KR101928274B1 - Natural flavoring substance from microorganism producing deacetyl and kefir grain and process for the preparation thererof - Google Patents

Abstract

The present invention relates to a natural seasoning material using a diacetyl-high-production strain and a kefir grain, and a method for manufacturing the same. The natural seasoning material according to the present invention imparts cheese and butter flavor through the increase of diacetyl, Due to its salty and rich taste, it can be widely used as a natural seasoning material which can replace synthetic sodium glutamate (MSG).

Description

FIELD OF THE INVENTION [0001] The present invention relates to a natural flavoring substance using a diacetyl high-yielding strain and a kefir grain, and a method for producing the natural flavoring substance from a microorganism producing microcrystalline kefir grain and process for the preparation thererof,

The present invention relates to a lactic acid-tolerant Lactococcus lactis subsp. Crmoris LRCC5306 (Deposit No. KCCM11733P) strain having excellent ability to produce diacetyl, which is isolated from a mature milk, and a powdered natural seasoning material obtained by drying fermented milk fermented by adding kefir grain to milk And a method for producing the same. The natural seasoning material according to the present invention imparts cheese and butter flavor through the increase of diacetyl, and the synthetic sodium glutamate (MSG), etc., due to the salty taste and the rich taste produced by yeasts and strains in the kefir grain Can be widely used as a natural seasoning material to replace.

The causes of deaths in Korea by age group are as follows: traffic accidents in 20 ~ 30s, liver disease in 40s, vascular diseases after 50s, hypertension and stroke. In particular, as the life expectancy increases, the number of people suffering from adult diseases such as hypertension, diabetes, and paralysis is increasing, and the number of people suffering from adult diseases in their thirties and forties is increasing. With regard to this phenomenon, the food culture has also been greatly changed as interest in health and well-being due to the improvement of living standards has rapidly increased in recent years. As a result, efforts to find health-oriented foods such as natural foods, health foods, and functional foods have greatly increased, and many people have changed to such eating habits.

According to this tendency, recently, not only the safety and nutritional aspects of processed foods are satisfied but also consumers' awareness of the risk has increased, and thus natural seasonings and functional natural food materials utilizing natural taste have been attracting attention.

Korean Patent Publication No. 2012-76107 discloses a seasoning material containing amino acids and peptides using Bacillus subtilis (KFCC 11492P), a glutamic acid-producing strain, and Korean Patent No. 10-0572029 discloses a seasoning composition containing Hun- The present invention relates to a natural seasoning material using a residue and a method for manufacturing the same, and efforts are being made to replace artificial seasoning materials.

Generally, sodium glutamate (MSG) is used as a method of improving the taste of foods in seasonings. These artificial seasonings are composed of only materials that exhibit seasoning performance and are widely used because of their excellent seasoning characteristics per unit weight. However, the artificial seasoning alleviates salty, sour, and bitter tastes, but it delivers its own irritating taste or strong irritation to the tongue. If the artificial seasoning is tended to be stimulated, it becomes difficult to feel the taste of the food without adding the artificial seasoning, and the state of being unable to feel the natural silver and the more complex taste. In addition, artificial seasonings are considered to be more toxic to humans than natural materials, and excessive use can actually cause problems for the human body. Especially for people with weak stomach or intestines, even using a small amount causes problems.

L-glutamic acid, which constitutes sodium glutamate, is known to be one of the major metabolites produced by yeast in microorganisms. Considering this point, in the natural-oriented seasoning material market, the expansion of seasoning materials utilizing yeast extract . However, seasonings based on natural extracts have a lower intensity and a higher market price than those of MSG, making it difficult to apply them to the food industry.

On the other hand, kefir fermented milk originating from the Soviet Union's Caucasus region is fermented by adding milk, goat milk, horse oil, etc. with kefir grain as a starter. Most of them are cultured in the conventional manner, so the kind of milk used for cultivation, Time, culture container, etc. are unified, the kind and amount of the microorganisms contained in the fermented milk are also various. In particular, it has been known that regular fermented milk products of more than 50 kinds of lactic acid bacteria, acetic acid bacteria, and yeast are natural fermented milk products that can obtain a variety of health beneficial effects such as regular action, anti-obesity action, anti-inflammatory action and antimicrobial action . Kefir dairy products manufactured using these kefir seeds are rich in lactic acid bacteria and yeast fermentation, resulting in a refreshing and refreshing taste with unique cooling sensations and flavors.

Since the kefir contains yeast, L-glutamic acid is expected to be produced as a metabolite in the production of kefir fermented milk, and this L-glutamic acid gives a rich taste. In addition, as one of the species forming the kefir grain, Lactococcus lactis strain produces diacetyl as a metabolite, which produces a unique cheese or butter flavor with a cooling sensation in the kefir fermented milk.

Therefore, there is still a discovery of natural seasoning materials that can replace artificial seasoning materials by utilizing various materials.

1. Korean Patent Publication No. 2012-76107 2. Korean Patent No. 10-0572029

Therefore, the present inventors have found that, in the case of a powder obtained by drying a lactic acid fermented milk complexed with Lactococcus lactis subspecies crmoris LRCC5306 (accession number KCCM11733P), which is a novel strain having excellent ability to produce diacetyl, and kefir grains, And a buttery flavor, and it has a salty taste and a rich flavor, and thus it can be substituted for synthetic sodium glutamate (MSG). Thus, the present invention has been accomplished.

Accordingly, the present invention relates to a method for producing lactose, which is excellent in the ability to produce diacetyl Lactococcus lactis subspecies crmoris LRCC5306 lactis subsp . cremoris , accession number KCCM11733P) , and drying the fermented milk fermented by adding the kefir grain to the milk.

It is another object of the present invention to provide a method for producing the natural seasoning material.

The object of the present invention is not limited to the above-mentioned object. The objects of the present invention will become more apparent from the following description, which will be realized by means of the appended claims and their combinations.

In order to achieve the above object, the present invention includes the following configuration.

The present invention provides a powdered natural seasoning material obtained by drying a lyophilized powder of Lactococcus strain having excellent diacetyl production ability and a fermented milk fermented by adding kefir grain to milk.

The culture medium for the culture of the Lactococcus strain includes peptone, sodium acetate, ammonium citrate, magnesium sulfate, manganese sulfate, and dipotassium phosphate ), And tween (Tween). This is a helpful factor for the growth of the strain.

The lyophilized powder of the culture broth of Lactococcus strain prepared in the above-mentioned culture medium can be obtained by cultivating a strain of a strain selected from the group consisting of glucose, malto-dextrin and starch, Or more of the above additives, and this is a factor that contributes to the stability of the lactic acid bacteria in the lyophilized powder as an excipient during lyophilization of the strain.

Preferably, the Lactococcus strain is selected from the group consisting of Lactococcus lactis cremoris ) LRCC5306 (Accession No. KCCM11733P). This is particularly advantageous in that the amount of diacetyl produced is particularly high, giving a high cheese and butter flavor.

Further, the kefir grain of the present invention is composed of strains such as yeast, lactococcus lactis, leuconostach mesenteroid, streptococcus thermophilus and the like, which is obtained by adding glucose and maltodextrin and lyophilized It is added to milk in the form of powder and left to stand at 25 ~ 35 ℃ for 9 ~ 15 hours to become fermented milk.

The fermented milk according to the present invention is obtained by fermenting a mixture of 0.1 to 0.5 g of lyophilized powder of Lactococcus lactis subspecies crmoris strain and 0.1 to 0.5 g of kefir grains on the basis of 250 ml of milk, 0.1 to 0.3 g of lyophilized powder of the Tissue subspecies crmoris strain and 0.1 to 0.3 g of kefir grains can be used. The natural seasoning material according to the present invention is obtained by spray-drying the fermented milk and has a powder form.

The present invention also relates to a pharmaceutical composition comprising (a) Lactococcus lactis subspecies crurus LRCC5306 subsp . Cremoris , Accession No. KCCM11733P) was prepared and lyophilized to prepare a lyophilized powder of lactic acid bacteria; (b) adding the lactic acid bacteria lyophilized powder and kefir grain to milk and culturing to produce a fermented milk; And (c) spray drying the fermented milk to obtain a natural seasoning material.

The step (a) is a step of preparing a lyophilized powder of lactic acid bacteria, wherein the dry powder further comprises at least one selected from the group consisting of glucose, malto-dextrin and starch. This is a factor that helps the stability of lactic acid bacteria in freeze-dried powder as an excipient during lyophilization of strain.

Next, in step (b), 0.1 to 0.5 g of the dried lactic acid bacterium powder and 0.1 to 0.5 g of the kefir grains are mixed on the basis of 250 ml of milk, preferably 0.1 to 0.5 g of the lyophilized powder of Lactococcus lactis subsp. To 0.3 g of the mixture and 0.1 to 0.3 g of kefir grains are mixed and allowed to stand at a temperature of 25 to 35 ° C for 9 to 15 hours to be cultured. In the case of the fermented milk obtained in the step (b), the cheese, buttery, Richness and the like are given.

The fermented milk of the step (c) may further contain 2 to 6% by weight of at least one additive selected from the group consisting of glucose, malto-dextrin and starch, based on the total weight of the fermented milk . This helps the growth stability of lactic acid bacteria, so it increases the flavor and diacetyl production and helps to improve the flavor of chitin.

The natural seasoning material according to the present invention is provided as a natural seasoning capable of replacing synthetic sodium glutamate (MSG) and the like due to the salty taste and the richness taste produced by the yeast and the like in the kefir grain, .

The kefir fermented milk prepared by the combined fermentation of Lactococcus lactis crmorris strain and the kefir grain according to the present invention has a calorie sensation and a rich flavor inherent to kefir together with a cheese and butter flavor peculiar to diacetyl, It is possible to preserve the characteristics and flavor, and it can be applied as a natural seasoning material to foods and the like.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a graph showing flavor components analyzed from 1 to 3 for comparison and an electronic tongue for fermented milk of Example 1. FIG.
Fig. 2 is a graph showing taste components analyzed by an electronic tongue for a seasoning material obtained by spray-drying the fermented milk of Example 1. Fig.

Hereinafter, the present invention will be described in detail by way of examples. The embodiments of the present invention can be modified into various forms as long as the gist of the invention is not changed. However, the scope of the present invention is not limited to the following embodiments.

In the following description, well-known functions or constructions are not described in detail since they would obscure the invention. As used herein, " comprising " means that other elements may be included unless otherwise specified.

Manufacturing example  One: LRCC5306 strain  Preparation of powder

The Lactococcus lactis subspecies crmoris LRCC5306 strain was prepared in powder form for inoculation into fermented milk.

Specifically, a strain in a vial form at -80 ° C. was inoculated on a MRS-BCP solid medium and cultured at 30 ° C. for 24 hours to form a colony. Then, the strain was inoculated into 50 ml of the MRS liquid medium and cultured under the same conditions. .

Next, 50 ml of the prepared seed culture was inoculated into a 3 L MRS liquid medium, cultured under the same conditions as above, and centrifuged (10,000 rpm, 10 minutes) to collect only the cells. 5% by weight of maltodextrin was added to the total weight of the recovered cells, followed by freeze-drying to complete the lactic acid bacterium powder. The finished lactic acid bacteria powder was about 230 g and the number of live bacteria in the powder was 3.1 x 10 ¹⁰ CFU / g.

Comparative Example  And Example : Preparation of fermented milk

The kefir fermented milk was prepared using the lyophilized powder of Lactococcus lactis subspecies crmoris LRCC5306 strain and the kefir grain prepared above.

Specifically, the kefir grains were commercially available from DuPont (CHOOZIT DT, Dupont, USA). 0.1 g of lyophilized powder of LRCC5306 strain and 0.1 g of kefir grain were added to 250 ml of commercially available milk, and the mixture was allowed to stand in an incubator maintained at 30 캜 for 12 hours to prepare fermented milk (Example 1).

Fermented milk fermented with LRCC5306 strain powder alone (Comparative Example 2) and Kefirgrain alone (Comparative Example 3) was prepared as a control.

Experimental Example  1: pH, Appearance, and Diacetyl  Content analysis (1) and Electronic tongue  Analysis of flavor components through (2)

The fermented milk prepared in the above Comparative Examples and Examples were analyzed for pH, flavor and diacetyl content, and the results are shown in Table 1. The flavorings were measured with an electronic tongue (Taste Sensing System SA402B, Insent Co., Ltd., Japan) To compare the taste profile and compare the values obtained when the milk before fermentation (Comparative Example 1) was taken as a zero point (FIG. 1).

Item Comparative Example 1
(Blank) Comparative Example 2
(LRCC5306 strain alone) Comparative Example 3
(Kefir grain
Exclusive) Example 1 pH 6.4 5.2 4.3 3.8 Appearance Liquid phase Semi-solid elegance elegance Diacetyl
(mg / kg) 0.0 17.6 1.7 23.7

The results of [Table 1] and [Figure 1] show that when the LRCC5306 strain alone fermented (Comparative Example 2), the pH did not fall to the general fermented milk level although the pH was slightly decreased, And salty taste were slightly increased. The amount of diacetyl produced was about 17.6 mg / kg.

In addition, when fermented with Kefir grain alone (Comparative Example 3), the pH was 4.3, indicating that general fermented milk was produced, the sourness was slightly increased, the salty taste and the richness (abundant) were rapidly increased but the diacetyl production was 1.7 mg / kg.

On the other hand, when the LRCC5306 strain and the kefir grain were combined (Example 1), the pH was 3.8, and it was confirmed that the sour taste, salty taste, and full flavor (national) increased to a higher level than that of the kefir grain alone fermentation. The amount of diacetyl produced at this time was 23.7 mg / kg, and it was found that diacetyl was produced at a higher level than LRCC5306 alone or when it was fermented by Kefir grain alone. In the case of complex fermentation, It is presumed that the products complement each other.

Experimental Example  2: Comparison of amino acid components

In order to compare amino acid components of the fermented milk prepared in the above Comparative Examples and Examples, amino acid composition was analyzed by an UPLC-MS / MS (Ultra Performance Liquid Chromatography-Tandem Mass Spectrometer) system.

amino acid Comparative Example 1
(Blank) Comparative Example 2
(LRCC5306 strain
Exclusive) Comparative Example 3
(Kefir grain
Exclusive) Example 1 Histidine 1.71 2.16 2.94 3.51 Asparagine 2.55 5.31 11.27 13.37 Serine 4.16 8.23 12.11 14.27 Arginine - 0.36 1.01 1.63 Glycine 1.11 1.65 1.89 2.16 Aspartic acid 1.31 4.72 13.18 13.84 Glutamic acid 0.07 0.15 3.14 4.27 Threonine - - - - Alanine 0.57 1.13 2.28 3.25 Proline 3.53 4.01 4.39 4.44 Cysteine 7.52 9.45 29.95 35.96 Lysine 2.73 3.52 10.82 12.74 Tyrosine 2.15 2.27 9.17 12.9 Methionine - - - - Valine 1.89 1.93 2.03 1.98 Isoleucine 3.42 5.32 6.94 8.8 Leucine 7.13 7.23 7.93 8.39 Phenylalanine 0.78 1.15 5.66 6.32 Tryptophan - - 1.11 1.76

The result of the above [Table 2] shows that when the fermentation of the strain and the kefir grain is performed in a similar manner to the analysis of the electronic tongue of [Fig. 1], richer amino acids are produced .

Experimental Example  3: For fermented milk dry powder Electronic tongue  Analysis of flavor components through

The LRCC5306 strain prepared in Example 1 and fermented milk complexed with kefir grain were spray dried to prepare seasonings.

Specifically, 4% by weight of maltodextrin was added to the fermented milk of Example 1 and then dried at 220 by using a spray dryer. The flavor component profile of the dried powder was analyzed and shown in Fig.

In the result of FIG. 2, it was confirmed that the sour taste, the salty taste, the full-bodied taste (overall) and the like of the Example 1 according to the present invention were increased, and the richness (tail) was slightly increased. Also, the content of diacetyl was 461.5 mg / kg, and it was confirmed that flavor of cheese was felt in addition to fermented milk flavor.

Therefore, through the above experimental example, the natural seasoning material according to the present invention is provided with a cheese and butter flavor through the increase of diacetyl, and the synthetic sodium glutamate (MSG) and the like It is natural seasoning that can be done.

The present invention has been described in detail. However, the scope of rights of the present invention is not limited thereto, but is defined by the following claims.

Claims (8)

A lyophilized powder of Lactococcus lactis subsp. Cremoris strain Lactococcus lactis subsp. Cremoris with an accession number KCCM11733P, which is excellent in diacetyl production ability, and powder form obtained by drying fermented milk fermented by adding kefir grain to milk Natural seasoning material.
The fermented milk according to claim 1, wherein the fermented milk is obtained by mixing 0.1 to 0.5 g of a lyophilized powder of Lactococcus lactis subspecies crmoris strain and 0.1 to 0.5 g of a kefir grain on the basis of 250 ml of milk, Seasoning material.
The fermented milk according to claim 1, wherein the fermented milk further comprises 2 to 6% by weight of at least one additive selected from the group consisting of glucose, malto-dextrin and starch, based on the total weight of the fermented milk A natural seasoning material that features.
(a) preparing a lyophilized powder of lactic acid bacteria by lyophilizing a culture solution obtained by culturing Lactococcus lactis subsp. cremoris strain having an accession number KCCM11733P, which is superior in diacetyl production ability, to Lactococcus lactis subsp. cremoris ;
(b) adding the lactic acid bacteria lyophilized powder and kefir grain to milk and culturing to produce a fermented milk; And
(c) spray drying the fermented milk to obtain a natural seasoning material;
≪ RTI ID = 0.0 > 1, < / RTI >
5. The method of claim 4, wherein the freeze-dried powder of step (a) comprises at least one additive selected from the group consisting of glucose, malto-dextrin, and starch, And 2 to 6% by weight based on the weight of the seasoning.
[6] The method of claim 4, wherein the step (b) is performed by mixing 0.1 to 0.5 g of the lactic acid bacteria-dried powder and 0.1 to 0.5 g of the kefir grain on the basis of 250 ml of milk.
[6] The method of claim 4, wherein the fermented milk of step (c) comprises at least one additive selected from the group consisting of glucose, malto-dextrin and starch in an amount of 2 to 6 wt% % ≪ tb >< tb >< / TABLE >
[6] The method of claim 4, wherein the cultivation in step (b) is performed at a temperature of 25 to 35 DEG C for 9 to 15 hours.

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