KR20150080312A - Red pepper paste comprising a black rice and seaweed - Google Patents

Abstract

The present invention relates to a functional red pepper paste that enhances antioxidant, anti-cancer and inflammation inhibiting effects of a red pepper paste. More specifically, the invention relates to a functional red pepper paste containing a black rice and a functional red pepper paste containing a black rice and seaweed. The red pepper paste containing a black rice and seaweed of the present invention contains a large amount of sugar and an amino acid and contains a large amount of fucoidan and therefore, has a flavor and is beneficial to health. Accordingly, the invention can be effectively used as a functional red pepper paste.

Description

[0001] The present invention relates to a red pepper paste comprising a black rice and a seaweed,

The present invention relates to a functional kochujang and a method for producing the same.

Kochujang, which was produced in the form of traditional domestic handicrafts, is a kind of traditional soup made by adding red pepper powder as main ingredient and adding malt and sub ingredient. Depending on the type of cereals to be added, it is divided into glutinous rice kochujang, flour kochujang, barley kochujang, sorghum kochujang, and red bean kochujang, etc. Also, it can be made into fruit juice kochujang, Recently, three major domestic companies have started production of kochujang, which accounts for 37% of domestic kochujang market, and kochujang occupies the largest market among 80% of total kochujang market. Therefore, It is a standardized form according to production method.

 Domestic kochujang market is estimated to have grown by 1% compared to the previous year, but the amount of exports has increased more than 128% compared to the previous year as of February 2010. While the market share of Japanese smile, which is a fermented soybean which has been selling well in major importing countries such as the US, China and Japan, has declined, the market share of Korean varieties has increased from $ 27,999 thousand in 2007 to $ 29,252 thousand in 2010, In addition to the spread of Korean food, which is recognized as a health food, it is recognized that the interest of kochujang increases in foreign countries. However, in the case of kochujang, which is manufactured in various forms, it is manufactured in a uniform form by large corporations. Therefore, it is necessary to develop various products in order to strengthen long - term product competitiveness in saturation of the domestic market and overseas advance.

In order to solve this problem, the addition of Baekryikcho extract and enzyme-treated amino acid powder increased the production of kochujang with increased GABA content, the establishment of solid kochujang manufacturing method with increased simplicity, portability and processability, the development of kochujang containing sensitive syrup (KR10-2013 -0128682A), and the development of kochujang for prevention of hypertension using germinated barley. Various studies are underway to improve quality characteristics and functionality by adding various natural herb extracts to kochujang.

It is reported that 400 seaweed species are native to Korea, and about 50 species of seaweed are used for edible purposes, including seaweed, seaweed, kelp, tortoise, and palaea. Seaweeds are rich in common nutrients and minerals such as proteins, lipids, and carbohydrates, and are characterized by high content of essential trace elements such as calcium, magnesium, iodine, iron, and zinc. In addition, it has been reported that it is effective for prevention of adult disease and obesity. Recently, it has been attracting attention as a health food, but the case applied to kochujang is insignificant. In addition, black rice has been reported as a kind of colored pigments showing black purple color and various physiological activities have been reported. Therefore, various researches on the development of high value-added foods have been tried, but the utilization rate is very small.

Therefore, in order to increase the utilization of local biomass, which has the highest market share as a major producer of seaweeds and grains distributed throughout the country, The present invention has been accomplished while studying the possibility of application of seaweed and black rice.

Korean Patent Publication 10-2013-0128682 Abstract and claims

It is an object of the present invention to provide a functional kochujang having an antioxidant effect, an anti-cancer effect and an inflammation-inhibiting effect.

In order to achieve the above object, one embodiment of the present invention provides a functional red pepper paste containing black rice. At this time, the black rice may be germinated black rice. In addition, the functional kochujang may further include saccharified liquid, red pepper powder, meju powder, and salt.

Here, black rice means rice in which a part or whole of the rice grain is black. In addition, germinated black rice means rice spreading in black rice. The bud may be about 1 to 5 mm, and the germinated black rice used means that the black rice is germinated in a germinated state of 50% or more, 80% or more, preferably 90% or more of the total black rice used.

The saccharified liquid may be a liquid prepared by mixing water and malt flour, removing the malt flour, and adding black rice. The powdered malt refers to a powder prepared by pulverizing malt or malt. The black rice may be contained in the saccharified liquid thus prepared and contained in the red pepper paste.

The saccharified liquid may include 10% to 90% of the total weight of the kochujang. Further, it may include 15% to 85%, 20% to 80%, 25% to 75%, 30% to 70%, 35% to 65%, 40% to 60%, or 45% to 50%.

The red pepper powder means that the red pepper is pulverized by drying or freeze drying. The red pepper powder may include 10 wt% to 20 wt%, and may include 14 wt% to 18 wt%, and may include 16 wt%, based on the weight of the saccharified liquid.

Meju flour means that Meju is pulverized by drying or freeze-drying. The meju powder may include 5 wt% to 15 wt%, 6 wt% to 10 wt%, and 8 wt% of the saccharified liquid.

In addition, the salt may contain 5 wt% to 15 wt%, 6 wt% to 10 wt%, and 8 wt% of the saccharified liquid.

Another embodiment is to provide a functional red pepper paste containing black rice and seaweed. At this time, the black rice may be germinated black rice. In addition, the functional kochujang may further include saccharified liquid, red pepper powder, meju powder, and salt. In addition, the seaweed may be any one selected from the group consisting of kelp, sea bass, seaweed, maize, marigold, spinach, pauliai and top.

In addition, the above-mentioned seaweeds may be contained in the form of seaweed powder or seaweed hot-water extract. The powder form means that the seaweed is dried or lyophilized and then pulverized to produce a powder form.

In addition, the seaweed hot-water extract can be extracted by any extraction method known in the art, but dry seaweed can be extracted by pulverization and then immersed in an extraction solvent. The extraction solvent may be water, alcohol, or a mixture thereof. The alcohol may be a C ₁ to C ₃ lower alcohol, preferably ethanol or methanol, but is not limited thereto. The extraction solvent may be added by 2 to 10 times the total amount of the seaweed, and the extraction solvent may be added by 3 to 8 times, but is not limited thereto. The extraction temperature may be from 30 to 100 ° C and from 45 to 85 ° C, but is not limited thereto. This is because the maintenance of the temperature is effective for preventing corruption, discoloration and detachment. The extraction time may be 5 to 20 hours, and may be 10 to 15 hours, but is not limited thereto. In addition, the number of times of extraction can be repeated 1 to 5 times, and can be repeated 2 to 3 times, but is not limited thereto. Extraction methods such as hydrothermal extraction, hot water extraction, ultrasonic extraction, Soxhlet extraction, and reflux cooling extraction can all be used in the conventional extraction methods.

The algae may contain 1 to 10 wt% or 3 to 5 wt% based on the weight of the saccharified liquid. And preferably 3 wt%. In addition, the red pepper powder may include 10 wt% to 20 wt%, and may include 14 wt% to 18 wt%, and may include 16 wt%, based on the weight of the saccharified liquid. In addition, the meju powder may contain 5 wt% to 15 wt%, 6 wt% to 10 wt%, and 8 wt% of the saccharified liquid. In addition, the salt may contain 5 wt% to 15 wt%, 6 wt% to 10 wt%, and 8 wt% of the saccharified liquid.

The kochujang containing the black rice, the kochujang including the germinated black rice, and the kochujang containing the algae additionally in the kochujang have excellent antioxidative effect, anti-cancer effect and excellent anti-inflammatory effect and can be used as a functional kochujang.

The functional kochujang according to the present invention is excellent in sugar content and flavor, has excellent antioxidative effect, has anticancer effect and anti-inflammatory effect and is highly likely to be used as a high-functional kochujang.

Fig. 1 shows the cytotoxicity of hepatocarcinoma cell line of algae-free kochujang: A; General glutinous saccharification solution + no added seaweed, B; Black rice glycation solution + no added seawater, C; Germinated black rice glycation solution + no added seaweed.
2 shows the NO inhibitory effect of kochujang without added seaweeds: A; General glutinous saccharification solution + no added seaweed, B; Black rice glycation solution + no added seawater, C; Germinated black rice glycation solution + no added seaweed.
Figure 3 shows the iNOs inhibitory effect of kochujang without added seaweeds: A; General glutinous saccharification solution + no added seaweed, B; Black rice glycation solution + no added seawater, C; Germinated black rice glycation solution + no added seaweed.
Figure 4 shows the cytotoxicity of hepatocellular carcinoma cells of the black rice seaweed kochujang: I; Germinated black rice glycation solution + seaweed powder 1% before fermentation, II; Germinated black rice glycation solution + seaweed powder 3% pre-fermented foodstuffs, III; Germinated black rice saccharification solution + seaweed powder 5% pre-fermented foodstuff, IV; Germinated black rice glycation solution + seawater hot water 1% pre-fermented foodstuffs, V; Germinated black rice glycation solution + seawater hot water 3% pre-fermented foodstuffs, VI; Germinated black rice glycation solution + seawater hot water 5%. Germinated black rice glycation solution + seaweed powder 3% after fermentation, VIII; Germinated black rice glycation solution + seawater hydrothermal 3% added after fermentation, IX; Germinated black rice glycation solution + no added seaweed.
Figure 5 shows the NO inhibitory effect of the black rice seaweed kochujang: I; Germinated black rice glycation solution + seaweed powder 1% before fermentation, II; Germinated black rice glycation solution + seaweed powder 3% pre-fermented foodstuffs, III; Germinated black rice saccharification solution + seaweed powder 5% pre-fermented foodstuff, IV; Germinated black rice glycation solution + seawater hot water 1% pre-fermented foodstuffs, V; Germinated black rice glycation solution + seawater hot water 3% pre-fermented foodstuffs, VI; Germinated black rice glycation solution + seawater hot water 5%. Germinated black rice glycation solution + seaweed powder 3% after fermentation, VIII; Germinated black rice glycation solution + seawater hydrothermal 3% added after fermentation, IX; Germinated black rice glycation solution + no added seaweed.
Figure 6 shows the iNOs inhibitory effect of the black rice seaweed koji paste: I; Germinated black rice glycation solution + seaweed powder 1% before fermentation, II; Germinated black rice glycation solution + seaweed powder 3% pre-fermented foodstuffs, III; Germinated black rice saccharification solution + seaweed powder 5% pre-fermented foodstuff, IV; Germinated black rice glycation solution + seawater hot water 1% pre-fermented foodstuffs, V; Germinated black rice glycation solution + seawater hot water 3% pre-fermented foodstuffs, VI; Germinated black rice glycation solution + seawater hot water 5%. Germinated black rice glycation solution + seaweed powder 3% after fermentation, VIII; Germinated black rice glycation solution + seawater hydrothermal 3% added after fermentation, IX; Germinated black rice glycation solution + no added seaweed.
Fig. 5 shows a process for producing red pepper paste.

In order to select grains suitable for the production of kochujang, the analysis of physicochemical components and the functional tests of three types of kochujang prepared by using glutinous rice, which is generally used for making kochujang, All of the three kinds of kochujang showed pH 4.554.57, which was lower than pH 5, which is the general pH of kochujang. In case of acidity, samples using black rice or germinated black rice were found to be more advantageous for the production of organic acid than common glutinous rice.

The amino acid nitrogen content was higher in the black rice and the germinated black rice than in the general glutinous rice samples, and the germinated black rice was increased in the flavor enhancement of red pepper paste. It was judged to be advantageous.

The inhibitory effect of nitric oxide (NO) and the DPPH free radical scavenging activity on hepatocellular carcinoma cell lines were higher than those of black rice and ordinary glutinous rice. Was also found to be beneficial for the increase of the function of kochujang.

The sensory evaluation showed that the taste and flavor of kochujang made with black rice and germinated black rice was better than those with ordinary glutinous rice.

Therefore, germinated black rice was applied to the preparation of seaweed kochujang which was superior in overall composition, functional and sensory preference in the production of kochujang.

In order to select appropriate seaweed application method for the production of kochujang, the ginseng was prepared in the form of powder and hot water extracts, which had a high fucoidan content in the kochujang prepared by germination process, which had relatively good quality characteristics, functionality and sensory preference. %, 3%, and 5%, respectively, and analyzed the components, functionalities and sensory evaluation of nine kinds of kochujang prepared by adding them before and after fermentation.

As a result, pH, acidity, total sugar and reduction were different according to the samples but relatively similar results were obtained, whereas amino acid nitrogen content was higher than 1% and 5% at 3% , And the addition of pre - fermented soybean paste to fermented soybean paste before fermentation showed relatively high amino acid content after fermentation.

The fucoidan content of the 3% added group showed the highest value, while the 1% and 5% added groups showed lower values than the 3% added group. In general, fucoidan content was lower when fermented after fermentation than before fermentation. The fermentation rate of fermentation was 3% before fermentation.

The cytotoxicity, NO inhibitory activity and DPPH free radical scavenging activity of hepatocarcinoma cell line were higher than those of algae - free group. In general, the activity of the powdered and hydrothermal extracts increased to 1% 3%, while the activity of 5% of the powdered and hot-water extracts decreased to 3% It was judged to be appropriate.

The sensory preference was also higher in the 3% added group before fermentation than in the other samples, and 3% of the non - fermented powder was added before fermentation.

Hereinafter, the present invention will be described in more detail with reference to the following examples and test examples, but the present invention is not limited to the following examples and test examples.

Manufacturing example  One. Black rice , Germination Black rice  And kochujang containing seaweeds

1.1. Kochujang manufacturing

The glutinous rice cultivated in Jindo - gun in 2013 was used for the glutinous rice cultivation of red pepper paste, and the germinated black rice was used for the experiment.

1.2. Glutinous rice and Black rice , Germination  Manufacture of kochujang

2 Kg of malt flour was added to 20 L of water and immersed for 30 minutes. 15 L of malt juice was obtained by filtering the malt flour. 5 Kg of rice flour (glutinous rice, black rice, germinated black rice) was added to 15 L of malt juice, and saccharified liquid (20 Kg) was prepared at 60 for 2 hours. After adding 3.2 Kg of red pepper powder (equivalent to 16% by weight of saccharified liquid), 1.6 kg of meju powder (equivalent to 8% of saccharified liquid weight) and 1.6 kg of salt (equivalent to 8% of saccharified liquid weight) And fermented to produce kochujang (Table 1).

Production conditions of kochujang without added seaweed using glutinous rice, black rice, germinated black rice Manufacture of saccharified liquid Glycation Kochujang manufacturing Fermentation Sample number Malt juice + ordinary glutinous rice flour -> Red pepper powder + meju powder + salt -> A Malt juice + black rice flour -> Red pepper powder + meju powder + salt -> B Malt juice + germinated black rice flour -> Red pepper powder + meju powder + salt -> C

1.3. Germination black rice  And Kochujang using seaweed

(1%, 3%, 5%) and addition time (before and after fermentation of kochujang fermentation) were different in germinated black liquor glycyrrhiza in the preparation of kochujang using ginseng black rice and seaweed, And the preparation conditions of the black rice seaweed kochujang were shown in Table 2.

That is, 2 Kg of malt flour was added to 20 L of water and immersed for 30 minutes. Then, 15 L of malt juice was obtained by filtering the malt flour. After addition of 5 Kg of germinated black rice flour to 15 L of malt juice, saccharified liquid (20 Kg) was prepared by saccharification at 60 for 2 hours. After adding 3.2 Kg of red pepper powder (equivalent to 16% by weight of saccharified liquid), 1.6 kg of meju powder (equivalent to 8% of saccharified liquid weight) and 1.6 kg of salt (equivalent to 8% of saccharified liquid weight) And fermented to make kochujang. The sample was prepared by adding 0.2 Kg of powder (equivalent to 1% of the saccharified liquid, Sample I), 0.6 Kg (equivalent to 3% of the saccharified liquid, Sample II), 1.0 Kg (equivalent to 5% In the case of the hot-water extract, 3% (VII) and 3% (VI) of the powdered starch powder (IV), 3% % ≪ / RTI > The amount of salt added was adjusted to the same salt concentration by calculating the concentration of salt contained in the salt water.

Preparation Condition of Gypsophila Added Gypsum Using Germinated Black Rice sample
number Manufacture of saccharified liquid
Glycation Kochujang manufacturing Fermentation Addition after fermentation I Germinated black rice flour
+ Malt juice Almond powder (1%) -> + Red pepper powder + meju powder + salt -> - II Germinated black rice flour
+ Malt juice Powdered eggplant (3%) -> + Red pepper powder + meju powder + salt -> - III Germinated black rice flour
+ Malt juice Ground Powder (5%) -> + Red pepper powder + meju powder + salt -> - IV Germinated black rice flour
+ Malt juice Hot water extract
(Equivalent to 1% of powder) -> + Red pepper powder + meju powder + salt -> - V Germinated black rice flour
+ Malt juice Hot water extract
(Equivalent to 3% of powder) -> + Red pepper powder + meju powder + salt -> - VI Germinated black rice flour
+ Malt juice Hot water extract
(Equivalent to 5% of powder) -> + Red pepper powder + meju powder + salt -> - VII Germinated black rice flour
+ Malt juice - -> + Red pepper powder + meju powder + salt -> Seaweed powder (3%) VIII Germinated black rice flour
+ Malt juice - -> + Red pepper powder + meju powder + salt -> Seaweed hot water extract
(Equivalent to 3% of powder) IX Germinated black rice flour
+ Malt juice - -> + Red pepper powder + meju powder + salt -> -

1.4. Extract preparation

The supernatant and precipitate were obtained by centrifugation (3,000 rpm, 15 min) after immersing 10 g of the sample in 10 mL of distilled water and stirring at 90 for 1 hour. The supernatant was filtered and freeze-dried To obtain a hot water extract.

The solvent extracts were prepared by immersing 20 g of kochujang sample in 200 mL of 20% methanol, sonication for 1 hour, and filtering to obtain filtrate and residue. The extracted extract was extracted and concentrated under reduced pressure and freeze-dried at 37 using a vacuum evaporator equipped with a cooling aspirator (TOKYO RIKAKIKAI, JAPAN).

1.5. Reagents and media

The organic solvent used for the sample extraction was at least grade 1, and the organic solvent used for the analysis of the components was a tertiary distilled water (Fisher scientific korean Ltd.), which was a special solvent.

Reagents such as 1,1-diphenyl-2-picrylhydrazyl (DPPH), ascorbic acid, dimethyl sulfoxide (DMSO), thiazolyl blue tetrazolium bromide (MTT), lipopolysaccharide (LPS) and L- chemical co. (St. Louis, Mo., USA). In addition, the reagents used in the experiment were purchased from the first-class reagents.

(ATCC, Rockville, Md., USA) was used for the cell line Raw264.7 (macrophage, ATCC TIB-71, USA) and HepG2 Dulbecco's Modified Eagle Medium (DMEM), minimum essential medium (MEM), fetal bovine serum (FBS) and penicillin-streptomycin were purchased from Gibco-BRL (Grand Island, NY, USA).

Experimental Example  1. Physicochemistry of Kochujang Functional ingredient  analysis

1.1. pH  Measure

For pH measurement, 10 g of sample was sampled, placed in a triangular flask, diluted by adding 40 mL of distilled water, stirred for 30 minutes or more on a hot plate, and then measured using a pH meter.

1.2. Acidity measurement

Titratable acidity was measured by titration with 0.1 N NaOH solution until the pH reached 8.3, and the amount of 0.1N NaOH consumed was expressed as titratable acidity.

1.3. Total measurement

5 g of the sample was placed in a 500 mL triangular flask, and 180 mL of distilled water and 20 mL of 25% HCl were added to the reflux apparatus and hydrolyzed on a hot & stir plate for 3 hours. The hydrolysis solution was diluted with 500 mL of Mass Flask and tested by DNS.

1.4. Reduction sugar measurement

The reducing sugar was measured by DNS (dinitrosalicylic acid) method. Twenty milliliters of distilled water was added to 2 g of the sample, and the mixture was stirred and centrifuged at 3,000 rpm for 15 min to obtain a supernatant and a precipitate. To the supernatant was added 3 mL of dinitrosalicylic acid 90 water bath for 5 min. The ice bath was filled with ice. After cooling for 5 min, absorbance was measured at 550 nm and converted using a Glucose standard curve.

1.5. Amino acid  Nitrogen measurement

Amino nitrogen was measured by adding 20 mL of 36% Formaldehyde solution to the sample after the titration, and titrating the pH to 8.3 with 0.1 N NaOH (A). Blank test of 0.1N NaOH solution was carried out in the same manner and (B) Amino nitrogen content was calculated by the following equation.

< Equation  1>

Amino nitrogen (mg%) = (A-B) X 1.4 X F X 100 / S

1.6. Chromaticity measurement

Color difference meter to measure the color change of red pepper paste was measured using a (CR-400, Konica Minolta Sensing . INC., Japan) Hunter color value, that is ^{L * (Lightness), a *} (Redness), b * (Yellowness ) Values.

1.7. Fucoidan  analysis

One) Crude fucoidan  extraction

Crude fucoidan extraction was performed by centrifuging (3,000 rpm, 15 min) with 10 g of CaCl ₂ to remove alginic acid in 250 mL of each hot water extract. The supernatant and sediment were obtained, and the concentration of ethanol in the supernatant was 80% After putting it, let it stand. After allowing to stand, centrifugation (3, 00 rpm, 15 min) was performed. The resulting supernatant and the supernatant were discarded and the precipitate was lyophilized and used in the experiment.

2) HPLC / ELSD  analysis

Crude fucoidan analysis was performed using HPLC system (high performance liquid chromatography, Shimadzu 2010, Tokyo, Japan) under the same conditions as Table 3.

HPLC conditions for analysis of fucoidan                                                        Condition Instruments Shimadzu 2010 HPLC Column Imtakt Presto FF C18 (4.6150 mm, 2 m) Flow rate 0.3 mL / min ven temp. 37 ℃ ressure (N2 gas) 47 psi rift tube temp. 50 ℃ obile Phase  A (85%): Water: Triethylamine: Acetic acid (100: 1.1: 0.5)  B (15%): Acetonitrile

Experimental Example  2. In liver cancer cell line  To measure cytotoxicity against

2.1. Cell culture

HepG2 cell line (human hepatoma, ATCC - 77400, USA) has given cultured under the conditions of 37, 5% CO ₂ using 10% fetal bovine serum (FBS) Dulbecco's Modified Eagle Medium (DMEM) that contains, all experiments The cell lines used in this study were subcultured 37 times after thawing.

2.2. Cytotoxicity measurement

The cytotoxicity of 20% MeOH extracts from each of the kochujangs was measured using the HepG2 cell line (human hepatoma, ATCC - 77400, USA) MTT colormetric assay. HepG2 cell lines were seeded at a concentration of 110 ⁵ cells / mL in a 96-well plate at a concentration of 200 L / well, cultured for 24 hours at 37 ° C and 5% CO ₂ , and then mixed with 9 concentrations of 20% MeOH extract Respectively. Cells treated with the samples were cultured for 24 hours, and 20 L of the MTT reagent prepared at a concentration of 2 mg / mL was added thereto, and the cells were regenerated for 4 hours to form formazan. Formazan formed on the bottom of a 96 well plate was dissolved in 200 uL of dimethylsulfoxide (DMSO, Sigma, USA) and absorbance was measured at 540 nm using an ELISA reader. When the absorbance of the control was set at 100%, relative cell growth and inhibition rates were obtained.

Experimental Example  3. Measurement of inflammation inhibition effect using immune cells

3.1. Cell culture

Macrophage the Raw264.7 cell line (ATCC TIB-71 , USA) using a 10% fetal bovine serum (FBS) dulbecco's modified eagle medium (DMDM) gave containing the cultured under the conditions of 37, 5% CO _2, Cell lines used for all experiments were subcultured within 37 cycles after thawing.

3.2. Nitric oxide  ( NO ) And inducible nitric oxide synthase  (iNOs) inhibitory activity measurement

The inhibition of nitric oxide production by 20% MeOH extract of kochujang was measured by Raw264.7 cell line (ATCC TIB-71, USA) g riess reagent assay. The macrophage Raw264.7 cell line was seeded at a density of 510 ⁴ cells / mL in a 24-well plate and cultured at 37 ° C and 5% CO ₂ for 24 hours. Lipoprotein (LPS, Sigma, USA) and L-arginine (10 L / well) were treated at the concentration of 10 g / mL. After culturing for 24 hours at 37 ° C and 5% CO ₂ , 300 L of the culture was reacted with 300 L of griess reagent and the absorbance (540 nm) was measured by UV / Vis spectrophotometer. The percentage of nitric oxide production in the sample-treated group was expressed as a percentage (%). We also measured the amount of L-citrulline produced by Raw 264.7 in order to confirm the inhibition of iNOS activity by samples. The culture supernatant (300 L) was mixed with 300 L of chromogenic reagent and incubated at 100 for 10 min. The amount of L-citrulline in the reaction was measured at 530 nm using a UV / Vis spectrophotometer.

Experimental Example  4. Antioxidant activity measurement

The antioxidant activity of DPPH radical scavenging ability was measured by the method of Blois (1985) modified as follows. 400 L of 100 mM Tris-HCl buffer (pH 7.4) was added to 100 L of each prepared sample solution, and then 500 L of 200 mM DPPH in MeOH solution was added thereto. After mixing in a vortex mixer for 10 min in a dark place, the absorbance was measured at 517 nm. Scavenging activity was measured by the absorbance of the DPPH radical itself to the DPPH radical scavenging absorbance of the sample.

< Equation  2>

DPPH radical scavenging activity (%) = (1-A / B) X100

A: Absorbance value of sample

B: Absorbance value of distilled water

Experimental Example  5. Sensory Evaluation of Kochujang

Sensory evaluation of the prepared black rice seaweed kochujang was performed by 30 students of fermentation academy as a panel. The taste, aroma, color and general preference degree items were ranked according to the 10 point scale method.

Experimental Example  6. Selection of rice varieties suitable for kochujang

6.1. Glutinous rice and Black rice , Germination  Analysis of Physicochemical Composition of Kochujang Prepared Using

In order to select grains suitable for the production of kochujang, the physicochemical components of three kinds of kochujang prepared by using glutinous rice, which is generally used in the manufacture of kochujang, and those of kochujang produced by using black or germinated black rice, (Table 4). As a result, all three kinds of kochujang showed similar pH of 4.554.57, which is consistent with the report that the pH decreased due to the effect of organic acids produced by microorganisms in the general fermentation of kochujang, Respectively. On the other hand, acidity of sample B and sample C using black rice or germinated black rice was higher than that of sample A using common glutinous rice.

In the case of total sugar and reducing sugar contained in kochujang, all three samples showed similar results. In case of amino acid nitrogen content produced by decomposition of protein, sample B using black rice and germinated black rice and sample C, and Sample C showed higher amino acid content than Sample B, indicating that kochujang using germinated black rice as a raw material for protein degradation was relatively superior. Thus, it is advantageous to use germinated brown rice for improving the flavor of kochujang .

Physico-chemical composition of kochujang without added seaweed
Sample Physics & chemistry analysis pH Acidity Total sugar (mg / g) Reducing sugar (mg / g) Amino nitrogen (mg%) A 4.57 18.55 51.52 5.13 73.43 B 4.55 20.60 50.55 5.11 84.56 C 4.56 20.05 50.61 5.17 85.46

A; General glutinous saccharification solution + no added seaweed, B; Black rice glycation solution + no added seawater, C; Sprouted black rice glycation solution + no added seaweed

6.2. Glutinous rice and Black rice , Germination  The color analysis results of kochujang prepared by using

As a result of analyzing chromaticity, one of the sensory quality indicators of kochujang, redness and yellowness were higher in sample C using germinated black rice than in sample B using black rice.

Color of kochujang without added seaweed Sample No. Hunters value A
L 31.6 a 12.5 b 7.74 B

L 27.8 a 4.2 b 2.7 C

L 27.2 a 6.4 b 4.1

A; General glutinous saccharification solution + no added seaweed, B; Black rice glycation solution + no added seawater, C; Sprouted black rice glycation solution + no added seaweed

6.3. Glutinous rice and Black rice , Germination  Functional Test Results of Kochujang Prepared Using

One) In liver cancer cell line  To measure cytotoxicity against

In order to select the grains suitable for the manufacture of kochujang, the toxicity of hepatocellular carcinoma cells to three types of kochujang produced using glutinous rice, which is generally used in the manufacture of kochujang, and black or germinated black rice were compared (FIG. 1). As a result, the activity of the sample C prepared using germinated black rice was the highest, and the activity of the sample B using the black rice was slightly higher than that of the sample A using the general glutinous rice, but the difference was not large. Therefore, it was concluded that the method of using germinated brown rice, which is relatively available as an anticancer material, is more advantageous for increasing the functionality of kochujang.

2) Measurement of inflammation inhibition effect using immune cells

Inhibition of nitric oxide (NO) overexpressed in macrophages in order to compare the inhibitory effects of three kinds of kochujang prepared by using glutinous rice, which is generally used in the manufacture of kochujang, (FIGS. 2 and 3). As a result, the activity of the sample C prepared using germinated black rice was the most excellent, and the NO inhibition effect was shown in the order of the sample B> the sample A, in that order. In addition, in order to examine whether the NO inhibitory effect of the sample is an inhibitory effect on iNOs related to NO production, the results of comparing the inhibitory effects of iNOs production also showed the same tendency and the possibility of use as an inflammation suppressing material was confirmed. Therefore, it was concluded that the method of using germinated brown rice, which is relatively high in availability as an anti - inflammatory material, is advantageous for increasing the functionality of kochujang.

3) Antioxidant activity measurement

The activity of DPPH free radical scavenging was measured to determine the antioxidative activity of the prepared kochujang using common glutinous rice, black rice and germinated black rice. As a result, the activity of sample C prepared using germinated black rice was 5.36 g / mL, (5.63 g / mL) and sample A (6.89 g / mL) prepared by using germinated brown rice in terms of antioxidant activity.

DPPH free radical scavenging effect of kochujang without added seaweed Sample DPPH freeradical scavenging activity (SC _50, g / mL) A 6.89 B 5.63 C 5.36

A: general glutinous saccharification solution + no addition of seaweed, B: saccharification solution of black rice, + no added seaweed, C: germinated black rice saccharified solution +

6.4. Glutinous rice and Black rice , Germination  Sensory Evaluation Results of Kochujang Prepared Using

In order to select grains suitable for the production of kochujang, the sensory evaluations of three kinds of kochujang prepared by using glutinous rice, which is generally used in the manufacture of kochujang, and black rice or germinated black rice, The taste and flavor were relatively better than those of the ordinary glutinous rice, and the preference of the sample C using the germinated black rice was evaluated to be the best in the general preference degree and taste and flavor.

Sensory evaluation results of kochujang without added seaweed Sample flavor incense color Comprehensive preference A 6.000.36 ^a 6.800.55 ^a 6.000.59 ^a 6.100.74 ^a B 7.000.40 ^b 8.200.38 ^b 4.900.92 ^a 5.800.35 ^a C 7.800.25 ^c 9.400.28 ^b 5.100.42 ^a 6.600.57 ^a

A; General glutinous saccharification solution + no added seaweed, B; Black rice glycation solution + no added seawater, C; Sprouted black rice glycation solution + no added seaweed

Experimental Example  7. Germination  Setting method of applying algae of kochujang prepared by using

7.1. Kochujang Fucoidan  Content analysis

In order to select the algae to be added in the production of kochujang, the hot-water extract of each seaweed was prepared and the content of fucoidan was analyzed by HPLC / ELSD (Table 8). As a result, the content of fucoidan contained in the seaweed was the highest. In general, when the sea tangle and seaweed used in fucoidan extraction were compared, the fucoidan content of seaweed was 469.181 mg / g and the seaweed was 351.024 mg / g The fucoidan content of the seaweed was higher than that of the seaweed by about 1.8 times higher than that of the seaweed. The result showed that the application of the fucoidan to the preparation of kochujang was beneficial for the addition of fucoidan .

Results of fucoidan content analysis of seaweed extracts. Sample Fucoidan component (mg / g) Kelp 469.191 Seaweed 351.024 Seamount stem 282.955 Imperial 518.347 Mysterious 91.010 Root 163.712 Blue 0.000 Top 180.968

8.2. Germination  Analysis of Physico-chemical Components of Seaweed Kochujang Prepared Using

In order to select appropriate seaweed application method for the preparation of kochujang, the ginseng was prepared by using germinated brown rice, which had relatively good quality characteristics, functional and sensory preference, 3%, and 5%, respectively, were analyzed for physicochemical properties of nine kinds of kochujang prepared before and after fermentation. As a result, pH of all nine kinds of kochujang were similar to pH 4.12 ~ 4.62, and acidity was 19.12 ~ 22.60. It was concluded that the fermentation proceeded normally in accordance with the report that the pH decreased to below pH 5 due to the influence of the organic acid generated by microorganisms in the general fermentation of kochujang.

In case of total sugar and reducing sugar contained in kochujang, 9 kinds of samples showed similar results. Amino nitrogen content produced by decomposition of protein was relatively higher than 1% and 5% at 3% , And the addition of fermented seaweeds to the kochujang before fermentation showed a relatively high content of amino nitrogen at pre - fermentation stage after fermentation.

In case of chromaticity, the addition of seaweed before fermentation showed a relatively higher lightness (L) than that after fermentation. In the case of addition of powder, there was no difference in L, a, (A) and yellowness (b) tended to increase with increasing addition ratio (Table 10).

Physico-chemical composition of black rice seaweed kochujang
Sample Physics & chemistry analysis pH Acidity Total sugar (mg / g) Reducing sugar (mg / g) Amino nitrogen (mg%) I 4.51 19.61 48.43 4.93 81.76 II 4.47 20.39 50.21 4.92 97.68 III 4.51 19.87 47.66 4.30 93.55 IV 4.36 19.12 46.19 4.73 86.53 V 4.12 22.60 47.15 4.92 97.22 VI 4.31 20.82 45.38 4.51 94.58 VII 4.58 21.81 49.67 4.60 92.01 VIII 4.61 21.22 49.89 4.85 87.47 IX 4.56 20.05 50.61 5.17 85.46

I; Germinated black rice glycation solution + seaweed powder 1% before fermentation, II; Germinated black rice glycation solution + seaweed powder 3% pre-fermented foodstuffs, III; Germinated black rice saccharification solution + seaweed powder 5% pre-fermented foodstuff, IV; Germinated black rice glycation solution + seawater hot water 1% pre-fermented foodstuffs, V; Germinated black rice glycation solution + seawater hot water 3% pre-fermented foodstuffs, VI; Germinated black rice glycation solution + seawater hot water 5%. Germinated black rice glycation solution + seaweed powder 3% after fermentation, VIII; Germinated black rice glycation solution + seawater hydrothermal 3% added after fermentation, IX; Sprouted black rice glycation solution + no added seaweed

Black rice Color of seaweed red pepper paste Sample No. Hunters value
I
L 27.68 a 6.14 b 4.08
II
L 26.84 a 6.43 b 4.51
III
L 26.31 a 6.58 b 4.62
IV
L 31.49 a 5.83 b 4.07
V
L 27.5 a 7.83 b 5.44
VI
L 25.98 a 9.06 b 6.14
VII
L 22.79 a 5.53 b 3.62
VIII
L 23.48 a 5.16 b 3.48
IX
L 27.19 a 6.36 b 4.05

I; Germinated black rice glycation solution + seaweed powder 1% before fermentation, II; Germinated black rice glycation solution + seaweed powder 3% pre-fermented foodstuffs, III; Germinated black rice saccharification solution + seaweed powder 5% pre-fermented foodstuff, IV; Germinated black rice glycation solution + seawater hot water 1% pre-fermented foodstuffs, V; Germinated black rice glycation solution + seawater hot water 3% pre-fermented foodstuffs, VI; Germinated black rice glycation solution + seawater hot water 5%. Germinated black rice glycation solution + seaweed powder 3% after fermentation, VIII; Germinated black rice glycation solution + seawater hydrothermal 3% added after fermentation, IX; Sprouted black rice glycation solution + no added seaweed

8.3. Germination  Analysis of Functional Components of Seaweed Kochujang Prepared Using

The fucoidan content of the 3% added group was higher than that of the other group, and the fucoidan content of the 3% added group was higher than that of the other group. 1% and 5% groups showed lower values than 3% groups. In general, fucoidan content was lower when fermented than fermented before fermentation. Fucoidan content was increased in 1% and 3% fermented group, while fucoidan content in 5% fermented group was lowered in both powder and hot water extract And the addition rate of algae was 3% before fermentation.

Fucoidan content of black rice seaweed koji paste Sample No. Fucoidan contents (g / mg) I 421.12 II 462.28 III 441.05 IV 434.87 V 450.65 VI 423.83 VII 383.11 VIII 397.55

I; Germinated black rice glycation solution + seaweed powder 1% before fermentation, II; Germinated black rice glycation solution + seaweed powder 3% pre-fermented foodstuffs, III; Germinated black rice saccharification solution + seaweed powder 5% pre-fermented foodstuff, IV; Germinated black rice glycation solution + seawater hot water 1% pre-fermented foodstuffs, V; Germinated black rice glycation solution + seawater hot water 3% pre-fermented foodstuffs, VI; Germinated black rice glycation solution + seawater hot water 5%. Germinated black rice glycation solution + seaweed powder 3% after fermentation, VII; Germinated black rice saccharification solution + seawater hot water 3% after fermentation

8.4. Germination  Functional Test Results of Seaweed Kochujang Prepared Using

One) In liver cancer cell line  To measure cytotoxicity against

As a result of comparing the cytotoxic effect of Kochujang samples on the liver cancer cell line by the addition of seaweed, the activity of seaweed added group was higher than that of the seaweed free group IX. The cytotoxicity of hepatocellular carcinoma cells was higher than that of the other samples regardless of the addition ratio, and increased to 1% to 3% in both powder and hot water extracts. And the addition ratio was 3%. Also, the method of adding before fermentation showed higher activity in both powder and hot water extracts than after fermentation (see Figure 4).

2) Measurement of inflammation inhibition effect using immune cells

The inhibitory effect of NO on the kochujang samples by macrophage addition method was higher than that of seaweed without added seaweed. The inhibitory activity of NO was higher than that of other samples regardless of the addition ratio, and the activity was increased up to 1% 3% and 5% for both powder and hot water extracts in the sample group added with pre-fermented powder And the addition ratio was 3%. Also, the method of adding before fermentation showed higher activity in both powder and hot water extracts than after fermentation (see FIG. 5).

3) Antioxidant activity measurement

As a result of comparing DPPH free radical scavenging activity of Kochujang samples by seaweed addition method, DPPH free radical scavenging activity was higher than that of seaweed free group IX when fermented pre - fermented seaweed was added as powder or hot water extract, Showed similar or decreased activity. In the case of pre-fermentation group, the addition of non-seaweed powder showed a higher DPPH free radical scavenging activity than that of hot-water extract, and activity of up to 1% 3% in both powder and hot water extracts was increased The activity tended to decrease at 5% and the addition ratio of 3% was considered to be suitable.

DPPH free radical scavenging activity of black rice kochujang Sample DPPH freeradical scavenging activity (SC _{50 ,} g / mL) I 4.18 II 3.43 III 4.45 IV 5.48 V 4.43 VI 4.65 VII 5.12 viii 6.51 IX 5.36

I; Germinated black rice glycation solution + seaweed powder 1% before fermentation, II; Germinated black rice glycation solution + seaweed powder 3% pre-fermented foodstuffs, III; Germinated black rice saccharification solution + seaweed powder 5% pre-fermented foodstuff, IV; Germinated black rice glycation solution + seawater hot water 1% pre-fermented foodstuffs, V; Germinated black rice glycation solution + seawater hot water 3% pre-fermented foodstuffs, VI; Germinated black rice glycation solution + seawater hot water 5%. Germinated black rice glycation solution + seaweed powder 3% after fermentation, VIII; Germinated black rice glycation solution + seawater hydrothermal 3% added after fermentation, IX; Sprouted black rice glycation solution + no added seaweed

4) Germination  Sensory Evaluation Results of Seaweed Kochujang Prepared Using

As a result of comparing the sensory characteristics of Kochujang samples according to the addition of seaweeds, the overall sensory preference was higher than that of seaweed - free group IX when fermented pre - fermented soybean powder was added as a powder or hot water extract. The pre - fermented group had lower sensory preference than the non - additive group (IX) before the fermentation. The pre-fermentation group had higher preference than the hot-water extract group in the addition of the non-inoculum powder, and the preference was increased to 1% to 3% in both powder and hot-water extracts. And the addition ratio was 3%.

Sensory Evaluation Results of Black Rice Seaweed Kochujang Sample flavor incense color Comprehensive preference I 6.601.90 ^c 6.301.89 ^bc 6.102.18 ^b 6.101.45 ^c II 7.201.14 ^c 8.801.14 ^d 3.401.51 ^a 7.101.10 ^c III 6.301.64 ^c 5.701.34 ^b 3.201.40 ^a 3.702.26 ^a IV 6.001.49 ^c 5.501.65 ^b 6.202.20 ^b 6.101.79 ^c V 6.701.34 ^c 7.601.58 ^c 5.601.96 ^b 6.401.26 ^c VI 5.802.20 ^b 5.201.55 ^b 3.401.78 ^a 5.501.84 ^b VII 4.401.71 ^ab 3.902.13 ^ab 6.602.55 ^b 3.702.26 ^a VIII 4.001.89 ^a 4.001.76 ^ab 6.501.51 ^b 3.702.06 ^a IX 3.501.78 ^a 3.102.18 ^a 6.601.84 ^b 4.002.00 ^ab

I; Germinated black rice glycation solution + seaweed powder 1% before fermentation, II; Germinated black rice glycation solution + seaweed powder 3% pre-fermented foodstuffs, III; Germinated black rice saccharification solution + seaweed powder 5% pre-fermented foodstuff, IV; Germinated black rice glycation solution + seawater hot water 1% pre-fermented foodstuffs, V; Germinated black rice glycation solution + seawater hot water 3% pre-fermented foodstuffs, VI; Germinated black rice glycation solution + seawater hot water 5%. Germinated black rice glycation solution + seaweed powder 3% after fermentation, VIII; Germinated black rice glycation solution + seawater hydrothermal 3% added after fermentation, IX; Sprouted black rice glycation solution + no added seaweed

The kochujang of the present invention is excellent in flavor, excellent in physicochemical properties, and excellent in antioxidative, anti-cancer and anti-inflammatory effects, and can be utilized as a functional kochujang instead of the conventional kochujang.

Claims (13)

Functional red pepper paste containing black rice. The functional red pepper paste according to claim 1, which is germinated black rice. The functional red pepper paste according to claim 1, wherein the functional red pepper paste comprises a saccharified liquid, red pepper powder, meju powder and salt. The functional red pepper paste according to claim 3, which is prepared by mixing the saccharified liquid and the malt flour, removing the malt flour, and then adding black rice. [6] The method of claim 3, wherein the red pepper powder comprises 10 wt% to 20 wt% of the saccharified liquid, and the meju powder contains 5 wt% to 15 wt% of the saccharified liquid, And 5 wt% to 15 wt%. The method of claim 5, wherein the red pepper powder comprises 16 wt% based on the weight of the saccharified liquid, the meju powder includes 8 wt% based on the weight of the saccharified liquid, and the salt includes 8 wt% Functional red pepper paste. Functional red pepper paste containing black rice and seaweed. 8. The red pepper paste according to claim 7, wherein the black rice is germinated black rice. [Claim 7] The red pepper paste according to claim 7, wherein the seaweed is any one selected from the group consisting of kelp, seaweed, seaweed, maize, marbling, spinach, [Claim 7] The red pepper according to claim 7, wherein the seaweed is seaweed powder or seaweed hot water extract. The functional red pepper paste according to claim 7, wherein the functional red pepper paste comprises a saccharified liquid, red pepper powder, meju powder and salt. 12. The red pepper paste according to claim 11, wherein the algae are 1 to 10 wt% based on the weight of the saccharified liquid. [12] The method of claim 11, wherein the red pepper powder comprises 10 wt% to 20 wt% based on the weight of the saccharified liquid, wherein the meju powder contains 5 wt% to 15 wt% And 5 wt% to 15 wt%.

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