KR100845032B1 - Rice peptide containing antioxidant activity and radical scavenging activity and the method thereof - Google Patents

Abstract

The present invention relates to a rice peptide having an antioxidant activity and a radical scavenging activity, and also added to the rice protein powder of 60 to 80% by weight of water 3 to 15 times the weight of the rice and endopeptidase and exopeptidase Hydrolysis using agents sequentially or simultaneously; Dehydrating endopeptidase and exopeptidase by heating at 90-100 ° C. for 10-20 minutes after the hydrolysis; And separating the hydrolyzate into a solid phase and a liquid phase, and then, centrifuging and filtering the liquid phase, to a method of preparing a rice peptide having antioxidant activity and radical scavenging activity.

According to the present invention, since most of the existing antioxidants have problems with toxicity and thermal stability and are limited in their use as fat-soluble, rice protein containing amino acids very similar to breast milk without causing allergy. By using the above, a water-soluble rice peptide having a strong radical scavenging activity can be obtained.

Rice Protein, Rice Peptide, Radical Scavenging Activity

Description

Rice peptide containing antioxidant activity and radical scavenging activity and preparation method thereof {Rice peptide containing antioxidant activity and radical scavenging activity and the method}

Figure 1 shows the results of analyzing the molecular weight distribution of the rice peptide using HPLC.

The present invention relates to a rice peptide having an antioxidant activity and a radical scavenging activity, and also added to the rice protein powder of 60 to 80% by weight of water 3 to 15 times the weight of the rice and endopeptidase and exopeptidase Hydrolysis using agents sequentially or simultaneously; Dehydrating endopeptidase and exopeptidase by heating at 90-100 ° C. for 10-20 minutes after the hydrolysis; And separating the hydrolyzate into a solid phase and a liquid phase, and then, centrifuging and filtering the liquid phase, to a method of preparing a rice peptide having antioxidant activity and radical scavenging activity.

Antioxidant action is to cause damage to cells by generating harmful oxygen in the body to injure the wound. This action to remove the harmful oxygen, it is helping to prevent skin aging and various adult diseases. In detail, our bodies constantly use oxygen to obtain energy through respiration, which is absolutely necessary for life support, but the stable molecular state of the ground state triplet oxygen is the enzyme, reductive metabolism, chemical superoxide radicals by for various physical and chemical factors, such as drugs, pollutants, photochemical reactions ^{_{(superoxide radical, O 2 -)}} , hydroxy radicals (hydroxyl radical), hydrogen peroxide (H ₂ O _2), singlet oxygen (singlet oxygen When converted to highly reactive reactive oxygen species, such as ¹ O ₂ ), it has a double sided effect that causes fatal toxicity to the living body. These reactive oxygen species have non-selective and irreversible destructive effects on cell components such as lipids, proteins, sugars, deoxyribohexane (DNA), and brain diseases such as cancer, stroke, Parkinson's disease, heart disease, ischemia and skin diseases. It is known to cause various diseases and aging such as gastrointestinal diseases, inflammation, rheumatism (B. Halliwell, Drugs 42: 569, 1991; K. Fukuzawa and Y. Takaishi, J. Act. Oxyg. Free Rad. 1:55 , 1990; J. Neuzil, and JM Gebicki, J. Med. 320: 915, 1989).

Recently, studies on the development of antioxidants that can regulate such active oxygen species are actively conducted, such as superoxide dismutase (peroxide dismutase), peroxidase (peroxidase), catalase (catalase), glutathione peroxidase (glutathione peroxidase) Studies on the Antioxidant Effect of Enzymes and Small Molecules Derived from Natural Products, such as Tocopherol, Ascorbate, Carotenoid, Glutathione, etc. (SS Chang et al., J. Food Sci 42: 1102, 1977; PA Hammerschmidt and DE Pratt, J. Food Sci. 43: 556, 1977; DE Pratt and BW Watts, J. Food Sci. 29:17, 1964). The development of effective antioxidants is of great importance in that they can lead to the development of new medicines as well as food and cosmetic supplements.

However, most of the existing antioxidants have problems with toxicity and thermal stability, and their use is limited due to fat soluble. Therefore, the present inventors determined that the peptide may exhibit antioxidant activity even in a non-toxic, water-soluble hydrolyzed rice protein, and the rice peptide having strong radical scavenging activity was determined as a result of measuring the functionality after preparing the rice peptide. Invented to complete the present invention.

It is an object of the present invention to economically provide a rice peptide containing a large amount of factors for enhancing antioxidant activity and radical scavenging activity by using rice protein powder, which is a plant protein source that we consume everyday.

According to the present invention, a rice peptide having antioxidant activity and radical scavenging activity is provided.

In addition, according to the present invention, by adding water of 3 to 15 times the weight of the rice to the protein protein of 60 to 80% by weight of protein and hydrolyzing using endopeptidase and exopeptidase sequentially or simultaneously step; Dehydrating endopeptidase and exopeptidase by heating at 90-100 ° C. for 10-20 minutes after the hydrolysis; And separating the hydrolyzate into a solid phase and a liquid phase, and then, a method of preparing a rice peptide having antioxidant activity and radical scavenging activity, which comprises the step of centrifuging and filtering the liquid phase.

Preferably, the endopeptidase is protease NP, and the exopeptidase is provided with a method for producing a rice peptide having antioxidant activity and radical scavenging activity, which is Promod 279.

Further preferably, the protease and the promod 279 (Promod 279) are each provided with a method for producing a rice peptide having an antioxidant activity and a radical scavenging activity of the concentration of 0.1 to 10% by weight.

In addition, the hydrolysis is provided with a method for producing a rice peptide having antioxidant activity and radical scavenging activity made for 2 to 10 hours at 45 ~ 60 ℃.

Hereinafter, the content of the present invention will be described in detail.

For the production of rice peptides, protein hydrolyzate containing 60 to 80% by weight of water is added to the water by 3 to 15 times the weight of rice, followed by the proteases Promod 279 (Promod 279) and Protease (Protease). NP) was enzymatically decomposed for 2-10 hours at 45-60 ° C using 0.1-10% by weight, and then heated at 90-100 ° C for 10-20 minutes to inactivate the enzyme, followed by filtration by centrifugation and vacuum concentrator. It was obtained by concentrating to 30 ~ 40 Brix (Brix) using. The rice peptide concentrate thus obtained was powdered by spray drying, and a rice peptide containing 90% or more low molecular peptide was prepared. In order to measure the antioxidant activity, the free radicals DPPH ((1,1-diphenyl-2-picrylhydrazyl) and rice peptides were reacted to observe the color change of the purple color of DPPH. As a result, the rice protein did not have a radical scavenging activity, while the rice peptide was found to increase the radical scavenging activity as the concentration increases.

Hereinafter, the present invention will be described in detail through examples. However, these examples are only for illustrating the present invention, and the present invention is not limited to these examples.

Example 1 Analysis of Enzyme Optimal Temperature Conditions

Protease NP as endopeptidase and promod 279 as exopeptidase were simultaneously treated to check the degree of hydrolysis of the peptide. The degree of hydrolysis upon hydrolysis for 5 hours was relatively compared, that is, the degree of hydrolysis at 55 ° C. was 100.

Degree of hydrolysis = AN (free amino acid nitrogen) / TN (total nitrogen)

TABLE 1 Peptide Hydrolysis Degree

Temperature (℃) 30 40 45 50 55 60 70 Relative degree of hydrolysis (%)  38  75  80  95  100  88  40 Enzyme titers - + + ++ +++ ++ -

Table 1 shows that the degree of hydrolysis is high in the temperature range of 45 ~ 60 ℃. Thus, the optimal temperature range for Protease NP and Promod 279 was 45-60 ° C.

Example 2 Analysis of Optimal Decomposition Time of Enzymes

When the protease NP and promod 279 were treated simultaneously, the high hydrolysis degree was checked by time zone to analyze the optimal hydrolysis time of the protease NP and promod 279.

TABLE 2

Hours (h) One 3 5 7 10 15 Relative comparison of hydrolysis degree (%)  75  94  97  99  100  98

Table 2 is a relative comparison value when the degree of hydrolysis for 10 hours at 55 ℃ as 100, according to Table 2, the optimum hydrolysis time was 2 to 10 hours.

Example 3 Optimal Concentration Analysis of Enzymes

When the protease NP (Protease NP) and promod 279 were simultaneously treated for each concentration, the degree of hydrolysis was analyzed.

TABLE 3

density(%) 0.05 0.1 0.5 One 3 5 10 20 Relative degree of hydrolysis (%) 55 70 74 80 85 90 100 95

Table 3 is a relative comparison value when the degree of hydrolysis at 100% is 100. According to Table 3, the optimum concentration of enzyme was 0.1 to 10%.

Example 4 Preparation of Rice Peptides Using Enzymatic Degradation

To prepare rice peptides, 10 times of water was added to 1 kg of rice protein powder, followed by warming up to 50 ° C, and 70 g of protease, Promod 279 and Protease NP, were added. After enzymatic degradation for 10 hours, the enzyme was inactivated for 15 minutes at 100 degrees. The product was filtered by centrifugation and then concentrated to 30-40 Brix using a vacuum concentrator to obtain a concentrate. Spray drying was used to powder the powder, and the powdering conditions were 165 degrees blowing air temperature and 95 degrees blowing air temperature, and the atomizer was 10,000 rpm.

<Comparative Example 1> Preparation of soybean peptide using enzymatic digestion

Soybean peptide was prepared in the same manner as in Example 1 using skim soybean powder.

Comparative Example 2 Preparation of Silk Peptides

Silk peptide was prepared in the same manner as in Example 1 using silk fibroin.

<Comparative Example 3> Preparation of casein peptide using enzymatic digestion

Casein peptide was prepared in the same manner as in Example 1 using casein protein powder.

Test Example 1 Peptide Molecular Weight Distribution Analysis

In order to confirm the molecular weight distribution of the rice peptide obtained in Example 4, the analysis was performed under the same conditions as in Table 4 below.

[Table 4] Peptide distribution analysis conditions of rice peptides

Way SE-HPLC (BIO-RAD Method) ¹⁾ column Bio-Sil SEC-125 column (300 × 7.8 mm) Buffer A 0.05 M NaH ₂ PO ₄ + 0.15 M NaCl Buffer B 0.05 M Na ₂ HPO ₄ + 0.15 M NaCl Flux 1.0 mL / min Absorbance (UV) 280 nm Appliance system Waters 510 HPLC Pump, Waters 486 Detector

¹⁾ Borneo, R. and Khan, K . : Cereal Chem , 76, 5, 711-717, 1999.

Experimental Example 2 Radical Scavenging Activity of Rice Peptides

Antioxidant Activity of the Rice Peptide and Rice Protein Powder Prepared in Example 1 by Antioxidant Activity Measurement Using Free Radical DPPH (1,1-diphenyl-2-picrylhydrazyl) (MS Blois., Nature, 181: 1199, 1958; BS Yun.J. Antibiotics, 53: 114, 2000).

First, 100 ml of each sample compound dissolved in ethanol was added to 4900 ml of 150 mM DPPH ethanol solution. The sample used the rice peptide of the present invention as an experimental group, and the rice protein powder as a comparative group. Each sample mixture was mixed by vortex and left at room temperature for 30 minutes, and then absorbance was measured at 517 nm. The results are shown in Table 1 below. The ED ₅₀ value represents the amount of sample (mg / ml) needed to reduce the absorbance value of the control by 50%.

TABLE 5

sample Antioxidant activity (ED ₅₀ , mg / ml) Example 1 1.1 X 10 ³ Rice protein powder none

As shown in Table 5, the rice protein powder of the present invention did not have a radical scavenging activity, while the rice peptide was found to have a high radical scavenging activity.

<Test Example 3> investigation of radical scavenging activity by peptide type

 Relative radical scavenging activity was measured using DPPH at the same concentration of 3 mg / ml using the rice peptide powder prepared in Example 1 and the soybean peptide, silk peptide, and casein peptide prepared in Comparative Examples 1, 2 and 3 Measured by. At this time, when the radical scavenging activity at the rice peptide concentration of 3 mg / ml was 100, the relative radical scavenging activity was shown for each peptide type.

TABLE 6

Example 1 Comparative Example 1 Comparative Example 2 Comparative Example 3 Relative radical scavenging activity (%) 100 33.2 62.2 34.7

As shown in Table 6, as a result of analyzing the relative radical scavenging activity by peptide type, the peptide was prepared through the same enzymatic digestion process, but the peptide prepared using rice was about 3 times higher than the soybean peptide and casein peptide. It was found that it showed 1.6 times higher radical scavenging activity than silk peptide.

As mentioned above, although this invention was demonstrated by the limited embodiment, this invention is not limited by this and it will be described below by the person of ordinary skill in the art, and the following. Of course, various modifications and variations are possible within the scope of the claims.

According to the present invention, using a rice protein was able to obtain a water-soluble rice peptide powder having a strong radical scavenging activity to solve the problems of the existing antioxidants. In addition, it can be seen that it shows a high radical scavenging activity compared to soybean, casein, silk peptide.

Claims (5)

delete Adding hydrolyzed protein to rice protein powder having a protein content of 60 to 80% by weight 3 to 15 times the weight of rice and using endopeptidase and exopeptidase sequentially or simultaneously to hydrolyze; Inactivating the endopeptidase and exopeptidase by heating at 90 to 100 ° C. for 10 to 20 minutes after the hydrolysis; And After separating the hydrolyzate into a solid phase and a liquid phase, the method of producing a rice peptide having antioxidant activity and radical scavenging activity, characterized in that the liquid phase comprises centrifugation and filtration. delete The method of claim 2, The endopeptidase and the exopeptidase is a method for producing a rice peptide having antioxidant activity and radical scavenging activity, characterized in that the concentration is 0.1 to 10% by weight, respectively. The method of claim 2, The hydrolysis is a method for producing a rice peptide having antioxidant activity and radical scavenging activity, characterized in that for 2 to 10 hours at 45 ~ 60 ℃.

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