KR102586229B1 - Method for preparing hydrolyzate of Tenebrio molitor protein using NURUK and protease, and hydrolyzate of Tenebrio molitor protein with enhanced antioxidant content - Google Patents

Abstract

The present invention relates to a method for producing a hydrolyzate of brown mealworm (mealworm) protein using yeast and protein hydrolytic enzymes, and to a hydrolyzate of brown mealworm protein with increased antioxidant content prepared therefrom, and to the yeast and protein singer of the present invention. The method for producing hydrolyzate of brown mealworm protein using a decomposition enzyme can recover brown mealworm protein hydrolyzate in high yield compared to treatment with protease alone, and also contains antioxidant functional components such as water-soluble peptides and amino acids from the production method of the present invention. Since brown mealworm protein hydrolyzate with increased content is provided, there is a useful effect of providing a health functional food composition with improved antioxidant properties.

Description

Method for preparing hydrolyzate of brown mealworm protein using yeast and protease, and hydrolyzate of brown mealworm protein with increased antioxidant content {Method for preparing hydrolyzate of Tenebrio molitor protein using NURUK and protease, and hydrolyzate of Tenebrio molitor protein with enhanced antioxidant content}

The present invention relates to a method for producing a hydrolyzate of brown mealworm ( Tenebrio molitor ) protein using yeast and proteolytic enzymes, and to a hydrolyzate of brown mealworm protein with increased antioxidant content prepared therefrom.

The edible insect market size is expected to increase from 11 trillion won in 2007 to 38 trillion won in 2020. Korea is also starting to pay attention to the insect industry, and the government is fostering the insect industry through easing policies and regulations after announcing the 5-year comprehensive plan to foster the insect industry in 2011. The Ministry of Agriculture, Food and Rural Affairs is reforming 32 key regulations on agricultural and food products for edible insects in 2014. It was selected as one of the tasks. The domestic market size grew from 157 billion won in 2009 to 200 billion won in 2014, and is expected to grow to 700 billion won in 2020.

The brown mealworm ( Tenebrio molitor ) is an insect of the family Coleoptera and is also called “brown rice mealworm.” It is currently distributed throughout the world, including Korea. It has strong adaptability, has a short metamorphosis period, and is easy to breed, making it easy for industrialization. , It was registered as a high-protein edible insect raw material in the Korea Food Industry and Energy Agency in 2016.

Brown mealworm is a highly utilized raw material with high protein content, and brown mealworm protein hydrolyzate is known to increase the content of antioxidant functional ingredients such as water-soluble peptides and amino acids. In one study, an attempt was made to develop a food material by treating brown mealworms with a commercial protein hydrolyzate to improve the antioxidant content of brown mealworm protein hydrolyzate (Public Patent No. 10-2018-0010067). However, single treatment with a commercial protease hydrolyzes proteins with specificity due to substrate specificity, so the hydrolysis effect is sufficient for complex protein foods containing multiple proteins, such as brown mealworm protein. Even if the protease treatment amount is increased, the yield of the hydrolyzate cannot be increased beyond a certain level, and it causes problems such as off-flavor. Accordingly, treatment alone with a commercial proteolytic enzyme does not sufficiently extract the antioxidant functionality of brown mealworm protein.

Against this background, the present inventors confirmed that by combined treatment with nuruk, a traditional Korean alcohol fermentation agent, and a commercial proteolytic enzyme, hydrolyzate can be recovered at a higher yield from brown mealworm protein raw material compared to treatment with a commercial proteolytic enzyme alone. In addition, it was confirmed that brown mealworm protein hydrolyzate with increased antioxidant content could be provided, and the present invention was completed.

The purpose of the present invention is to provide a brown mealworm protein hydrolyzate that can solve the problem of low yield of protein hydrolyzate, which cannot be increased beyond a certain level by treatment with commercial protease alone, and the problem of insufficient extraction of antioxidant functional components of brown mealworm protein. A method for producing decomposed products is provided.

Another object of the present invention is to provide a brown mealworm protein hydrolyzate with increased antioxidant content.

In order to achieve the above purpose,

The present invention includes the steps of treating brown mealworm ( Tenebrio molitor ) with yeast and proteolytic enzyme to hydrolyze brown mealworm protein; and

It provides a method for producing brown mealworm protein hydrolyzate, including the step of recovering protein hydrolyzate from the brown mealworm treated with the yeast and proteolytic enzyme.

Additionally, the present invention provides a hydrolyzate of brown mealworm protein prepared from the above production method.

Furthermore, the present invention provides an antioxidant health functional food composition containing a hydrolyzate of the brown mealworm protein.

The method for producing brown mealworm protein hydrolyzate of the present invention can recover brown mealworm protein hydrolyzate in high yield by combined treatment with yeast and proteolytic enzyme, and can also increase the content of antioxidant functional ingredients, the present invention The brown mealworm protein hydrolyzate prepared from the manufacturing method has the useful effect of being provided as a health functional food composition with improved antioxidant functionality.

Figure 1 shows the hydrolysis process for brown mealworm raw materials.
Figure 2 shows the measurements of the free amino acid content of brown mealworm protein hydrolyzate according to the conditions of treatment with protease alone, treatment with yeast alone, or treatment with combination of yeast and protease.
Figure 3 confirms the molecular weight composition range of brown mealworm protein hydrolyzate according to the conditions of brown mealworm protein raw material and protease treatment alone, yeast only treatment, or yeast and protease combined treatment conditions.
Figure 4 shows the ABTS antioxidant activity test results of brown mealworm protein hydrolyzate according to treatment conditions with protease alone, yeast alone, or yeast and protease combined treatment conditions, showing peptides of 3 kDa or less or more than 3 kDa. It shows the antioxidant activity of each fraction.
Figure 5 shows the ABTS antioxidant activity test results of brown mealworm protein hydrolyzate according to the treatment conditions of protease alone, yeast alone, or yeast and protease combined treatment conditions, 10 kDa or less, 10-30 kDa or This shows the antioxidant activity of each peptide fraction exceeding 30 kDa.

Hereinafter, the present invention will be described in detail.

The present invention includes the steps of treating brown mealworms with yeast and proteolytic enzymes to hydrolyze brown mealworm proteins; and

It provides a method for producing brown mealworm protein hydrolyzate, including the step of recovering protein hydrolyzate from the brown mealworm treated with the yeast and proteolytic enzyme.

The conventional method of treating a commercial protease alone mainly hydrolyzes proteins with specificity due to the substrate specificity of the protease, and the hydrolysis effect is not sufficient for complex proteins such as brown mealworm protein. There is a problem, and even if the treatment amount is increased, the time of the hydrolysis process is shortened, the yield cannot be increased beyond a certain level, and if it is used beyond the treatment amount recommended by the manufacturer, it causes problems such as off-flavor. In addition, treatment with commercial protease alone has the problem of not being able to sufficiently extract the various water-soluble peptides and amino acid components of brown mealworm protein due to substrate specificity.

The method for producing brown mealworm protein hydrolyzate of the present invention is a method of producing brown mealworm protein hydrolyzate by complex processing nuruk, a traditional Korean alcohol fermentation agent, and a commercial protein hydrolytic enzyme, and the bacteria present in nuruk and the bacteria contained therein. Through the action of various proteolytic enzymes, it is possible to hydrolyze proteins for which commercial proteolytic enzymes do not have specificity, thereby improving the yield of hydrolyzate, which could not be increased beyond a certain level with conventional proteolytic enzyme treatment alone. Yield can be increased, and the type and content of water-soluble peptides and amino acids extracted from brown mealworm protein raw materials can be increased.

In one aspect of the present invention, NURUK is a traditional Korean liquor fermentation agent, made by propagating Aspergillus mold on grains. The yeast is, for example, Aspergillus oryzae and Aspergillus niger. , it may be obtained using any one or more of Monascus anka, Monascus purpureus, Monascus barkeri, and Aspergillus luchuensis. Meanwhile, the yeast includes commercially available yeast in addition to yeast produced by traditional methods, and improved yeast can also be used.

In another aspect of the present invention, the proteolytic enzyme is an enzyme that has hydrolytic activity on brown mealworm protein, and the proteolytic enzyme is, for example, Flavorzyme, Protamax, It is one or more types selected from the group consisting of Alcalase and Neutrase. In addition, the hydrolytic enzyme treatment can be performed by combining Flavorzyme, Protamax, Alcalase, and Neutrase, for example, Neutrase, Al It can be treated with Kalase, Protamax, and Flavorzyme, and it can be treated with Alkalase and Flavorzyme.

The yeast and proteolytic enzyme may be added in an amount of 0.1 to 20% by weight, for example, 0.2 to 18% by weight, 0.3 to 15% by weight, or 0.5 to 10% by weight, based on the weight of the brown meal raw material. It can be.

In the case of the yeast, it can be added at 1 to 20% by weight, for example, at 2 to 18% by weight, 3 to 15% by weight, or 5 to 15% by weight, based on the weight of the brown meal raw material. , for example, 5 to 15% by weight or 10% by weight of commercial yeast may be used, or 5 to 15% by weight or 10% by weight of improved yeast may be used.

In one embodiment of the present invention, the proteolytic enzyme may be added in an amount of 0.1 to 10% by weight, for example, 0.1 to 8% by weight, 0.1 to 5% by weight, based on the weight of the brown mealworm raw material. %, 0.1 to 3% by weight, 0.2 to 2% by weight, or 0.5 to 1.5% by weight, for example, 0.5 to 1.5% by weight of alkalase or 0.5 to 1.5% by weight of fleverzyme may be used, Alternatively, a mixed enzyme of Alkalase and Flavorzyme can be used in an amount of 0.5 to 1.5% by weight.

On the other hand, the treatment time of the yeast and protease is 30 minutes to 30 hours, 40 minutes to 25 hours, 60 minutes to 20 hours, 2 to 20 hours, 5 to 20 hours, 10 to 20 hours, or 15 to 20 hours. hours, 17 to 19 hours, or about 18 hours.

The treatment time for the yeast may be 5 to 20 hours, 10 to 20 hours, or 15 to 20 hours, 17 to 19 hours, or about 18 hours.

The protease treatment time may be 30 minutes to 5 hours, 40 minutes to 3 hours, 60 minutes to 2 hours, 70 minutes to 110 minutes, 80 minutes to 100 minutes, or about 90 minutes.

On the other hand, when treated with the yeast and proteolytic enzyme, the pH is 4 to 8 or 4.5 to 7.

When processing the yeast, the pH is 4 to 6, 4.2 to 5, or about 4.5.

When treated with the proteolytic enzyme, the pH is 6 to 8, 6.5 to 7.5, or about 7.

On the other hand, the temperature during treatment of the yeast and protease is 20 to 80°C, 30 to 70°C, 40 to 60°C, 45 to 55°C, or about 50°C.

The temperature during the yeast treatment is 20 to 50°C, 30 to 40°C, 35 to 40°C, or about 36°C.

The temperature during treatment with the proteolytic enzyme is 30 to 80°C, 40 to 70°C, 40 to 60°C, or about 50°C.

In the step of recovering the brown mealworm protein hydrolyzate, known methods can be used without limitation. For example, the brown mealworm protein hydrolyzate can be recovered by centrifuging the hydrolysis reaction to separate the layers and taking the supernatant. there is.

In the brown mealworm protein hydrolysis step, the brown mealworm may further include the steps of drying, powdering, and sterilizing the brown mealworm, and the powdered brown mealworm may be used as a raw material and hydrolyzed. The drying, powdering, and sterilization steps may be performed using known methods without limitation.

After the protein hydrolysis step, an enzyme deactivation step may be further included to terminate the protein hydrolysis reaction. The enzyme deactivation step can be performed using any known method without limitation. For example, the activity of the proteolytic enzyme can be deactivated by adjusting pH or temperature. In one embodiment of the present invention, the reaction system temperature is set to 70 to 90 C. ℃ or about 80℃, and the enzyme reaction is deactivated by treatment for 5 to 30 minutes, 10 to 20 minutes, or about 15 minutes.

In another aspect of the present invention, the method for producing the brown mealworm protein hydrolyzate may further include the step of obtaining a fraction by fractionating the recovered brown mealworm protein hydrolyzate. The fraction may be used without limitation as long as it is a known protein fractionation method. For example, the recovered brown mealworm protein hydrolyzate may be classified and fractionated according to molecular weight size. Additionally, the fraction obtained from the above fractionation step may be a fraction of 30 kDa or less, 20 kDa or less, or 10 kDa or less, or may be a 3 to 30 kDa, 3 to 20 kDa, or 3 to 10 kDa fraction.

In one embodiment of the present invention, the present inventors evaluated the quality characteristics of each brown mealworm protein hydrolyzate obtained by treating brown mealworm protein raw material with proteolysis alone, yeast alone, or a combination of yeast and protease. As a result of the comparison, superior quality characteristics were confirmed in the yeast and proteolytic enzyme combination treatment group compared to the proteolysis treatment alone. Specifically, as a result of checking the yield of brown mealworm protein hydrolyzate obtained for each treatment condition, it was confirmed that the yield was improved to 47% in the yeast and protease complex treatment group compared to the protease alone treatment group (see Table 2). ). In addition, as a result of analyzing the free amino acid content contained in the brown mealworm protein hydrolyzate obtained under each treatment condition, the amino acid content of the brown mealworm protein hydrolyzate in the yeast and protein hydrolyzase combination treatment group was higher than that in the proteinase-only treatment group. It was confirmed that there was an increase (see Figure 2). In addition, as a result of analyzing the organic matter content of the brown mealworm protein hydrolyzate obtained under each treatment condition, it was confirmed that more than 77% of the protein components contained in the brown mealworm raw material were extracted through the combined treatment of yeast and protein hydrolyzate (Table 3).

In addition, as a result of analyzing the antioxidant activity of the brown mealworm protein hydrolyzate obtained under each treatment condition, superior antioxidant activity was confirmed in the yeast and protease complex treatment group compared to the protease-only treatment group. In addition, as a result of analyzing the antioxidant activity of the obtained protein hydrolyzate by size, it was confirmed that the antioxidant activity of low-molecular-weight peptides of 3 kDa or less was superior to that of high-molecular-weight proteins of more than 3 kDa (see Figure 4), and in the peptide fraction of less than 10 kDa, the protein While the antioxidative activity of the group treated with hydrolytic enzyme alone was 65%, the antioxidative activity of peptides less than 10 kDa was more than 70% in the group treated with the combination of yeast and proteolytic enzyme, confirming that the content of antioxidant functional ingredients increased ( see Figure 5).

Additionally, the present invention provides a hydrolyzate of brown mealworm protein prepared from the above production method.

The hydrolyzate of brown mealworm protein obtained through the treatment of yeast and protein hydrolytic enzyme of the present invention is a hydrolyzate obtained by extracting about 77% or more of the protein component contained in the brown mealworm raw material, and is obtained through treatment with a conventional protein hydrolytic enzyme alone. Since the obtained brown mealworm protein has a higher free amino acid content compared to the hydrolyzate, it can be used in food compositions and feed compositions rich in amino acid content.

Furthermore, the present invention provides an antioxidant health functional food composition containing a hydrolyzate of the brown mealworm protein.

The hydrolyzate of brown mealworm protein obtained through treatment with yeast and protease of the present invention has a higher content of peptide fractions with antioxidant activity compared to the hydrolyzate of brown mealworm protein obtained through treatment with conventional protease alone. As it exhibits antioxidant activity, a health functional food composition with improved antioxidant functionality is provided.

Hereinafter, examples and experimental examples of the present invention will be described in detail below. However, the examples and experimental examples described below only illustrate a part of the present invention and are not limited thereto.

<Example 1> Preparation of brown mealworm protein hydrolyzate treated with yeast and protease

<1-1> Preparation of brown mealworm raw materials

The brown mealworm (Tenebrio molitor L.) larvae used in the present invention, produced from an insect farming farm located in Damyang, Jeollanam-do, were fasted for 72 hours, washed, sterilized, dried with hot air, and pulverized using a household grinder. Brown mealworm raw material powder was prepared.

<1-2> Preparation of brown mealworm protein hydrolyzate

After adding an appropriate amount of water to the prepared brown mealworm raw material powder, 0.5% by weight of Alcalase and 1% by weight of Flavourzyme were added based on the weight of the raw material powder and treated for 90 minutes at pH 7 and 50°C, followed by commercial use based on the weight of the raw material powder. 10% by weight of yeast was added and treated for 18 hours at pH 4.5 and 36°C to perform a hydrolysis reaction. After hydrolysis, the layers were separated through centrifugation, and the supernatant was recovered as brown mealworm protein hydrolyzate. The sediment was classified as an insoluble residue derived from the brown mealworm outer shell (chitin), and the supernatant and sediment were freeze-dried and sampled. It was used as.

<Comparative Example 1> Preparation of brown mealworm protein hydrolyzate treated with yeast alone

After adding an appropriate amount of water to the brown mealworm raw material powder prepared in <1-1> above, 10% by weight of commercial yeast based on the weight of the raw material powder was added and treated for 18 hours at pH 4.5 and 36°C to perform a hydrolysis reaction. It was carried out. After hydrolysis, the layers were separated through centrifugation, and the supernatant was recovered as brown mealworm protein hydrolyzate. The sediment was classified as an insoluble residue derived from the brown mealworm outer shell (chitin), and the supernatant and sediment were freeze-dried and sampled. It was used as.

<Comparative Example 2> Preparation of brown mealworm protein hydrolyzate treated with protease alone

After adding an appropriate amount of water to the brown mealworm raw material powder prepared in <1-1> above, 0.5% by weight of Alcalase and 1% by weight of Flavourzyme were added based on the weight of the raw material powder and treated for 90 minutes at pH 7 and 50°C. A hydrolysis reaction was performed. After hydrolysis, the layers were separated through centrifugation, and the supernatant was recovered as brown mealworm protein hydrolyzate. The sediment was classified as an insoluble residue derived from the brown mealworm outer shell (chitin), and the supernatant and sediment were freeze-dried and sampled. It was used as.

The hydrolysis reaction conditions performed in Example 1 and Comparative Examples 1 and 2 are shown in Table 1 below.

experimental group Applied enzyme and usage (% by weight) Reaction conditions (pH, temperature, time) Example 1 MW-CN-AF CN(10), A(0.5) F(1) After treatment at pH 7, 50℃, 90 minutes,
pH 4.5, 36℃, 18 hours treatment Comparative Example 1 MW-CN CN(10) pH 4.5, 36℃, 18 hours treatment Comparative Example 2 MW-AF A(0.5) F(1) pH 7, 50℃, 90 minutes treatment

(In the above table, MW: Mealworm A: Alcalase 2.4L, F: Flavourzyme 100L, C: celluclast, V: viscozyme, CN: commercial yeast)

<Experimental Example 1> Yield of brown mealworm protein hydrolyzate

The brown mealworm protein hydrolyzate supernatant and insoluble precipitate obtained after centrifugation in Example 1 and Comparative Examples 1 and 2 were separated from each other, the weight of the sample recovered after freeze-drying was precisely measured, and the hydrolysis yield was derived using the formula below. and the results are shown in Table 2 below.

experimental group Hydrolysis yield (%) Example 1 MW-CN-AF 47 Comparative Example 1 MW-CN 30.18 Comparative Example 2 MW-AF 41.08

(In the above table, MW: Mealworm A: Alcalase 2.4L, F: Flavourzyme 100L, C: celluclast, V: viscozyme, CN: commercial yeast)

Looking at Table 2, the hydrolyzate yield of the group treated with protease alone was found to be about 41%, and the hydrolysis yield of the group treated with the combination of yeast and protease was confirmed to have improved to 47%.

<Experimental Example 2> Free amino acid content of brown mealworm protein hydrolyzate

The AccQ-Tag method was used to measure the free amino acid content of the brown mealworm protein hydrolyzate obtained in Example 1 and Comparative Examples 1 and 2, and for amino acid derivatization, a Waters AccQ-Tag Ultra Derivatization kit (Waters, Milford, MA) was used. , USA) was used. For analysis, the Waters ACQUITY UPLC system (Waters, USA) and 2.1 Measurements were made using the gradient provided in the kit under conditions of 20°C, column 55°C, detector 260 nm, and flow rate of 0.7 mL/min. Amino acid standard H (Thermo Fisher Scientific Inc., Rockford, IL, USA) was used as the amino acid standard, and the results are shown in Figure 2.

Looking at Figure 2, it is confirmed that the free amino acid content of the yeast and protease complex treatment group of the present invention was improved compared to the free amino acid content of the protease-only treatment group.

<Experimental Example 3> Organic matter content of brown mealworm protein hydrolyzate

In order to analyze the change in the organic element (carbon, nitrogen) content of the brown mealworm protein raw material and the brown mealworm protein hydrolyzate obtained in Example 1 and Comparative Examples 1 and 2, the supernatant and insoluble residue after the hydrolysis reaction were fractionated and placed in a freeze dryer ( Using MCFD8518, Ilshin Lab Co. Lat, Seoul, Korea), samples freeze-dried for 120 hours under the conditions of -80°C condenser temperature and 5 mmTorr pressure were uniformly ground and used as analysis samples. For carbon and nitrogen analysis, N ₂ O was reduced to N ₂ and the volume of N ₂ gas was measured and quantified using the Dumas method. After weighing 0.2 g each, an elemental analyzer (Elementary, vario MAX cube., Germany) was used, and crude protein The content was expressed by multiplying the nitrogen concentration by the brown mealworm protein conversion coefficient of 4.76, and the results are shown in Table 3 below.

experimental group Nitrogen content (%) protein content supernatant insoluble residue supernatant Example 1 MW-CN-AF 7.1441 9.21 34.0 Comparative Example 1 MW-CN 5.2353 10.10 24.9 Comparative Example 2 MW-AF 6.25 10.839 29.7 Raw MW
(Brown mealworm protein raw material) 9.27 44.1

For brown mealworm protein, the protein component reduced to low molecular weight through hydrolysis under each treatment condition dissolves in the aqueous solution component. Therefore, to check the water-soluble protein content in the sample, the nitrogen content was checked and converted to protein content.

Looking at Table 3, it was confirmed that the protein content of 44.1% contained in the brown mealworm raw material was contained at 29.7% in the hydrolyzate of the protease-only treatment group, and about 67% was recovered, and the yeast and protease complex of the present invention It was confirmed that the hydrolyzate of the treated group contained 34.0% and that approximately 77% was recovered.

Meanwhile, there is a research report that the reason the nitrogen content in the insoluble residue is measured as high as 10% is because chitin, which makes up the outer wall of edible insects such as brown mealworms, is composed of unit substances containing non-protein nitrogen such as glucosamine, Accordingly, it was judged that the content was high.

<Experimental Example 4> Peptide molecular weight range of brown mealworm protein hydrolyzate

<4-1> Measurement of composition range of water-soluble peptide fraction using FPLC to confirm molecular weight of brown mealworm protein hydrolyzate

The experimental conditions for confirming the molecular weight composition range of the brown mealworm protein raw material and the brown mealworm protein hydrolyzate obtained in Example 1 and Comparative Examples 1 and 2 are as follows. A column dedicated to protein size exclusive chromatography (SEC, size exclusive chromatography; Superdex 30 increase 10/300 GL,) was mounted on an FPLC (AKTA explorer 100, GE healthcare, Sweden) analysis device, and the supernatant was analyzed after the hydrolysis reaction for each condition. Using the same amount of freeze-dried powder, a solution dissolved in the same unit volume was injected at 10 mg/mL, and eluent buffer (phosphate saline buffer) was passed through the column at a flow rate of 0.5 ml/ml. After hydrolysis reaction under each condition, the supernatant was obtained. The molecular weight of the water-soluble peptide contained in was analyzed, and the results are shown in Figure 3. At this time, in the case of the SEC method, since the polymer material passes through the column first, it can be assumed that the material eluted at 10 minutes is more polymeric than the material eluted at 60 minutes.

<4-2> Water-soluble peptide fraction molecular weight composition range results of brown mealworm protein hydrolyzate

Looking at Figure 3, the brown mealworm protein raw material was confirmed to be composed of water-soluble peptide substances that pass between 10-20 minutes and 55-60 minutes, and the substance identified in the brown mealworm protein raw material at 15 minutes was brown mealworm protease alone. It was confirmed that the hydrolyzate of the treated group changed into a substance that eluted within 30-40 minutes. On the other hand, in the hydrolyzate of the yeast-only treatment group, it was confirmed that there were few substances eluted at 10-20 minutes, and that characteristic substances were hydrolyzed at 38 minutes, 45 minutes, and 56 minutes. In addition, the chromatogram results of the hydrolyzate of the yeast and protease complex treatment group showed that the content of water-soluble peptides constituting the same amount of sample increased, as indicated by the peaks eluting at 40 minutes, 44 minutes, 58 minutes, and 60 minutes. It was possible to infer from the area, and it was confirmed that the effect of the combined treatment result was excellent.

<Experimental Example 5> Antioxidant activity of brown mealworm protein hydrolyzate

<5-1> Measurement of antioxidant activity in fractions below 3 kDa or above 3 kDa of brown mealworm protein hydrolyzate

To confirm the antioxidant activity of brown mealworm protein hydrolyzate, ABTS antioxidant activity was tested. At this time, in order to determine the size of the hydrolyzed peptide that is effective for antioxidant activity, a centricon-type membrane filter was used to divide the peptides into fractions less than 3 kDa or more than 3 kDa, and the antioxidant activity was quantified.

The total antioxidant activity of the brown mealworm digest was expressed by ABTS (2,2-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt) radical scavenging activity. After reacting ABTS with potassium persulfate to prepare radical cations, 1 mL of ABTS solution was reacted with the sample for 6 minutes, and the percent radical scavenging ability was calculated and shown in FIG. 3.

Looking at Figure 3, compared to the group treated with protease alone, the yeast and protease combined treatment group of the present invention was confirmed to have better antioxidant activity in both peptide fractions of 3 kDa or less or more than 3 kDa.

<5-2> Measurement of antioxidant activity in fractions below 10 kDa, 10-30 kDa, or above 30 kDa of brown mealworm protein hydrolyzate

In the case of the antioxidant activity of the peptide exceeding 3 kDa confirmed in <5-1> above, the value may be due to the effect of concentrating relatively large molecular weight substances during the molecular weight size fractionation process of ultrafiltration, so the brown mealworm protein hydrolyzate After filtering through a 30 kDa size filter, the fraction was again filtered through a 10 kDa size filter, divided into fractions below 10 kDa, 10-30 kDa, or above 30 kDa, and analyzed for antioxidant activity. The results are shown in Figure 4. It was.

Looking at Figure 4, in the case of the peptide fraction of less than 10 kDa, the group treated with protease alone had an antioxidant activity of about 65%, whereas the group treated with the yeast and protease combination of the present invention had an antioxidant activity of less than 10 kDa peptides. Activity is confirmed to be approximately 70% or more.

Claims (10)

In the method for producing brown mealworm protein hydrolyzate, comprising the step of recovering protein hydrolyzate from brown mealworm,
The step of hydrolyzing brown mealworm protein by treating yeast and proteolytic enzymes,
1) Treating brown mealworms with proteolytic enzyme at 0.1 to 5% by weight based on the weight of brown mealworm raw materials for 60 to 120 minutes at pH 6 to 8 and temperature 40 to 60°C; and
2) treating the proteolytic enzyme-treated brown mealworm with yeast at 1 to 20% by weight based on the weight of the brown mealworm raw material for 10 to 20 hours under conditions of pH 4.2 to 5 and temperature 30 to 40°C; A method for producing brown mealworm protein hydrolyzate, comprising:
delete According to paragraph 1,
The proteolytic enzyme is characterized in that it is one or more proteolytic enzymes selected from the group consisting of Flavourzyme, Protamax, Alcalase, and Neutrase. , Method for producing brown mealworm protein hydrolyzate.
delete delete delete delete Brown mealworm protein hydrolyzate prepared from the production method of claim 1.
The brown mealworm protein hydrolyzate according to claim 8, wherein the brown mealworm protein hydrolyzate is a peptide fraction having a molecular weight of 10 kDa or less.
An antioxidant health functional food composition containing the brown mealworm protein hydrolyzate of claim 8.