KR20140108965A - Preparation method of lactic acid bacteria-containg rice wine which has high content of probiotics lactic bacteria and improved stability - Google Patents

Abstract

The present invention relates to a method for manufacturing lactobacillus-containing rice wine having high content of probiotic lactobacillus and improved safety. The method for manufacturing lactobacillus-containing rice wine having improved safety comprises the steps of: a first stage fermentation which mixes and ferments a starch material, water, malt, and lactobacillus; a second stage fermentation which mixes and ferments a starch material, water, malt, and yeast with the first stage fermented product; homogenizing the second stage fermented product; and finishing the product.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a lactic acid bacteria having high content and high stability of probiotic lactic acid bacteria,

The present invention relates to a method for producing lactic acid bacteria rice wine having high content and stability of probiotic lactic acid bacteria.

Recently, interest in health has been increasing, and research has been conducted on foods containing probiotic Lactobacillus, which is known to secrete good lactic acid bacteria, that is, various useful physiologically active substances in the body. Accordingly, the development of makgeolli containing lactic acid bacteria (hereinafter referred to as lactic acid bacteria makgeolli) with the addition of probiotic lactic acid bacteria is also under way.

A known method for producing lactic acid bacteria makgeolli is to add lactic acid bacteria in the two-stage soaking step or to add lactic acid bacteria in the sifting step after completion of the two stage soaking step as in the registered patent 10-0954089. However, this is disadvantageous in that the function of probiotic lactic acid bacteria added at the two-stage immersion or the addition of the lactic acid bacteria at the time of the preparation is deteriorated by the lactic acid bacteria which are proliferated at the first stage, that is, naturally occurring lactic acid bacteria in the makgeolli. Furthermore, the conventional lactic acid bacteria makgeolli finally sterilize the lactic acid bacteria in order to enhance the stability of makgeolli, so it is hard to say that it is true lactic acid bacteria makgeolli.

Therefore, the inventors of the present invention found that probiotic Lactobacillus was added at a certain temperature and cultured at a specific temperature in a one stage fermentation while alcohol fermentation was carried out in a two stage fermentation while researching a high quality rice wine having a high probiotic lactic acid bacterium content The present invention has been accomplished by confirming that it is possible to produce lactic acid bacteria makgeol which does not need further roughening process because of high stability of makgeolli.

It is an object of the present invention to provide a process for producing lactic acid bacteria rice wine having high content and stability of probiotic lactic acid bacteria.

According to an aspect of the present invention,

A one stage fermentation step in which starch material, water, yeast and lactic acid bacteria are mixed and fermented;

A two stage fermentation step in which fermentation is carried out by mixing the first stage fermentation product with a starch material, water, yeast and yeast;

Homogenizing the two-stage coating material; and

The method comprising the steps of: preparing a lactic acid bacteria having increased stability.

Further, according to the present invention,

A stable lactic acid bacteria rice wine produced by the production method of the present invention is provided.

The lactic acid bacteria makgeolli prepared by the method of the present invention has high content and stability of probiotic lactic acid bacteria.

Figures 1 and 2 show lactic acid efficacy according to the ethanol content in the medium of the strains isolated from the koji.
Fig. 3 shows the 16S rDNA of Wysselacvia KSD-914.
Figure 4 shows the blue colonies generated after incubation of Wiselsa cervaria KSD-914 in MRS medium containing BPB.
Fig. 5 shows the results of glucose tolerance test of Wysselacvia KSD-914.
Fig. 6 shows changes in acidity of lactic acid bacteria rice wine according to the present invention at room temperature.
Fig. 7 shows changes in the lactic acid concentration according to the storage of the lactic acid bacteria rice wine of the present invention at room temperature.
Fig. 8 shows changes in alcohol content of the lactic acid bacteria of the present invention at room temperature.
9 shows changes in the number of lactic acid bacteria according to the storage of the lactic acid bacteria of the present invention at room temperature.

According to the present invention,

A one stage fermentation step in which starch material, water, yeast and lactic acid bacteria are mixed and fermented;

A two stage fermentation step in which fermentation is carried out by mixing the first stage fermentation product with a starch material, water, yeast and yeast;

Homogenizing the two-stage coating material; and

And a method for producing lactic acid bacteria rice having increased stability, which comprises a step of forming a lactic acid bacteria.

Also,

A one stage fermentation step in which starch material, water, yeast and lactic acid bacteria are mixed and fermented;

A two stage fermentation step in which fermentation is carried out by mixing the first stage fermentation product with a starch material, water, yeast and yeast;

Homogenizing the two-stage mortar;

Sterilizing the homogenized two-stage plating solution;

Adding lactic acid bacteria and prebiotics to the sterilized two-step smoking article;

Injecting carbonic acid; and

And a method for producing lactic acid bacteria rice having increased stability, which comprises a step of forming a lactic acid bacteria.

Further, according to the present invention,

A stable lactic acid bacteria rice wine produced by the production method of the present invention is provided.

Hereinafter, the present invention will be described in detail.

Lactobacillus

In the present invention, lactic acid bacteria added in the production of lactic acid bacteria rice wine are probiotic lactic acid bacteria which are known to produce healthier physiologically active substances. Therefore, the probiotic lactic acid bacteria of the present invention are distinguished from the unspecified lactic acid bacteria that naturally exist in the makgeolli during the production of general makgeolli. The lactic acid bacterium of the present invention may be a general probiotic lactic acid bacterium, preferably Wisceras cyboria KSD-914 deposited under accession number KCTC12106BP. The lactic acid bacteria may be added in a form capable of proliferation, and the form of addition is not limited. The lactic acid bacteria may be added in the form of a culture solution or a powder, but the present invention is not limited thereto.

Starch material

The starch material of the present invention is highball rice, steamed rice, rice crumbs and the like. . At this time, it is preferable to use rice. The rice pulverized product is prepared by immersing rice in which water is not added and pulverizing it

One-step soak

The one stage fermentation is carried out at a temperature suitable for proliferation of the added probiotic lactic acid bacteria. That is, the one-stage immersion is performed at 30 to 40 ° C, preferably at 32 to 38 ° C.

The yeast is not included in the one-stage immersion step. When the yeast is included in the first stage, the alcohol content is increased due to the yeast activity, so that the added probiotic lactic acid bacteria can not sufficiently propagate.

1st stage Depot

There is a lot of probiotic lactic acid bacteria added in the first stage soaked in the first stage soak. This is because the probiotic lactic acid bacteria sufficiently proliferated in the stage 1 immersion step. The probiotic lactic acid bacterium as the live bacteria is present in the first stage fermentation product in an amount of 1.0E + 07E cfu / ml or more, preferably 1.0E + 08E cfu / ml or more, more preferably 1.0E + 09E cfu / ml or more exist.

Two-stage soaking

The two-stage immersion is performed at a two-stage immersion temperature, that is, at room temperature, in a general makgeolli production process. For example, the two-stage immersion is carried out at 20 to 35 ° C, preferably 20 to 30 ° C, but is not limited thereto.

Since the probiotic lactic acid bacteria added in the first stage of the present invention are present in the first stage of the present invention, the fermentation by the probiotic lactic acid bacteria proceeds as well as the alcohol fermentation of the yeast during the two stage fermentation.

Homogenization step

The homogenization refers to a process of pulverizing or separating a solid component contained in a two-stage substrate. Specifically, the second-stage substrate is treated with a homogenizer according to a conventional method to remove the solid component contained in the fermentation column It refers to the process of pulverizing or separating the solid component using a fine sieve. The homogenizer refers to a pulverizer that pulverizes the solid material contained in the liquid. In the experimental example of the present invention, the two-stage mortar was homogenized by filtering with a sieve.

Sterilization

The present invention may further comprise a sterilization process. The sterilization refers to a process in which various enzymes including saccharification enzymes are inactivated by applying heat to a homogenized two-stage substrate. Specifically, the sterilization preferably refers to heat-treating the homogenized raw material in a third step at 60 to 80 ° C for 10 to 30 seconds using a commonly used heat exchanger.

The saccharifying enzyme refers to an enzyme which is converted into a sugar by hydrolyzing a starch raw material with an enzyme contained in an enteral or nuruk, and the saccharifying enzyme contained in the homogenized two-stage charcoal is inactivated through the heat treatment, You can block the party from switching. Thus, the degree of alcohol fermentation by the yeast can be constantly derived through this step.

Additional addition of lactic acid bacteria

The present invention may further comprise the step of adding probiotic lactic acid bacteria to the two-stage plating solution prepared through the two-stage soaking. In this case, since the makgeolli is highly stable due to the probiotic lactic acid bacteria contained in a high content in the second step, additional addition of the probiotic lactic acid bacteria promotes the production of useful bioactive materials without affecting the stability of the makgeolli . The form of the probiotic lactic acid bacteria to be added at this time is not limited and may be a culture solution or a powder, preferably a powder.

Prebiotics  adding

The present invention may further include a step of adding the prebiotics to the two-stage fermentation product prepared through the two-stage immersion. This helps the growth of the lactic acid bacteria contained in the second stage fermentation product in a high content. The prebiotics may be conventional prebiotics used for probiotic lactic acid bacteria, and are not particularly limited. For example, the prebiotics of the present invention can be, but are not limited to, oligosaccharides, dietary fibers or gluconic acid.

Injection of carbonic acid

The present invention may further include the step of adding carbonic acid to the two-stage fermentation product prepared through the two-stage immersion. As a result, the two-step curing agent is sterilized, resulting in sterilized makgeolli.

Remedy

The preparation of the present invention may further include the step of adding an additive such as water, saccharides, acids, etc. in accordance with the standard of the makgeolli product. As the acids, citric acid, lactic acid, succinic acid, malic acid and the like may be added. As the saccharides, one or more selected from glucose, fructose, sugar, dextrin and oligosaccharide may be added.

Lactic acid rice wine

The lactic acid bacteria rice wine produced in the present invention has a number of lactic acid germ cells of 1.0E + 07E cfu / ml or more, preferably 1.0E + 08E cfu / ml or more in the production of makgeolli.

The lactic acid bacteria of the present invention inhibits the growth of other lactic acid bacteria due to the proliferation of the specific probiotic lactic acid bacteria added at the first stage soaking, thereby preventing the acidic bacteria caused by the growth of other lactic acid bacteria when stored at room temperature. Therefore, the stability is high and the shelf life is increased.

In addition, the lactic acid bacteria of the present invention is good for health because it is rich in specific probiotic lactic acid bacteria added at the first stage.

Stability of lactic acid bacteria rice wine

The acidity change of the lactic acid bacteria of the present invention is within ± pH 1 when stored at room temperature for 8 days. For example, if the pH of the lactic acid bacteria of the present invention measured immediately after preparation is pH 4.5, the acidity measured after storage at room temperature for 8 days is 3.5 to 5.5.

In addition, the lactic acid bacteria of the present invention has a lactic acid concentration of 2.5 g / L or less when stored at room temperature for 8 days.

In addition, the number of lactic acid bacteria living organisms according to the present invention is 0.001 Na <Nb <1000 Na, preferably 0.01 Na <Nb <100 Na. In this case, Na is the number of lactic acid bacteria (cfu / ml) in makgeolli at the time of making makgeolli, and Nb is the number of lactic acid bacteria (cfu / ml) in makgeolli after preparing makgeolli and storing at room temperature for 8 days.

BRIEF DESCRIPTION OF THE DRAWINGS The advantages and features of the present invention, and the manner of achieving them, will be apparent from and elucidated with reference to the embodiments described hereinafter in conjunction with the accompanying drawings. It should be understood, however, that the invention is not limited to the disclosed embodiments, but is capable of many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, To fully disclose the scope of the invention to those skilled in the art, and the invention is only defined by the scope of the claims.

<Experimental Example 1> Isolation of Wisella cibaria KSD-914

Manufacture of yeast

Commercially available wheat bran was purchased and pulverized using a pulverizer. 8 kg of crushed wheat bran were mixed with 7.5 liters of water to make a milky lotion. The cotton cloth was spread in a nutskin, spreaded in a bowl, and then put into a bowl. At that time, the mold was shaped to have a barrel shape of 5 to 20 cm in diameter and 1 to 5 cm in thickness. The rug was laid with rice straw, pine needle and wormwood leaves, % Humidity for 2 ~ 4 weeks and then pulverized to prepare yeast.

Isolation of strain from yeast

1 g of the yeast was dissolved in sterilized distilled water and diluted 10-fold. Then, it was spread on MRS-agar (0.04% BCP) and cultured at 37 ° C, and 43 kinds of strains were identified by dilution step. In order to select heterotrophic strains with low acid production, only a single blue colony was isolated on the plate surface. As a result, a total of 9 strains were selected (strains 1-2, 2-1, 3- 1, 3-2, 3-3, 4-1, 4-3, 4-13, 4-16) (Table 1).

Alcohol sensitivity test

The isolated lactic acid bacteria were inoculated into MRS medium (Difco) supplemented with 0%, 5%, 7% and 10% of alcohol, respectively, and cultured at 30 ~ 37 ℃. Samples were taken according to the number of days of culture, and the supernatant was measured for lactate using a biochemical analyzer.

As a result, it was confirmed that the strain 2-1 inhibits growth as the alcohol concentration increases (Figs. 1 and 2).

Identification of the strain isolated from the modified yeast

The 16s rDNA nucleotide sequence of the strain No. 2-1 was analyzed and identified. The result of identification is 99% homology to Weisella cibaria and confirmed as Wyssela cervaria. The results of the 16s rDNA sequence analysis of the strain 2-1 (hereinafter, referred to as Wysselacia bilata KSD-914) are shown in SEQ ID NO: 1 (FIG. 3)

<Experimental Example 2> Bacteriological properties of Wisella cibaria KSD-914

Morphological characteristics

The blue colonies formed after culturing the strain No. 2-1, ie, Wysselacia bilata KSD-914, on an MRS medium containing 0.002% BPB (Bromophenol blue) for 16 hours at 37 ° C. are shown in FIG.

Biochemical property

The biochemical characteristics of Wisella cibaria KSD-914 are shown in Table 2 below.

Glucose utilization

49 sugar-bearing experiments were carried out on Wysselacivaria KSD-914 using API50 CHL kit (API BioMerieux, France). The results are shown in the following Table 3 and FIG. 5, and it was judged that the strains were novel strains because their mycological properties were different from those of the existing strains of Wysselacs.

&Lt; Experimental Example 3 > Properties of Wisella cibaria KSD-914

Acid resistance

The cells were harvested from the culture medium and inoculated with MRS (1N HCl) adjusted to pH 2, pH 3, pH 6.5 (control) and incubated at 30 ° C for 1 hr. , And cultured for 2 or 3 hours. Then, 1 ml of each was taken and diluted with physiological saline for dilution. 100 의 of diluted solution was plated on MRS agar plate medium and cultured at 30 캜 for 48 hours to compare the number of colonies present.

As a result, it was confirmed that Wysselacivaria KSD-914 was well grown even at pH 2 and pH 3, and was found to have acid resistance (Table 4).

My bile

100 占 퐂 / ml of Wiselachibaria KSD-914 was plated on a MRS solid medium supplemented with 100 占 퐂 / ml, 300 占 퐂 / ml and 500 占 퐂 / ml of sterilized bile acid and cultured at 30 占 폚 for 24 hours. The viability of the lactic acid bacteria was measured by comparing the number of colonies after that. As a result, it was confirmed that Wiselsa cvaria KSD-914 had good bile growth due to good growth in the medium supplemented with bile acid (Table 5).

<Experimental Example 4> Production of lactic acid bacteria rice wine

One kilogram of rice was immersed for 5 hours and water was removed for about 1 hour. The rice without water was pulverized with a blender and then 1.5 kg of purified water was added. 15 g of WASELASHIBARIA KSD-914 was added as lactic acid bacterium, and 9.5 g of yeast was added, followed by overnight fermentation at 30 to 40 DEG C to carry out single-stage immersion.

2kg of rice was immersed for 5 hours and then water was removed for about 1 hour. The rice without water was pulverized with a blender and 3 kg of purified water was added. To this was added 19 g of modified Nuruk, 30 g of yeast, and the above-prepared one-stage charcoal, followed by fermentation at 27 ° C for 7 days to obtain a two-stage soaking process to produce lactic acid rice wine.

The two-step curing agent was sieved with a sieve to separate and solidify the solid components, and then the mixture was homogenized.

At this time, the above-mentioned Wyse Clavaria KSD-914 was cultured after inoculating a single colony of lactic acid bacteria into MRS medium, and the cultured cells were expanded on MRS medium and added at one stage of immersion.

<Experimental Example 5> Stability test of lactic acid bacteria rice wine

The stability of the lactic acid bacteria rice wine prepared in Experimental Example 4 was tested. The rice wine prepared in the same manner as in Experimental Example 4 was used as a control, except that 15 g of yeast was added instead of the lactic acid bacteria at the first stage of immersion.

<5-1> Number of lactic acid bacteria in the first stage

The number of lactic acid bacteria in the first stage digestion water before the two stage digestion was measured in the lactic acid bacteria rice wine and the control group prepared in Experimental Example 4 were measured. As a result, Lactobacillus acidophilus was confirmed to be live bacteria of 1.0E + 09E cfu / ml or more. However, in the control group, the number of lactic acid bacteria in the first stage was 1.0E + 05 cfu / ml. Therefore, it was confirmed that the probiotic lactic acid bacteria added to the lactic acid bacteria of the present invention during the one stage fermentation were sufficiently proliferated.

<5-2> Evaluation of shelf life at room temperature

The acidity, lactate content, alcohol content and number of lactic acid bacteria were measured during storage at the room temperature (25 ° C), respectively, of the lactic acid bacteria rice wine and the control group prepared in Experimental Example 4.

As a result, in the control group, acidity and lactic acid levels started to increase rapidly from the 4th day of storage, reaching the limit of drinking time at 6 ~ 7 days. However, in case of lactic acid bacteria rice wine, And drinking was possible. In case of control group, the number of lactic acid bacteria increased rapidly when stored at room temperature, while the lactic acid bacteria rice wine of the present invention did not change abruptly even when stored at room temperature. This suggests that proliferation of other lactic acid bacteria was inhibited by the probiotic lactic acid bacteria that were proliferated in the process of producing the lactic acid bacteria of the present invention, thereby preventing acidic lung disease caused by the proliferation of other lactic acid bacteria (FIGS. 6 to 9).

Therefore, it was confirmed that the stability of rice wine was increased at room temperature when probiotic lactic acid bacteria were added.

Claims (16)

A one stage fermentation step in which starch material, water, yeast and lactic acid bacteria are mixed and fermented;
A two stage fermentation step in which fermentation is carried out by mixing the first stage fermentation product with a starch material, water, yeast and yeast;
Homogenizing the two-stage coating material; and
Wherein the lactic acid bacteria is a lactic acid bacteria.
The method according to claim 1,
Wherein the lactic acid bacteria are probiotic lactic acid bacteria.
The method according to claim 1,
Wherein the one-stage immersion step is performed at 30 to 40 占 폚.
The method according to claim 1,
Wherein the one-stage immersion step is carried out without containing yeast.
The method according to claim 1,
Wherein the starchy material is a rice flour prepared by immersing steamed rice, rice gruel or rice pulverized rice in water without grinding it and then pulverizing it.
The method according to claim 1,
Wherein said lactic acid bacterium is Weissella cibaria KSD-914 deposited with Accession No. KCTC12106BP.
The method according to claim 1,
Wherein the two-stage immersion step is performed at 20 to 35 占 폚.
The method according to claim 1,
Wherein the lactic acid bacteria rice wine has a number of lactic acid germ cells measured at the time of production of 1.0E + 07E cfu / ml or more.
The method according to claim 1,
Wherein the acidic rice wine has an acidity change within a range of pH 1 when stored at room temperature for 8 days after the preparation.
The method according to claim 1,
Wherein the lactic acid bacteria rice cake has a lactic acid concentration of 2.5 g / L or less when stored for 8 days at room temperature after production.
The method according to claim 1,
The number of lactic acid bacteria in the above mentioned lactic acid bacteria rice wine was
0.001 Na < Nb < 1000 Na,
At this time, Na is the number of lactic acid bacteria (cfu / ml) in makgeolgi at the time of making makgeolli,
And Nb is the number of lactic acid bacteria (cfu / ml) in makgeolli after the makgeolli is prepared and stored at room temperature for 8 days.
The method of claim 1, wherein
Characterized in that lactic acid bacteria and prebiotics are added to the homogenized two-stage mortar and then the preparation step is carried out
The method according to claim 1,
Wherein the homogenized two-stage mortar is sterilized, and then the lactic acid bacteria and the prebiotics are added, and the preparation step is carried out.
The method according to claim 1,
And sterilizing the homogenized two-stage mortar and then injecting carbonic acid.
Wherein the lactic acid bacteria and the prebiotics are added to the homogenized two-stage mortar, then carbonic acid is injected, and the preparation step is carried out.
16. A stabilized lactic acid bacteria rice wine produced by the method of any one of claims 1 to 15.

Images