KR102723128B1 - Yogurt using rice bran ferment and process for preparing the same - Google Patents

Abstract

The present invention relates to yogurt using a rice bran culture solution and a method for producing the same, and more specifically, to yogurt using a rice bran culture solution which not only exhibits rich nutrients and taste but also has various functions such as antioxidant function, hangover relief function, liver function improvement function, and fatigue recovery function, and a method for producing the same. The rice bran yogurt according to the present invention can be produced by appropriately utilizing lactic acid bacteria derived from fermented foods and a medium suitable for exhibiting functionality and palatability, so that consumers can readily select and use it. In addition, by utilizing rice husk and rice bran, which are recyclable resources, as a nutrient source for the medium, the material cost is low, so that a high value-added product can be provided.

Description

Yogurt using rice bran ferment and process for preparing the same {Yogurt using rice bran ferment and process for preparing the same}

The present invention relates to yogurt using rice bran culture and a method for producing the same, and more specifically, to yogurt which not only exhibits rich nutrients and taste, but also has various functions such as antioxidant function, hangover relief function, liver function improvement function, and fatigue recovery function, and a method for producing the same.

Yogurt is a type of fermented milk. It is a curdled culture that is fermented by inoculating lactic acid bacteria into milk. It is relatively acidic and has a refreshing flavor. Whole milk or skim milk is used as the raw material, and skim milk powder, sweetened condensed milk, agar, etc. are added for flavor and appearance, or sugar, honey, etc. are added as sweeteners, and after homogenization, it is heat-sterilized (95℃, 5 minutes) and used.

The lactic acid bacteria used in making yogurt are Lactobacillus bulgaricus , Streptococcus thermophilus , and Lacidophillus . They are cultured at low temperatures and two or three types are mixed appropriately to give the product a unique flavor. Consumption of fermented milk with lactic acid bacteria has the functions of promoting digestion and improving intestinal health.

In the case of yogurt, texture is a very important quality variable in fermented milk. It is very important to provide a smooth mouthfeel, taste, and an appropriate level of viscosity.

In addition, high viscosity of fermented milk products can be obtained by using lactic acid bacteria cultures that produce exopolysaccharides, and at the same time, the density of viscosity is also very important.

Recently, scientific research is being conducted on the health promotion effects of lactic acid bacteria supplied by yoghurt rather than the nutritional value of yoghurt and the effect of promoting digestion and absorption. In the intestines of infants, intestinal microorganisms centered on Bifidobacterium, a type of lactic acid bacteria, form a flora, which suppresses the settlement and proliferation of harmful microorganisms and maintains intestinal health. In addition, it is said that there are antibacterial, anticancer, and immune-enhancing effects obtained by lowering the pH of intestinal contents by lactic acid produced as yoghurt grows, and the production of antibiotics and neutralizing toxins.

These yogurts are dairy products with improved nutrition and digestibility of milk, and their unique flavor and various physiological functions have led to a steady increase in demand worldwide. In Korea, the supply of fermented milk and lactic acid bacteria has gradually expanded, and not only has the functional food material been strengthened by enhancing dietary fiber, oligosaccharides, vitamins, and minerals, but the fermented milk market is becoming more active by developing products that emphasize health aspects such as improving constipation, stomach, and liver function.

As a technology related to yogurt, Korean Publication No. 10-1999-0047128 (July 5, 1999) discloses a process for manufacturing a wolfberry extract; a process for preparing a yogurt medium and sterilizing it; and a yogurt manufactured by a yogurt culture and homogenization process.

In addition, Korean registration number 10-1328362 (November 5, 2013) discloses a method for providing highly functional yogurt beneficial to the human body by using makgeolli, which is rich in physiologically active substances such as various lactic acid bacteria, yeast, vitamin B group, essential amino acids, and glutathione, as a raw material.

Korean Registration No. 10-1408974 (June 11, 2014) discloses a method for producing yogurt using a fermentation promoting composition and yogurt produced by the method, which comprises the steps of: mixing milk and sugar to produce a yogurt mix (A100); sterilizing and cooling the yogurt mix (A200);

A method for manufacturing yogurt, comprising: a step (A300) of inoculating a strain into a yogurt mix that has gone through the above A200 step; and a step (A400) of fermenting the yogurt mix that has gone through the above A300 step;

In the above A400 step, a fermentation promoting composition is added,

The above fermentation promoting composition is added in an amount of 2 to 7 parts by weight per 100 parts by weight of the yogurt mix that has gone through the A300 step.

The above fermentation promoting composition applies a hydroxy radical generating composition in the form of a ball, flake or powder, or a superoxide generating composition in the form of a ball, flake or powder,

The above hydroxy radical generating composition is formed by a structure in which a shell (200) is coated on the surface of a core (100), and the core (100) is formed by a superoxide generating compound (11) being fixed to the surface of a first silica precursor (10), and the shell (200) is formed by a transition metal compound (21) being fixed to the surface of a second silica precursor (20).

The above superoxide generating composition is formed by a structure in which a shell (200') is coated on the surface of a core (100'), and the core (100') is formed by a superoxide generating compound (11') being fixed to the surface of a first silica precursor (10'), and the shell (200') is formed by a calcium compound (21') being fixed to the surface of a second silica precursor (20').

The above first silica precursor (10, 10') is a silica sol, which is a mixture of 20 to 40 wt% of powdered silicon oxide (SiO2) having a particle size of 02 to 10 and 60 to 80 wt% of water.

The above superoxide generating compound (11,11') is used alone or in combination of two or more of silver nitrate (AgNO3), gold chloride (AuCl3, HAuCl4), or platinum chloride (PtCl4).

The above second silica precursor (20, 20') is used alone or in combination of two or more kinds from among tetraethoxyorthosilicate (TEOS), methyltrimethoxysilane (MTMS), tetramethoxyorthosilicate (TMOS), tetrapropoxyorthosilicate (TPOS), tetrabutoxyorthosilicate (TBOS), tetrapentoxyorthosilicate (TPEOS), tetra(methylethylketooxymo)silane, vinyloxymosilane (VOS), phenyl tris(butanoneoxime)silane (POS), and methyloxymosilane (MOS).

The above transition metal compound (21) is characterized in that it is used alone or in combination among iron salt compounds or copper salt compounds, and the above calcium compound (21') is characterized in that it is used alone or in combination among calcium oxide or calcium hydroxyoxide.

Korean Publication No. 10-2019-0052193 (May 16, 2019) includes: a step of washing and soaking barley rice to prepare and wash barley rice to be fermented, and then soaking the barley rice in water to make the barley rice soaked (S100); a step of mixing barley rice and fermentation bacteria (S200) to remove water from the soaked and soaked barley rice and then mix it with fermentation bacteria; a step of culturing barley rice and fermentation bacteria (S300) to place the barley rice mixed with the fermentation bacteria in a sealed container and then multiply yeast to make fermented barley nuruk; a step of mixing and soaking non-glutinous rice and glutinous rice (S400) to prepare non-glutinous rice and glutinous rice, wash them, mix them at a constant weight ratio, and soak them in water to make the barley rice and glutinous rice porridge to make the porridge (S500); A method for producing a functional fermented beverage is disclosed, which includes a porridge cooling step (S600) of cooling porridge manufactured using the above non-glutinous rice and glutinous rice to control the moisture content contained in the porridge; a porridge fermentation and maturation step (S700) of mixing the barley malt and porridge at a certain weight ratio and maturing them; and a processing step (S800) of processing the fermented and maturated porridge to produce a product.

Korean registration number 10-1846277 (April 2, 2018) discloses a natural healthy sweet potato yogurt and a method for producing the same, which not only preserves the natural color of sweet potatoes but also does not contain any additives such as flavoring or coloring.

Korean Publication No. 10-2020-0066985 (June 11, 2020) discloses a method for producing milk tea yogurt with enhanced antioxidant efficacy, comprising the steps of: 1) mixing milk, black tea leaf powder, and sugar, sterilizing the mixture, and cooling to remove sediment; 2) filtering the mixture to produce a milk tea composition; 3) mixing a rosemary extract into the composition and inoculating it with lactic acid bacteria; and 4) culturing the lactic acid bacteria to ferment the milk tea composition into yogurt.

Recently, with the development of various fermentation technologies and the improvement of living standards, various functional products are being developed one after another, and in addition to these functions, consumers' eyes may be drawn to products that combine taste, nutrition, and palatability.

Korean Publication Number 10-1999-0047128 (July 5, 1999) Korean Publication Number 10-2009-0048903 (May 15, 2009) Korean Publication No. 10-2020-0066985 (June 11, 2020) Korean registration number 10-1328362 (November 5, 2013) Korean Registration No. 10-1846277 (April 2, 2018)

The inventors of the present invention have conducted extensive research to develop a yogurt that not only exhibits rich nutrients and taste, but also has various functions such as antioxidant function, hangover relief function, liver function improvement function, and fatigue recovery function. As a result, they discovered that yogurt manufactured using a rice bran culture medium suitable for the growth of starter as a medium can satisfy the above requirements, and thus completed the present invention.

Accordingly, the purpose of the present invention is to provide a yogurt and a method for producing the same, which not only exhibits rich nutrients and taste but also has various functions such as antioxidant function, hangover relief function, liver function improvement function, and fatigue recovery function by producing the yogurt using a rice bran culture medium suitable for the growth of starter as a medium.

The purpose of the present invention as described above is, on one hand,

a) A step of preparing a rice bran medium by adding rice bran or rice bran extract and arginine to distilled water and sterilizing at 121°C for 20 minutes;

b) A step of activating the lactic acid bacteria strain isolated from fermented food by repeatedly culturing it in MRS medium;

c) Step of inoculating a lactic acid bacteria strain confirmed to form ornithine into MRS medium containing 1% arginine;

d) A step of primary culturing the medium inoculated with the lactic acid bacteria strain at 37°C for 48 hours;

e) A step of centrifuging the cultured medium at 3,500 RPM for 20 minutes to separate the lactic acid bacteria and the medium;

f) A step of washing the lactic acid bacteria separated from the medium by adding sterilized water;

g) Step of adding rice bran medium to the washed lactic acid bacteria and performing secondary culture at 37℃ for 48 hours;

h) A step of manufacturing rice bran yogurt by adding sugar and milk to a rice bran medium cultured with lactic acid bacteria and fermenting at 37°C for 6 hours;

i) a step of packaging the manufactured rice bran yogurt into a certain unit; and

j) A method for producing rice bran yogurt, characterized by including a step of refrigerating the packaged rice bran yogurt.

The rice bran yogurt according to the present invention is manufactured by appropriately utilizing lactic acid bacteria derived from fermented foods and a medium suitable for demonstrating functionality and palatability, so that consumers can easily select and use it. In addition, by utilizing rice husk and rice bran, which are recyclable resources, as a nutrient source for the medium, the material cost is low, so a high value-added product can be provided.

Figure 1 is a process diagram for manufacturing yogurt using rice bran extract according to the present invention.
Figure 2 is a photograph of yogurt fermented by adding the rice bran culture solution of the present invention.
Figure 3 is a drawing confirming the growth of lactic acid bacteria using BCP medium.
Figure 4 is a graph showing the growth-confirmed ornithine activity confirmed by TLC.
Figure 5 is a graph showing the antioxidant activity of yogurt manufactured using rice bran culture solution.
Figure 6 is a graph showing the polyphenol content of yogurt manufactured using rice bran culture solution.
Figure 7 is a photograph showing the ornithine production rate of lactic acid bacteria using a TLC plate.
Figure 8 is a graph showing the ornithine standard.
Figure 9 is a photograph showing microbial deposition information for the lactic acid bacteria strain of the present invention.

The present invention, in one aspect,

a) A step of preparing a rice bran medium by adding rice bran or rice bran extract and arginine to distilled water and sterilizing at 121°C for 20 minutes;

b) A step of activating the lactic acid bacteria strain isolated from fermented food by repeatedly culturing it in MRS medium;

c) Step of inoculating a lactic acid bacteria strain confirmed to form ornithine into MRS medium containing 1% arginine;

d) A step of primary culturing the medium inoculated with the lactic acid bacteria strain at 37°C for 48 hours;

e) A step of centrifuging the cultured medium at 3,500 RPM for 20 minutes to separate the lactic acid bacteria and the medium;

f) A step of washing the lactic acid bacteria separated from the medium by adding sterilized water;

g) Step of adding rice bran medium to the washed lactic acid bacteria and performing secondary culture at 37℃ for 48 hours;

h) A step of manufacturing rice bran yogurt by adding sugar and milk to a rice bran medium cultured with lactic acid bacteria and fermenting at 37°C for 6 hours;

i) a step of packaging the manufactured rice bran yogurt into a certain unit; and

j) A method for producing rice bran yogurt is provided, characterized by including a step of refrigerating packaged rice bran yogurt.

The present invention, in a further aspect,

The above lactic acid bacteria

a1) Step of diluting fermented food with peptone water;

a2) Step of culturing the diluted solution in MRS medium;

a3) Step of obtaining colonies from the medium in which culture has been completed;

a4) A step of isolating a strain that is dark yellow and does not form a ring among the colonies; and

a5) A method for producing rice bran yogurt is provided, characterized by confirming the ornithine production ability of the separated strain by TLC.

The present invention, in a further aspect, provides a method for producing rice bran yogurt, characterized in that the lactic acid bacteria strain is Pediococcus pentosaceus MAC-11 (KCCM11359P) lactic acid bacteria strain.

The present invention, in another additional aspect, provides rice bran yogurt characterized by being manufactured by the above method.

Hereinafter, the rice bran yogurt of the present invention and its manufacturing method will be described in more detail through preferred examples.

Hereinafter, specific details for implementing the present disclosure will be described in detail with reference to the attached drawings. However, in the following description, specific descriptions of widely known functions or configurations will be omitted if there is a risk of unnecessarily obscuring the gist of the present disclosure.

In the attached drawings, identical or corresponding components are given the same reference numerals. In addition, in the description of the embodiments below, the description of identical or corresponding components may be omitted. However, even if the description of a component is omitted, it is not intended that such a component is not included in any embodiment.

The advantages and features of the disclosed embodiments, and the methods for achieving them, will become apparent with reference to the embodiments described below together with the accompanying drawings. However, the present disclosure is not limited to the embodiments disclosed below, but may be implemented in various different forms, and these embodiments are only provided to make the present disclosure complete, and to fully inform a person having ordinary skill in the art to which the present disclosure belongs of the scope of the invention.

Hereinafter, terms used in this specification will be briefly described, and the disclosed embodiments will be described in detail. The terms used in this specification have been selected from widely used general terms as much as possible while considering the functions in this disclosure, but this may vary depending on the intention of engineers engaged in the relevant field, precedents, the emergence of new technologies, etc. In addition, in certain cases, there are terms arbitrarily selected by the applicant, and in this case, the meanings thereof will be described in detail in the description of the relevant invention. Therefore, the terms used in this disclosure should be defined based on the meanings of the terms and the overall contents of this disclosure, rather than simply the names of the terms.

In this specification, singular expressions include plural expressions unless the context clearly specifies that they are singular. In addition, plural expressions include singular expressions unless the context clearly specifies that they are plural. When a part throughout the specification is said to "include" a certain element, this does not mean that other elements are excluded, but rather that other elements can be included, unless otherwise stated.

The inevitable production of ROS (reactive oxygen species) by aerobic organisms is a health threatening factor, and although antioxidants exist in the human body, the activity of antioxidants decreases due to aging, lifestyle habits, etc. The present invention has a technical feature in that it provides a popular yogurt that helps maintain health by providing various functionalities including antioxidant function.

FIG. 1 is a process diagram for manufacturing yogurt using rice bran extract according to the present invention, FIG. 2 is a photograph of yogurt fermented by adding the rice bran culture solution of the present invention, FIG. 3 is a drawing for confirming the growth of lactic acid bacteria using BCP medium, FIG. 4 is a graph for confirming ornithine activity with confirmed growth by TLC, FIG. 5 is a graph showing the antioxidant activity of yogurt manufactured using the rice bran culture solution, FIG. 6 is a graph showing the polyphenol content of yogurt manufactured using the rice bran culture solution, FIG. 7 is a photograph for confirming the ornithine production rate of lactic acid bacteria using a TLC plate, FIG. 8 is a graph showing an ornithine standard, and FIG. 9 is a photograph of a microbial deposit certificate for the lactic acid bacteria strain of the present invention.

The method for manufacturing yogurt using rice bran extract according to the present invention largely includes a medium manufacturing step, an activation step, an inoculation step, a primary culture step, a separation step, a washing step, an addition step, a secondary culture step, a tertiary culture step, a packaging step, and a storage step, as shown in Fig. 1. Hereinafter, the individual steps will be described in detail.

a) Badge manufacturing step

This is a step for preparing rice bran (rice bran) or rice bran extract for culturing lactic acid bacteria (lactic acid bacteria) in distilled water. 80 to 120 g of rice bran and 8 to 12 g of arginine are added to 1 L of distilled water, and then sterilized at 121°C for 20 minutes to create a medium for culturing lactic acid bacteria.

Rice bran contains a large amount of gamma-oryzanol, which has an antioxidant power approximately 200 times that of natural tocopherol (vitamin E), and can provide antioxidant and various other functional properties by using it as a nutrient source for microorganisms.

It may be desirable for the content of the above rice bran to be included in an amount of 80 to 12 parts by weight based on 100 parts by weight of distilled water for proper proliferation of lactic acid bacteria.

When sterilizing rice bran extract, it may be desirable to stop operation when the temperature of the autoclave reaches 90 to 100℃ and add the rice bran extract to sterilize.

The above arginine (L-Arginine) is a component necessary for lactic acid bacteria to produce ornithine (L-Ornithine). The taste of arginine may vary depending on the production site and process, and a high arginine content may result in a strong bitter taste. Therefore, it may be desirable to include 0.8 to 1.2 parts by weight of arginine based on 100 parts by weight of distilled water.

b) Activation phase

Separately, lactic acid bacteria strains isolated from fermented foods (e.g., cheonggukjang) or Pediococcus pentosaceus MAC-11 lactic acid bacteria strains were repeatedly cultured on MRS [DE MAN, ROGOSA and SHARPE (MRS) agar medium (Difco, Detroit , MI, USA) supplemented with 1% arginine at 37°C for 48 hours to activate the bacteria.

The above MRS medium contains 10 g/L isomalto-oligosaccharides, 40 g/L lactose, 10 g/L peptone, 10 g/L beef extract, 5 g/L yeast extract, 7 g/L sodium acetate, 2 g/L disodium phosphate, 2 g/L ammonium citrate, 1 g/L tween 80, 005 g/L MgSO4, 005 g/L maleic acid, 05 g/L MnSO4, 05 g/L L-cysteine HCl.

The activation of the above lactic acid bacteria

a1) Step of diluting fermented food with peptone water;

a2) Step of culturing the diluted solution in BCP medium;

a3) a step of isolating a colony that is dark yellow and does not form a ring from the cultured medium; and

a4) It may be desirable to perform the step of confirming the ornithine production ability of the separated strain by TLC.

Meanwhile, in the isolation of beneficial lactic acid bacteria from fermented foods (e.g., cheonggukjang), it may be desirable to isolate lactic acid bacteria capable of producing ornithine as an indicator of the production capacity of physiologically active substances.

Specifically, in the step of diluting the fermented food with peptone water, the fermented food sample is diluted with peptone water in the range of 10 ^-1 to 10 ^-8 .

The above fermented foods are not limited to these, and representative examples include cheonggukjang, meju, and doenjang.

Next, it may be desirable to dispense 0.1 mL of the diluted solution and culture it 2 to 5 times on MRS medium supplemented with 0.002% Bromocresol purple (BCP) (Sigma Chemical Co. St. Louis, USA) to activate the bacteria, and culture it at 37°C for 48 hours to obtain colonies.

The above medium has the characteristic of changing color according to the change in pH, and can be used to check for contamination and activation of lactic acid bacteria. When lactic acid bacteria grow, the color around the colony in the purple medium changes to yellow.

In the colony separation step, the colonies that do not form rings and are dark yellow are selected as the primary candidate lactic acid bacteria strains and separated.

In the next step of confirming the ornithine production ability of the isolated strain by TLC, the colonies selected as the primary candidate are cultured in MRS (Difco, Detroit, MI, USA) broth supplemented with 1% arginine at 37°C for 48 hours, and then the ornithine production ability is examined using TLC.

TLC may be preferably performed using silica gel F ₂₅₄ and standard ornithine (Merck, Damstadt, Germany), and a mobile phase (Butanol: Acetic acid: Dichoromethanol: water = 5:3:3:3).

In order to check the ornithine production ability (including production time efficiency) of lactic acid bacteria, it may be desirable to perform TLC at 24-hour intervals. As confirmed in the examples described below, the measurement results showed that lactic acid bacteria cultured for 48 hours produced more ornithine.

After confirming the activity through TLC, it was confirmed to have good activity and was finally selected and used as a lactic acid bacteria for producing yogurt.

A strain isolated in a natural state from a fermented food through an activation step as described above can be preferably used.

In addition, Pediococcus pentosaceus MAC-11 strain, a lactic acid bacteria strain isolated from cheonggukjang, can be more preferably used.

The above lactic acid bacteria strain is a lactic acid bacteria with the metabolic ability to convert arginine into ornithine, and ornithine can provide beneficial functions such as relieving hangovers, improving liver function, and reducing fatigue after exercise.

The above strain is a strain of the genus Pediococcus , known as one of the important dominant species that determines the flavor of Cheonggukjang. In order to isolate lactic acid bacteria that produce ornithine from Cheonggukjang, among the colonies grown on a medium supplemented with bromocresol purple, those that did not form rings and were dark yellow were selected as candidate strains. The ornithine production potential of the candidate strains was investigated using TLC, and four candidate strains with excellent ornithine production ability were selected through TLC.

Among the selected strains, the morphological and biochemical characteristics of MAC-11 were investigated. The isolated strain was a gram-positive coccus that produced carbon dioxide under anaerobic conditions. The isolated lactic acid bacteria strain produced acids by decomposing L-arabinose, ribose, D-xylose, and glucose, but did not decompose adonitol, sorbitol, or starch (see Table 1). The selected strain grew well in MRS medium at 37℃ and pH 6.5, and the temperature and pH for growth were 37℃ and pH 6.5, respectively.

In Table 1 above, + means positive and - means negative.

The above strain is a strain disclosed in Korean Publication No. 10-2017-0073839 (June 29, 2017), and the deposit information is as follows.

d) Inoculation stage

This is a step for inoculating a lactic acid bacteria strain confirmed to form ornithine into an MRS medium containing 1% arginine. It may be desirable to inoculate 1 mL of lactic acid bacteria per 50 mL of MRS medium.

e) Primary culture stage

The medium inoculated with the lactic acid bacteria strain is cultured at 37°C for 24 to 48 hours. A culture time of 48 hours may be more desirable.

f) Separation stage

Centrifuge the cultured medium at 2,500 to 5,000 RPM for 20 minutes to separate the lactic acid bacteria and the medium (MRS broth).

g) Washing step

Wash the lactic acid bacteria separated from the medium with sterilized water.

h) Secondary culture stage

Next, 50 g of rice bran extract is added to 1 g of washed lactic acid bacteria and secondary culture is performed at 37°C for 48 hours.

i) Fermentation stage

In a sterilized glass bottle, add 40 to 50 g of secondary cultured rice bran extract, 2 g of sugar, and the remaining amount of milk (skimmed milk may be preferable) to make 100.

Next, rice bran yogurt is manufactured by fermenting at 37℃ for 6 hours. At this time, it may be more preferable to perform the fermentation under shaking conditions at 90 to 100 rpm.

Meanwhile, when making yogurt by directly adding arginine to milk, there is a problem of poor taste, so instead of directly adding arginine to milk, milk is added during the process of culturing lactic acid bacteria in a rice bran medium, thereby providing an improved method through fermentation of lactic acid bacteria, which constitutes another feature of the present invention.

j) Packaging stage

The rice bran yogurt manufactured in the above step is filled into packaging containers according to specifications and packaged.

k) Storage phase

Store packaged rice bran yogurt refrigerated (4-6℃) until use.

Next, after testing for appearance, foreign substances, moisture, bacterial count, E. coli, etc. in accordance with the standards and specifications, only suitable products are shipped.

The rice bran yogurt of the present invention may contain, in addition to its effective ingredients, a sweetener, a flavoring agent, a physiologically active ingredient, minerals, etc.

Sweeteners may be used in an amount that provides the food with an appropriate sweetness, and may be natural or synthetic. Preferably, natural sweeteners are used, and examples of natural sweeteners include sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose, and maltose.

Flavoring agents can be used to improve taste or aroma, and both natural and synthetic ones can be used. It is preferable to use natural ones. When using natural ones, in addition to flavor, the purpose of enhancing nutrition can also be used. Natural flavoring agents can be obtained from apples, lemons, tangerines, grapes, strawberries, peaches, etc., or from green tea leaves, dandelions, bamboo leaves, cinnamon, chrysanthemum leaves, jasmine, etc. In addition, those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, ginkgo nuts, etc. can be used. Natural flavoring agents can be liquid concentrates or solid extracts. In some cases, synthetic flavoring agents can be used, and synthetic flavoring agents can use esters, alcohols, aldehydes, terpenes, etc.

As physiologically active substances, catechins such as catechin, epicatechin, gallogatechin, and epigallocatechin, and vitamins such as retinol, ascorbic acid, tocopherol, calciferol, thiamine, and riboflavin can be used.

Minerals that can be used include calcium, magnesium, chromium, cobalt, copper, fluoride, germanium, iodine, iron, lithium, magnesium, manganese, molybdenum, phosphorus, potassium, selenium, silicon, sodium, sulfur, vanadium, and zinc.

In addition, the rice bran yogurt of the present invention may contain an acidulant, etc., in addition to the sweetener, etc., as needed.

It is preferable that these acidulants, etc. be added and used in extremely small amounts that can achieve the purpose for which they are added. When expressed numerically, the extremely small amount means a range of 0.0005 wt% to about 0.5 wt% based on the total weight of the food composition.

Preservatives that can be used include sodium calcium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, and EDTA (ethylenediaminetetraacetic acid).

Emulsifiers that can be used include acacia gum, carboxymethyl cellulose, xanthan gum, and pectin.

Acidulants that can be used include lactic acid, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid, etc. In addition to the purpose of enhancing taste, these acidulants can be added to the food composition to have an appropriate acidity for the purpose of inhibiting the growth of microorganisms.

Examples of viscosifiers that can be used include suspending agents, sedimentation agents, gelling agents, and bulking agents.

In a preferred embodiment, the rice bran yogurt according to the present invention can increase preference or taste by adding one or more ingredients selected from the group consisting of honey, malt, glycerin, red ginseng, cornelian cherry, poria, rehmannia glutinosa, scutellaria baicalensis, cordyceps sinensis, grapefruit extract, licorice, barley herb leaf powder, hawthorn extract, St. John's wort, selenium yeast, vitamin C, citric acid, nicotinic acid, sodium benzoate, aspartame, saccharin, pectin, malitol, sorbitol, xylitol, guar gum, skimmed milk powder, and oligosaccharides. It is appropriate to use these ingredients in an amount of about 50 to 80% based on the total weight of the composition of the present invention.

(Example)

Hereinafter, the present invention will be described in more detail based on the following examples. These examples are presented for the purpose of more easily explaining the present invention, and it will be apparent to those skilled in the art that the present invention is not limited to these examples, and that substitutions and equivalent other examples can be made without departing from the technical spirit of the present invention.

Example 1: Isolation of lactic acid bacteria from Cheonggukjang

The commercially available Cheonggukjang was diluted with peptone water to a range of 10 ^-1 to 10 ^-8 , and 0.1 mL of the diluted solution was dispensed into BCP medium [MRS medium supplemented with 0.002% Bromocresol purple (BCP) (Sigma Chemical Co. St. Louis, USA)]. The solution was cultured twice at 37°C for 48 hours to activate the bacteria. Among the colonies formed, those that did not form rings and were dark yellow were selected as the primary candidate for lactic acid bacteria strains and isolated. The characteristics of the grown bacteria are shown in Fig. 3. Fig. 3 is a drawing confirming the growth of lactic acid bacteria using BCP medium.

The colonies selected as the primary candidates were cultured in MRS (Difco, Detroit, MI, USA) broth containing 1% arginine at 37°C for 48 hours, and ornithine production ability was examined using TLC, and the results are shown in Fig. 4. Fig. 4 is a graph confirming ornithine activity (yellow) where growth was confirmed by TLC.

TLC was performed using silica gel F ₂₅₄ and standard ornithine (Merck, Damstadt, Germany), and mobile phase (Butanol: Acetic acid: Dichoromethanol: Water = 5:3:3:3). The results showed that lactic acid bacteria cultured for 48 hours produced more ornithine.

Example 2: Preparation of rice bran yogurt

100 g of rice bran and 10 g of arginine were added to 1 L of distilled water and sterilized at 121°C for 20 minutes to obtain a rice bran extract. When the temperature of the sterilizer (autoclave) reached 90–100°C, operation was stopped and the rice bran extract was added for sterilization.

In Example 1, 1 mL of the lactic acid bacteria strain confirmed to form ornithine was inoculated into 50 mL of MRS medium supplemented with 1% arginine. The medium inoculated with the lactic acid bacteria strain was first cultured at 37°C for 48 hours, and the cultured medium was centrifuged at 3,500 RPM for 20 minutes to separate the lactic acid bacteria and the medium (MRS broth). The lactic acid bacteria separated from the medium were washed with sterilized water, and 50 g of rice bran extract was added per 1 g of the washed lactic acid bacteria, followed by second culture at 37°C for 48 hours. 40 and 50 g of the second-cultured rice bran extract were each placed in two sterilized glass bottles, and 2 g of sugar and the remaining skim milk were added to make 100 g, and fermented at 90 to 100 rpm for 6 hours at 37°C to manufacture rice bran yogurt (see Fig. 2).

Example 3: Preparation of rice bran yogurt

Rice bran yogurt was prepared according to Example 2, but Pediococcus pentosaceus MAC-11 (KCCM11359P) isolated from cheonggukjang was used as the lactic acid bacteria.

Test Example 1: Analysis of antioxidant capacity of rice bran yogurt

DPPH and amino acid extraction were performed on the yogurt manufactured in Example 2 to confirm the antioxidant and ornithine content in the yogurt.

Commercial yogurt, yogurt containing 40% sugar and rice bran starter, and yogurt containing 50% sugar and rice bran starter were freeze-dried for use as samples, and vitamin C was used as a control. 5 mg of each sample and vitamin C were quantified in an EP tube, and diluted with DW according to the concentration used. After labeling in a 96-well plate, 200 ㎕ of 100% EtOH was dispensed for color correction, and 50 ㎕ of 100% EtOH was dispensed to the control group. 50 ㎕ of sample was dispensed for color correction and digestion, and 200 ㎕ of 0.2 mM DPPH was dispensed to the control group and specimen, and left in the dark at room temperature for 30 minutes. The absorbance was measured at 517 nm using TECAN, and the results are shown in Fig. 5. Fig. 5 is a graph showing the antioxidant activity of yogurt manufactured using rice bran culture.

As confirmed from the results of Fig. 5, the rice bran yogurt of the present invention was found to have superior antioxidant capacity compared to commercial products.

Test Example 2: Polyphenol Analysis of Rice Bran Yogurt

Freeze-dried samples (commercial yogurt, yogurt containing 2% sugar and 40% rice bran starter, yogurt containing 2% sugar and 50% rice bran starter) were individually quantified and diluted with DW. 50 ㎕ of each concentration sample was quantitatively dispensed into EP tubes, followed by 900 ㎕ of 1% Na ₂ CO ₃ solution, and then vortexed.

After mixing 50㎕ of Folin & Ciocalteu’s phenol regent, leaving it in the dark for 30 minutes, measuring the absorbance at 760 nm using TECAN, and quantifying the amount of polyphenol contained in the sample using a standard graph diluted by concentration, the results are shown in Fig. 6. Fig. 6 is a graph showing the polyphenol content of yogurt manufactured using rice bran culture solution.

From the results of Fig. 6, the rice bran yogurt of the present invention showed a higher polyphenol content than the commercially available comparative product.

Test Example 3: Analysis of ornithine content in rice bran yogurt

Amino acids were extracted from the freeze-dried rice bran yogurt of Example 3 and the ornithine content was analyzed. 200 mg of the sample was placed in an EP tube, 800 μL of free amino acid (methanol:chloroform:water=12:5:3) reagent was added, and centrifuged at 12,000 RPM for 15 min at 4°C, and the supernatant was separated.

After separating the supernatant, 200 μL of chloroform and 400 μL of 3DW were added to the solid to extract the excess amino acids, centrifuged at 12,000 RPM for 15 min at 4℃, then the supernatants were combined, filtered, and developed with TLC to check the ornithine content, and after developing the ornithine standard in advance, the size of the spot was measured to create a standard, and the amount of ornithine in the sample was calculated using the created standard. The calculation was performed using a graph with an R² value of 0.9 or higher, and the results are shown in Figs. 7 and 8. Fig. 7 is a photograph showing the ornithine production rate of lactic acid bacteria using a TLC plate, and Fig. 8 is a graph showing the ornithine standard.

As a result, yogurt containing 40% of 2% sugar and brown rice starter had 29 mg/mL, and yogurt containing 50% of 2% sugar and brown rice starter had 38 mg/mL (based on 200 mg of freeze-dried yogurt sample).

Test Example 4: Hangover Relief Test

As panel members who participated in the test, 50 male subjects were selected as experimental subjects. The experimental group (5 people) was asked to drink the yogurt of the examples and comparative examples 1 hour after drinking. The control group (5 people) was not given the product. In order to increase the accuracy of the experiment, they were prohibited from drinking alcohol for 10 days before the experiment. In addition, in order to ensure the same level of intoxication, they were asked to drink half a bottle (120 ml) of soju over 1 hour. The hangover evaluation items were evaluated on a 5-point scale based on the degree of relief from a total of 5 hangover symptoms: headache, nausea, thirst, fatigue, and nausea. The results are shown in Table 2.

Evaluation items Example 2 Example 3 Comparative Example 1 headache 4.8±0.5 4.8±0.44 3.2±0.4 Nausea 4.8±0.5 4.8±0.6 2.5±0.5 thirst 4.8±0.5 4.8±0.4 3.5±0.5 Tiredness 4.8±0.5 4.8±0.5 2.9±0.4 Nausea 4.8±0.6 4.8±0.6 3.1±0.3

As a result, it was observed that the rice bran yogurt according to the present invention is effective in relieving hangover symptoms that appear after drinking. In particular, hangover symptoms did not appear compared to usual after drinking, and although the hangover made it difficult to eat breakfast due to an upset stomach, it was found that the next day after taking the yogurt according to the present invention, the stomach felt comfortable even after eating breakfast.

Test Example 5: Measurement of alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) activities

For the products of the above examples and comparative examples, alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) activities were evaluated.

Alcohol dehydrogenase (ADH) is an enzyme produced in the liver that breaks down alcohol to create acetaldehyde, and acetaldehyde dehydrogenase (ALDH) is an enzyme that breaks down acetaldehyde, a toxic substance, into acetic acid and water. The most important thing in relieving a hangover is to activate the enzymes that break down alcohol. In addition, it is important to quickly restore liver function so that the liver, which produces alcohol dehydrogenase, can play its normal role.

The activity evaluation of the above alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) was performed by measuring the alcohol decomposition activity of the samples of Examples 2 to 3 and Comparative Examples using an alcohol and acetaldehyde analysis kit. The treatment group and the control group (distilled water) of each sample were dissolved in DMSO (dimethyl sulfoxide) at a concentration of 10 ㎍/㎕ and used in the experiment, and the activity of alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) in all experimental groups was measured for absorbance at 340 nm using an ultraviolet spectrophotometer. The activity of each sample was measured using an in vitro assay system and expressed as a relative rate to the control group. The experimental results are shown in Table 3 below.

Relative enzyme activity (%) Control group Example 2 Example 3 Comparative example ALD 100 159.71 158.35 86.43 ALDH 100 187.54 185.36 98.43

As a result, the samples of the examples showed higher ADH and ALDH activities than those of the comparative examples compared to the control group (distilled water), confirming that the samples had a liver function protective effect when consumed before and after alcohol consumption.

Test Example 6: Sensory Test

A sensory test was conducted on the yogurts of Examples 2 to 3 and Comparative Examples. The panel consisted of 30 trained inspectors who evaluated the appearance, aroma, taste, and overall preference using the following 5-point scale, and the average of each value was calculated and the results are shown in Table 4 below. The evaluation criteria were set to 5 for very good, 4 for slightly good, 3 for average, 2 for poor, and 1 for very poor.

Evaluation items Example 2 Example 3 Comparative example Appearance color 4.8±0.5 4.8±0.44 3.2±0.3 Incense 4.9±0.3 4.8±0.6 2.3±0.5 taste 4.8±0.5 4.8±0.33 3.2±0.4 Comprehensive evaluation 4.8±0.3 4.8±0.4 2.7±0.5

As can be confirmed from the results in Table 4 above, it was confirmed that the rice bran yogurt product of the example of the present invention had an overall superior effect in appearance, taste, and overall preference compared to the comparative example.

As described above, the rice bran yogurt according to the present invention not only increases the ornithine content but also reduces the amount of milk used, thereby reducing manufacturing costs. In addition, since the product is made without discarding the rice bran solids, the nutrients remaining in the rice bran can be efficiently consumed.

Although the present invention has been described with reference to preferred embodiments thereof, it will be understood by those skilled in the art that various modifications and changes may be made therein without departing from the spirit and scope of the invention as set forth in the claims below.

Korean Culture Center of Microorganisms (KCCM) KCCM11359P 20130129

Claims (4)

a) A step of preparing a rice bran medium by adding rice bran or rice bran extract and arginine to distilled water and sterilizing at 121°C for 20 minutes;
b) A step of activating the lactic acid bacteria strain isolated from fermented food by repeatedly culturing it in MRS medium;
c) Step of inoculating a lactic acid bacteria strain confirmed to form ornithine into MRS medium containing 1% arginine;
d) A step of primary culturing the medium inoculated with the lactic acid bacteria strain at 37°C for 48 hours;
e) A step of centrifuging the cultured medium at 3,500 RPM for 20 minutes to separate the lactic acid bacteria and the medium;
f) A step of washing the lactic acid bacteria separated from the medium by adding sterilized water;
g) Step of adding rice bran medium to the washed lactic acid bacteria and performing secondary culture at 37℃ for 48 hours;
h) A step of manufacturing rice bran yogurt by adding sugar and milk to a rice bran medium cultured with lactic acid bacteria and fermenting at 37°C for 6 hours;
i) a step of packaging the manufactured rice bran yogurt into a certain unit; and
j) A method for producing rice bran yogurt, characterized by including a step of refrigerating packaged rice bran yogurt. In claim 1,
The above lactic acid bacteria
a1) Step of diluting fermented food with peptone water;
a2) Step of culturing the diluted solution in BCP medium;
a3) a step of isolating a colony that is dark yellow and does not form a ring from the cultured medium; and
a4) A method for producing rice bran yogurt, characterized in that activation is confirmed by a step of confirming the ornithine production ability of the separated strain by TLC. A method for producing rice bran yogurt according to claim 1, characterized in that the lactic acid bacteria strain is Pediococcus pentosaceus MAC-11 (KCCM11359P) lactic acid bacteria strain. Rice bran yogurt characterized by being manufactured by a method according to any one of claims 1 to 3.