KR102161126B1 - Extraction method of protein hydrolysate from horsemeat and protein hydrolysate manufactured by the same - Google Patents

Abstract

The present invention includes a pretreatment step of preparing raw meat by crushing horse meat and removing impurities; Hydrolysis step of adding water and enzyme to the raw meat and reacting at a temperature range of 40 to 75°C; A post-treatment step of heat-treating the hydrolyzate obtained through the hydrolysis step at a temperature range of 85 to 105°C; A separation step of extracting a liquid protein mixture by centrifuging the hydrolyzate obtained through the post-treatment step; Filtration to remove foreign substances from the protein mixture; A concentration step of concentrating the filtrate obtained through the filtration step; And a drying step of drying the concentrate obtained through the concentration step; comprising, a method for extracting a protein hydrolyzate using horse meat, which has a characteristic off-flavor of horse meat and a bitter taste generated during protein production, and has a low molecular weight. A protein hydrolyzate having a high absorption rate to the human body can be prepared, including amino acids.

Description

Extraction method of protein hydrolysate from horsemeat and protein hydrolysate manufactured by the same}

The present invention relates to a method of extracting a protein hydrolyzate using horse meat that can be used as food or health food, and to a protein hydrolyzate prepared accordingly, and more specifically, a pretreatment step, a hydrolysis step, a post treatment step, and a separation step. It relates to a method for extracting a hydrolyzed protein comprising a.

Cognitive meat is the fainting of livestock in the slaughterhouse to bleed, the skin and intestines are removed to remove the head, limbs, and tail. The lean meat is used for butchery and bones are used for bone.

Horse meat is being used as a food resource in various countries such as Europe and Japan, and in Donguibogam of Heo Jun, “The meat of white horse is cold in nature and tastes spicy and bitter. It nourishes tendons and bones, and strengthens the waist and spine. And makes the body healthy.” The carcass of horses is 61% of the living body, and the lean meat of the carcass excluding bone and fat is 43.3% of the living body, and is used for roasting, soup, and stew depending on the part.

Horse meat has more protein and less fat than beef and pork. Specifically, it contains 68.34-76.83% moisture, 19.80-22.31% protein, 0.12-6.63% intramuscular fat, and 0.98-4.03% minerals.

Among them, protein is a component of body tissues such as muscle and connective tissue, and is necessary for the composition of enzymes, hormones, and antibodies, and transport and storage of essential nutrients or active substances in the body, maintenance of body fluids, acid-base balance, energy, glucose , As an essential nutrient necessary for the synthesis of lipids, protein is classified into essential amino acids and non-essential amino acids, and the lean meat of horses contains both essential and non-essential amino acids, increasing its value as a high protein health food.

Horse meat contains higher levels of essential amino acids lysine, leucine, and arginine than beef, pork, and chicken. Lysine helps growth and development, muscle formation, improves osteoporosis, and enhances immunity, while leucine prevents muscle weakness and reduces camellia sclerosis. It contributes to reducing, and arginine is effective in improving erectile dysfunction or sperm movement, preventing alliance sclerosis, recovering from fatigue, and beautifying the skin.

In addition, horse meat is rich in non-essential amino acids glutamine, aspartic acid, and alanine than beef, pork, and chicken. Glutamine contributes to preventing muscle loss, skeletal muscle formation, and promoting fat metabolism in the liver, and aspartic acid is used to recover from fatigue and liver. It helps in detoxification activities, and alanine is effective in recovering from fatigue and improving high intensity exercise capacity.

In addition, among the lean meats of horses, intramuscular fat is characterized by a higher content of unsaturated fatty acids than other meats, and is a health functional food due to its high content of inorganic nutrients such as phosphorus (P), iron (Fe), retinol, and niacin, and vitamins. It is very suitable as

In general, health functional foods containing proteins should be prepared by separating and purifying proteins from beans, oils, eggs, fish and shellfish, meat, nuts, grains, and edible insects, or by decomposing them with proteolytic enzymes or autolytic enzymes. However, until now, a method for manufacturing a protein product having a high amino acid content while removing off-flavor of horse meat has not been developed, and thus development is required.

Registered Patent No. 10-1850064 (registered on April 12, 2018)

In the present invention, it is intended to provide a method for extracting a protein hydrolyzate using horse meat having a low off-flavor and a high protein content, including a pretreatment step, a hydrolysis step, a post treatment step, and a separation step, and a protein hydrolyzate prepared accordingly.

An embodiment of the present invention for achieving the object as described above, the pre-treatment step of preparing raw meat by crushing horse meat and removing impurities; Hydrolysis step of adding water and enzyme to the raw meat and reacting at a temperature range of 40 to 75°C; A post-treatment step of heat-treating the hydrolyzate obtained through the hydrolysis step at a temperature range of 85 to 105°C; A separation step of extracting a liquid protein mixture by centrifuging the hydrolyzate obtained through the post-treatment step; Filtration to remove foreign substances from the protein mixture; A concentration step of concentrating the filtrate obtained through the filtration step; And a drying step of drying the concentrate obtained through the concentration step. It relates to a method of extracting a protein hydrolyzate using horse meat.

The pretreatment step may include a grinding step of grinding horse meat; And an immersion step of immersing the pulverized horse meat in water or an aqueous sodium solution for a predetermined time.

In the hydrolysis step, an enzyme activity aid may be further added.

The enzyme may include at least one or more of an endoprotease type enzyme and an exopeptidase type enzyme.

The separation step may include two centrifugation processes.

In the filtering step, the protein mixture may be filtered using an activated carbon filter.

The concentration step may be a step of heating in a vacuum state.

Another embodiment of the present invention relates to a protein hydrolyzate prepared by the above manufacturing method and a food product containing the protein hydrolyzate.

According to the method of extracting protein hydrolysates of the present invention, the red color and fishy odor inherent in horse meat, and the bitter taste generated during protein production are reduced to increase consumer preference, and include low-molecular-weight amino acids. There is this.

In addition, the protein hydrolyzate according to the present invention has effects such as strengthening muscles and increasing bone density, and is suitable for middle-aged men and women, the elderly, athletes, youth, etc. who need smooth metabolism.

1 is a flow chart of a method for producing a protein hydrolyzate according to an embodiment of the present invention.
2 is a photograph of raw meats of Example 1 and Comparative Example 1 taken in Experimental Example 1.
3 is a result of measuring the molecular weight distribution of a protein hydrolyzate according to an embodiment of the present invention.
4 is a photograph of a protein singer according to an embodiment of the present invention.

It should be noted that terms or words used in the claims should not be interpreted as being limited to a conventional or dictionary meaning, but should be interpreted as meanings and concepts consistent with the technical idea of the present invention.

Throughout this specification, when a certain part "includes" a certain component, it means that other components may be further included rather than excluding other components unless otherwise stated.

Throughout this specification, "%" used to indicate the concentration of a specific substance is (weight/weight)% for solid/solid, (weight/volume)% for solid/liquid, and liquid unless otherwise stated. /Liquid means (volume/volume)%.

Hereinafter, an embodiment of the present invention will be described. However, the scope of the present invention is not limited to the following preferred embodiments, and those skilled in the art can implement various modified forms of the contents described in the present specification within the scope of the present invention.

The present invention relates to a method for producing a protein hydrolyzate having a high content of amino acids having a low molecular weight by processing horse meat, and to a protein hydrolyzate prepared accordingly. Horse meat used in the present invention includes lean meat of horses, and may include slaughter by-products or meat by-products.

1 is a flow chart of a method for producing a protein hydrolyzate according to an embodiment of the present invention.

First, a method for producing a protein hydrolyzate according to an embodiment of the present invention includes a pretreatment step of preparing raw meat by grinding horse meat and removing impurities; Hydrolysis step of adding water and enzyme to the raw meat and reacting at a temperature range of 40 to 75°C; A post-treatment step of heat-treating the hydrolyzate obtained through the hydrolysis step at a temperature range of 85 to 105°C; A separation step of extracting a liquid protein mixture by centrifuging the hydrolyzate obtained through the post-treatment step; Filtration to remove foreign substances from the protein mixture; A concentration step of concentrating the filtrate obtained through the filtration step; And a drying step of drying the concentrate obtained through the concentration step.

The pre-treatment step is a step of pre-treating the horse meat to remove foreign substances and off-flavor of horse meat and facilitate hydrolysis, and a grinding step of pulverizing the horse meat; And an immersion step of immersing the pulverized horse meat in water or an aqueous sodium solution for a predetermined time.

The crushing step is a step performed to increase the processing efficiency in a subsequent processing step by increasing the surface area of the horse meat, and is a step of cutting the horse meat into a predetermined size and pulverizing it using a grinder.

The immersion step is performed to reduce off-flavor of pulverized horse meat, remove foreign substances, and brighten color, and is a step of immersing the horse meat in water or sodium aqueous solution as an immersion liquid for a predetermined time.

At this time, the immersion liquid is preferably used to the extent that the horse meat can be completely immersed, and when using a sodium aqueous solution as the immersion liquid, the sodium content is preferably 0.1 to 0.5 parts by weight based on 100 parts by weight of the horse meat, and the immersion time is 1 minute. To 1 hour.

In order to increase the efficiency of removing off-flavors and foreign substances in the immersion step, it is preferable that stirring is performed during immersion, the foreign substances that have risen to the top must be removed, and the immersion liquid is preferably replaced at least two times.

The raw meat obtained through the pretreatment step is decomposed to the amino acid level through the hydrolysis step.

The hydrolysis step is a step in which water and enzymes are added to the raw meat and reacted at a temperature of 40 to 75°C. At this time, water is added in an amount of 50 to 150 parts by weight based on 100 parts by weight of crushed horse meat, and an enzyme is added in an amount of 0.1 to 4.0 parts by weight.

When the content of the enzyme is less than 0.1 parts by weight, the hydrolysis efficiency is insignificant, and when it exceeds 4.0 parts by weight, the effect of improving the hydrolysis efficiency according to the amount of the added enzyme is insignificant. Do.

At this time, the enzyme may include at least one or more of an endoprotease type enzyme and an exopeptidase type enzyme. Preferably, both types of enzymes may be included, and endoprotease-type enzymes are enzymes that randomly degrade proteins and have high hydrolysis efficiency, but if only endoproteases are used, bitter taste due to protein degradation occurs. Since there is a problem, in order to prevent this, it is preferable to use an exopeptidase-type enzyme that breaks down into amino acid units at the end of the protein.

For example, alcalase may be used as an endoprotease-type enzyme, and flavozyme may be used as an exopeptidase-type enzyme, but are not limited thereto, and various types of endoprotease-type enzymes and Exopeptidase type enzymes can be used.

In this step, in order to increase the efficiency of enzyme hydrolysis, an enzyme activating aid may be further added to a mixture containing raw meat, water, and enzyme.

Enzyme activation aids are added to increase the pH of the mixture so that both endoprotease-type enzymes and exopeptidase-type enzymes are activated, and enzyme activation aids include potassium polyphosphate, sodium metaphosphate, sodium polyphosphate, polymix-CS and pyrolysis. At least any one or more phosphate salts of sodium phosphate may be used.

The enzyme activation aid may be included in an amount of 0.01 to 1.5 parts by weight based on 100 parts by weight of pulverized horse meat, and if it is contained in an amount less than 0.01 parts by weight, the effect of improving the pH is insignificant and the enzyme activity is not improved. Since there is no effect of improving the pH, there is a problem that the manufacturing cost is improved, and saltiness and off-flavor due to an increase in the content of the enzyme activating agent may be excessively increased, making it difficult to intake protein hydrolysates. It is desirable.

The hydrolysis step may be performed for 4 hours to 16 hours at a temperature range of 40 to 75°C, and it is preferable to stir as necessary to increase the hydrolysis efficiency. When the hydrolysis temperature and time are less than the above range, sufficient hydrolysis does not occur, resulting in poor protein extraction efficiency, and when the temperature and time exceed the above range, additional hydrolysis does not occur and thus only the working time is prolonged. It is preferable that the hydrolysis proceeds within the range of conditions.

The post-treatment step is a step of heating the hydrolyzate obtained through the hydrolysis step as described above to a high temperature to induce a Maillard reaction, wherein the heating temperature is 85 to 105°C, and the heating time is 5 minutes. To 30 minutes.

The Maillard reaction is a chemical reaction between amino acids and reducing sugars, and the post-treatment step is performed to enhance the flavor of the protein product through the Maillard reaction.

Next, the separation step is a step of extracting a liquid protein mixture by centrifuging the hydrolyzate obtained through the post-treatment step. This step can be carried out at 30 ~ 80 ℃.

As an example, this step may be a step of extracting only the liquid protein mixture by separating the hydrolyzate into a solid content, a liquid protein mixture, and a suspension through a three-phase centrifugation.

As another example, this step may be a step involving two centrifugation processes. Specifically, a first separation step of performing two-phase centrifugation; And a second separation step of performing three-phase centrifugation. Since the hydrolyzate contains a large amount of solid content, such as about 10-50%, when the two centrifugation processes are performed, a significant amount of solid content is removed in the first separation step, and the separation efficiency in the second separation step is remarkably You can get an improved effect.

Through the second separation step, solids, liquid protein mixtures and suspensions are separated from each other from the bottom of the centrifuge, myoglobin is removed together with the solids, and oil extracted from intramuscular fat is removed as a suspension to obtain a liquid protein mixture. I can.

Horse meat is rich in unsaturated fatty acids, which can cause unpleasant odors due to oxidation of unsaturated fatty acids in the process of transporting, storing, processing and distributing raw materials.The freezing point of unsaturated fatty acids is variously distributed from -49 to 0°C. Since it is difficult to completely remove the fat component through, it is preferable to remove the fat component through such a three-phase centrifugation.

When the separation step is performed through a two-stage centrifugation process, centrifugation may be performed at a speed of 1,800 to 3,500 rpm in the first separation step, and centrifugation at a speed of 4,000 to 24,000 rpm in the second separation step. Separation can be performed.

In centrifugation, the radius, rotational speed, and mass of a rotating body are key factors that accelerate sedimentation separation. Since the horseyuk hydrolyzate centrifuged through the first separation step is fine particles, if the diameter of the rotating body is 75mm to 300mm in the second separation step, if the rotation speed is less than 4,000 rpm, the solid content or liquid that could not be separated in the first separation step Since the three-phase separation of the protein mixture and the suspension is not completely performed, foreign matters in the liquid protein mixture increase and the amino acid content decreases. Therefore, it is desirable to maintain a high speed of 4,000 rpm or more.

Next, a filtration step is performed to remove foreign substances contained in the liquid protein mixture obtained through the separation step.

This step may be a step of filtering the protein mixture through a filter having pores of a predetermined size, and an activated carbon filter may be preferably used as a filter used at this time. When the activated carbon filter is used, off-flavor of the protein mixture is additionally removed, and the color of the protein mixture can be kept constant by removing the exotic color.

Next, the filtrate obtained through the filtration step is concentrated through a concentration step.

This step is a step for concentrating the filtrate to facilitate subsequent drying, and may be a step of heating the filtrate to 38 to 65° C. in a vacuum state. It is preferable to concentrate so that the solid content is 40 to 90% by weight through this step.

Next, a drying step of drying the concentrate obtained through the concentration step is performed, and the drying step may be performed using a spray drying or freeze drying method.

The protein hydrolyzate obtained through such a step has an excellent small intestine absorption rate because the ratio of low molecular weight proteins of 900 Da or less is 99% or more, and the ratio of 500 Da or less is 85% or more. In addition, since various foreign substances and off-flavor removal steps are performed several times, and flavor is improved through the Maillar reaction step, consumer preference can be improved.

Another embodiment of the present invention relates to a protein hydrolyzate prepared according to the above manufacturing method, and such a protein hydrolyzate may be used as a food, or as a food additive to be used as a tablet, powder, granule, capsule, or liquid. It can be used in beverages, bars, powders, confectionery, and baking in the form of gels, jelly, etc., and can also be used as animal feed.

Hereinafter, specific actions and effects of the present invention will be described through an embodiment of the present invention. However, this is presented as a preferred example of the present invention, and the scope of the present invention is not limited according to the embodiment.

[ Manufacturing example ]

First, horse meat was prepared, cut into 5-10 cm size, and then crushed. Next, 100 g of pulverized horse meat was immersed in water for 3 minutes and stirred, and then the floating matter and blood that floated on the water were removed. Thereafter, water was added to adjust the total weight of the pulverized horse meat and water to 200 g, and 0.6 g of alkaline and 1.2 g of flavozyme were added, followed by heating at 50° C. for 10 hours under the condition of pH 5.7 and atmospheric pressure to proceed with hydrolysis. Then, under the same pressure, the temperature was raised to 95° C. and heated for 15 minutes to carry out a Maillard reaction.

Thereafter, the hydrolyzate was centrifuged at 45° C. at 3,000 rpm for 4 minutes to remove solids, and only the supernatant was obtained, and the supernatant was centrifuged at 5,250 rpm for 5 minutes to separate the remaining solids and the fat, which is a floating matter above the supernatant. The liquid component containing the protein hydrolyzate was collected.

Thereafter, the collected liquid component was filtered using an activated carbon filter, and the filtrate was heated to 50° C. in a vacuum, and concentrated to a solid content of 75% by weight. Thereafter, the protein hydrolyzate of Example 1 in powder form was obtained through freeze drying.

[ Experimental example One]

A protein hydrolyzate was prepared using the same method as in Preparation Example, but the immersion step was omitted from the pretreatment step to obtain the protein hydrolyzate of Comparative Example 1. Example 1 and Comparative Example 1 Each raw meat was photographed before the hydrolysis reaction during the manufacturing process and shown in FIG. 2, and the protein content of the obtained protein hydrolyzate was tested by Kjeldahl quantitative method according to the Health Functional Food Code.

As a result of the test, in the case of Comparative Example 1, 14.5 g per 100 g of horse meat, and 11.2 g per 100 g of horse meat in Example 1 were found to decrease the protein yield when the immersion step was performed.

However, when going through the immersion step, the color becomes bright and off-flavor due to impurities is significantly reduced.

[ Experimental example 2]

A protein hydrolyzate was prepared using the same method as in Preparation Example.

At this time, 0.1 g of alkaline, 0.1 g of flavozyme, and sodium pyrophosphate were added in an amount of 0.44 to 1.7 g in the hydrolysis step, and the pH was measured for 11 hours after adding these additives, and the results are shown in Table 1. . Thereafter, the remaining steps including the hydrolysis step were performed in the same manner as in Preparation Example. The pH measurement was performed in 1 hour increments using HANNA edge.

pH Sodium pyrophosphate content (g) - 0.44 0.87 1.30 1.70 Elapsed time 0h 5.64 7.21 7.43 7.82 7.84 1h 5.57 6.02 6.22 6.48 6.50 2h 5.56 5.86 6.01 6.26 6.28 3h 5.55 5.86 5.89 6.16 6.17 4h 5.53 5.84 5.89 6.13 6.14 5h 5.52 5.73 5.76 6.10 6.11 6h 5.51 5.66 5.75 6.07 6.08 7h 5.51 5.65 5.74 6.06 6.07 8h 5.50 5.64 5.73 6.04 6.05 9h 5.50 5.63 5.72 6.00 6.01 10h 5.50 5.62 5.71 5.99 6.00 11h 5.49 5.61 5.70 5.96 5.98

Looking at Table 1, when sodium pyrophosphate was not included, the pH was found to be as low as about 5.7 or less, and when sodium pyrophosphate was included as 0.44 to 1.30 g, the initial pH appeared as high as about 7.2 to 7.8 and time passed. It was confirmed that it decreases as it increases.

Alkalase, an enzyme used for hydrolysis, has an optimal pH of 7-8, in the case of flavozyme, it is 5.5-7.5, and the alkalinase is active at a relatively high pH, and flavozyme is weakly acidic to neutral. Shows activity in acidity.

In addition, since the alkaline agent reacts first during hydrolysis and the flavozyme reacts slowly, the pH must be kept high at the beginning of the hydrolysis reaction.

As shown in Table 1, when sodium pyrophosphate is included, it can be seen that the pH of the reaction product at the initial stage of the reaction is shown to be as high as about 6 or more, so that sodium pyrophosphate helps enzyme activity.

However, when the content of sodium pyrophosphate is 1.7 g, the range of pH change is insignificant compared to when the content of sodium pyrophosphate is 1.3 g. Since it may be difficult, the content of sodium pyrophosphate is preferably included in an amount of 1.5 parts by weight or less based on 100 parts by weight of crushed horse meat.

[ Experimental example 3]

A protein hydrolyzate was prepared using the same method as in Preparation Example, but 0.44 g of sodium pyrophosphate was used together during the hydrolysis reaction step, and a Minolta chromameter (Model CR-400, Minolta Camera co. Osaka., Japan) was used to prepare a protein hydrolyzate. Lightness, which is the L* value of the horse meat crushed during the manufacturing process, the hydrolyzate that has gone through the post-treatment step, and the filtrate that has gone through the filtration step, the redness representing the a* value, and the yellowness representing the b* value Was measured, and the results are shown in Table 2.

Crushed horse meat Post-processing steps
Coarse hydrolyzate Filtration step
Coarse filtrate L*(lightness) 38.60±0.90 52.54±0.80 59.33±0.44 a*(redness) 22.86±0.31 10.37±0.42 6.68±0.13 b*(yellowness) 10.88±0.39 22.84±0.84 17.05±0.30

It was confirmed that the pulverized horse meat increased in lightness, redness decreased, and had a light off-white color as each step went through. In addition, it was found that yellowness increased when the Maillard reaction was carried out through the post-treatment step.

[ Experimental example 4]

The molecular weight distribution of the protein hydrolyzate prepared in Experimental Example 3 was measured using a Q-TOF (Quadrupole time of flight) mass spectrometer, and the result is shown in FIG. 3, and the prepared protein hydrolyzate was photographed and shown in FIG. Shown.

Referring to Figure 3, the protein hydrolyzate according to the embodiment of the present invention showed a molecular weight distribution of less than 1 kDa 99% or more, and a molecular weight of 500 Da or less showed 87.61%. From these results, it can be predicted that the protein hydrolyzate according to the embodiment of the present invention mainly contains amino acids having a low molecular weight, and thus has a high human absorption rate.

The present invention is not limited to the specific embodiments and description described above, and any person with ordinary knowledge in the technical field to which the present invention pertains without departing from the gist of the present invention claimed in the claims can implement various modifications And, such modifications are within the scope of the present invention.

Claims (9)

A pretreatment step of preparing raw meat by crushing horse meat and removing impurities;
Hydrolysis step of adding water and enzyme to the raw meat and reacting at a temperature range of 40 to 75°C;
A post-treatment step of heat-treating the hydrolyzate obtained through the hydrolysis step at a temperature range of 85 to 105°C;
A separation step of extracting a liquid protein mixture by centrifuging the hydrolyzate obtained through the post-treatment step;
Filtration to remove foreign substances from the protein mixture;
A concentration step of concentrating the filtrate obtained through the filtration step; And
Including; a drying step of drying the concentrate obtained through the concentration step,
The enzyme includes an endoprotease-type enzyme, Alcalase, and an exopeptidase-type enzyme, Flavozyme,
In the hydrolysis step, at least one or more enzyme activating aids selected from potassium polyphosphate, sodium metaphosphate, sodium polyphosphate, polymix-CS, and sodium pyrophosphate are further added by 0.01 to 1.5 parts by weight based on 100 parts by weight of horse meat. Characterized in that, the extraction method of protein hydrolyzate using horse meat. The method of claim 1, wherein the pretreatment step,
A crushing step of crushing horse meat; And
An immersion step of immersing the pulverized horse meat in water or an aqueous sodium solution for a predetermined period of time for a predetermined period of time. delete delete The method of claim 1,
The separation step, characterized in that it comprises two centrifugation processes, the method of extracting a protein hydrolyzate. The method of claim 1,
In the filtration step, the protein mixture is filtered using an activated carbon filter. The method of claim 1,
The concentration step, characterized in that the step of heating in a vacuum state, the protein hydrolyzate extraction method. A protein hydrolyzate prepared by the method of any one of claims 1, 2, and 5 to 7. Food containing the protein hydrolyzate of claim 8.

Images