KR20130000922A - Kochujang containing gaba and having skin care effect - Google Patents

Abstract

PURPOSE: Red pepper paste with a skin care effect is provided with a function improving wrinkle condition using extracts of white lotus and amino acid powder treated with enzymes. CONSTITUTION: Malt 30-60 parts by weight are mixed with purified water 1,200-1,700 parts by weight. The mixed malt liquid is saccharified at 40-70 deg. C for 2-5 hours. The saccharified malt is mixed with 1-4 parts by weight of amino acid, 40-60 parts by weight of fermented soybean powder, 150-350 parts by weight of red pepper paste, and 80-150 parts by weight of salt. The mixed ingredients are packed and stored in a packing container. 1,200-1,700 parts by weight of white lotus extracts are mixed with 30-60 parts by weight of malt. The white lotus and malt liquid is saccharified for 2-5 hours at 40-70 deg. C. The saccharified white lotus and malt liquid is mixed with 40-60 parts by weight of fermented soybean powder, 150-350 parts by weight of red pepper powder, and 80-150 parts by weight of salt. The mixed ingredients are packed and stored in a container. The white lotus extracts are produced through the static extraction for 30-60 hours at 30-70 deg. C after adding 25-70 parts by weight of white lotus leaves into 700-1,200 parts by weight of pure water. 1-4 parts by weight of enzyme treated amino acid powder is additionally mixed with the red pepper paste.

Description

Kochujang containing GABA and having skin care effect

The present invention relates to red pepper paste, and more particularly, to a red pepper paste containing GABA having antioxidative and cancer cell proliferation inhibitory effects by using white lotus extract and enzyme-treated amino acid (GABA-derived GABA) powder.

Gochujang is a source of protein derived from soybeans, sweet taste, and carbohydrate foods such as glutinous rice, rice, and barley rice.It is not only nutritious, but also sweet and salty from red pepper powder and salty from soy sauce and salt used to match the liver. It is one of the unique and unique traditional Jangjang, which is indispensable on our table.

Kochujang has been scientifically proven to have as many nutrients as soy sauce and miso. Kochujang contains many nutrients that are beneficial to our body, such as protein, fat, vitamin B2, vitamin C, and carotene. Capsaicin, a red pepper seed-containing ingredient, has antimicrobial activity, and some red pepper powders containing beta-carotene and vitamin C have been anti-mutated, anti-inflammatory and anti-cancer. In addition, the pepper-flavored capsaicin ingredient not only reduces body fat, but also red pepper powder, which is known to burn body fat from meju or ripened red pepper paste, has been popular in some countries.

Such kochujang is traditionally manufactured in each household and limited to family consumption. However, as the complexity of manufacturing process, women's participation in social activities, and nuclear familyization, the demand for commercialized kochujang is increasing. Kochujang market is more popular than doenjang and soy sauce. As the scale is large and the market manufacturing and sales industry matures, consumers' preferences are also increased, and the development of products with emphasis on high quality and functionality is required. In order to satisfy the consumer's needs, the manufacturing method with different red pepper varieties, low salted red pepper paste preparation, a method of improving the functionality by adding plum and Schizandra chinensis extract, sweet potato, etc., red pepper paste with red ginseng and functional kelp powder Red pepper paste, red pepper paste with tuna porridge solution, red pepper paste with garlic porridge, red pepper paste with juice, red pepper paste with pumpkin, licorice, red pepper paste with mustard and chitosan were prepared, and natural substances containing large amounts of functional substances were prepared. Research on adding kochujang is ongoing.

On the other hand, lotus ( Nelumbo nucifera ) is a perennial aquatic plant of the water lily family, widely distributed in India, China, tropical, temperate eastern Asia, Korea, Japan, etc. It has been eaten. These lotuses are characterized by white color of flowers and white color of red flowers, and red color of red flowers. The roots and leaves of Nelumbo nucifera Gaertner are used in folk medicine, such as diabetes, hyperlipidemia and hypertension. It has been used for filamentous diseases and is known to be effective in diarrhea, pain, dizziness, bleeding and detoxification.

These leaves of alkaloids contain alkaloids and are known to be effective in diabetic and hyperlipidemia as well as sedative and antipyretic effects. Lotus leaf has been used as a folk remedy for bleeding gastric ulcers, gastritis, hemorrhoids, bleeding, diarrhea, headache and dizziness, hemorrhage, postpartum bleeding, enuresis and detoxification. In addition, lotus leaf contains a biologically active ingredient is not only used as a natural drug and dietary supplement, but also recently drink tea.

These studies on lotus root mainly include the study of the composition of free sugars, fatty acids, amino acids and dietary fiber, the study of lipid composition and health improvement function, and the effect of powder addition on the aptitude of baking. In addition, research on the seed of lotus has a study on the composition of fats and oils, and research on the lotus leaf has an antioxidant activity and the manufacture of lotus leaf tea and its quality characteristics.

Furthermore, it has recently been suggested that white lotus extract contains gamma-aminobutyl acid (aka "GABA", hereinafter referred to as "GABA") and has a sleeping effect through animal experiments, which may help insomnia. It also became.

GABA is a nonprotein amino acid that is the major excitatory neurotransmitter in the central nervous system of humans and animals. In plants, GABA has the effect of improving the adaptability to environmental stress. GABA is widely distributed in foods such as mulberry leaves, green tea, germinated brown rice, soybeans, mandarin oranges, vegetables, fruits, etc., and it improves blood flow, increases oxygen supply, and promotes metabolic function of brain cells. It is known to be effective in lowering blood pressure, reducing depression, preventing stroke and dementia, and treating insomnia and alleviating insomnia. In addition, it has recently been researched that it will be effective in improving wrinkles, and thus the use of functional cosmetics is increasing. Although GABA has been approved as a drug for improving cerebral blood flow in the United States and Japan, GABA ingredients are mainly used for medicinal purposes. However, in Korea, various attempts have been made to use food, food additives, and cosmetics. Increasingly, the use as a cosmetic material for.

On the other hand, one of the causes of skin wrinkles is known as lack of collagen, collagen is a major protein constituting the dermis of the skin, maintains the structure and elasticity of the skin, decreases the production and increases the decomposition with age It is known to induce the depression of the dermal layer of the skin to produce wrinkles of the skin. Retinol (vitamin A) and adenosine, which are conventionally used to improve wrinkles, are widely used as substances involved in skin regeneration by increasing the synthesis of collagen and normalizing the epidermal keratinization process, but are unstable to light and heat and It is known to have a stimulus. Wrinkle improvement studies include many studies such as increasing collagen production and suppressing degradation of 17b-estradiol cells, promoting and inhibiting elastin production, increasing hyaluronic acid production, promoting aging cell regeneration, and removing reactive oxygen radicals. As cosmetic materials are being replaced by natural materials according to naturalism, research on whitening and wrinkle improvement by using wormwood fraction, research on cosmetic wrinkle improvement effect with black ginseng, cosmetic application research on chestnut leaf extract, anti-hwang extract Development of cosmetic materials derived from natural products and herbal ingredients, such as research on aging effects, whitening of metabolites of natural products such as mushrooms and medicinal plants, and research on cosmetic materials for wrinkle improvement.

An object of the present invention is to provide a kochujang having the functionality including the wrinkle improvement effect by adding GABA content and white lotus extract and enzyme-treated amino acid (amino acid-derived GABA) powder to the traditional kochujang prepared kochujang.

The method for preparing red pepper paste containing GABA according to the present invention for achieving the above object and having a skin-cosmetic effect includes mixing 30 to 60 parts by weight of malt oil in 1200 to 1700 parts by weight of purified water; Saccharifying the mixed malt liquid at 40 to 70 ° C. for 2 to 5 hours; Mixing the raw materials with 1 to 4 parts by weight of enzyme-treated amino acid (amino acid-derived GABA) powder, 40 to 60 parts by weight of meju powder, 150 to 350 parts by weight of red pepper powder, and 80 to 150 parts by weight of salt to the saccharified malt solution; Characterized in that it comprises a step of packaging and storing the mixed raw material in a packaging container.

According to another aspect of the present invention, a method of preparing kochujang containing GABA and having a skin-cosmetic effect includes mixing 30 to 60 parts by weight of malt oil to 1200 to 1700 parts by weight of white lotus extract; Saccharifying the mixed white lotus-starch oil solution at 40 to 70 ° C. for 2 to 5 hours; Mixing raw materials with saccharified white lotus-starch oil solution with 40 to 60 parts by weight of meju powder, 150 to 350 parts by weight of red pepper powder, and 80 to 150 parts by weight of salt; Characterized in that it comprises a step of packaging and storing the mixed raw material in a packaging container.

In addition, Kochujang according to an aspect of the present invention is made in any one of the above two methods, the white lotus extract is put in the water of 25 ~ 70 parts by weight of dried white lotus leaf to 700 ~ 120 parts by weight of purified water 30 ~ 70 ~ 30 ~ It can be made by 60 hours stationary extraction, the second method can further mix 1 to 4 parts by weight of enzyme-treated amino acid (GABA derived from amino acid) powder.

The present invention is prepared by using kochujang using white lotus extract and enzyme-treated amino acid (amino acid-derived GABA) powder to provide kochujang with high amino nitrogen content and GABA content, antioxidant activity and cancer cell proliferation inhibitory ability, wrinkle improvement by ingesting kochujang Efficacy evaluation (hyaluronidase inhibitory activity, elastase inhibitory activity, melanoma cell proliferation inhibitory effect can be obtained.

In addition, kochujang prepared by adding white lotus extract and enzyme-treated amino acid (amino acid-derived GABA) powder contains GABA and not only has a skin beauty effect, but also has an anticancer effect, which can help to increase the value added of kochujang and promote health. There is also an effect.

1 is a manufacturing process chart of gochujang according to the present invention,
Figure 2 is a diagram showing the change in the water content of kochujang during the ripening period of kochujang according to the present invention,
3 is a chart showing the total nitrogen content change of kochujang according to the ripening period,
Figure 4 is a chart showing the pH content change of kochujang during the ripening period,
5 is a graph showing the change in the total acid content of kochujang during the ripening period,
6 is a graph showing the change in the total sugar content of kochujang during the ripening period,
7 is a chart showing the change in reducing sugar content of kochujang during the ripening period,
8 is a chart showing the amino nitrogen content change of kochujang during the ripening period,
9 is a chart showing the alcohol content of kochujang during the ripening period,
10 is a chart showing the change of GABA content of kochujang with the aging period,
11 is a chart showing the antioxidant effect of kochujang at day 0 of aging,
12 is a chart showing the antioxidant effect of kochujang at 30 days of aging,
13 is a chart showing the antioxidant effect of kochujang at 60 days of aging,
14 is a chart showing the antioxidant effect of kochujang at 90 days of aging,
15 is a chart showing the inhibitory effect of THP-1 cell line (human acute monocyte leukemia) proliferation of kochujang,
16 is a chart showing the effect of inhibiting the growth of HL60 cell line (human acute promyelocytic leukemia) of kochujang,
17 is a diagram showing the growth inhibition effect of U937 cell line (tissue lymphoma) of kochujang,
18 is a table showing the effect of inhibiting Hyaluronidase activity of 70% ethanol extract of kochujang at 30 days of aging,
19 is a diagram showing the effect of inhibiting Hyaluronidase activity of 70% ethanol extract of kochujang at 60 days of aging,
20 is a chart showing the elastase inhibitory effect of Kochujang 70% EtOH extract on the 30th day of aging,
21 is a chart showing the elastase inhibitory effect of Kochujang 70% EtOH extract at 60 days of aging,
22 is a chart showing the growth inhibition effect of Melanoma cell (B16F10 cell) of kochujang,
23 is a chart of sensory evaluation results of kochujang.

Hereinafter, the present invention will be described in more detail with reference to the accompanying drawings. However, the accompanying drawings are only examples for easily explaining the contents and scope of the technical idea of the present invention, whereby the technical scope of the present invention is not limited or changed. In addition, it will be apparent to those skilled in the art that various modifications and changes are possible within the scope of the technical idea of the present invention based on these examples.

Kochujang containing GABA according to the present invention and having a skin-cosmetic effect can be made by two methods. That is, 1) mixing 30 to 60 parts by weight of malt oil to 1200 to 1700 parts by weight of purified water; Saccharifying the mixed malt liquid at 40 to 70 ° C. for 2 to 5 hours; Mixing the raw materials with 1 to 4 parts by weight of enzyme-treated amino acid (amino acid-derived GABA) powder, 40 to 60 parts by weight of meju powder, 150 to 350 parts by weight of red pepper powder, and 80 to 150 parts by weight of salt to the saccharified malt solution; A method comprising the steps of packaging and storing the mixed raw materials in a packaging container; and (2) mixing 30 to 60 parts by weight of malt oil to 1200 to 1700 parts by weight of white lotus extract; Saccharifying the mixed white lotus-starch oil solution at 40 to 70 ° C. for 2 to 5 hours; Mixing raw materials with saccharified white lotus-starch oil solution with 40 to 60 parts by weight of meju powder, 150 to 350 parts by weight of red pepper powder, and 80 to 150 parts by weight of salt; It can be made by the method consisting of the steps of packaging and storing the mixed raw materials in a packaging container.

① method is to produce only the action of amino-reacted GABA by using only enzyme-treated amino acid (amino acid-derived GABA) powder, the method of ② is to obtain only the action of white lotus by the method of manufacturing using only white lotus extract. . However, it is preferable that it can provide both the action of the enzyme-treated amino acid (GABA) powder and white lotus, so that 1 to 4 parts by weight of the amino-reacted GABA powder is added to the raw material mixed in the process of making ②. It is preferable to prepare red pepper paste containing both the treated amino acid (amino acid-derived GABA) powder and white lotus.

The white lotus extract used in the red pepper paste prepared by the method as described above may be made by putting the dried white lotus leaf 25 ~ 70 parts by weight to 700 ~ 120 parts by weight of purified water after 30-60 hours stationary extraction at 30 ~ 70 ℃. .

In addition, the raw material is mixed, and the red pepper paste is matured in the process of packaging and storing, thereby enriching the taste and aroma, and increasing the effective efficacy of the enzyme-treated amino acid (amino acid-derived GABA) powder and white lotus. Optimum ripening temperature is room temperature and ripening time is 20 ~ 100 days.

Hereinafter, an experimental example of gochujang according to the present invention will be described in detail. Reference may be made to particular trademarks or material suppliers in the description of the following examples, which are intended to aid the present invention only, and thus the claims are not limited thereto.

[Example]

1. Material

The red pepper powder, glutinous rice flour, malt and meju powder used in this study were provided by Munokrye-ga, a farming company in Sunchang, and the salt was used as salt. White lotus extract used as gochujang material was prepared by purchasing white lotus ( Nelumbo nucifera Gaertner) from the ear farmers. 50 g of dried white lotus leaf and 1 L of water (purified water) were mixed, and after extracting at 50 ° C. for 48 hours, the mixture was squeezed with gauze and the compressed liquid was used as an extract. Enzymatically treated amino acid (amino acid-derived GABA) powder was purchased from MH 2 bio.

2. Gochujang manufacture

To prepare kochujang, kochujang was prepared by the addition of white lotus extract and enzyme-treated amino acid (amino acid-derived GABA) powder, as shown in Table 1 and FIG. 1. White lotus extract Kochujang was prepared using white lotus extract instead of water, and the addition of enzyme-treated amino acid (amino acid-derived GABA) powder Kochujang was prepared by saccharifying Sikhye solution and adding 2g of enzyme-treated amino acid (amino acid-derived GABA) powder. Kochujang prepared with water is made from CK (common chili pepper), kochujang prepared with water and enzyme-treated amino acid (GABA) powder, GK (GABA pepper paste), and kochujang prepared with white lotus extract is WK (baekyeon pepper paste) , Kochujang prepared with white lotus extract and enzyme-treated amino acid (GABA derived from amino acid) powder was prepared by dividing it into WGK (GABA white lotus pepper paste).

Sikhye was prepared by mixing water or white lotus extract with malt and glutinous rice powder for 3 to 4 hours at 60 ℃. After saccharification, GK and WGK kochujang were prepared by adding enzyme-treated amino acid (amino acid-derived GABA) powder, mixed with meju powder, red pepper powder, and salt in the order.

3. Analysis of Physicochemical Properties of Kochujang

1) Determination of moisture content

Moisture was analyzed twice by 105 ° C atmospheric pressure drying according to the AOAC method and expressed as an average value.

2) Determination of total nitrogen and amino nitrogen content

The total nitrogen content was analyzed by an automatic Kjeldahl analyzer (B-339, Buchi Co., Swiss). The amino nitrogen content was 2 g of the sample in a beaker, 100 mL of distilled water was added and stirred for 1 hour, then dissolved sufficiently. The NaOH solution was titrated to pH 8.4. 20 mL of neutral formalin solution was added thereto, followed by neutralization titration to pH 8.4 with 0.1 N NaOH solution.

3) pH and titration acidity measurement

50 g of kochujang was added to 50 mL of distilled water, stirred at 37 ° C. for 2 to 3 hours, and the pH was measured by repeated experiments using a pH meter (SevenEasy ™ pH meter S20, Mettler toledo AG, Switerland). Was taken. It was titrated with 0.1N NaOH solution until pH 8.4, and the consumption of this solution was made into titratable acidity.

4) Measure per total

The sugar content of kochujang was measured using a sample used for pH measurement. 0.5 mL of the sample was repeatedly measured three times with a sugar meter (ATAGO pocket reactometer PAL-1, Tokyo, Japan) and expressed as an average value.

5) Reducing Sugar Content

The reducing sugar content was tested by DNS (Dinitrosalicylic acid) method with kochujang filtrate measured at the titratable acidity. After diluting the sample solution 10 times, 3 mL of DNS was added to 1 mL of the filtered sample solution using a 0.45 μm PVDF filter (Millipore) and mixed with a Vortex mixer, and 1 mL of 40% Rochell salt was added while heating in a water bath. After heating for 10 minutes in boiling water, the tube was cooled in running tap water, and then absorbance was measured at 550 nm using a UV-VIS Spectrophotometer (UV1600 PC, Shimadzu). In the same way, a calibration curve was prepared using D-Glucose. The reducing sugar content of kimchi was expressed as D-Glucose equivalent per mL of kochujang sample.

6) Determination of Free Sugar Content

5.0 g of free sugar was added to the sample, and 50 mL of 70% methanol was added thereto, followed by extraction with a pretreatment device (Mars X, CEM Co., USA) at 75 ° C. for 1 hour and filtered. Take 10 mL of the sample above, centrifuge, filter the supernatant through a Sep-pak C ₁₈ cartridge, filter with a 0.2 μm membrane filter, and use HPLC (NS-2004GP, Futecs) equipped with ELSD (Model 2000, Softa Co., USA). Co., Korea). At this time, the column was Asahipak NH2P-504E (4.6 mm 250 mm), the column temperature was 35 ℃, the mobile phase was 75% acetonitrile, the flow rate was 1.2 mL / min.

7) Microbial count

1g of the sample collected every 30 days during the maturation period was added homogenized at room temperature with 9ml of sterile water, diluted with sterile water according to the decimal method, and bacteria in plate medium were tryptic soy agar (37 ℃, 2 days), yeast Was YM (25 ℃, 2 days), lactic acid bacteria were cultured in MRS (Difco) (37 ℃, 2 days) in a medium to measure the colony.

8) Alcohol content measurement

For alcohol content, 5 g of the sample was dissolved in 100 mL of distilled water, and 1 g of lime carbonate was distilled into a distillation flask to distill the distillate to 100 mL. Then, add 10 mL of 0.2 NK ₂ Cr ₂ O ₇ solution and 10 mL of concentrated H ₂ SO _{4 to} 10 mL of distillate, mix, cool for 1 hour in a cool dark place, add 200 mL of distilled water, 6.5 mL of 8% KI and 0.5 starch solution. Mix mL and titrate with 0.1 N Na ₂ S ₂ O ₃ solution for analysis.

9) Measurement of mineral content

The mineral content was analyzed by preparing a sample by wet decomposition according to the Food Code. In other words, take 3.0 g of the sample in a 250 mL digestion flask, add 10 mL of HNO ₃ , heat and dry the contents, and then add 10 mL of HNO ₃ solution (HNO ₃ : H ₂ O = 1: 1) and 10 mL of 60% HClO ₄ . Put and heat until colorless. Then dilute with a small amount of distilled water, transfer to an evaporating dish and heat again to remove HClO ₄ . After evaporation, 10 mL of HCl solution (HCl: H ₂ O = 1: 1) and the same amount of distilled water were added, completely dissolved in a water bath, and used as a sample for mineral analysis by diluting to 100 mL. At this time, Ca, K, Mg, Na, Fe, Cu, Zn and the like were measured by atomic absorption spectrophotometer (Solar-M5, Thermo elemental Co., England), P is a spectrophotometer (UV-1601, Shimadzu according to molybdenum blue color method) Co., Japan) at 650 nm.

10) Chromaticity Measurement

Kochujang's chromaticity was measured by using a colorimeter (CM-3500D, Minolta Co., Ltd., Osaka, Japan) to measure L (lightness), a (redness) and b (yellowness) values. and b values were 0.69, 0.00 and -0.24, respectively. The experiment was repeated twice and represented as an average value.

4. Determination of GABA and Amino Acid Content in Kochujang

GABA and amino acid content was carried out according to the method of Oh and Oh. To 1 g of kochujang, a mixture of methanol: chloroform: water (12: 5: 3) was added, and 9 mL was added to the mixture, followed by centrifugation (13,000 ㅧ g, 15 min, 4 ° C.) to obtain a supernatant, which was used for amino acid extraction. The solids were divided into supernatants and precipitates to extract amino acids, and then the supernatants and precipitates were mixed. Amino acid extraction method was added to 200mg or 200μL of sample, 800μL of a mixture of methanol: chloroform: water (12: 5: 3) was added and mixed, and the supernatant was obtained by centrifugation (13,000 ㅧ g, 15min, 4 ℃). 200 μL of chloroform and 400 μL of water were added to the precipitate, and the remaining amino acid was extracted secondarily, and the supernatant obtained from the first and second centrifugation was combined and freeze-dried. Then dissolved in a small amount of water and filtered by 0.45㎛ PVDF filter (Millipore) was used for analysis. AccQ · Fluor Reagent was analyzed for fluorescence derivatization of amino acids, and column was analyzed by 3.9 ㅧ 150mm AccQ · Tag ^TM (For hydrostat amino acid analysis, water, USA). Standard GABA (Sigma, St. Louis, MO., USA) and standard amino acid (Amino acid Standard, waters, USA) were used and the content was calculated using the autochro WIN program (Young-Lin, Korea).

5. Functional exploration of kochujang

1) Evaluation of antioxidant effect (Electron donating abilities, DPPH radical scavenging ability)

Kochujang samples according to the ripening period were extracted with water (48 hours 50 ℃), separated from the primary supernatant and solids by filter paper (Wathman No. 2), and then lyophilized to powder to measure antioxidant activity. Each powdered kochujang sample was dissolved in sterile water, dissolved in a constant concentration, and filtered with a 0.45㎛ PVDF filter (Millipore) to use as a sample. Antioxidant experiments of samples were measured by modifying Blois's method (1958) for electron donating abilities (EDA, scavenging ability of DPPH radicals). 40 μl of a sample that was constantly dissolved and 0.2 mM 2,2-diphenyl-1-picrylhydrazyl (DPPH, simga, St. Louis, MO., USA) dissolved in 160 μl of EtOH were added to a 96 wall plate and reacted at 37 ° C. for 30 minutes. I was. After fully reacting the reaction solution reacted for 30 minutes, it was measured using an ELISA reader (Molecular Devices, USA) at 517 nm. At this time, ascorbic acid was dissolved in sterile water as a sample for active comparison, and the electron donation was measured. The results were calculated by using EDA (Electron donating abilities,%), and the difference between the absorbance of the sample added group and the absorbance of the non-added group as a percentage (%) as shown below.The average value and standard deviation were calculated using the SPSS program. Calculated.

(1)

(One)

2) cancer cell proliferation inhibitory effect

Adjust the number of cells to be 5 × 10 ⁴ cells / well in 96 well culture plates for passage of human acute monocytic leukemia (THP-1), human acute promyelocytic leukemia (HL-60), and U937 (human acute histiocytic lymphoma) cells Then, 10 times, 100 times diluted kochujang sample was added and incubated in a 5% CO ₂ incubator at 37 ° C. for 24 hours, and measured by MTT method by Mosmann (1983). The sample used Kochujang sample extracted with water of CK, WK, GK, and WGK kochujang for 30 days of aging.

3) Evaluation of wrinkle improvement effect

① Determination of Hyaluronidase activity inhibitory effect

Hyaluronidase activity inhibitory effect was modified by the Morgan-Elson assay method using hyaluronic acid as a substrate, and the amount of N-acetylglucosamine produced was colorimetrically determined at 585 nm using p-dimethylaminobenzaldehyde (DMAB). 50 μl of hyaluronidase (214 unit / mL, simga, St. Louis, Mo., USA) dissolved in 0.1 M acetate buffer (pH 3.5) and 20 μl of the sample were mixed and reacted at 37 ° C. for 20 minutes. As a negative control, water was added instead of the sample solution and reacted under the same conditions. Thereafter, 100 µl of 12.5 mM CaCl ₂ was added thereto, and the reaction was again performed at 37 ° C. for 20 minutes. Inactive hyaluronidase is activated by Ca ²⁺ and 250 μl of hyaluronic acid (2.4mg / mL, simga, St. Louis, MO., USA) dissolved in 0.1M acetate buffer (pH3.5) in activated hyaluronidase solution After the reaction was performed at 37 ° C. for 40 minutes, 100 μl of 0.4N NaOH and 100 μl of 0.4M potassium tetraborate solution were added to the reaction solution, followed by heating for 3 minutes in boiled water, followed by cooling at room temperature. After adding 3 mL of DMAB reagent (4 g of p-dimethylaminobenzaldehyde, 350 mL of 100% acetic acid, 50 mL of 10N NaOH) to the cooled reaction solution, the reaction solution was reacted at 37 ° C. for 20 minutes using an ELISA reader (Molecular Devices, USA) at 585 nm. It was measured by. Inhibitory activity was determined by the following Equation 2.

(2)

(2)

ODc is the absorbance of the control group without the sample solution at 585 nm, and ODs represents the absorbance value measured by adding the sample solution. The positive control group was compared with tannic acid (Simga, St. Louis, Mo., USA) and averaged by three replicates.

② Elastase activity inhibition effect

Elastase activity inhibitory effect was measured as follows. Substrate 1.0 mM N-succinyl- (Ala) ₃ -p-nitroanilide (Sigma, St. Louis, MO., USA) in 200 μL of 20 μL of kochujang sample and 3 unit Elastase (from Porcine pancreas, Sigma, St. Louis, MO., USA) 10μL was added and reacted at 25 ° C. for 10 minutes, and the absorbance was measured at 410 nm using an ELISA reader. Elastase activity inhibition rate was calculated by the following Equation 3.

(3)

(3)

The positive control group was compared with ursolic acid (Sigma, St. Louis, MO., USA).

③ Inhibitory effect of mouse-derived Melanoma cell (B16F10 cell) proliferation

Adjust the cell number of the passaged B16F10 cells (human melanoma cells) to a 96 well culture plate to be 3 ㅧ 10 ⁴ cells / well, and add kochujang sample diluted 1-100 times with Kochujang filtrate for 24 hours. Incubation was carried out in a 5% C0 ₂ incubator at < RTI ID = 0.0 > C, < / RTI > As the sample, CK, WK, GK, and WGK kochujang samples extracted with water for 30 days of ripening were used.

5. Sensory Evaluation

The sensory evaluation of Gochujang was conducted by selecting 13 undergraduate and graduate students from Woosuk University who were recognized for their interest in this study and their suitability as an inspector. Then, sensory tests were conducted according to the experimental design method. The evaluation items were color, odor (salty, salty, sukhumhan), taste (salt, sweet, sour, hot, spicy), and overall preference in the five-point scale method. It evaluated by 1 point.

Results and Discussion

1. Analysis of Physicochemical Properties of Kochujang

1) Determination of moisture content

The change in moisture content of kochujang during the ripening period is shown in FIG. 2. The moisture content of CK, WK, GK and WGK pepper paste is 35.10 ~ 44.90%. The water content of CK, GK, and WGK kochujang decreased at 30 days of fermentation, then slightly increased after 30 days of fermentation, and then decreased at 90 days of fermentation. It was similar to the water content of traditional gochujang, and the decrease in moisture as fermentation proceeded is believed to be due to evaporation of water by aging.

2) total nitrogen content

The total nitrogen content of kochujang is the same as that of Figure 3 and CK, WK, GK. WGK kochujang showed total nitrogen contents of 1.40%, 1.33%, 1.2%, and 1.36% at 30 days of fermentation, respectively. This is generally similar to the total nitrogen content during the ripening period of kochujang.

3) pH and titratable acidity

The results of examining the change in pH and titratable acidity during kochujang ripening are shown in FIGS. 4 and 5. The pH change of each kochujang ranged from pH 5.05-4.88 to pH 5.00-4.97 at 90 days of fermentation. WK kochujang was generally lower than the other bulbs. The titratable acidity of WGK kochujang and WK kochujang prepared with white lotus extract increased from 30 to 60 days, and the acidity was higher than that of CK and GK kochujang.

4) Measure per total

The change in the total sugar content of red pepper paste during ripening is shown in FIG. 6. Changes in total sugar in kochujang are mostly influenced by microbial starch hydrolase. The change in total sugar in each kochujang increased until 30 days of fermentation and then slowly decreased. WK kochujang and WGK kochujang showed higher total sugar content than other bulbs. Changes in total sugar in kochujang are mostly influenced by starch hydrolase, which appears to be due to not only sugar being used as an energy source for microorganisms but also carbohydrate degradation by enzyme action.

5) Reducing Sugar Measurement

As shown in Figure 7, the reducing sugar content according to the ripening period is 25.22 ~ 27.15mg / mL CK Kochujang, 24.54 ~ 26.35mg / mL for WK Kochujang, 23.56 ~ 27.85mg / mL for GK Kochujang, 24.39 ~ 26.48 for WGK Kochujang In mg / mL, the reducing sugar content of each kochujang increased around 30 days of aging and then gradually decreased.

6) Organic Sugar Content Determination

Free sugar content of kochujang during aging period is shown in Table 3. In the free sugar content of CK kochujang, glucose decreased from 5.88% at the early stage of fermentation to 4.29% at the end of fermentation, while fructose and maltose increased to 3.42% and 11.84% compared to the early and middle fermentation period. Glucose, fructose and maltose of WK kochujang increased with fermentation period, especially maltose increased from 1.52% in early fermentation to 5.93% in later stage. Glucose and maltose of GK kochujang decreased with increasing fermentation except fructose. WGK kochujang increased glucose, fructose and maltose similarly to WK kochujang. In particular, maltose content increased from 1.31% at the beginning of fermentation to 5.01% as in WK kochujang. The maltose content of CK kochujang was higher than that of other bulbs.

7) amino nitrogen measurement

Amino nitrogen of kochujang is a protein that breaks down into free amino acid form during the fermentation process of kochujang and gives a delicious taste. The higher the content, the better the taste, and is used as a general quality index of kochujang. The amino nitrogen content change during the ripening process of kochujang is shown in FIG. 8. The amino nitrogen content of each kochujang increased with the fermentation period. During the aging period, amino nitrogen content was 84.99 to 93.54 mg% for CK kochujang, 88.84 to 103.32 mg% for WK kochujang, 103.01 to 118.44 mg% for GK kochujang, and 113.29 to 123.19 mg% for WGK kochujang, respectively. Kochujang showed the highest amino nitrogen content at 60 days of fermentation, but after 60 days of fermentation, the content was maintained or slightly decreased. Among them, GK and WGK kochujang added with enzyme-treated amino acid (GABA) powder had higher amino nitrogen content than CK and WK kochujang, which is a direct result of the addition of enzyme-treated amino acid (GABA) powder.

8) Microbial count

Microbial changes during the fermentation of kochujang decreased from 5.58 log CFU / g in CK kochujang to 30 days of fermentation at 30 days of fermentation, and then slightly increased to 5.26 log CFU / g at 90 days of fermentation. WK and WGK kochujang increased from 4.64 log CFU / g and 4.92 log CFU / g to 60 days of fermentation, and then decreased. The number of bacteria decreased in the latter stages of fermentation, similar to the study of traditional kochujang. In addition, the initial microbial count in GK kochujang was similar to that of WK and WGK kochujang, but it was constant with no change in microorganisms as the ripening period increased.

During the ripening period, the number of lactic acid bacteria was 4.61∼5.11 log CFU / g for CK kochujang, 39∼4.81 log CFU / g for GK kochujang, and each kochujang increased at 60 days of fermentation and then remained constant. The number of lactic acid bacteria in WK and WGK kochujang was 4.00 to 4.15 log CFU / g, and 4.16 to 4.86 log CFU / g. WGK kochujang increased after 60 days of lactic acid bacteria and then decreased. The number of yeast in kochujang was initially 4.38∼4.66 log CFU / g, and CK kochujang increased until 60 days of fermentation and then became constant.

WK kochujang decreased gradually over the ripening period, and GK kochujang remained constant over the ripening period, and increased up to 60 days in WGK kochujang and gradually decreased.

9) Alcohol content measurement

Alcohol produced during the fermentation of kochujang seems to affect the flavor of kochujang, but there was no difference between kochujang of CK, WK, GK, and WGK kochujang, and the alcohol content of each kochujang was maintained or slightly from 30 days to 90 days. It was a decreasing level (FIG. 9). The alcohol content in the fermentation properties of kochujang meju and red pepper varieties increased from 0.1% at the beginning of fermentation to 3.6% at 100 days of fermentation. This is known to reduce the reducing sugar in kochujang by using sugar.

10) Measurement of mineral content

The results of measuring the contents of Ca, P, Mg, Na, K, Fe, Cu, Zn as minerals during the ripening time of kochujang are shown in Table 5. The Ca content of each kochujang decreased from 30 to 60 days of aging and then increased. The content of P was minimal during fermentation in each kochujang except WGK, and WGK kochujang decreased after 60 days of fermentation. The content of Mg did not change with CK and GK kochujang ripening periods, but it decreased after 30 to 60 days of fermentation in WK kochujang and increased afterwards, similar to that of P in WGK kochujang. For example, at 90 days of aging, the Mg content decreased and then increased. The Na content of kochujang was higher in CK and WK kochujang than in other GK and WGK kochujang, and then decreased in 30 and 60 days of CK and WK kochujang. WGK kochujang decreased after 60 days of fermentation and then increased. There was no significant difference in K content, and Fe content decreased with aging of CK, WK, GK and WGK kochujang. Cu content was similar without any significant difference between kochujang and there was no change according to the ripening period. After fermentation period, Zn content in CK was decreased, whereas in WGK kochujang, it increased slightly at 30 and 60 days of fermentation and then decreased.

11) Chromaticity Measurement

Changes in chromaticity of CK, WK, GK, and WGK kochujang are shown in Table 6. CK, WK and WGK kochujang decreased the lightness (L) at 30 days of fermentation and gradually increased until 90 days. GK kochujang decreased slightly after 30 days of fermentation. The redness of the a value (redness) of CK kochujang gradually decreased with the aging period, and GK kochujang decreased after 30 days of fermentation and increased after 60 days and then decreased again at 90 days. WK kochujang appeared to be similar to CK kochujang, but a value was lower during fermentation than CK kochujang. WGK kochujang showed 18.53 ~ 21.17 values similar to WK kochujang. The yellowness of b showed a slight decrease in CK kochujang at 30 days of fermentation and then increased to a higher value of 33.74 than other bulbs at 60 days of fermentation. GK kochujang was similar to CK kochujang, but lower than CK kochujang. WK and WGK kochujang increased slightly at 60 days of fermentation and then slowly decreased at 90 days. The color change of kochujang during fermentation increased with the increase of L and the decrease of a and b values. Kochujang is a food with high sugar and amino acid content. HMF (5-hydroxymethyl furfural) and its oxidized polymer by Maillard reaction contribute to the discoloration of kochujang.The higher the storage temperature, the lower the L value and a value. Proportional to storage temperature.

2. Gochujang GABA  And free amino acid content determination

The GABA and amino acid contents of kochujang according to ripening period are shown in Table 7. The GABA content at 0 days of fermentation was CK kochujang 7.27mg / 100g, GK kochujang 139.90mg / 100g, WK kochujang 7.74mg / 100g, WGK kochujang 104.61mg / 100g. The rate of aging was 7.87mg / 100g and decreased slowly as the ripening period increased. The GABA content of GK kochujang added with enzyme-treated amino acid (GABA) powder decreased on the 30th day of fermentation (87.48mg / 100g) and then increased on the 60th day (113.78mg / 100g) and decreased again on the 90th day of fermentation (69.69mg / 100g ). During the ripening period of WK kochujang, GABA content increased on the 30th day (13.63mg / 100g) and then gradually decreased to the 30th day, and the change of GABA content of WGK kochujang showed a similar pattern to the GABA content of GK kochujang (Fig. 10). . CK kochujang and WK kochujang showed higher GABA contents at 30 days of fermentation, while GK kochujang and WGK kochujang showed higher GABA contents at 60 days of fermentation.

The content of free amino acids in kochujang increased with the maturation period, and the content of Glutamate (Glu) was 18.00-25.75mg / 100g when Kochujang ripening was 0, but the content of 21.03∼61.59mg / 100g, Aspartic acid (Asp) at 90 days ) The content increased from 7.08 to 10.05mg / 100g at 0 Kochujang fermentation and from 50.96 to 91.18mg / 100g at 90 days of fermentation. Serine (Ser) content in free amino acid increased from 7.00 to 8.51mg / 100g at 0 and increased from 40.66 to 128.75mg / 100g at 90 days of aging.

Proteins in kochujang are converted into free amino acids by the action of fermentative microorganisms to give a delicious taste.

3. Benefits of Gochujang

1) Antioxidant Activity Evaluation

The red pepper paste extract sample powder was prepared in sterile water at 4, 40, 400, and 4,000 ㎍ / mL concentration (final concentration) to modify the electron donating ability of Blosis (EDA, DPPH radical scavenging ability). Was evaluated. For activity comparison, ascorbic acid was treated with the sample to measure electron donation. Antioxidant activity of CK and GK kochujang at all concentrations was concentration-dependent, but did not change as the ripening progressed. WK and WGK kochujang were 56.61% and 61.69% at 30 days of fermentation at 4,000 ㎍ / mL (Fig. 12). ), At 60 days of aging, with 56.64% and 62.92%, respectively (Figure 13), showed higher antioxidant capacity than CK kochujang and GK kochujang. In CK and GK kochujang, GK kochujang showed good antioxidant activity. In general, it was confirmed that WK kochujang and WGK kochujang were higher in antioxidant activity than CK kochujang and GK kochujang from before ripening until 90 days (FIG. 14). This may be because the white lotus extract has its own high antioxidant capacity.

2) Measurement of cancer cell proliferation inhibitory effect

In order to measure the growth inhibitory effect on cancer cells, human acute mononuclear leukemia (THP-1), human acute promyelocytic leukemia (HL60), and histiocytoma (U937) were measured by MTT assay. The sample used Kochujang at 30 days of aging was excellent in antioxidant capacity and generally superior in GABA content. 10 g of CK, GK, WK, and WGK kochujang samples were mixed with 50 mL of sterile water, stirred at 37 ° C. for 2 hours, homogenized, filtered, and used as MTT assay samples. Experiments were performed with sterile water without addition of the sample as a control. The inhibitory effect of kochujang on THP-1 cells at 2,000µg / mL showed 42% inhibition of cancer cell proliferation in GK and WGK kochujang than CK kochujang (13.2%) and WK kochujang (29%) (Fig. 15). . The inhibitory effect of kochujang on HL60 cells was 62% in WK kochujang and 91.9% in WGK kochujang compared to CK pepper (11%) and GK kochujang (28.2%) at 2,000㎍ / mL. 16), the inhibition of proliferation of kochujang on U937 cells confirmed 77.9% of cancer cell proliferation inhibitory effect in WGK kochujang (FIG. 17). Overall, the anticancer effect is better in WGK kochujang, white lotus kochujang prepared by adding enzyme-treated amino acid (amino acid-derived GABA) powder than CK, GK and WK kochujang.

3) Evaluation of wrinkle improvement effect

① Determination of Hyaluronidase activity inhibitory effect

Hyaluronic acid is a polymer polysaccharide with glucuronic acid and glucosamine repeatedly connected. Hyaluronic acid inhibits the phagocytic ability of marcophage, an important factor in inflammation formation. On the other hand, hyaluronic acid degradation products or hyaluronic acid can increase anti-inflammatory and wrinkle improvement by increasing inflammation, fibrosis and collagen deposition in wound healing process, and hyaluronidase inhibition that decomposes hyaluronic acid can anti-inflammatory and wrinkle improvement. The hyalurinidase inhibitory effect was measured by searching for anti-wrinkle agents. Kochujang used in the experiment was extracted with 70% ethanol extract of kochujang at 30 and 60 days of fermentation, which had high GABA content and antioxidant effect, and tannic acid as a reference material.

Hyaluronidase inhibitory activity of kochujang at 30 days of aging with 70% ethanol was found to be concentration dependent in all samples. The inhibitory activity of the sample was 38.15 ~ 28.94% below 50% at 540㎍ / mL, and hlyaluronidase inhibitory activity was higher than CK, WK, and GK kochujang. 18). In the 70% ethanol extract of 60-day-old kochujang, CK kochujang (30.47%), WK kochujang (40.08%), and WGK kochujang (40.73%), except for GK kochujang (22.35%), showed high hyaluronidase inhibition at 540㎍ / mL. Appeared (FIG. 19).

Inhibitors of known hyaluronidase activity in plants are known as flavonoids, glycyrrhizin, tannins, iridoid glycosides or unsaturated fatty acids. The inhibitory activity of WK kochujang and WGK kochujang ethanol extracts containing white lotus extracts is known to affect the components derived from white lotus. It may be due to the possibility.

② Elastase activity inhibition effect

Matrix metalloproteinases (MMPs), which are present in the dermal layer of skin caused by UV rays and free radicals, are closely related to skin aging, that is, wrinkle formation, and the main components of MMPs are collagenase, gelatinase, and elastase, which reduce skin elasticity and prevent wrinkle formation. Decreased elastase activity is very important. Therefore, the improvement of wrinkles of kochujang was evaluated by measuring elastase activity inhibition and ursolic acid was used as a standard. Inhibition rate of kochujang by using ethanol extracts of 30 and 60 days of aging was high.

The results of the elastase inhibition rate measured by concentration (1.7 ㎍ / mL, 17 ㎍ / mL, 170 ㎍ / mL, 1,700 ㎍ / mL) by ethanol extraction of 30 and 60 days of kochujang samples were aged. The 70-day ethanol extract of kochujang at 30 days of fermentation showed elastase inhibition rate, and the inhibition rates of CK, WK, GK, and WGK kochujang were 12.31 ~ 33.79%, 17.28 ~ 48.69%, 19.50 ~ 40.32%, and 17.21 ~ 45.88%, respectively. Among them, WK kochujang and WGK kochujang extract with white lotus extract showed 48.69% and 45.88% elastase inhibition at 1,700㎍ / mL. The elastase inhibition rate of each kochujang in the 70% ethanol extract of 60-day-old kochujang was 42.79 ~ 23.93% for CK kochujang, 50.45 ~ 18.50% for WK kochujang, 52.53 ~ 28.62% for GK kochujang, and 44.35 ~ 20.83% for WGK kochujang. Inhibition rate was shown (FIG. 21). The elastase inhibition rate of CK and GK kochujang was higher than that of 30 days of fermentation, and WK pepper and WGK kochujang were similar to 30 days of fermentation.

③ Inhibitory effect of mouse-derived Melanoma cell (B16F10 cell) proliferation

22 shows the results of measuring the cytotoxic effect (MTT assay) on mouse-derived melanoma cells of 30 days of kochujang water extract, which had high GABA content and antioxidant function.

The cell activity was decreased due to the increase in the concentration of the extract due to the toxicity or adhesion interference of the extract. At 20,000㎍ / mL of the sample, the cell activity was 48.8% for CK kochujang, 48.3% for WK kochujang, and GK compared to the control group without the sample. Kochujang inhibited the growth of melanoma cells in 61.9% and WGK kochujang in 71.4%.

4. Sensory Evaluation

The sweetness, sourness, savory taste, and spicy taste of kochujang were similar in salty flavor, and the color of kochujang was slightly lower in GK, WK, WGK kochujang than CK kochujang, and the saltiness was different in GK kochujang and WGK kochujang. It was higher than that of CK and WK kochujang. There was no significant difference between WGK Kochujang (2.88 points), CK Kochujang (2.75 points), GK Kochujang (2.71 points), and WK Kochujang (2.71 points) (Table 8, Figure 23). Overall, the taste of kochujang is different because it adds starch syrup and other recipes. Kochujang manufacturing method used in this experiment may be somewhat unfamiliar to the general public based on the standard recipe, but it is a control group containing kochujang using white lotus extract and kochujang added with enzyme-treated amino acid (GABA) powder. Compared to kochujang, there is no difference in taste and flavor.

Claims (6)

In the manufacturing method of kochujang,
Mixing 30 to 60 parts by weight of malt oil in 1200 to 1700 parts by weight of purified water;
Saccharifying the mixed malt liquid at 40 to 70 ° C. for 2 to 5 hours;
Mixing the raw materials with 1 to 4 parts by weight of the enzyme-treated amino acid (amino acid-derived GABA) powder, 40 to 60 parts by weight of meju powder, 150 to 350 parts by weight of red pepper powder, and 80 to 150 parts by weight of salt to the saccharified malt solution;
It is a step of packing and storing mixed raw materials in a packing container.
A method for producing red pepper paste containing GABA, characterized in that the skin beauty effect.
In the manufacturing method of kochujang,
Mixing 30 to 60 parts by weight of malt oil to 1200 to 1700 parts by weight of white lotus extract;
Saccharifying the mixed white lotus-starch oil solution at 40 to 70 ° C. for 2 to 5 hours;
Mixing raw materials with saccharified white lotus-starch oil solution with 40 to 60 parts by weight of meju powder, 150 to 350 parts by weight of red pepper powder, and 80 to 150 parts by weight of salt;
It is a step of packing and storing mixed raw materials in a packing container.
A method for producing red pepper paste containing GABA, characterized in that the skin beauty effect.
The method of claim 2,
The white lotus extract contains GABA, characterized in that it is made by putting in the 25 ~ 70 parts by weight of dried white lotus leaves in 700 ~ 1200 parts by weight purified water 30 ~ 60 hours at 30 ~ 70 ℃, and extracts with a skin-cosmetic effect Manufacturing method.
The method according to claim 2 or 3,
The kochujang contains a GABA, characterized in that 1 to 4 parts by weight of the enzyme-treated amino acid (amino acid-derived GABA) powder is further mixed, and has a skin-cosmetic effect.
Kochujang prepared by the method of any one of claims 1 to 3.
Kochujang made by the method of claim 4.

Images