KR101433239B1 - Preparation method for water soluble rice peptide - Google Patents

Abstract

The present invention provides a process for producing rice peptides having excellent water solubility, reduced bitter taste and excellent fatigue recovery activity after exercise from rice syrup foil, which is a by-product of rice processing, (GMO) or allergic reaction of soybean or milk-derived protein which is being used as a functional food material that can be easily applied to beverages because of its excellent solubility in water Show your surname.

Description

Preparation method for water soluble rice peptides [

More particularly, the present invention relates to a method for producing rice peptides having excellent water solubility and reduced bitter taste from rice syrup foil, and having excellent endurance exercise ability or fatigue inhibiting activity.

According to the National Statistical Office (NSO) survey, domestic rice production increased by 4.7% in 2008 compared to 2007, but domestic consumption of rice decreased by 2.4% in 2007 from 76.9 kg in 2007.

Since the consumption of rice is decreasing every year while the amount of rice is increasing, the remaining rice is used to make and sell Takju, etc. The development and production of edible oil and phytoremediation using rice are gradually increasing, Is generated. Processing of byproducts of rice is costly and environmental problems are followed, so it is mostly used for feed or compost.

Therefore, there is a great need for research that can reuse by-products to develop new high value added materials and products and utilize byproducts.

Rice proteins are now attracting worldwide attention because they are free of allergenic problems and are rich in nutritional value compared to soy protein. In the case of soybean-based materials currently in widespread use, there is a problem related to GMO, while in the case of rice, there is no problem of GMO.

Although a number of overseas companies including Sunrich USA are conducting research on rice proteins, there have been no specific developmental achievements so far, and researches using byproducts have been actively conducted in various countries including Southeast Asia where rice is a main source, Research on rice-derived proteins is limited to the United States and Pakistan, and there is a limit to commercialization due to technical limitations.

In addition, considering the present domestic market situation, the commercial rice protein products have problems related to the water-solubilization of proteins, problems in raw material supply and taste, and therefore consumers are very concerned about the use of materials. However, In fact, it has not been revived in its demand.

Korean Patent No. 10-0503100 discloses a method for producing a low-molecular-weight peptide by using a vegetable protease from rice bran. However, in the case of rice bran, the content of the raw protein is less than 20% There is a limitation that separation is difficult, and when the enzyme treatment is carried out from a raw material having a low protein content, it is difficult to solve the problems of browning and already producing odor, and further, the vegetable proteolytic enzyme is more expensive than its potency and commercialization is limited.

Japanese Patent No. 4663001 discloses a method for producing a rice bran protein extract in which rice bran is treated with sodium hydroxide to extract proteins and then treated with hydrochloric acid to precipitate proteins and then washed with water, And saccharides contained in rice bran are browned to darken the color of the protein.

An object of the present invention is to provide a method for producing rice peptides excellent in water solubility from rice syrup foil, reduced bitter taste and excellent fatigue recovery activity after exercise.

The present invention relates to a process for the treatment of alpha-amylase which comprises reacting a rice syrup foil with an alpha-amylase; Removing the water-soluble material to wash the rice syrup foil treated with the alpha-amylase; And a proteolytic enzyme treatment step of reacting the washed rice syrup foil with a proteolytic enzyme.

In the method for producing rice peptides of the present invention, the alpha-amylase is characterized by being Temamyl.

The method for producing rice peptides of the present invention is characterized in that the above termamyl is reacted at 80 to 100 캜.

In the method for producing rice peptides according to the present invention, the water-soluble substance removing step may include dehydrating the α-amylase-treated rice syrup foil, mixing 2 to 30 times of the water content of the dehydrated rice syrup solids, Is repeated 1 to 5 times.

In the method for producing a rice peptide according to the present invention, the protease is characterized by being an alkalase or a nitrogenase.

In the method for producing rice peptides according to the present invention, the step of degrading with the protease comprises adding 0.01 to 1% by weight of a protease, reacting at 50 to 70 ° C for 0.5 to 3 hours, adjusting the initial pH to 8 to 9 , And the reaction is completed at a pH of 7 to 7.5.

In the method for producing rice peptides of the present invention, the method further comprises a step of recovering the peptide having a molecular weight of 1000 Da or less through ultrafiltration after the proteolytic enzyme treatment step.

In the rice peptide production method of the present invention, the rice peptide has a molecular weight of 300 to 700 Da of at least 80% by weight of the entire peptide.

In the method for producing a rice peptide according to the present invention, the content of amino nitrogen of the rice peptide is 10 to 1000 mg%.

In the rice peptide production method of the present invention, the content of glutamic acid in the rice peptide is 15 to 25% by weight of the total amino acids, and the content of branched amino acids is 12 to 20% by weight of the total amino acids.

In the method for producing a rice peptide according to the present invention, the water solubility of the rice peptide at pH 4.6 to pH 12 is 90% or more.

In the method for producing rice peptides according to the present invention, an alpha-amylase treatment step in which rice syrup foil and Termamyl are reacted at 80 to 100 ° C; A water-soluble substance removing step of dehydrating the rice syrup foil treated with the alpha-amylase, mixing the water with the dehydrated rice syrup solids content of 2 to 30 times the amount of water, and repeating the filtering step 1 to 5 times; And a proteolytic enzyme treatment step of adding the washed rice syrup foil in an amount of 0.01 to 1% by weight of an alkalase or a nitrogenase and reacting at 50 to 70 ° C for 0.5 to 3 hours, wherein the molecular weight of the rice peptide is Wherein the amino acid nitrogen content of the rice peptide is 10 to 1000 mg%, the content of glutamic acid in the rice peptide is 15 to 25% by weight of the total amino acids, The branched amino acid content is 12 to 20 wt% of the total amino acid, and the water solubility of the rice peptide at pH 4.6 to pH 12 is 90% or more.

In the method for producing rice peptides of the present invention, the rice peptides are characterized in that they have fatigue recovery activity after exercise.

The water-soluble rice peptides having a post-exercise fatigue recovery activity of the present invention may contain 9 to 12 parts by weight of aspartic acid, 3 to 5 parts by weight of threonine, 5 to 7 parts by weight of serine, 19 to 22 parts by weight of glutamic acid, 3 to 5 parts by weight of proline, 4 to 6 parts by weight of glycine, 5 to 7 parts by weight of alanine, 5 to 7 parts by weight of valine, 1 to 3 parts by weight of methionine, 3 to 5 parts by weight of isoleucine, 7 to 9 parts by weight of leucine, 5 to 7 parts by weight of phenylalanine, 2 to 4 parts by weight of lysine, 1 to 3 parts by weight of histidine and 7 to 10 parts by weight of arginine.

The present invention provides a process for producing rice peptides having excellent water solubility, reduced bitter taste and excellent fatigue recovery activity after exercise from rice syrup foil, which is a by-product of rice processing, (GMO) or allergic reaction of soybean or milk-derived protein which is being used as a functional food material that can be easily applied to beverages because of its excellent solubility in water Show your surname.

Fig. 1 is a MS chromatogram obtained by confirming the water-soluble rice peptides of the examples in Example 8 on a molecular weight fraction basis.
Fig. 2 is a mass spectrum of a water-soluble rice peptide having a molecular weight of 300 to 1000 Da in Experimental Example 8. Fig.
Fig. 3 is a graph showing the water solubility of the water-soluble rice peptides according to Examples in Experimental Example 9 according to pH.
FIG. 4 is a graph showing the water solubility of the water-soluble rice peptides of the examples in Experimental Example 9 with heating temperature and time.

The present invention relates to a process for the treatment of alpha-amylase which comprises reacting a rice syrup foil with an alpha-amylase; Removing the water-soluble material to wash the rice syrup foil treated with the alpha-amylase; And a proteolytic enzyme treatment step of reacting the washed rice syrup foil with a proteolytic enzyme.

In the present invention, the rice peptides are preferably prepared from syrup foil. As a byproduct of rice processing, it is possible to use remaining skim milk, for example, makgeolli and remaining skim milk, remaining rice bran, or rice bran oil remaining therefrom, but the protein content Is preferably at least 40% by weight of rice syrup. As a by-product of the rice syrup-containing rice syrup-containing rice syrup-producing process, the rice syrup itself can be used as a raw material to produce rice peptides or hot air dried at 60 to 70 ° C at a moisture content of 10% It is preferable to use them.

In the present invention, first, alpha-amylase treatment is performed to react rice syrup foil, which is a raw material, with alpha-amylase.

In the present invention, when the rice syrup foil is directly reacted with the proteolytic enzyme for the production of rice peptides, browning and insoluble precipitates can be formed by the starch or its decomposition products remaining in the rice syrup foil. If the protein content exceeds 80 wt% High purity products can not be produced.

As the alpha-amylase in the present invention, Termamyl of novozyme is preferred. In the case of tamarimel, the starch decomposition ability is excellent within a short time, and the optimum reaction temperature is 93 ° C. At the reaction at 80 to 100 ° C, the starch decomposition and sterilization of the raw material can proceed simultaneously.

Next, in the present invention, alpha-amylase treatment is performed to react rice syrup foil with alpha-amylase, and rice silage foil treated with alpha-amylase is washed to remove water-soluble substances.

A rice syrup foil treated with alpha-amylase is a liquid reaction mixture in which rice syrup bulk solids, enzyme-degraded water-soluble carbohydrates (mainly monosaccharides, disaccharides or oligosaccharides), and enzymes are mixed. This liquid reaction mixture is centrifuged, Dehydrated using a decanter or the like.

However, since the water-soluble carbohydrate can not be sufficiently removed by simply dehydrating the enzyme reaction solution, the α-amylase treated rice syrup foil is dehydrated and the amount of the dehydrated rice syrup solids is 2 to 30 times, preferably 5 To 15 times of water is mixed and sufficiently stirred, followed by dewatering is repeated 1 to 5 times, preferably 3 times or more, so that the water-soluble carbohydrate is removed and the protein content is increased.

Next, in the present invention, a proteolytic enzyme treatment step of reacting washed rice syrup foil with a proteolytic enzyme is performed in order to remove a water-soluble substance.

The protease of the present invention is preferably novozyme Alcalase or Neutrase. The proteinase degrades the rice syrup protein by decomposing the rice protein into a protein having a molecular weight of 1000 Da or less, preferably 300 to 700 Da, of 80% or less by weight of the whole peptide, By weight or more, preferably 90% by weight or more, so that it is possible to produce a water-soluble rice peptide having a bitter taste and a low water content. When treated with the proteolytic enzyme, the amino acid nitrogen content is 1000 mg% or less, preferably 200 to 800 mg%, and the bitterness is low enough without removing the amino acid or short residue peptide through separate ultrafiltration, This high peptide can be produced.

In the present invention, proteolytic enzyme is added in an amount of 0.01 to 1% by weight, preferably 0.1 to 0.5% by weight, and the mixture is reacted at 50 to 70 ° C for 0.5 to 3 hours.

When the concentration of the protease is less than the lower limit, the protein degradation efficiency is not sufficient. Therefore, the yield of the water-soluble rice peptide is insufficient relative to the amount of rice syrup used and the protein degradation efficiency is increased up to 0.1 wt% Do not increase anymore.

In addition, the longer the reaction time of the proteolytic enzyme, the higher the protein degradation rate, but the increase in the degradation rate after 3 hours is not clear. On the contrary, if the pH is decreased over 3 hours, Loses.

In addition, when the pH is adjusted to 8 to 9 at the initial stage of the enzymatic degradation by the protease, and the reaction is completed at pH 7 to 7.5, there is less concern about the occurrence of side patches and no need for neutralization after completion of the reaction, The salt content of the rice peptides can be prevented from increasing.

In the present invention, in order to obtain the water-soluble rice peptides in the enzymatic reaction solution, the enzyme reaction solution is first subjected to a proteolytic enzyme treatment step, and then the enzyme reaction solution is first heated at 75 DEG C or higher, preferably 85 to 100 DEG C for 10 minutes to 1 hour, After the enzyme is inactivated by heating for 15 to 30 minutes, the solids are removed by centrifugation or filter press in the enzyme reaction solution, and the liquid portion is concentrated and dried to prepare a water-soluble rice peptide.

The water-soluble rice peptides of the present invention can be produced by subjecting most of the peptides, for example, not less than 70% by weight, preferably not less than 80% by weight, more preferably not less than 90% Or less. However, in the present invention, in order to remove the peptide having a molecular weight of more than 1000 Da and to further increase the concentration of the peptide in the desired molecular weight range, the liquid portion of the enzyme reaction solution is further passed through an ultrafiltration membrane to concentrate a portion having a molecular weight of 1000 Da or less .

The water-soluble rice peptide of the present invention has a molecular weight of at least 80% by weight, preferably at least 90% by weight, more preferably at least 95% by weight of the peptide of 300 to 700 Da in total peptide.

The content of amino nitrogen of the rice peptides of the water-soluble rice peptides of the present invention is 10 to 1000 mg%, preferably 800 mg% or less, more preferably 700 mg% or less. Amino nitrogen content is advantageous as it contains less amino acid or short residue peptide causing bitter taste and bad odor, but is usually about 400 to 700 mg% in the peptide of the present invention.

The glutamic acid content of the water soluble rice peptides of the present invention is 15 to 25% by weight of the total amino acids. The branched amino acid content is 12 to 20% by weight of the total amino acids, and the content of branched amino acids known to help improve the raw material and exercise capacity of glutathione Is high.

The water-soluble rice peptides of the present invention preferably contain 9 to 12 parts by weight of aspartic acid, 3 to 5 parts by weight of threonine, 5 to 7 parts by weight of serine, 19 to 22 parts by weight of glutamic acid, 3 to 5 parts by weight of proline, 5 to 7 parts by weight of alanine, 5 to 7 parts by weight of valine, 1 to 3 parts by weight of methionine, 3 to 5 parts by weight of isoleucine, 7 to 9 parts by weight of leucine, 4 to 6 parts by weight of tyrosine, 7 to 10 parts by weight of arginine, 2 to 4 parts by weight of lysine, 1 to 3 parts by weight of histidine, and 7 to 10 parts by weight of arginine.

The water-soluble rice peptides of the present invention have a water solubility of 90% or more, preferably 95% or more, at pH 4.6 to pH 12, and are very highly water-soluble even at pH 4.6, which is an acidic region unlike the conventionally available rice proteins. It can be easily applied to acid drinks.

In addition, the water-soluble rice peptides of the present invention exhibit increased endurance exercise capacity or post-exercise fatigue inhibitory activity by increasing blood glucose concentration and reducing plasma ammonia concentration upon oral ingestion.

Hereinafter, the present invention will be described more specifically with reference to the following examples. However, the following examples are provided only to aid understanding of the present invention, and the scope of the present invention is not limited to the following examples.

Experimental Example 1: Analysis of general components of rice processing by-products

As a byproduct of rice processing, sake bran, takju bran, skim rice bran and rice syrup bran were used. Cheongju, takju and rice syrup were used after drying.

However, in the case of the sake oil (moisture content 91.56 wt%) having a relatively high moisture content, when the hot air drying was performed, the sample could not be analyzed due to the bad smell and the mold could not be used because of the bad odor. Wt%) was dried at 50 캜 for 72 hours because hot air drying at a temperature of 60 캜 or higher failed to use the sample due to carbonization.

The components were analyzed by protein, fat, ash, water, and carbohydrate. The components were analyzed by AOAC method. The results were analyzed by Kjeldahl Nitrogen Analysis, Soxhlet method, Dry ashing and Air oven method. Carbohydrates were calculated as the fraction excluding protein, fat, ash and water.

 division rice Takju Defatted rice bran Rice syrup foil Crude protein (% by weight) 6.12-8.67 25.3 ± 0.6 18.7 ± 0.1 45.1 ± 0.9 Crude fat (% by weight) 0.35-0.45 4.3 ± 0.0 4.3 ± 0.0 8.1 ± 0.2 Views min (% by weight) 0.59-0.63 0.4 ± 0.0 11.2 ± 0.2 0.8 ± 0.0 Water (% by weight) 13.22-14.23 6.4 ± 0.2 7.2 ± 0.3 6.3 ± 0.3 Carbohydrates (% by weight) 76.6-79.12 63.6 ± 0.4 58.6 ± 0.3 36.7 ± 0.1

In the case of takju bean, the carbohydrate fraction decreased by yeast degradation, and that of the defatted rice bran increased compared to that of whole rice. Also, in the case of rice syrup leaves, the carbohydrate fraction was markedly decreased due to the carbohydrate saccharification reaction.

Experimental Example 2: Determination of Optimum Reaction Time by Enzyme

In order to further decompose carbohydrates remaining in the rice syrup foil, the optimal reaction time was checked for carbohydrase such as starch, cellulose, and hullulan. The properties, composition, activity, density, optimum pH and optimum temperature of the six enzymes used in the experiments are summarized in Table 2.

division Appearance Constitutive enzyme Minimal activity density Optimum pH Optimum temperature
(° C) Celluclast Brown liquid cellulase 700 EGU / g 1.22 g / mL 4.5-6.0 (4.5) 50-60 (60) Fungamyl 800L brown liquid α-amylase 800 FAU / g 1.25 g / ml 5 (5) 55-60 (60) Liquozyme supra amber liquid α-amylase 135 KNU / g 1.26 g / ml 5.5 to 7.0 (6.5) 50 (50) Promosy D2 brown liquid pullulanse 400 PUN / ml 1.25 g / ml 5 (5) 60 (60) Termamyl type L brown liquid α-amylase 120 KNU / g 1.20 to 1.25 g / ml 6.5 (6.5) 93 (90) Viscozyme L brown liquid β-glucanase
arabanase, cellulase, hemicellulase, xylanase 120 FBG / ml 1.2 g / ml 4.6 (4.6) 44 (44)

To establish the optimal reaction time for each enzyme, 5 g of rice syrup was added to distilled water (28.3 mL) to prepare a 15% suspension, and the environment was adjusted to the optimum pH and optimum temperature of each enzyme. Were reacted at 1.5, 3, 4, 5, 6, 7.5, 9, 10.5, 12 and 24 hours respectively. The optimum reaction time for each enzyme was determined by using the weight loss amount and it is shown in Table 3.

division Celluclast Fungamyl 800L Liquozyme supra Promosy D2 Termamyl type L Viscozyme L Optimum reaction time (hrs) 24 9 4.5 9 1.5 24

Among the α - amylases, the optimum reaction time was the shortest for the tammaril, so it was selected as the enzyme for starch decomposition of rice syrup. In addition, Celluclast and Viscozyme, a complex enzyme, were selected as a cellulolytic enzyme which is expected to have a synergistic effect when used together with tamarind, and their carbohydrate decomposing effects were confirmed by their respective and combined use.

Experimental Example 3: Confirmation of carbohydrate degradation by alpha-amylase treatment

The weight loss of carbohydrates was calculated as the amount of degradation due to the water-solubilization of the carbohydrate fractions using the weight difference between the precipitates before and after the enzyme reaction. The addition amount of the enzyme was 1 wt% of the content of rice syrup solids. In the case of single enzyme treatment, the optimum pH and temperature of the enzymes used in Table 2 were 1.5 hours for teramylase, 24 hours for viscose, And in the case of complex enzyme treatment, the enzyme treatment was performed sequentially from the highest to the lowest temperature. The reaction solution was subjected to centrifugation (3000 g) for 20 minutes, and the weight and precipitate of the precipitate thus obtained were analyzed to compare and analyze changes in protein content.

 division Carbohydrate decomposition amount
( mg / g) Crude protein
(weight%) Rice syrup foil ^- 43.06 Termamyl 85.69 ± 0.60 ^A 70.68 Viscozyme 61.74 ± 7.12 ^C 74.01 Celluclast 68.43 ± 5.13 ^BC 75.00 T + V 68.03 + - 1.54 ^BC 73.69 V + C 64.03 + - 2.40 ^BC 74.62 T + C 69.70 ± 0.90 ^BC 72.40 T + V + C 72.04 + - 4.75 ^B 72.11

In Table 4, T stands for Termamyl, V stands for Viscozyme, and C stands for Celluclast.

It was found that termamyl (85.69 mg / g) had the greatest effect on weight change with single enzyme. This is a larger change in weight than the treatment of all three enzymes (72.04 mg / g). In addition, the protein content of syrup oil increased from 43.06% by weight to more than 70% by treatment with tamarisk.

In addition, it was confirmed that the starch was decomposed in the carbohydrates through the treatment of tammaril, and the protein content was relatively increased due to the weight reduction.

On the other hand, the amount of carbohydrate degradation was lower in biscozyme or cellulase than that of teramil, but the crude protein content could be increased. However, the increase in the crude protein content was small compared to the increase in the reaction time for the enzyme treatment.

Experimental Example 4: Determination of Protein Content Changes by Washing Number after Alpha-Amylase Treatment

0.1% by weight of turmeric was added to the rice syrupy solid content, and the mixture was allowed to react at 90 DEG C for 1.5 hours. Then, the supernatant was removed and dehydrated through a horizontal decanter to obtain a solid product (Production Example 4-1) 10 times the amount of water was added and mixed with stirring for 20 minutes. Then, again dehydrated solid (Preparation Example 4-2: once washing) and 10 times water were further added to Sample 2 and mixed with stirring for 20 minutes (Example 4-3: twice of washing) and Production Example 4-4 of this washing three times, Production Example 4-5 of four times of washing, Production Example 4-6 of five times of washing And the increase of the protein content was confirmed. The results are shown in Table 5.

division Production Example 4-1 Production example 4-2 Production Example 4-3 Production Example 4-4 Production Example 4-5 Production Example 4-6 Crude protein
(weight%) 67.6 72.69 74.14 78.01 78.45 78.61

After the enzymatic reaction, the protein content of the dehydrated solid was 67.6% by weight, and the protein content was increased with each washing of the water soluble substance to 78.01% by weight and the protein content was increased by about 10% by weight. However, even after washing 4 times or more, no increase in protein content could be confirmed.

Experimental Example 5: Determination of Protein Degradation Rate and Bitter Taste According to Protease

In order to confirm the proteolytic enzyme suitable for proteolysis of rice syrup fiber, protein degradation rate using the proteolytic enzyme shown in Table 6 was confirmed.

 division manufacturer Optimum pH Optimum temperature (℃) Protamex novozyme 5.5-7.5 (7) 35-60 (40) Neutrase novozyme 5.5-7.5 (7) 45-55 (45) Flavourzyme novozyme 5-7 (7) 50 (50) Alcalase novozyme 6.5-8.5 (7) 60 (60) Protease M amano 4.5 (4.5) 50 (50) Protease N amano 7 (7) 55 (55) Protease A amano 7 (7) 50 (50) Protease F kikoman 3 (3) 50 (50)

 To 5 g of the washed rice syrup oil of Preparation Example 4-4 was added distilled water (28.3 ml) to prepare a 15% suspension. Using 1 N HCl or 1 N NaOH, The pH was adjusted, and 0.1% enzyme was added to the solid content of the sample at the optimum temperature, followed by reaction for 4 hours. The reaction suspension was centrifuged (3000 g) for 20 minutes, and the protein content of each of the obtained supernatants was compared with each other.

 division protein (%) Protamex 75.43 + 1.81 Flavourzyme 78.60 + - 0.56 Neutrase 32.73 + - 1.75 Alcalase 82.46 ± 1.24 Protease M 83.32 + - 0.33 Protease N 86.43 + - 2.12 Protease A 62.73 + - 0.35 Protease F 58.07 ± 0.56

The sensory evaluation was performed on the sensory evaluation of bitterness and acceptability of the 17 sensory test panels, which were obtained by centrifuging the supernatant containing the water-soluble rice peptides obtained in the above reaction, using the 9-point rating method. The results are shown in Table 8 Respectively.

 division bitter Likelihood Protamex 7.2 ± 0.5 2.4 ± 0.6 Flavourzyme 5.7 ± 0.4 3.4 ± 0.5 Neutrase 4.1 ± 0.2 4.2 ± 0.3 Alcalase 3.6 ± 0.4 5.5 ± 0.5 Protease M 7.7 ± 0.4 1.9 ± 0.3 Protease N 8.0 ± 0.4 1.3 ± 0.2 Protease A 7.4 ± 0.4 2.3 ± 0.3 Protease F 4.4 ± 0.4 5.1 ± 0.5

From the above results, it has been found that when a water-soluble rice peptide is prepared from rice syrup foil, it is possible to produce a rice peptide having high palatability by using a nutrient or an alkalase. In particular, when alkalase is used, Respectively.

Experimental Example 6: Reaction conditions of proteolytic enzyme

The protein degradation rate, pH, and sensory properties of the protease selected in Experimental Example 5 and the protein content of the enzymolytic degraded product according to the addition amount of protease were analyzed according to the reaction time of the alkaline protease.

In order to determine the optimum reaction time of the alcalase, distilled water (28.3 ml) was added to 5 g of the washed rice syrup foil of Production Example 4-4 of Experimental Example 4 to prepare a 15% suspension, and 1 N HCl or 1 N NaOH was used to adjust the pH to 7, and 1% of enzyme was added to the solid content of the sample at an optimum temperature of 60 ° C. The protein content of the enzyme hydrolyzate at 1, 2, 3, The pH was measured and the strength of the noodle patch was examined by sensory evaluation using the 9-point scale method for 17 sensory panels.

Reaction time (h) protein (%) pH Strength of side patch One 71.5 6.5 2.4 2 77.3 6.3 2.6 3 81.5 6.1 2.7 5 85.2 5.4 4.2 24 90.3 4.5 5.8

Protein degradation rate gradually increased with reaction time. However, after 3 hours, the pH rapidly decreased and the patches started to occur, and it was determined that there was a quality problem, and the optimum reaction time was determined to be 3 hours.

In order to determine the optimum amount of enzyme to be added to the alkaline protease, the washed rice syrup foil of Production Example 4-4 of Experimental Example 4 was determined to be 0.005, 0.01, 0.2 and 1% by weight based on the substrate. After fixed for 3 hours, the protein was quantified by the lowry method.

Amount of enzyme added (% by weight) 0.005 0.01 0.1 0.5 One protein (%) 42.5 71.2 82.2 83.4 83.5

When the content of protease was more than 0.01 wt%, it was enough to decompose the rice syrup. At 0.1 wt%, the protein degradation rate was increased with increasing amount of enzyme, but the protein decomposition rate was not increased any more.

Example:

0.1% by weight of Termamyl SC was added to a rice syrup foil having a moisture content of 45% by weight and a moisture content of 45% by weight in dry solid matter, and the mixture was reacted at 90 캜 for 1.5 hours. The reaction supernatant was removed and dehydrated through a horizontal decanter. Water was added to the solid material 10 times as much as the solids content, mixed with stirring for 20 minutes, and then dehydrated again. The dehydrated solid obtained by repeating the washing process three times was added with a solid amount 10 times of water was mixed and adjusted to pH 8.5. Then, 0.1 wt% of alkalase was added to the solid amount, and the mixture was reacted at 60 ° C for 3 hours. The final pH of the enzyme reaction solution was 7.2, and the enzyme was inactivated by heating at 85 ° C for 15 minutes. The inactivated enzyme reaction solution was dehydrated through a horizontal decanter, and the liquid phase was concentrated to 20 wt% Dried at 60 ° C for 12 hours, and pulverized to obtain a water-soluble rice peptide powder.

Experimental Example 7: General composition and amino acid composition of the water-soluble rice peptide powder

Crude protein, crude fat, crude protein, and carbohydrate content of the water-soluble rice peptide powder of the above Example were analyzed in the same manner as in Experimental Example 1. [

 division Water-soluble rice peptide powder Crude protein (% by weight) 82.1 Crude fat (% by weight) 2.2 Views min (% by weight) 3.1 Water (% by weight) 6.4 Carbohydrates (% by weight) 6.2

A quantitative test was conducted on 16 kinds of constituent amino acids of the water - soluble rice peptides. The analysis was carried out by ninhydrin HPLC analysis and the results are shown in Table 12.

Constituent amino acid Brown rice
(Korean Journal of Agricultural Chemistry, 1977) Soluble rice peptides
(%) Aspartic acid 9.5 10.45 Threonine 3.9 3.41 Serine 5.2 5.72 Glutamic acid 19.4 21.64 Proline 4.6 4.34 Glycine 4.7 5.10 Alanine 6.5 5.91 Valine 6.2 5.71 Methionine 2.1 1.86 Isoleucine 4.0 3.68 Leucine 8.5 7.59 Tyrosine 4.4 4.98 Phenylalanine 5.7 5.73 Lysine 3.6 2.90 Histidine 2.5 2.38 Arginine 9.3 8.60 total 100.1 100 * BCAA 18.7 16.98

The constituent amino acids of the rice peptides were almost identical to those of the starting rice, and it was confirmed that the content of glutamic acid was contained in a high proportion of about 20%. It was also confirmed that the content of Branched Chain Amino Acid (BCAA), which has been reported to improve exercise performance and fatigue recovery, is as high as 16.98%.

Experimental Example 8: Molecular weight distribution and amino nitrogen content of water-soluble rice peptide powder

HR-MS analysis was performed to confirm the molecular weight distribution of the water-soluble rice peptides of the above examples. The concentration of the sample was 2 mg / mL. Distilled water and acetonitrile were used as solvents, and formic acid was used as a reagent. Experimental methods and instrument conditions are shown in Tables 13 and 14, respectively.

One Chromatography Thermo Accela UPLC 2 Mass spectrometry Thermo LTQ-Orbitrap XL 3 Colum ACQUITY BEH130C18, 1.7um, 100 * 2.1mm 4
Solvent
A: DW (0.1% formic acid) B: Acetonitrile (0.1% Formic acid) 5






Elution condition






Time A B 0.0 99 One 3.0 99 One 53.0 50 50 53.5 0 100 56.0 0 100 56.5 99 One 60.0 99 One 6 Flow rate 0.3 ml / min 7 Injection 3 μl

One Detection ion mode Positive ion ((M + H] +) 2 Scan range MS: m / z 300-1600 3 Spray voltage 4.5 kV 4 Capillary voltage 35 V 5 Capillary Temp 300 6 CID energy 35 7 Software Xcalibur

MS chromatogram confirmed by molecular weight fractions under the above conditions is shown in Fig. According to FIG. 1, most of the molecular weight is 1000 Da or less, and among these, the molecular weight of about 300 to 700 Da is 95% or more.

A mass spectrum of a molecular weight of 300 to 1000 Da is shown in Fig.

Amino nitrogen content was also measured to determine the degree of decomposition of the water soluble rice peptides of the examples into amino acids. According to the Formol titration method, 2 ml of the eluate was taken in a 100 ml measuring flask and mixed with 100 ml of distilled water. 25 ml of the diluted eluate was taken in a 100 ml beaker and 0.1 N NaOH was added and the pH was adjusted to 8.4 with a pH meter. To this, 20 ml of 35% formaldehyde solution (v / v) (Samchun Pure Chemical) was added and neutralized to pH 8.4 with 0.1 N NaOH. Amino nitrogen value was calculated as "[(A-B) x1.4xFxDx100] / S". A is the consumption of 0.1 N NaOH, B is the coarse experimental consumption of 0.1 N NaOH, F is the factor value of 0.1 N NaOH, D is the dilution factor, S is the sample weight (g) (Mg) equivalent to 1 ml.

The amino nitrogen content of the water-soluble rice peptides of the examples was 600 mg%, indicating that the degree of decomposition into amino acid units was not large.

EXPERIMENTAL EXAMPLE 9: Change of Solubility According to pH, Heat and Storage of Water-soluble Rice Peptide Powder

The protein content of the aqueous solution was measured at each pH in order to confirm the stability of the water-soluble rice peptide powder of the examples according to pH. The results are shown in FIG. In general, proteins exhibit an isoelectric point near pH 4 and precipitate. Albumin, a representative milk protein, exhibits an isoelectric point at pH 4.6. The water soluble rice peptides were very stable in aqueous solutions ranging in pH from 5 to 12 and exhibited gradual precipitation from pH 4.8 and maximum precipitation at pH 4.2. and the water solubility was 95% or more at pH 4.6 or higher.

In order to confirm the thermal stability, the water-soluble rice peptide powder was diluted 100 times with distilled water, and then 10 ml of the solution was dispensed into a 15 ml test tube and treated at 60, 80, 100 and 120 ° C for 0.5 h and 1 h, respectively. 4. As a result, the decrease of protein started gradually at 80 ℃ and 1h treatment condition, and loss of about 2% at 100 1h and loss of protein of about 6% at 120 1h were observed. Therefore, it was confirmed that the water-soluble rice peptide powder of the examples was a relatively stable substance in heat, and it was confirmed that it could be sterilized in the usual general beverage manufacturing process and stable at the process temperature.

Water soluble rice peptides were prepared by dissolving 3% by weight of water in water, and then sterilized at 100 ℃ for 15 minutes. In order to confirm the hygienic stability according to the storage period, the microorganisms were tested simultaneously during the storage period after sterilization. There was no significant difference in protein content between 0, 1, 2, 3, 4, 8, and 12 weeks after the sterilization. After 12 weeks from the sterilization, general bacteria, E. coli and E. coli were all not detected and sterilized for 12 weeks .

Experimental Example 10: Fatigue recovery activity after exercise of water-soluble rice peptide powder

Continued exercise reduces the glycogen content of the muscle and increases the amount of blood glucose into the muscle, which reduces blood glucose and results in fatigue.

In addition, aspartic acid is a major constituent of the purine nucleotide circuit in muscle, which regenerates the ATP pool during extreme high-intensity exercise, providing energy to the muscle and forming free ammonia during this process. It is known that the ammonia produced as a result of the decomposition of amino acids during exercise is released from the muscles into the blood, and the blood ammonia is transmitted to the brain to increase the central fatigue. Ammonia accumulated in the muscle cells stimulates the afferent nerves associated with muscle pain sensation, inhibits the TCA cycle and your biological function, and causes muscle fatigue by causing lactic acid production.

Therefore, in order to confirm whether or not the water-soluble rice peptides of the present invention can affect the exercise-restoring fatigue activity, the rats were orally administered for 4 weeks, and then the blood glucose and plasma ammonia concentrations were changed.

The experiment was carried out in Chemon, a GLP (Good Laboratory Practice) certification body, and the blood analysis of rats was conducted through Green Cross Medical Foundation. The water soluble rice peptides of the present invention were administered at low dose, medium dose and high dose, respectively. As a comparative group, isolated soybean protein was administered at an intermediate dose, and negative control was administered only with distilled water. Each dose was 67 mg / kg / day, 335 mg / kg / day, and 670 mg / kg / day, respectively, based on the protein content of the low dose group.

No deaths were observed for 4 weeks, and no significant weight change was observed in all groups compared to the control group.

The blood glucose concentrations after 4 weeks in each experimental group are shown in Table 15. [

division Average( mg / Dl) Standard Deviation Control group 70.80 11.81 Example (Low Capacity) 88.00 12.30 Example (intermediate capacity) 91.71 12.15 Example (high capacity) 98.80 10.52 Isolated soy protein (medium dose) 87.37 11.60

It was found that the concentration of glucose in blood gradually increased as the dose of the water soluble rice peptide of the Example was increased.

Plasma ammonia concentrations after 4 weeks in each experimental group are shown in Table 16.

division Mean (/ / ㎗) Standard Deviation Control group 753.57 199.33 Example (Low Capacity) 694.29 160.32 Example (intermediate capacity) 638.00 140.19 Example (high capacity) 600.71 67.27 Isolated soy protein (medium dose) 705.43 192.78

The plasma ammonia concentration of the control group was 753.57 / / dl. As the concentration of the water soluble rice peptides of the present invention was increased, the plasma ammonia concentration was decreased and the plasma ammonia reduction effect was superior to that of the comparative soybean protein isolate.

Therefore, the water soluble rice peptides of the present invention can be interpreted as having an effect on the fatigue lowering effect of rats after exercise.

Claims (15)

delete delete delete delete delete delete delete delete delete delete delete An alpha-amylase treatment step in which rice syrup foil having a protein content of 40% by weight or more and teramyl (Termamyl) are reacted at 80 to 100 ° C;
The rice syrup foil treated with the alpha-amylase is dewatered, and 2 to 30 times of the water content of the dewatered rice syrup solids is mixed and stirred. Thereafter, the process of dewatering is repeated 1 to 5 times ; And
Treating the washed rice syrup foil with a 0.1 to 1 wt% alcalase solution at 50 to 70 ° C for 0.5 to 3 hours,
The rice peptide obtained by treating the rice syrup foil with a proteolytic enzyme has a molecular weight of 300 to 700 Da of at least 90% by weight of the total peptide, an amino nitrogen content of 10 to 700 mg% of the rice peptide, Wherein the content of glutamic acid in the rice peptide is 15 to 25% by weight of the total amino acid, the content of the branched amino acid is 12 to 20% by weight of the total amino acid, the water solubility of the rice peptide at pH 4.6 to pH 12 is 90%
Wherein the amino acid composition of the rice peptide is 9 to 12 parts by weight of aspartic acid, 3 to 5 parts by weight of threonine, 5 to 7 parts by weight of serine, 19 to 22 parts by weight of glutamic acid, 3 to 5 parts by weight of proline, 4 to 6 parts by weight of glycine, 5-7 parts by weight of alanine, 5-7 parts by weight of valine, 1-3 parts by weight of methionine, 3-5 parts by weight of isoleucine, 7-9 parts by weight of leucine, 4-6 parts by weight of tyrosine, 5-7 parts by weight of phenylalanine, 2 to 4 parts by weight of lysine, 1 to 3 parts by weight of histidine and 7 to 10 parts by weight of arginine. delete delete delete

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