to the fermentation medium of rice kefer, sterilized medium containing 0.1%, 0.5%, and 1% of peptone in rice 5% saccharified liquid, and then, Lactobacillus plantarum ( Lactobacillus) plantarum MG207) and Lactobacillus bulgaricus MG515) was inoculated at 1% each and fermented at 37 ℃ measurement results of the pH is as shown in FIG. Lactobacillus plantarum as shown in Figure 5 ( Lactobacillus plantarum MG207) shows the lowest pH change at 0.1% peptone.

≪ Example 1 >

Lactobacillus ( Lactobacillus) for the Selection of Yeast Strains Suitable for Rice Kefer Preparation by Mixed Fermentation Experiment of Lactic Acid Bacteria and Yeast plantarum MG207) and yeast ( Saccharomyces) exiguus KFRI138 and S. cerevisiae MG111) were inoculated at 1% each for 1st fermentation at 30 ° C for 24 hours and 2 days of fermentation at 15 ° C and 25 ° C for 2 days. 6 and 7 as shown in FIG.

As shown in FIG. 7, L. plantarum and S. exiguus showed lower pH than L. plantarum and S. cerevisiae , while L. plantarum and S. exiguus showed higher pH. . It can be estimated that more acid is produced in the mixed fermentation of L. plantarum and S. exiguus and the fermentation ability is superior.

As a result of varying the secondary fermentation temperature to 15 ℃ and 25 ℃, there was no difference in pH, viable cell number, and acidity by temperature, but in sensory test of taste and aroma of each sample after fermentation was completed. The sensory beauty of the 15 ℃ fermentation test group was slightly better than the 25 ℃ fermentation test group. In addition, the mixed fermentation of L. plantarum and S. exiguus resulted in a little less thickening of carbon dioxide due to the mixed fermentation of L. plantarum and S. cerevisiae . Therefore, the secondary fermentation temperature was 15 ℃, mixed fermentation strain was determined by L. plantarum and S. cerevisiae .

<Example 2>

Lactic acid bacteria and the established determining a primary fermentation temperature suitable for the growth of lactic acid bacteria in the initial rice kepeo prepared by mixed fermentation of the yeast rice liquid medium (5% Rice Hydrolyzate, peptone 0.1%) in the lactic acid bacteria (Lactobacillus plantarum ) and yeast ( Saccharomyces) cerevisiae MG111) was inoculated in 1% increments. And after primary fermentation for 8 hours at 30 ℃, 25 ℃, 20 ℃, the result of measuring the number of viable cells, pH and acidity while fermenting at 15 ℃ for 2 days is as shown in FIG. As shown in FIG. 8, the pH change at 20 ° C. was slow compared to the primary fermentation temperatures of 30 ° C. and 25 ° C., and the growth was markedly slow at 20 ° C. in the viable cell number. In acidity, there was no significant difference according to the primary fermentation temperature. Therefore, the primary fermentation temperature was determined to be 30 ℃ to minimize the fermentation time.

<Example 3>

Inoculated 1%, 3% and 5% of lactic acid bacteria and yeast in established rice liquid medium (rice 5% saccharification solution, peptone 0.1%) to determine the suitable inoculation amount in the production of rice kefir by mixed fermentation of lactic acid bacteria and yeast After 8 hours of incubation at 30 ℃, fermented at 15 ℃ for 2 days while measuring the number of viable cells and pH is as shown in Figure 9. As can be seen in Figure 9, pH and acidity did not show a significant difference according to the inoculation amount, but in the number of viable bacteria lactic acid bacteria 1% and yeast 1% when both the lactic acid bacteria and yeast showed a high number of viable bacteria to promote growth Appeared. Therefore, the inoculation amount was determined to be 1% lactic acid bacteria and 1% yeast.

<Example 4>

Experimental results for producing lyophilized products using rice kefer fermentation products are shown in Table 1 below. As shown in Table 1, the best survival rate was obtained when sucrose was used as a cryoprotectant. Table 1 summarizes the survival rate according to the change in the number of viable cells before and after lyophilization and the use of a cryoprotectant.

Viable Count (Survival Rate)

Freeze drying progress Lactobacillus
(Survival rate) Yeast
(Survival rate) Before lyophilization 0.95 billion CFU / ml 0.19 billion CFU / ml After lyophilization No additives 0.021 billion CFU / g
(0.11%) 0.0001 billion CFU / g (0.0026%) sucrose group 500 million CFU / g
(79%) 0.12 billion CFU / g
(9.5%) sucrose, trehalose, maltose group 320 million CFU / g
(50%) 0.064 billion CFU / g
(5.0%)

Although the present invention has been described in connection with the specific embodiments of the present invention, it is to be understood that the present invention is not limited thereto. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. In addition, the materials of each component described herein can be readily selected and substituted for various materials known to those skilled in the art. Those skilled in the art can also omit some of the components described herein without changing the performance or add components to improve performance. In addition, those skilled in the art may change the order of the method steps described herein depending on the process environment or equipment. Therefore, the scope of the present invention should be determined by the appended claims and equivalents thereof, not by the embodiments described.

Claims (6)

Lactic acid bacteria Lactobacillus plantarum MG207 strain and yeast Saccharomyces cerevisiae strain MG111 After inoculating, primary fermentation at 25 ° C. to 30 ° C. for 4 hours to 10 hours, and secondary fermentation at 15 ° C. to 25 ° C. for 21 hours to 27 hours. According to claim 1, The Lactobacillus Lactobacillus plantarum MG207 strain is inoculated 1 to 10wt% relative to the total weight of the medium, The yeast Saccharomyces cerevisiae MG111 strain is inoculated 1 to 10wt% relative to the total weight of the medium Method of producing a rice keffer, characterized in that. delete delete Rice keper prepared by the method of claim 1 or 2. 6. The rice hopper of claim 5, wherein the rice hopper is lyophilized using sucrose as a cryoprotectant.

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