KR20060062917A - Black Soybean Peptide Composition with Inhibition of Increasing Blood Lipid Level and Weight Loss Effect - Google Patents

Abstract

The present invention relates to a peptide composition derived from black soybean protein, comprising: at least 10 mg of arginine, at least 10 mg of aspartic acid, and at least 20 mg of glutamic acid per 1 g of the peptide composition, wherein the low molecular weight peptide having a molecular weight of 10,000 or less in the peptide composition is 70 It relates to a black soybean peptide composition having an effect of inhibiting the increase in the concentration of blood lipids of more than% and weight loss effect.

The black soybean peptide composition according to the present invention is a general food by using the black soybean extract commonly ingested diet foods that suppress the increase in serum lipid concentration and weight loss effect (relative to water, etc.) without relatively allergic side effects. This can be useful for development.

Black Bean, Diet, Anti-Obesity, Weight Loss, Peptide

Description

Black soybean peptide which has an effect on weight reduction

1 is a graph showing the change in weight according to the diet.

2 is a graph showing changes in triglyceride and cholesterol levels in blood according to diet {TG (triglyceride), TC (total cholesterol), HC (HDL-cholesterol), LC (LDL-cholesterol), VLC (VLDL-cholesterol) )}.

Figure 3 is a graph showing changes in total fat, triglycerides and total cholesterol levels of the liver according to the diet.

Figure 4 is a graph showing the change in total fat, triglyceride and total cholesterol level of feces according to the diet.

The present invention relates to a peptide composition derived from black soybean protein, comprising: at least 10 mg of arginine, at least 10 mg of aspartic acid, and at least 20 mg of glutamic acid per 1 g of the peptide composition, wherein the low molecular weight peptide having a molecular weight of 10,000 or less in the peptide composition is 70 It relates to a black soybean peptide composition having an inhibitory effect on the increase in the concentration of blood lipids of more than% and weight loss.

In recent years, the incidence of obesity has increased significantly in our country due to the improvement of living standards and the effects of dietary life such as developed countries. The cause of obesity is not yet clear, but body fat accumulation due to an imbalance between energy intake and consumption is known (Grundy: Am, J. Clin. Nutr. 67 (suppl), s563-s572, 1998). And multiple factors, including genetic and genetic factors, may be the cause (Albu et al .: Nutr. Rev. 55: 150-156, 1997).

The seriousness of obesity is closely related to various adult diseases (hypertension, diabetes, arteriosclerosis, etc.), so the problem is more serious. In particular, triacylglyceride, which is known to cause obesity, is known to be the cause of coronary arteries along with LDL-cholesterol. Increased triglyceride levels in the blood have been reported to cause diseases such as hypertension and atherosclerosis. (Jung Sun Hwa, et al .: Korean J. Food Sci. Technol ., 31, 4, 1101-1108. 1999).

Currently, in order to inhibit the increase in the concentration of triglycerides in the blood, dietary control, intake of diet foods and the administration of various drugs are prevalent.

Drugs used include lipoproteins in the blood, dextrin sulfate to increase lipase activity, nicocom to inhibit lipid absorption, and crofibrate and pravastatin to promote lipid metabolism. (Patent Publication 1999-21880, Patent Publication 2003-0015537).

However, dietary control is a very difficult method in modern dietary cultures, and drug administration can also have serious side effects. Therefore, there is an urgent need for the development of new functional materials without side effects for improving and preventing obesity.

Peptides are intermediates between amino acids and proteins in molecular weight. As a third nitrogen source together with amino acids, peptides have recently attracted attention for bioregulatory functions and methods for producing soy proteinases containing blood pressure lowering factors (Patent Publication 2000) -14663), peptides having an antimicrobial insecticidal effect (Patent Publication No. 195-31476, Patent Publication No. 199-700087), Peptides production method having a platelet aggregation inhibitory effect (Patent Publication No. 1997-6318) and mineral absorption Promoting peptide production method (Patent Publication No. 1996-022556) and the like have been reported, in addition to the situation that a peptide having a variety of physiological activity is being developed.

In particular, a method for manufacturing a peptide for lowering blood lipid concentration in relation to weight control has been reported (Patent Publication No. 2003-0015537), but the peptide has been reported to increase the weight of the lipid but improve the blood lipid.

It is an object of the present invention to provide a black soybean peptide composition having a neutral fat improvement effect and a weight loss effect in blood from black soybean-derived protein hydrolysates commonly consumed as a general food and a black soybean peptide drink using the same.

In order to achieve the above object, the present invention provides a peptide composition derived from black soybean protein, comprising at least 10 mg of arginine, at least 10 mg of aspartic acid, and at least 20 mg of glutamic acid per 1 g of the peptide composition, wherein the peptide composition has a molecular weight of 10,000 or less. It provides a black soybean peptide composition having an effect of inhibiting the increase in the concentration of blood lipids with a low molecular weight peptide of more than 70% by weight and weight loss.

The black soybean peptide composition may be obtained by separating the solid phase and the liquid phase obtained by sequentially hydrolyzing black soybean with an endopeptidase and exopeptidase enzyme, and then filtration of the liquid phase to powder.

In addition, the present invention provides a black soybean peptide drink having an inhibitory effect on the increase in blood lipid concentration and weight loss, including a black soybean peptide composition, high fructose, citric acid, juice and purified water.

The black soybean peptide composition of the present invention and a method for producing a beverage are as follows.

Black soybean concentrate was pulverized black soybean, hydrolyzed 3 to 15 times of water, and then enzymatically decomposed at 45 ° C to 55 ° C for 2 to 5 hours using protease, and then at 90 ° C to 100 ° C. The enzyme was inactivated by heating for 10 to 20 minutes, filtered through 60 to 120 mesh, and concentrated to 45 to 55 Brix using a vacuum concentrator.

For the pretreatment of the black soybean concentrate, add 5 to 10 times the water of the black soybean concentrate, stir and raise the temperature to 100 ° C. for 5 minutes to denature the protein, and then change the temperature again. Adjust to ° C.

After slowly adding 5 normal sodium hydroxide to adjust the pH to 8.0, 0.4 to 0.8% by weight of an enzyme, an endopeptidase, was added to the raw protein, followed by stirring for 2 hours. After the reaction was completed, the temperature was adjusted to 55 ° C., and then pH was adjusted to 5.0 by adding 6 normal hydrochloric acid. Then, the enzyme exopeptidase (0.4-0.8% by weight) of the raw protein is added and stirred for 4 hours to react. After the reaction is completed, the temperature is raised to 100 ° C. and maintained for 5 minutes to inactivate the enzyme.

The resulting enzyme reactant is added to diatomaceous earth and separated into a solid and liquid phase using a filter press, and then a liquid phase containing particles having a diameter of 70 μm or less is separated and recovered using a mesh. Transfer to a concentrator was concentrated to 50% by weight solid content at 50 ± 5 ℃.

The black soybean peptide thus obtained was powdered, and the animal experiments using rats showed the effect of inhibiting the increase of blood lipid concentration, reducing total cholesterol and reducing LDL-cholesterol, and in particular, the weight loss effect was remarkable.

In addition, peptide drink was prepared using the black soybean peptide diet effect was verified.

Add 5-20% of black soybean peptide to 100 mL of water and mix sweetener (high fructose) to give sweetness and refreshment, acidulant (citric acid) and fruit flavor to lower the harmonious taste and pH of beverage After filling with heat to prepare a peptide beverage.

(Example)

Hereinafter, the present invention will be described in more detail with reference to Examples and the accompanying drawings. The following examples are merely provided to more easily understand the present invention, but the present invention is not limited to the following examples.

Example 1 Functional Peptide Preparation Method Using Black Soybean

After grinding 1kg of black soybeans and adding 6L of water in a 10L reactor, 1.0g of protease was added, followed by enzymatic digestion at 50 ° for 3 hours and heating at 100 ° for 5 minutes to deactivate the enzyme. After filtration at 60 mesh, it was concentrated to 50 brix using a vacuum concentrator to obtain a black bean concentrate.

Pretreatment of this black soybean concentrate 375 g of black soybean concentrate (50 Brix) was added to a reaction tank with a 10 L capacity, 2 L of water was added thereto, followed by stirring to raise the temperature to 100 ° C. The temperature was kept for 5 minutes, then cooled to adjust the temperature to 60 ° C., and then the pH was adjusted to 8.0 by the addition of 45 mL of 5 normal sodium hydroxide. To this, 1.5 g of an endopeptidase [Promod 278] enzyme was added to make an aqueous solution, followed by stirring for 2 hours. After the reaction was completed, the temperature was lowered to 55 ° C. and pH was adjusted to 5.0 by adding 65 mL of 6 normal hydrochloric acid. Here, 3.0 g of an exopeptidase [Promod 279] enzyme was added to an aqueous solution, and then reacted with stirring for 4 hours. After the reaction was completed, the enzyme was inactivated by raising the temperature to 100 ° C. and maintaining it for 5 minutes.

4 kg of diatomaceous earth was added to the preparation of the reaction tank, the mixture was stirred, filtered through a filter press, and the filtrate was transferred to a concentrator and concentrated at 50 ± 5 ° C. while maintaining a solid content of 50% by weight.

Test Example 1 Peptide and Amino Acid Analysis

In order to confirm the peptide distribution and amino acid composition of the black soybean peptide obtained in Example 1, the analysis was performed under the same conditions as in Tables 1 and 2 below.

Table 1: Peptide distribution analysis conditions of black soybean 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

As a result of analyzing the peptide distribution of the black soybean peptide, it was confirmed that the low molecular weight peptide having a molecular weight of 10,000 or less is 78.6% by weight.

Table 2: Amino acid composition analysis conditions of black soybean peptide

Way Pico-Tag Method column Pico-Tag column (150 × 3.9 mm, S / N: T12641G) Buffer A Pico-Tag Eluent A (Waters, S / N: 909979) Buffer B 60% CH ₃ CN Flux Gradient (1.0 ~ 1.5 mL / min) Absorbance (UV) 254 nm Appliance system Waters 510 HPLC Pump, Waters 486 Detector

As a result of analyzing the amino acid of black soybean peptide, glutamic acid, aspartic acid and arginine showed the highest contents, and the results are shown in Table 3 below.

Table 3: Amino acid analysis of black soybean peptides

amino acid Content (mg / g) amino acid Content (mg / g) Alanine 1.72 Lysine 4.19 Arginine 10.37 Methionine 2.55 Aspartic acid 13.50 Phenylalanine 6.21 Cystine - Proline 6.12 Glutamic acid 22.36 Serine 5.25 Glycine 4.42 Threonine 4.10 Histidine 3.31 Tryptophan - Isoleucine 5.78 Tyrosine 4.34 Leucine 7.73 Valine 5.78

Example 2 Demonstration of Weight Loss Effect of Black Soy Peptides through Animal Experiments

1. Experimental diet and experimental animals

        The experimental animals were purchased for 3 weeks and Sprague-Dawley rats were adapted for 1 week, and then fed with high fat diet for 4 weeks to 6 weeks. Black beans peptide diet is supplied for 4 weeks from 10 weeks after obesity induction. The normal group fed the AIN-93G diet during the experimental period from 4 weeks to 11 weeks, and the high-fat diet group (the control group) used AIN-93G high-fat diet using beef tallow as a fat source for 40% of the total calories. To supply fat. The composition of the fed diet is shown in Table 4.

Table 4: Experimental Dietary Composition

ingredient Experimental diet group (g / 100g diet) Control Experimental Group 1 Experimental Group 2 Experimental Group 3 Casein 20.0 18.0 14.0 10.0 Peptide of Example 1 - 2.0 6.0 10.0 L-Cystine 0.3 0.3 0.3 0.3 Corn starch 41.95 41.95 41.95 41.95 Sucrose 10.0 10.0 10.0 10.0 Cellulose 5.0 5.0 5.0 5.0 Soybean oil 7.0 7.0 7.0 7.0 Beef tallow 10.0 10.0 10.0 10.0 Cholesterol 1.0 1.0 1.0 1.0 Mineral Mix (AIN-93G: INC Co. USA) 3.5 3.5 3.5 3.5 Vitamin Mix (AIN-93VX: INC Co. USA) 1.0 1.0 1.0 1.0 TBHQ (t-Butylhydroquinone) 0.0014 0.0014 0.0014 0.0014 Choline bitartrate 0.25 0.25 0.25 0.25

Experimental animals are divided into five groups of 10 by egg mass according to body weight, and water and diet are supplied without restriction. The animals are fasted for 16 weeks, anesthetized with ether, and opened to collect blood from the abdominal aorta. Blood is collected by heart puncture at the time of sacrifice and centrifuged at 300 × g for 15 minutes to obtain plasma and stored at -70 ° C. To determine cholesterol and triglycerides in the liver after blood collection, the liver is removed, washed with 0.9% NaCl, weighed and stored at -70 ° C until analysis. The stools are collected for 4 days during the experimental period, measured the dry weight of the stool, stored at -70 ℃ and dried for 24 hours in an 80 ℃ dry oven before analysis to weigh the dry and then used in the experiment.

        2. Dietary Intake and Weight Measurement

Dietary intake and weight are measured twice a week. The body weight of the test animals was measured by starting weight after 6 weeks of induction of obesity, and weighing the final body weight after supplying the experimental formula containing black bean peptide (test group) for 4 weeks. Dietary efficiency is calculated by dividing body weight gain by dietary intake for 4 weeks. The results are shown in Table 5 and FIG.

TABLE 5

Dietary Intake (g / day) Weight gain (g / day) Control 18.85 ± 1.27 6.80 ± 0.25 Experimental Group 1 20.12 ± 1.07 6.07 ± 0.22 Experimental Group 2 18.05 ± 0.30 5.32 ± 0.62 Experimental Group 3 18.00 ± 2.31 3.62 ± 0.81

In the case of weight gain, there was no difference in dietary intake between the control group and the experimental group, but there was a weight loss effect in the experimental group fed the black soybean peptide, especially the highest weight loss effect in the experimental group 3 fed the 10% black bean peptide. Indicated.

3. Lipid analysis in the blood

Triglyceride measurement in blood was calculated by measuring the absorbance at 505nm using a standard enzyme colorimetric kit to calculate the triglyceride concentration of blood. Total cholesterol in blood was measured by measuring absorbance at 500 nm using a standard enzyme colorimetric kit to calculate total cholesterol concentration in blood. In order to measure high-density lipoprotein (HDL) cholesterol, a solution was added to the precipitate using a kit, and the absorbance was measured at 555 nm using the supernatant as a sample. Low-density lipoprotein (LDL) and very low-density lipoprotein (VLDL) cholesterol were calculated using the formula of Fidewald et al. (Friedward, WT et al., Clin. Chem ., 18, 499, 502, 1972). . The results are shown in Table 6 and FIG.

TABLE 6

Triglyceride (mg / dl) Cholesterol (mg / dl) Arteriosclerosis index Total cholesterol HDL cholesterol LDL cholesterol VLDL Cholesterol Control 94.28 ± 22.85 126.90 ± 17.72 20.73 ± 3.87 87.55 ± 18.89 18.62 ± 4.92 5.36 ± 1.80 Experimental Group 1 89.86 ± 18.35 118.28 ± 17.41 26.8 ± 7.85 73.51 ± 18.75 17.97 ± 3.67 3.71 ± 1.62 Experimental Group 2 77.72 ± 33.99 109.45 ± 13.50 29.23 ± 1.35 64.68 ± 10.53 15.54 ± 6.80 2.74 ± 0.38 Experimental Group 3 69.58 ± 11.45 94.96 ± 14.71 46.15 ± 6.23 34.89 ± 11.95 13.92 ± 2.29 1.07 ± 0.30

The total cholesterol in serum was decreased in proportion to the concentration of black soybean peptide in the experimental group compared to the control group. It was confirmed that significantly suppressed.

In addition, HDL-cholesterol, known as beneficial cholesterol, was also increased in the experimental group fed with the black soybean peptide, and showed the highest increase in the experimental group 3 fed the 10% black bean peptide. On the other hand, LDL-cholesterol, which is known to be an unhealthy cholesterol, was all decreased in the experimental group, especially in the experimental group 3 in which 10% of the black soybean peptides were fed.

The arteriosclerosis index was found to be much lower than 5 times in the experimental group ingested black soybean peptides compared to the control group.

4. Liver Tissue and Lipid Analysis

The analysis of total cholesterol and triglycerides in the liver was carried out using Folch et al. (Folch, J. et al .: J. Biol. Chem ., 226, 497-509, 1957). It was measured using the same kit, the results are shown in Table 7 and FIG.

TABLE 7

Liver weight (g) Total fat (mg / g liver) Triglycerides (mg / g liver) Total cholesterol (mg / g liver) Control 13.24 ± 1.13 557.27 ± 71.05 106.20 ± 4.95 140.99 ± 5.37 Experimental Group 1 12.05 ± 0.39 511.12 ± 47.27 106.63 ± 3.32 144.50 ± 10.01 Experimental Group 2 10.67 ± 0.22 469.05 ± 43.20 109.08 ± 3.36 133.25 ± 9.03 Experimental Group 3 10.06 ± 0.26 451.97 ± 71.14 94.30 ± 11.99 131.79 ± 8.22

The liver weight was lower than that of the control group in all groups. Especially, in the experimental group 3 fed 10% of the black soybean peptide, total fat, triglyceride, and total cholesterol were significantly reduced.

5. Lipid analysis of feces

For analysis of total cholesterol and triglyceride of feces , Folch et al. (Folch, J. et al .: J. Biol. Chem ., 226, 497-509, 1957) analyzed the various substrates in the same way as the liver Table 8 and Figure 4 are shown.

TABLE 8

Dry weight of feces (g / day) Total fat (mg / g fecal) Triglycerides (mg / g feces) Total cholesterol (mg / g fecal) Control 1.86 ± 0.10 203.33 ± 45.09 7.58 ± 1.47 15.48 ± 0.78 Experimental Group 1 1.15 ± 0.30 245.00 ± 49.50 6.87 ± 0.36 17.73 ± 0.35 Experimental Group 2 1.00 ± 0.00 265.00 ± 35.36 6.34 ± 0.14 16.83 ± 0.11 Experimental Group 3 1.92 ± 0.14 290.00 ± 17.32 7.41 ± 0.49 18.00 ± 2.13

There was no difference in dry weight of feces, but the triglyceride and total cholesterol contents were increased in the experimental group fed black soybean peptide. Compared with the control group, the total fat content of feces increased by 21, 31 and 43% in experimental group 1, experimental group 2 and experimental group 3, respectively.

Example 3 Preparation of Drinking Peptide Drink Using Black Soy Peptide with Diet Effect

The black soybean peptide drink using the black soybean peptide having a dietary effect obtained in Example 1 was prepared.

10 mL of the black soybean peptide prepared in Example 1 was added to 100 mL of water and sweeteners (high fructose) to give sweetness and refreshing taste, acidulants (citric acid) and fruit flavor (flavor) to lower the harmonious taste and pH of the beverage Was prepared in Table 9 below, followed by heating with HTST to prepare a peptide drink.

TABLE 9

Raw material name Compounding amount (g) Raw material name Compounding amount (g) Black Bean Peptide 20.0 Sodium citrate 0.09 High fructose 9.0 crush 10.0 Baekbaekdan 3.0 Fruit Flavor 0.1 Citric acid 0.35 Purified water 57.46

                                            (Total: 100 g)

According to the present invention, the black bean-derived peptides that can be routinely consumed as general foods can suppress the increase in the concentration of triglycerides in the blood and liver, improve lipid metabolism, and excrete triglycerides through defecation to reduce weight. It works.

In addition, there is an advantage that easier to consume by preparing a beverage using black soybean peptide.

Claims (3)

As a peptide composition derived from black soy protein, At least 10 mg of arginine, at least 10 mg of aspartic acid, and at least 20 mg of glutamic acid per gram of peptide composition, The black soybean peptide composition having the effect of inhibiting the increase in the concentration of lipids in the blood having a molecular weight of 10,000 or less in the peptide composition of 70% by weight or more and weight loss effect. The method of claim 1, The black soybean peptide composition is characterized by separating the solid and liquid obtained by sequentially hydrolyzing black soybeans with endopeptidase and exopeptidase enzyme, and then the liquid phase is filtered and powdered to inhibit the concentration increase of blood lipids and body weight Black soybean peptide composition having a reducing effect. Claims 1 or 2 of the black soybean peptide composition, high fructose, citric acid, juice and purified water containing black lipid peptide drinking water having an effect of inhibiting the increase in blood lipid concentration.

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