KR20200050483A - An enzyme treatment method to activate antioxidants from wheat embryos containing antioxidants - Google Patents

Abstract

The present invention relates to an enzyme treatment method of wheat germ containing antioxidants, wherein water is added to wheat germ which is then added with celluclast to induce a reaction in a double boiler, followed by filtering, freezing and drying. The present invention activates antioxidants that suppress the generation of excess oxygen, that is a free (active) oxygen radical, that causes cancer and degenerative diseases, and thus can be added to be applied as a material for functional health foods, anti-cancer foods, and the like.

Description

Enzyme treatment method to activate antioxidants from wheat embryos containing antioxidants

The present invention is an antioxidant that suppresses the generation of excess oxygen, ie, free (active) oxygen radicals (free radicals), which cause cancer and degenerative diseases, through the addition of celluclast to wheat germ, a by-product of flour milling. Activated to enable the application of supplementary ingredients such as health functional foods and anti-cancer foods. Also, through the high value-added food production of undervalued wheat germs, antioxidants are used to increase public health and increase farmers 'income, such as increasing wheat farmers' income. It relates to an enzyme treatment method for activating antioxidants from this contained wheat germ.

Recently, as consumers around the world as well as Korean consumers have increased interest in safe, functional and nutritious foods, research to develop new functional materials is rapidly increasing. In particular, research is underway to identify high-functional substances among substances that were discharged as by-products during processing in food resources and to develop new health foods using them.

Corn, rice and wheat are the world's three largest crops, and wheat is the staple food in most countries except Asia, where rice is the staple food. Wheat has been milled and used in various food processing as well as bread and noodles. In the Asian countries, the amount of processed wheat products has recently increased, and in Korea, 33-35 kg, which is more than 1/2 of 61.8 kg of rice per year in 2017, consumes one meal a day. Yes (KOSTAT 2018).

Wheat refers to the grass or grains of the rice family (Gra 科 Gramineae) and the wheat (― 屬 Triticum). It is used to make bread (Triticum vulgare / T. Aestivum), and durum wheat (T) used to make pasta such as spaghetti or macaroni. .durum), cake cracker cookie pastry, etc. There are soft T. compactum used for gravity.

Wheat grown in dry fat is usually hard wheat, which contains 11 to 15% of protein and is rich in gluten, making it best for making bread, whereas wheat grown in wet fat is soft wheat. It contains 8-10% and has a low gluten content, making it suitable for cake cracker cookie pastries, home wheat flour, etc.

As a main energy source for humans, such wheat usually contains 12% water, 70% carbohydrates, 12% protein, 2% fat, 1.8% minerals, and 2.2% crude fiber. 100g of wheat can produce about 330 calories. A small amount of vitamin A is found in wheat, but it is common to remove it during milling.

Hereinafter, a typical wheat milling process will be described, consisting of wheat selection, foreign matter removal, hydrolysis, tempering, grinding, sieving, and packaging. That is, the process of selecting wheat or removing foreign substances is generally referred to as a selection process, and a process of maintaining an appropriate amount of water and proper moisture conditions to facilitate crushing / separation of the selected wheat as described above is called a hydrolysis process. When the process of separating the skin and the seed skin is subjected to a sieving or crushing process, and then the wheat is pressed and rubbed to grind into a fine flour, and then the crushed flour is separated by particles using a sieve or after a sieving process and packaged. Will be.

It is true that the flour produced in this way excludes embryos from the components of wheat and has a problem of discarding many of the nutrients of the actual wheat.

Wheat germ accounts for about 2-3% of wheat grains, and is a material that can be separated purely during the crushing process, but has a high content of unsaturated fatty acids and rapidly reduces the nutrition and functionality of wheat germ during storage due to lipolytic enzymes or oxidase. Its use has been severely reduced and its use is limited (Boukid F, etc. 2018). Wheat germ contains a significant amount of bioactive compounds. Contains about 10-15% lipid, 26-35% protein, 17% sugar, 1.5-4.5% fiber and 4% minerals (Brandolini A & Hidalgo A 2012). Wheat embryos contain antioxidants such as α-tocopherol, phytosterols and polycosanols, and vitamin B (thiamin), riboflavin and niacin (Gelmz N et al. 2009, etc.) Megahed MG 2011).

Wheat germ, which currently contains about 8-14% fat (average 10%), is used in some food, medical and cosmetic industries, but it is necessary to improve its functionality and antioxidant activity to increase consumption. Antioxidants contained in wheat germ include fat-soluble vitamins, tocophenols and phenolic compounds, flavonoids and carotenoids, so embryo oil obtained from embryos can be said to be a good food containing antioxidants and vegetable sterols. For this reason, studies on antioxidant activity from fractions of whole wheat and wheat husks and embryos have been conducted steadily (Liyana-Pathirana CM & Shahidi F, 2007; Vaher M et al., 2010; Zhou et al., 2004). No studies have been used.

Korean Patent Application No. 10-2014-0168153 Korean Patent Registration No. 10-0491362 Korean Patent Registration No. 10-1345729

The present invention activates antioxidants that suppress the production of excess oxygen, which is the cause of cancer and degenerative diseases, that is, free (active) oxygen radicals (free radicals) through the addition of cellulcrust to wheat germ, a by-product of flour milling. It is possible to apply it as a material for health functional foods and anti-cancer foods, and it also contains antioxidants to increase national health and increase farm income by increasing the import of wheat farms through the high value-added foodization of undervalued wheat germ. The aim is to provide an enzyme treatment method for activating antioxidants from wheat embryos.

The present invention is a specific solution means for achieving the above object,

"The method of enzyme treatment of wheat germ containing antioxidants, characterized in that water is added to the wheat germ and then added to the cellulose to react with a water heater, filtered, and then frozen and dried.

200 ml of water is added to 50 g of wheat germ, and then 1.5 L of cellulose is added, reacted in a water bath at 50 ° C. for 6 to 30 hours, filtered, frozen at -50 ° C., and dried, containing antioxidants. Wheat germ enzyme treatment method,

The boiling water reaction time is 6 hours, 12 hours, 18 hours, 24 hours, 30 hours, selected from any one of the wheat germ enzyme treatment method containing antioxidant, characterized in that made by the constitutional characteristics of the above You can achieve your purpose.

The present invention configured as described above suppresses the production of excess oxygen, which is a cause of cancer and degenerative diseases, that is, free (active) oxygen radicals (free radicals) by adding cellulcrust to wheat germ, a by-product of flour milling. Activating antioxidants, it is possible to apply it as a material for health functional foods and anti-cancer foods, and localization of wheat germ extracts, which currently rely on imports, strengthens price competitiveness and replaces imports, anti-cancer of wheat germ extracts With regard to functionality, more than 150 functionalities related to anti-cancer effects have been recognized abroad, which can help reduce the economic burden of treatment for cancer patients and contribute to the improvement of national health by utilizing it for prevention. The cost burden of wheat is offset by the added value of wheat germ. It can exert the effect of the increase.

1 is an analysis condition table of HPLC for α-tocopherol analysis from enzyme-treated wheat embryos by an enzyme treatment method for activating antioxidants from wheat embryos containing antioxidants according to the present invention,
Figure 2 is a result of measuring the total polyphenol compound, total flavonoids, α-tocopherol from wheat embryos enzymatically treated by an enzyme treatment method for activating antioxidants from wheat embryos containing the antioxidants of the present invention,
3 is an enzyme treatment method for activating antioxidants from wheat embryos containing the antioxidants of the present invention, (A) is fresh wheat germ, (B) wheat germ added with cellulast 1.5L, (C ) Is a graph showing the content of α-tocopherol contained in wheat germ treated as a standard by HPLC chromatogram,
Figure 4 is the present invention according to different reaction times for wheat embryos treated with the enzyme cellocluster 1.5L by the enzyme treatment method for activating antioxidants from wheat embryos containing antioxidants according to different reaction times ABTS (A) and DPPH (B ) Is a graph showing enzyme activation.

Specific structural or functional descriptions of the embodiments according to the concept of the present invention disclosed in this specification are exemplified only for the purpose of explaining the embodiments according to the concept of the present invention, and the embodiments according to the concept of the present invention Since it is possible to make various changes and have various forms, it includes all changes, equivalents, or substitutes included in the spirit and scope of the present invention, and terms or words used in the specification and claims are conventional or dictionary Not to be construed as being limited to meaning, the inventor can, on the basis that he can appropriately define the concept of terms to describe his or her invention in the best way, meanings and concepts consistent with the technical spirit of the present invention Should be interpreted as Therefore, the embodiments shown in the specification and drawings of the present invention are only the most preferred embodiments of the present invention and do not represent all the technical spirit of the present invention, and are replaced at the time of filing of the present invention It should be understood that various equivalents and variations that can be made are possible or possible.

In addition, unless defined otherwise in the specification of the present invention, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by a person skilled in the art to which the present invention pertains. Have Terms such as those defined in a commonly used dictionary should be interpreted as having meanings consistent with meanings in the context of related technologies, and should not be interpreted as ideal or excessively formal meanings unless explicitly defined herein. Does not.

Hereinafter, preferred embodiments of the present invention will be described with reference to the accompanying drawings.

1 is an analysis condition table of HPLC for α-tocopherol analysis from an enzyme-treated wheat embryo by an enzyme treatment method for activating an antioxidant from a wheat embryo containing the antioxidant material of the present invention, and FIG. 2 is an antioxidant of the present invention Total polyphenol compounds, total flavonoids, α-tocopherols are measured from enzyme-treated wheat embryos by an enzyme treatment method for activating antioxidants from wheat embryos containing substances, and FIG. 3 is an antioxidant of the present invention In the enzymatic treatment method for activating antioxidants from the wheat germ containing this, (A) is fresh wheat germ, (B) is wheat germ added with Cellulcrust 1.5L, and (C) is wheat treated as standard. It is a graph showing the content of α-tocopherol contained in embryos by HPLC chromatogram, respectively, and FIG. 4 is an antioxidant from wheat embryos containing the antioxidant substance of the present invention It is a graph showing enzyme activation using ABTS (A) and DPPH (B) according to different reaction times of wheat embryos treated with enzyme cellocluster 1.5L by an enzyme treatment method for activating substances.

First, in the present invention, water is added to wheat germ, and then added to the cellulite to react with a water bath, filtered, then frozen and dried.

Preferably, 200 mL of water is added to 50 g of wheat germ, and then 1.5 L of cellulose is added, reacted in a water bath at 50 ° C. for 6 to 30 hours, filtered, then frozen and dried at −50 ° C.,

More preferably, the hot water reaction time is made by selecting any one of 6 hours, 12 hours, 18 hours, 24 hours, and 30 hours.

Looking at the contents of the experimental analysis for the above configuration,

Wheat germ was used as a by-product wheat germ during the milling process at Sajo Dong-awon Co., Ltd. (Dangjin, Korea). Wheat embryos were stabilized and used as samples. Celluclast 1.5L is Novozymes Corp. (Bagsvaerd, Denmark). 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonate) (ABTS) were purchased from Sigama Aldrich Co. (St. Louis, MO, USA).

Hereinafter, a method for measuring the antioxidant substance contained in the enzyme-treated wheat germ, that is, the total polyphenol compound content, the total flavonoid content, and the α-tocopherol (alpha-tocopherol) content and the results will be described as follows. same.

1. Total polyphenol compound and total flavonoid content measurement

1) Total polyphenol compound content

The total polyphenolic compound content (TPC) was analyzed by modifying the colorimetric method (Chen L et al. 2015) using Folin-Ciocalteu'phenol reagent. In a 15 mL centrifuge tube, mix 10 mg of enzyme-treated wheat embryos with 2.5 mL 10% Folin-Ciocalteu reagent, react at room temperature for 5 minutes, and then mix 2 mL of 7.5% (v / v) Na ₂ CO ₃ Was added to. The mixed solution was reacted for 2 hours at room temperature, and separated by centrifugation (Union 5KR, Hanil, Incheon, Korea) for 2 hours at 10,000 × g. The absorbance of the supernatant was measured at 750 nm with a UV-vis spectrophotometer (Libra S22, Biochrom, Cambridge, England). As a standard, gallic acid was used, and the total phenolic compound content was expressed as mg gallic acid equivalents (GAE) / g dry weight.

2) Total flavonoid content

Total flavonoid content (TFC) was measured according to the method of Abozed SS et al. (2014). 2 mg of enzyme-treated wheat embryos were placed in a test tube, and added to 20 mL of 70% (v / v) ethanol, followed by shaking extraction for 15 hours. After centrifugation for 5 minutes at 2,500 × g, 0.5 mL (v / v) of the supernatant was placed in a test tube, and 1.5 mL of 95% (v / v) ethanol and 0.1 mL of 10% aluminum chloride solution were added. 100 mL of 1 M potassium acetate solution and 2.8 mL of distilled water were added. After standing at room temperature for 30 minutes, absorbance (Libra S22 UV-vis spectrophotometer, Biochrom, Cambridge, England.) Was measured at 415 nm. The standard curve was tested using rutin and the total flavonoid content was calculated from the standard curve. The results were repeated 3 times and expressed as mean and standard deviation.

2. Measurement of α-tocopherol (alpha-tocopherol) content in wheat embryos using HPLC

The content of α-tocopherol in the enzyme-treated wheat embryo is HPLC (Spectra System, Thermo Electron Co., consisting of columns (ZORBAX ODS 150 × 4.6 mm), as shown in FIG. 1 according to the method of Lee SM et al. (2006). Waltham, MA, USA) and a fluorescence detector (excitation wavelength 290 nm, emission wavelength 330 nm) were measured. The mobile phase was 1.3% isopropanol containing hexane, the flow rate was 1.0 mL / min, and the injection volume was 20 mL. 5 g of the enzyme-treated wheat germ was taken, placed in a cylindrical filter paper, and 100 mL of hexane containing 0.01% BHT was extracted with a solvent in Soxhlet at 70-80 ° C. for 4 h. After hexane extract was used as 100 mL, 2 mL of the sample was concentrated under nitrogen injection, and hexane containing 1.3% isopropanol was added and analyzed by HPLC.

3. Measurement of antioxidant activity of enzyme-treated wheat embryos

1) ABTS radical scavenging ability measurement

1 g of enzyme-treated wheat germ was placed in 20 mL of distilled water, and extracted with ultrasonic waves for 30 minutes (ST505 Solutech, Osan, Gyeonggido Korea), followed by centrifugation (4,500 rpm, 5 min, Union 5KR, Hanil Co., Incheon, Korea), and the supernatant. Used. Measurement of ABTS (2,2-azino-bis-3-ethylbenzo-thiazoline-6-sulfonate) radical scavenging ability was carried out by partially changing the experimental method of Park KR et al. (2009). ABTS radical solution was prepared by mixing 1.0 mM AAPH [(2,2 'and 2.5 mM ABTS reagent in 100 mL of phosphate-buffered saline (PBS, pH 7.4) solution and reacting in a 70 ° C constant temperature water bath. The absorbance was adjusted to 0.650 ± 0.020 absorbance at 734 nm by reacting 980 μL of ABTS radical solution and 20 μL of sample in a constant temperature water bath at 37 ° C. for 10 minutes to measure the decrease in absorbance at a wavelength of 734 nm and vitamin C as a standard. The antioxidant activity was calculated as mg vitamin C equivalents (VCE) / g dry weight.

2) Antioxidant activity by DPPH

DPPH radical scavenging ability was measured by modifying the method of Blois MS (1958). 1 g of the enzyme-treated wheat germ was placed in 20 mL of distilled water and extracted with ultrasonic waves for 30 minutes (ST505 Solutech, Osan, Gyeonggido Korea), followed by centrifugation (4,500 rpm, 5 min, Union 5KR, Hanil Co., Incheon, Korea). Used. After adding 0.5 mL of a 0.2 mM DPPH (1,1-diphenyl-2-picrylhydrazyl) solution to 0.5 mL of the sample, mixing and allowing to stand for 30 minutes, the absorbance was measured at 517 nm with a spectrophotometer. The control was measured in the same way by adding a solvent instead of the sample, and absorbance was measured in the same manner by adding methanol instead of 0.2 mM DPPH to correct the absorbance value for the color of the sample itself. DPPH radical scavenging capacity was calculated as follows.

DPPH radical scavenging effect (%) = (1-absorbance of sample addition / absorption of no addition) × 100

4. Analysis

The experiment was repeated three or more times and the results were expressed as the mean and standard deviation. As a result of the experiment, ANOVA was performed using Minitab (Minitab17, Minitab Inc., University Park, PA, USA), and the test was conducted by applying the Tukey method for the difference between means at the p <0.05 level.

5. Results and Discussion

1) Total polyphenolic compound content

In order to increase the elution of antioxidants of wheat germ isolated in the milling process, an enzyme (Celluclast 1.5L) that breaks down cellulose constituting the cell membrane was added and reacted for a certain time. After the reaction, the embryos were separated, lyophilized, and the results of measuring the total polyphenol compound (TPC) from the enzyme-treated wheat embryos are as shown in FIG. 2. TPC increased rapidly until 6 hours after enzyme treatment. That is, the TPC content of 9.35 mg / g increased by about 2.5 times to 22.66 mg / g, but the TPC content measured after the reaction every 6 hours increased from 24.57 mg / g to 37.63 mg / g after 30 hours of reaction. According to a report by Laroze L et al. (2010), when the raspberry by-product was enzymatically treated at 50 ° C, the antioxidant content of polyphenols increased by 35% compared to the control group. Kalcheva-Karadzhova K et al. (2014) observed that an increase in extract yield, an increase in the value of total polyphenols and anthocyanins and antioxidants was reacted by enzyme treatment. In addition, it was reported that enzymes that hydrolyze cellulose, such as Celluclast (1.5L), were highly effective in extraction by liberating polyphenols bound to proteins or carbohydrates in cells or in cell walls. The cell wall of the plant has a structure in which skeletal components such as cellulose, hemicellulose, and protein are surrounded by pectin, an interstitial cell (Carpiat NC & Gibeau DM 1993). Among them, phenolic substances do not show a constant distribution in cells, and insoluble phenolic compounds are known as cell wall components, and water-soluble phenolic compounds are known to exist in vacuoles (Wink M 1997). Therefore, it has been found that several studies have been conducted to extract useful substances in cells such as phenolic substances by decomposing substances surrounding the cells, such as cell walls, with enzymes. Li BB et al. (2006) used the enzymes that break down cell walls (Celluzyme CL, Celluzyme MX, Kleepase AFP 106L) in the case of citrus fruit, a tropical fruit, and when extracted with water, the highest phenolic compound was extracted when reacted for 6 hours. After showing a tendency to decrease slightly, it was reported that the effect of the initial 6 hours was great. In this experiment, similar results showed similar celluclast 1.5L, which can decompose cell walls, and various enzyme treatments facilitated the extraction of polyphenols from wheat embryos, which showed a more rapid increase in the initial reaction and tended to increase with increasing extraction time. I could see that.

2) Total flavonoid content

  The total flavonoid content (TFC) of wheat embryos treated with Celluclast 1.5L increased by about 3.0 times from 2.93 mg / g to 8.64 mg / g until the initial 6 hours of the reaction, as shown in FIG. By time, only 1.21 mg / g increased further. However, after a 30-hour reaction, the increase was increased to 12.78 mg / g. Chen S et al. (2011) used Penicillium decumbens cellulase to extract flavonoids from the bank, destroying the cell wall and increasing solubility, increasing 102% compared to the absence of enzyme use. In addition, a new enzyme was prepared to increase flavonoid extraction from grape skins of Tomaz I et al. (2016) to determine the optimal conditions. In general, it was found that the degree of extraction by enzyme treatment affects the optimum conditions of the raw material and the enzyme used, and also varies depending on the extraction solvent. In this experiment, it was confirmed that when the Celluclast 1.5L was reacted and extracted with water, the total yield of phenolic compounds or total flavonoids was highest in the initial 6 hours.

3) α-tocopherol content using HPLC

 Extraction and separation of α-tocopherol confirmed by HPLC were purely separated as shown in FIG. 3, and the content obtained from the area of the peak was as shown in FIG. 2.

The α-tocopherol content of wheat embryos treated with Celluclast 1.5L was 40.17 mg / 100 g without treatment, but decreased by 28% to 29.31 mg / 100 g after 6 hours after enzyme treatment, until 30 hours after reaction. There was no significant change. It has been reported that wheat embryos contain more α-tocopherol than any part of the grain of wheat (Gili RD et al. 2017). In particular, α-tocopherol is known to act as an antioxidant that removes radicals generated in the oxidation process of cell membranes and lipoproteins (Kamal-Eldin A 1996), which affects the extraction method, but does not contact oxygen with enzyme treatment. It was thought that it was the result of the increase. Oh et al. (2016) confirmed that α-tocopherol content was extracted more by supercritical extraction method than by hexane extraction, but reported that β-tocopherol was not detected in both solvent extraction and supercritical fluid extraction. Zou Y et al. (2018) reported that the tocopherol content of wheat germ decreases with time when roasting for stabilization, and the reason is that it is the result of thermal decomposition. Kumar GS et al. (2014) reported similar results, and it was thought that α-tocophenol was decomposed by heat or oxygen and thus the content decreased. In this study, for stabilization of wheat germ, it was thought that the temperature of the extrusion treatment increased and the contact of oxygen was easy through the extraction process after the enzyme treatment, so that the α-tocopherol value was lower than that of the untreated group. Therefore, it was found that α-tocopherol, an antioxidant as fat-soluble vitamin, may be reduced due to oxidation rather than increase extraction due to cell decomposition such as cell wall.

4) Antioxidant activity

Measurement of radical scavenging activity by ABTS is a colorimetric method that is frequently used for evaluating antioxidant activity (Adedapo AA et al. 2008). As a result of this experiment, the ABTS value was not increased until 24 hours of treatment with Celluclast 1.5L treated with enzyme-treated wheat embryos as shown in FIG. It showed a tendency to decrease slightly, but the antioxidant activity increased in wheat embryos that were treated for 30 hours. It is said that the measurement of radical scavenging activity by DPPH is sensitive enough to detect active ingredients at low concentrations and can identify many samples in a short time (Hseu YC et al. 2008). Chen D et al. (2016) reported that treatment of oat shells with cellulase for up to 80 minutes significantly increased ABTS and DPPH radical scavenging activity and FRAP values. In the present invention, as shown in FIG. 4, both ABTS and DPPH showed a slight decrease compared to the initial value, and when the enzyme was treated for more than 24 hours, the antioxidant activity tended to increase. This was thought to be because the total phenolic compound and flavonoid increase and α-tocopherol content were collectively considered. In order to increase the antioxidant activity, treatment with Celluclast 1.5L as a cell wall hydrolase requires more than 30 hours of treatment. Became. In this experiment, it was stabilized by an extrusion molding machine to inactivate hydrolyzate and oxidase of wheat embryo fat, but it showed a tendency to decrease the tocopherol content, so as to increase the antioxidant activity of wheat embryo, tocopherol as well as increase the extraction ability of phenolic compounds. It was confirmed that the treatment for the stabilization of the need.

As such, to increase the antioxidant activity of wheat embryos, Celluclast 1.5L is used to decompose cell walls to treat wheat embryos for a certain period of time, followed by enzymatically treated wheat embryos with total phenolic compounds (TPC) and total flavonoid content (TFC). α-tocopherol content was measured and antioxidant activity was compared with ABTS and DPPH radical scavenging activity. TPC and TFC showed a sharp increase after the initial 6 hours, but there was no significant change, and TPC increased 4.3-fold and TFC increased 3.0-fold after 30-hour treatment. The content of α-tocopherol was 40.17 mg / 100 g in stabilized wheat embryos, which decreased by 28% after 6 hours of enzyme treatment, and there was no significant change after that. When the antioxidant activity was compared with ABTS and DPPH radical scavenging ability, it decreased at the initial stage of enzyme treatment, but increased after 30 hours of enzyme treatment. In order to improve the antioxidant activity, treatment time with Celluclast 1.5L required 30 hours of treatment time. From the above results, it is possible to further improve the antioxidant activity by confirming the effect of the complex action of other enzymes that degrade cells and the causes of tocopherol reduction.

As described above, although the present invention has been described by limited embodiments and drawings, the present invention is not limited to the above embodiments, and the contents described above by those skilled in the art to which the present invention pertains Of course, various modifications and variations may be possible.

Therefore, the technical idea in the present invention should be grasped by the claims described below, but it will be apparent that all equivalent or equivalent modifications fall within the scope of the technical idea of the present invention.

Claims (3)

A method of enzymatic treatment of wheat germ containing antioxidants, characterized in that water is added to the wheat germ and then added to the cellulite to react with a water heater, filtered, and then frozen and dried.
According to claim 1,
200 ml of water is added to 50 g of wheat germ, and then 1.5 L of cellulose is added, reacted in a water bath at 50 ° C. for 6 to 30 hours, filtered, frozen at -50 ° C., and dried, containing antioxidants. Wheat germ enzyme treatment method.
According to claim 2,
The jungtanggi reaction time is 6 hours, 12 hours, 18 hours, 24 hours, 30 hours, wheat germ enzyme treatment method containing an antioxidant, characterized in that any one selected.

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