KR102325312B1 - Preparing method of food protein derived from edible insects and food derived from edible insects prepared thereby - Google Patents

Abstract

The present invention relates to a method for producing an insect-derived food protein using a grafting technique and to an insect-derived composition prepared accordingly, comprising the steps of: (A) pulverizing insects to degrease fat; (B) extracting salt-soluble protein from the defatted insect powder; and (C) forming a glycoprotein polymer by grafting technology by reacting the salt-soluble protein with the sugar and then spray-drying for the Maillard reaction. By including; It can be used for a variety of foods.

Description

A method for preparing an insect-derived food protein using grafting technology and an insect-derived composition prepared accordingly {Preparing method of food protein derived from edible insects and food derived from edible insects prepared thereby}

The present invention relates to a method for producing an insect-derived food protein using a grafting technique to increase the gel-forming and emulsifying power of insect protein, which is lower than that of edible meat protein, and an insect-derived composition prepared accordingly.

About 2,000 insect species are consumed in over 100 countries around the world, and in 2013, the International Food and Agriculture Organization (FAO) proposed insects as a high-protein future food resource to cope with climate change, livestock environmental problems, and food shortages.

In Korea, the nutritional value and composition of silkworms (Bombyx mori ) have been evaluated since the 1970s with respect to insects, and recent studies have been conducted to identify the quality characteristics of processed foods containing insects and the physiological effects of insects. However, due to the external characteristics of insects, there is a limitation in positioning it as a universal food. In particular, in Europe and North America, where the culture of eating insects is unfamiliar, there is a difference in perception of insect eating depending on the region, such as expressing insect consumption as a phobia.

In the food industry, insects are used in the form of adding simple dry matter or powder to existing processed foods, but the use of insects as a food material is very low because the addition amount of insects in processed foods is as low as 3 to 10%. In addition, varieties with established processing aptitude among insects are extremely limited, and according to previous research results, it is known that it is difficult to use them as raw materials for processed foods compared to existing food proteins (soy protein and meat protein).

Insects contain more fat and chitin than edible meat, and it has been reported that insect proteins have inferior functional properties such as gel formation and emulsification power compared to edible meat proteins. In particular, it is known that insects have limitations as processed food materials because when 20% or more is added to processed food, the viscosity and elasticity of the final product are lowered or oil separation phenomenon is significantly increased (Choi et al., 2017; Kim et al., 2015; Kim et al., 2016). Therefore, there is a need for research on improving the functional properties of insects for food materialization.

On the other hand, when protein and sugar are heated under alkaline conditions, a glycoprotein polymerization reaction occurs in which the amino group of the protein and the carbonyl group of the reducing sugar are combined. According to previous studies, glycoprotein polymers are formed by binding of protein and polysaccharide to proteins such as soy protein isolate, whey and gelatin (Etxabide et al., 2015; Guan et al., 2006; Sun et al., 2011), In particular, it is known that saccharides are bound to amino acids such as asparagine, serine and threonine among proteins to improve the functional properties of the protein.

It is known that various functional properties such as gel-forming ability, emulsifying power, thermal stability, antioxidant and antimicrobial effects of glycoprotein polymers are improved depending on the type and ratio of sugars bound to proteins and heating conditions, respectively. Insects have a higher protein content than existing commercially available protein sources, and although there are differences between varieties, the content of asparagine, serine and threonine, which are essential amino acids for the formation of glycoprotein polymers, is high or similar. Therefore, insects are useful protein materials for the formation of glycoprotein polymers, and in particular, glycoprotein polymers using insect proteins can be used in the manufacture of various processed foods as functional food materials with improved poor processing aptitude of existing insect proteins. However, studies on the formation of glycoprotein polymers using insect proteins so far are insufficient.

Republic of Korea Patent Publication No. 10-1493916 Republic of Korea Patent Publication No. 10-2015-0064392

It is an object of the present invention to provide a method for producing an insect-derived food protein using grafting technology to increase gel formation and emulsification power of insect protein, which is lower than that of meat protein.

In addition, another object of the present invention is to provide an insect-derived food composition prepared according to the above manufacturing method.

In addition, another object of the present invention is to provide a gelatinized product using the insect-derived food composition.

In addition, another object of the present invention is to provide a food composition using the gel.

In addition, another object of the present invention is to provide a feed composition comprising the gel.

In addition, another object of the present invention is to provide a cosmetic composition comprising the gel.

In order to achieve the above object, a method for producing an insect-derived food protein using the grafting technology of the present invention comprises the steps of (A) pulverizing an insect to degrease the fat; (B) extracting salt-soluble protein from the defatted insect powder; and (C) forming a glycoprotein polymer by grafting technology by reacting the salt-soluble protein with the sugar and then spray-drying for the Maillard reaction.

After the step (C), (D) gelling the spray-dried glycoprotein polymer; may further include.

The fat content of the insect powder defatted in step (A) may be 0.1 to 10 g/100 g.

The solvent used when degreasing the insect in step (A) may be hexane, distilled water, anhydrous methanol or anhydrous ethanol, preferably hexane.

In step (B), the salt-soluble protein is mixed with an insect and a solvent in a weight ratio of 1: 2-5, homogenized at 10000 to 15000 rpm for 10 to 50 minutes, and then left for 30 to 120 minutes, then at 10000 to 15000 rpm It may be obtained by homogenizing for 20 to 80 minutes.

The solvent may be at least one selected from the group consisting of saline solution, sodium hydroxide aqueous solution and sodium phosphate aqueous solution.

The extraction of step (B) may be performed under the conditions of pH 9-11.

In step (C), salt-soluble protein and sugar are mixed in a weight ratio of 1: 2-10, homogenized at 10000 to 15000 rpm for 30 to 120 minutes, and then bathed at 70 to 90 ° C. for 30 to 120 minutes, and then 1 to A glycoprotein polymer can be obtained by performing a Maillard reaction after recovering the supernatant by centrifugation at 2000 to 5000 xg at 5° C. for 5 to 50 minutes.

The saccharides may be monosaccharides, disaccharides, oligosaccharides or polysaccharides, preferably monosaccharides.

In the step (C), the salt-soluble protein reacted with the saccharide for the Maillard reaction during spray drying may be injected at a temperature of 140 to 170 °C.

The insect may be at least one selected from the group consisting of a grasshopper order, a coleopteran order, and a Lepidoptera order, and preferably a mealworm larva ( Tenebrio molitor , TM), a long beetle larva ( Allomyrina dichotoma , AD), Protaetia brevitarsis seulensis , PB), rice locust ( Oxya chinensis sinuosa Mistshenko ), twin cricket ( Gryllus bimaculatus ) and silkworm pupa ( Bombyx mori ) may be at least one selected from the group consisting of, more preferably, Tenebrio TM molitor larvae ( ) ), long- lived beetle larvae (Allomyrina dichotoma , AD), and larvae of white spot beetle (Protaetia brevitarsis seulensis , PB) may be at least one selected from the group consisting of.

In addition, the insect-derived food protein of the present invention for achieving the above other object may be prepared according to the above manufacturing method.

In addition, the gel product of the present invention for achieving the above another object may be prepared including the insect-derived food protein.

In addition, the food composition of the present invention for achieving the above another object may be prepared including the gel.

The food composition may be a fish cake composition, a sausage composition, a patty composition, a yanggaeng composition, or a konjac composition.

In addition, the feed composition of the present invention for achieving the above another object may be prepared including the gel.

In addition, the cosmetic composition of the present invention for achieving the above another object may be prepared including the gel.

The insect-derived food protein of the present invention can obtain high-quality insect food protein by preparing a glycoprotein polymer, which is difficult to prepare as an insect. In particular, the insect-derived food protein of the present invention has excellent gelation properties, so that it is possible to provide a gelled product and a food composition using the same.

In addition, it is possible to provide a feed composition including the gelled material, as well as improving the emulsification power by including the gelled material can be used as a cosmetic composition.

1 is a photograph of electrophoresis measurement of defatted insect powder prepared according to Preparation Examples 1 to 4;
Figure 2a is a graph measuring the foam capacity (foam capacity) for the salt-soluble protein powder prepared according to Preparation Examples 1 to 4, Figure 2b is the foam stability of the salt-soluble protein powder prepared according to Preparation Examples 1 to 4 (Foam stability) is a measured graph, Figure 2c is a graph measuring the emulsion capacity (Emulsion capacity) for the salt-soluble protein powder prepared according to Preparation Examples 1 to 4, Figure 2d is according to Preparation Examples 1 to 4 It is a graph measuring the emulsion stability of the prepared salt-soluble protein powder.
3 is a graph measuring protein solubility according to pH.

The present invention relates to a method for producing an insect-derived food protein using a grafting technique to increase the gel-forming and emulsifying power of insect protein, which is lower than that of edible meat protein, and an insect-derived composition prepared accordingly.

Hereinafter, the present invention will be described in detail.

The method for producing an insect-derived food protein of the present invention comprises the steps of (A) degreasing the fat by crushing the insect; (B) extracting salt-soluble protein from the defatted insect powder; and (C) forming a glycoprotein polymer by grafting technology by reacting the salt-soluble protein with the saccharide and then spray-drying for the Maillard reaction; It may further include; gelling the glycoprotein polymer.

First, in the step (A), the insects are crushed to degrease the fat.

Although the insect has a low fat content, in order to perform gelation, degreasing is performed so that the fat content is 0.1 to 10 g/100 g, preferably 1 to 6 g/100 g. The degreasing method may be performed in the same manner as the conventional degreasing method using distilled water, anhydrous methanol or anhydrous ethanol, but is preferably degreasing using hexane.

If the fat content of the insect after degreasing is less than the lower limit, degreasing must be performed for a long time, so the protein is denatured to lower the emulsification power and increase the hardness.

Insects used in the present invention may be at least one selected from the group consisting of locusts, coleoptera, and lepidoptera, preferably mealworm larvae ( Tenebrio molitor , TM), long- lived beetle larvae (Allomyrina dichotoma , AD), white spot It may be at least one selected from the group consisting of Protaetia brevitarsis seulensis (PB), rice locust (Oxya chinensis sinuosa Mistshenko ), Gryllus bimaculatus and silkworm pupa ( Bombyx mori), more preferably a mealworm larva ( Tenebrio molitor , TM), long- lived beetle larvae (Allomyrina dichotoma , AD), and white spot larvae ( Protaetia brevitarsis seulensis , PB) may be at least one selected from the group consisting of.

The mealworm larva is rich in minerals and dietary fiber, so it is a food suitable for diet, and by containing a large amount of unsaturated fatty acids, it is known that it is good for the prevention and treatment of heart disease. It is also known to be effective in the treatment of cough, sputum, and hematemesis, as well as in the treatment of paralysis and hemiplegia.

In addition, the long-lived beetle is an insect belonging to the order Coleoptera and the family Dynastidae, and as folk remedies that the long-lived beetle larvae longevity beetle is effective for liver disease are known, examples of using it as a health supplement are increasing. . In addition, it has been reported that the protein isolated from the long-lived beetle shows strong inhibition of activity against bacteria, and the lectin extracted from the long-lived beetle shows strong anticancer ability and can be expected to have an anticancer effect. Papers that confirmed the

In addition, according to 'Donguibogam', the larvae of the white-spotted flower radish larvae include diseases originating from the human liver, i.e., liver cancer, cirrhosis, hepatitis, and relieving accumulated fatigue, and loss of vision, cataracts, malignant boils, stomatitis, tetanus, and paralysis. It is said to be effective in treating adult diseases such as

Next, in step (B), a salt-soluble protein is extracted from the defatted insect powder to obtain an insect-derived protein.

Salt-soluble protein and water-soluble protein can be extracted from the defatted insect, respectively. Since the water-soluble protein has lower gelation properties (bubble power, foam stability, emulsification capacity, emulsification stability) compared to salt-soluble protein, salt-soluble protein It is preferable to extract

The solvent for extracting salt-soluble protein from the defatted insect is not particularly limited as long as salt-soluble protein can be extracted, but preferably from the group consisting of saline solution, sodium hydroxide aqueous solution and sodium phosphate aqueous solution. One or more selected types can be mentioned.

When extracting the salt-soluble protein, the extraction should be performed at a pH of 9-11, preferably, pH 9-10 using the solvent to improve protein solubility and subsequently form a glycoprotein polymer. For example, since the glycoprotein polymer is completed through the Maillard reaction under weak alkaline conditions, the pH is set to the above conditions when the salt-soluble protein is extracted.

Specifically, the salt-soluble protein is obtained by mixing the insect and the solvent in a weight ratio of 1: 2-5, preferably 1: 2-3 to satisfy the above pH, and then 10 to 50 at 10000 to 15000 rpm. Min, preferably 20 to 30 minutes, followed by standing for 30 to 120 minutes, preferably 50 to 60 minutes, followed by homogenization at 10000 to 15000 rpm for 20 to 80 minutes, preferably for 30 to 40 minutes is extracted through

When the salt-soluble protein is extracted, the yield of the glycoprotein polymer formed is low and the gelation characteristic may be lowered if the extraction is not performed in the order of primary homogeneity, stationary, and secondary homogenization, and homogenization is performed only once or the stationary process is not performed.

Next, in step (C), a glycoprotein polymer is formed by grafting technology by reacting the salt-soluble protein with the saccharide and then spray-drying for the Maillard reaction.

The formation of a glycoprotein polymer using the grafting technique is accomplished through the Maillard reaction under weak alkaline conditions after mixing the protein and sugar. A glycoprotein polymer is formed by grafting technology by carrying out the process of making and 2. spray-drying at a high temperature.

Specifically, the glycoprotein polymer comprises the steps of (a) mixing salt-soluble protein and sugar in a weight ratio of 1: 2-10, preferably 1: 5-8; (b) homogenizing the mixture in which the salt-soluble protein and sugar are mixed at 10000 to 15000 rpm for 30 to 120 minutes, preferably 40 to 60 minutes; (c) bathing the homogenized mixture at 70 to 90° C. for 30 to 120 minutes, preferably 50 to 60 minutes; (d) recovering the supernatant by allowing the mixture to cool and then centrifuging at 1 to 5 °C at 2000 to 5000 xg for 5 to 50 minutes, preferably 10 to 20 minutes; (e) forming a glycoprotein polymer by injecting the supernatant at a temperature of 140 to 170° C. for the Maillard reaction.

In steps (a) and (b), the salt-soluble protein and sugar are mixed and homogenized.

If the content of sugars based on the salt-soluble protein is less than the lower limit, the glycoprotein polymer may not be formed, and if it exceeds the upper limit, the gelation property may be deteriorated.

Examples of the saccharide include monosaccharides, disaccharides, oligosaccharides, and polysaccharides, but monosaccharides are preferably used in view of high yield and excellent gelation properties of the formed glycoprotein polymer.

In addition, when the following process is performed by simply mixing the salt-soluble protein and sugar without homogenizing them, the gelation property may be deteriorated.

Next, after adding a solvent of pH 7-8 to the homogenized mixture in step (c), a water bath may be performed to form a glycoprotein polymer. The solvent of pH 7-8 does not need to be added, but adding the solvent of pH 7-8 may help to form a glycoprotein polymer in a high yield and have excellent gelation properties.

If the homogenized mixture is processed at a high temperature of 100° C. or higher without a bath, the glycoprotein polymer may not be formed, so it is preferable to use a bath.

Next, in steps (d) and (e), the supernatant is recovered by cooling the hot water mixture and then centrifuging at 1 to 5 ° C. at 2000 to 5000 xg for 5 to 50 minutes, preferably for 10 to 20 minutes. After that, the supernatant is injected at a temperature of 140 to 170 ° C. using a spray dryer for the Maillard reaction to form a glycoprotein polymer.

Since the final glycoprotein polymer is completed through the Maillard reaction, the supernatant of the boiling water mixture is sprayed at a temperature of 140 to 170° C. to form a glycoprotein polymer. When spraying using a spray dryer, if the temperature is less than the lower limit, the Maillard reaction is not performed and the glycoprotein polymer cannot be formed.

The glycoprotein polymer thus obtained had a molecular weight of 120 to 250 kDa, preferably 140 to 250 kDa by SDS-PAGE (sodium-dodecyl sulfate-polyacrylamide gel-electrophoresis), and a high molecular weight glycoprotein polymer was obtained. .

Next, in step (D), the spray-dried glycoprotein polymer is gelled.

After diluting the spray-dried glycoprotein polymer to a concentration of 70 to 80% by weight, water bath at 50 to 70° C. to obtain a gelled product.

In addition, a food composition of a fish cake composition, a sausage composition, a patty composition, a yokan composition or a konjac composition can be provided by using the gelled product.

In addition, the present invention may provide a feed composition including the gelled product.

The 'feed composition' as an active ingredient, in addition to the gelled product, food ingredients that can be used as foods described in the standards and specifications of food ('Food Code'), food additives described in the Food Additives Code, and food that can be used as food Even if it is not a raw material or food additive, raw materials that fall within the range of single feed in Attached Table 1 of 'Standards and Specifications for Feed, etc.' and those that fall within the range of Supplementary Feed in Attached Table 2 may be used.

The 'feed composition' may be an extractant among supplementary feeds according to the "standards and specifications of feed, etc.", and may be a compound feed including the supplementary feed.

When preparing a feed composition including the gelled material, the content of the gelled material does not need to be particularly limited, but for example, 0.1 to 99% by weight, 0.5 to 95% by weight, 1 to 90% by weight, 2 to 80% by weight , 3 to 70% by weight, 4 to 60% by weight, may be included in 5 to 50% by weight.

The gelled material as an active ingredient in the feed composition varies depending on the condition, weight, presence, severity and duration of the ingested animal, but may be appropriately selected by those skilled in the art. For example, it may be 1 to 5,000 mg, preferably 5 to 2,000 mg, more preferably 10 to 1,000 mg, still more preferably 20 to 800 mg, and most preferably 50 to 500 mg based on the daily dose. And, the number of administration does not need to be particularly limited, but can be adjusted by a person skilled in the art within the range of 3 times a day to once a week. In the case of long-term intake for the purpose of health and hygiene or health control, it may be less than the above range.

In addition, the present invention may provide a cosmetic composition including the gelled product.

In the cosmetic composition of the present invention, if necessary, other ingredients commonly blended in cosmetics may be blended. Examples of these blending ingredients include oils and fats, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, UV absorbers, preservatives, disinfectants, antioxidants, pH adjusters, alcohols, pigments, fragrances, blood circulation promoters, cooling agents, limiting agents, purified water, water-soluble vitamins, fat-soluble vitamins, polymer peptides, polymer polysaccharides, sphingolipids and seaweed extracts, and the like. .

The cosmetic composition of the present invention may be prepared in the form of emulsified formulations and solubilized formulations commonly used in the art.

In addition, the components included in the cosmetic composition of the present invention may include components commonly used in cosmetic compositions in addition to the above components as active ingredients, for example, conventional adjuvants such as stabilizers, pigments and natural fragrances; It may further include a carrier.

Products to which the composition of the present invention can be added include, for example, mist, skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, sunscreen cream , moisture cream, hand cream, foundation, essence, nourishing essence, mask pack, press powder, loose powder, eye shadow, etc., and soap, cleansing foam, cleansing lotion, cleansing cream, body lotion and body cleanser.

When the formulation of the present invention is a paste, cream or gel, animal fiber, vegetable fiber, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component. can

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and in particular, in the case of a spray, additional chlorofluorohydrocarbon, propane , butane or propellants such as dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solvating agent or emulsifying agent is used as a carrier component, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butylglycol oil, glycerol fatty esters, fatty acid esters of polyethylene glycol or sorbitan.

When the formulation of the present invention is a suspension, as a carrier component, water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, microcrystalline Cellulose, aluminum metahydroxide, bentonite, agar or tracanth may be used.

Hereinafter, preferred examples are presented to aid the understanding of the present invention, but the following examples are merely illustrative of the present invention and it will be apparent to those skilled in the art that various changes and modifications are possible within the scope and spirit of the present invention, It goes without saying that such variations and modifications fall within the scope of the appended claims.

Comparison by degreasing solvent and water-soluble protein

Preparation Example 1. Degreasing with hexane

To prevent cannibalism from occurring due to the fasting period by purchasing raw white spot larvae, the population density was placed at 50 in a constant space of 13 cmX19 cmX10 cm (width X length X height), and water was fed. Thus, fasting was carried out in a refrigerator at 4 °C for 5 days. After removing the mouth and legs of the fasted white spot larvae, it was powdered by freeze-drying and used as a sample. After adding hexane corresponding to 5 times the weight of the sample, the fat was separated by homogenization at room temperature for 60 minutes. The homogenized sample was centrifuged at 3,000 X g (4° C.) for 15 minutes, the supernatant containing fat was removed, and the residue (degreased white spot larvae larvae) was then freeze-dried.

After mixing the freeze-dried defatted white spot larvae powder and 6 N NaOH, the pH was adjusted to 10, homogenized at 12000 rpm for 30 minutes, left for 60 minutes, and then homogenized at 15000 rpm for 30 minutes for salt-soluble protein was extracted.

Preparation Example 2. Degreasing with distilled water

It was prepared in the same manner as in Preparation Example 1, but was used for protein extraction using distilled water instead of hexane.

Preparation Example 3. Degreasing with anhydrous methanol

It was prepared in the same manner as in Preparation Example 1, but was used for protein extraction using anhydrous methanol instead of hexane.

Preparation Example 4. Degreasing with absolute ethanol

It was prepared in the same manner as in Preparation Example 1, but was used for protein extraction using absolute ethanol instead of hexane.

Preparation 5. Water-soluble protein

It was prepared in the same manner as in Preparation Example 1, but water-soluble protein was extracted by adding water instead of 6 N NaOH to the freeze-dried defatted white spot larvae powder.

<Test Example_Ⅰ>

Test Example 1. Measurement of protein concentration, pH and color

1-1. Protein concentration (mg/ml): Protein concentration was performed by referring to the Bradford method. In a 96-well plate, 50 μl of the sample and 200 μl of Bradford's reagent were reacted, and the absorbance was immediately measured at 595 nm.

1-2. Measurement of pH: The pH of the powder prepared in Preparation Example was measured using a pH meter.

1-3. Chromaticity measurement: Using a colorimeter (Chroma meter, CR 210, Minolta, Japan) for the powder prepared in Preparation Example, L ^{* -value indicating lightness, a *} -value indicating redness, and yellow ^b* -values representing yellowness were measured. At this time, a ^{calibration plate with an L* -value of 97.83, a*} -value of -0.43, and b ^* -value of +1.98 was used as the standard color.

division protein concentration pH L* a* b* Preparation Example 1 82.43±0.04 6.94±0.01 14.71±0.11 1.75±0.12 -0.04±0.08 Preparation 2 66.80±1.42 6.67±0.01 18.89±0.11 1.31±0.11 0.99±0.09 Preparation 3 14.90±6.96 7.02±0.01 15.39±0.03 2.67±0.08 1.55±0.03 Preparation 4 32.14±4.20 6.86±0.01 19.70±0.42 1.41±0.04 0.16±0.06 Preparation 5 37.61±3.77 6.84±0.01 18.65±0.21 1.61±0.03 1.02±0.07

As shown in Table 1 above, it was confirmed that the protein concentration was the highest in Preparation Example 1, degreased with hexane, and the lowest in Preparation Example 3, degreased with anhydrous methanol.

Test Example 2. SDS-PAGE measurement

1 is a photograph of electrophoresis measurement of defatted insect powder prepared according to Preparation Examples 1 to 4;

Protein patterns were analyzed using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) according to the method of Laemmli (1970). The protein fraction for analysis is mixed with 10 mL of a diluted solution (25 mM sodium phosphate (pH 7.2), 1% SDS, 3.5 M urea) in 1 g of the sample, and reacted at room temperature for one day at 3,000 X g for 15 minutes. It was centrifuged and filtered (Whatman No. 1). The filtered sample was diluted (25 mM sodium phosphate (pH 7.2), 1% SDS, 3.5 M urea) to a protein concentration of 8 mg/mL. The prepared sample was prepared with 5X Laemmli sample buffer (312.5 mM Tris-HCl (pH 6.8), 50% glycerol, 5% SDS, 5% β-mercaptoethanol, 0.05% bromophenol blue, EBA-1052, Elpisbiotech, Daejeon, Korea) and 4: The mixture was mixed in a ratio of 1 and heated at 100° C. for 5 minutes. In the concentration of acrylamide gel used for analysis, stacking gel was prepared at 4% and loading gel at 12%. 20 μl of the prepared sample was loaded at 100 V for about 2 hours. The loaded gel was stained with 0.25% Coomassie Brilliant Blue R250 (B7920, Sigma, USA) solution. For each sample, the molecular weight was estimated according to standard protein markers (pre-stained DokDo-MARK, EBM-1032, Elpisbiotech, Daejeon, Korea), and protein patterns between samples were compared.

As shown in FIG. 1, Preparation Example 1 does not form a band lower than 25 kDa and contains a polymer protein between 75 and 250 kDa, whereas Preparation Examples 3 and 4 have a band that is difficult to identify with the naked eye. but it was confirmed that it contains a low molecular weight protein.

That is, when anhydrous methanol and anhydrous ethanol were used for degreasing, denaturation occurred to such a degree that the band was difficult to identify with the naked eye.

Test Example 3. Measurement of gelation properties

Figure 2a is a graph measuring the foam capacity (foam capacity) for the salt-soluble protein powder prepared according to Preparation Examples 1 to 4, Figure 2b is the foam stability of the salt-soluble protein powder prepared according to Preparation Examples 1 to 4 (Foam stability) is a measured graph, Figure 2c is a graph measuring the emulsion capacity (Emulsion capacity) for the salt-soluble protein powder prepared according to Preparation Examples 1 to 4, Figure 2d is according to Preparation Examples 1 to 4 It is a graph measuring the emulsion stability of the prepared salt-soluble protein powder.

The foam capacity was measured in % by homogenizing the insect powder aqueous solution at 12,000 rpm for 2 minutes. In addition, the foam stability was measured by measuring the bubble layer for 60 minutes and comparing it with the initial bubble formation degree. In addition, the emulsion capacity was measured in % by measuring the emulsion layer by mixing oil with an edible insect powder aqueous solution and homogenizing it for 2 minutes at 18,000 rpm. It was mixed with a sulphate solution and the absorbance was measured at 500 nm.

As shown in FIGS. 2A to 2D , it was confirmed that Preparation Example 1 was superior to other groups in foam capacity, foam stability and emulsion stability, and emulsion capacity ) confirmed that Preparation Examples 1, 3, and 4 except for Preparation Example 2 showed similar values.

That is, those showing excellent values in all of the foam capacity, foam stability, emulsion capacity and emulsion stability that can confirm the gelation properties were degreased with hexane of Preparation Example 1. Since the larvae were extracted, the larvae of the larvae degreased with hexane were used in the subsequent experiment.

Comparison of protein solubility according to pH

Preparation Example 1. pH 10

The salt-soluble protein prepared in Preparation Example 1 was used.

Preparation Examples 5 to 12. pH 2-9

Prepared in the same manner as in Preparation Example 1, except that the degreased white spot flower larvae powder and 6 N HCl or 6 N NaOH were mixed to adjust the pH to 2, 3, 4, 5, 6, 7, 8, 9, respectively. Soluble protein was obtained.

<Test Example_Ⅱ>

Test Example 4. Protein solubility measurement

The protein solubility evaluation of defatted insect powder was conducted by applying the method of Kim et al. (2017). 9 mL of distilled water was added to 1 g of defatted insect powder, and the pH of each sample was adjusted to 2-10 with 6 N HCl or 6 N NaOH (Orion Star ^™ A211 pH Benchtop Meter, Thermo Scientific, USA). Then, distilled water was added so that the total amount of the solution was 10 mL, and the total volume of the sample was adjusted to be the same (1 g/10 mL). All samples were mixed for 10 seconds using a solution vortex mixer, reacted in a refrigerator at 2 °C for one day, and then centrifuged at 3,000X g for 15 minutes (4 °C) to measure the protein concentration (biuret method) of the supernatant (Gornall et al.) al., 1949).

3 is a graph measuring protein solubility according to pH.

As shown in Figure 3, it was confirmed that the defatted insect powder showed the lowest protein solubility ( p <0.05) between pH 2-4, whereas the highest protein solubility at pH 10 ( p <0.05). It is estimated that the isoelectric point of the powdery mildew larvae is pH 3-4, and it is judged that the protein solubility increases as the pH is further away from the isoelectric point.

The formation of the glycoprotein polymer (grafting reaction) is made through the Maillard reaction under weak alkaline conditions. It is judged that it is appropriate to extract the larvae protein for glycoprotein polymer formation at pH 10, which has excellent protein solubility in alkaline conditions.

Comparison according to saccharides in the formation of glycoprotein polymers

Preparation Example 1. Defatted white-spotted flower radish larva and salt-soluble protein

The defatted white-spotted white spot larvae prepared in Preparation Example 1 and salt-soluble protein extracted using the larvae were used.

Example 1. Use of glucose

The salt-soluble protein and glucose glucose prepared in Preparation Example 1 were mixed in a weight ratio of 1:6 and homogenized at 12000 rpm for 60 minutes. The supernatant was recovered by centrifugation at 3500 x g at ℃ for 15 minutes, and then the supernatant was injected using a spray dryer at a temperature of 150 ℃ (the temperature at which the Maillard reaction actively occurs) to form a glycoprotein polymer.

Example 2. Use of lactose

It was carried out according to Example 1, except that lactose was used instead of glucose to form a glycoprotein polymer.

Example 3. Use of maltose

It was carried out according to Example 1, except that maltose was used instead of glucose to form a glycoprotein polymer.

Comparative Example 1. Heating

The heated protein was prepared by injecting the salt-soluble protein prepared in Preparation Example 1 at a temperature of 150 °C (a temperature at which the Maillard reaction actively occurs).

Comparative Example 2. Use of the water-soluble protein of Preparation Example 5

It was carried out according to Example 1, but a glycoprotein polymer was formed using the water-soluble protein prepared in Preparation Example 5 instead of the salt-soluble protein.

<Test Example_Ⅲ>

Test Example 5. Chromaticity measurement

Chromaticity was measured in the same manner as in Test Example 1.

division L* a* b* Preparation Example 1
(Smudged white spotted flower larvae) 59.21±0.97 4.00±0.02 17.30±0.13 Preparation Example 1
(salt-soluble protein) 33.40±0.17 6.29±0.02 17.86±0.16 Example 1 37.35±1.86 5.68±0.30 13.34±0.84 Example 2 51.12±1.71 5.73±0.05 17.15±0.25 Example 3 48.35±1.05 5.90±0.03 17.66±0.24 Comparative Example 1 50.28±0.79 5.46±0.04 16.58±0.15 Comparative Example 2 53.45±0.46 5.11±0.02 18.96±0.27

As shown in Table 2 above, it was confirmed that the glycoprotein polymer prepared according to Example 1 had superior chromaticity compared to Examples 2, 3, and Comparative Examples 1 and 2.

That is, in the salt-soluble protein of Preparation Example 1, the brightness was significantly lowered and the redness significantly increased, compared to the defatted white spot larvae powder of Preparation Example 1, but there was no change in the yellowness ( p >0.05). ). Compared to Examples 1 to 3, the simple heating protein of Comparative Example 1 had a significantly increased brightness, a significantly decreased redness, and a slightly lowered yellowness, but no significant difference was recognized. Among glycoprotein polymers, Examples 2 and 3, which were treated with lactose and maltose, did not show a significant difference in color compared to Comparative Example 1, but Example 1, which was treated with glucose, showed brightness compared to simple heating protein. and a tendency to decrease in yellowness ( p <0.05).

Test Example 6. Measurement of glycoprotein polymerization

The evaluation of the degree of glycoprotein polymerization is a measure capable of estimating the formation of glycoprotein polymers of proteins used in the reaction.

Glycoproteinization of insect proteins was analyzed using ortho-phthaldialdehyde (OPA) assay (Mu et al., 2010). For the OPA reagent used in the experiment, 200 mg of OPA was dissolved in 5 mL of methanol, and 12.5 mL of 20% (w/w) sodium dodecyl sulfate (SDS), 125 mL of 100 mM sodium tetraborate and 0.5 mL of β-mercaptoethanol were mixed. and distilled water was added to finally prepare 250 mL. The test solution was reacted by adding 0.2 mL of sample solution and 4 mL of OPA solution. The reaction solution was reacted at 35° C. for 2 minutes, and absorbance was measured at 340 nm using a UV/VIS spectrophotometer (Libra S22, Biochrom, UK). The standard curve was obtained using 0.25-2 mM L-leucine. The result value for glycoprotein polymerization of insect protein was expressed as a relative % of insect protein and sugar to simple mixed powder, and was calculated according to the following [Equation 1].

[Equation 1]

Degree of glycation (%) = (FA _{0 -} FA ₁ )/FA ₀ X 100

FA ₀ : The levels of free amino group in mixture (before heating)

FA ₀ : The levels of free amino group in protein-saccharide conjugates

division Example 1 Example 2 Example 3 Comparative Example 1 Comparative Example 2 Glycoprotein polymerization (%) 59.8 53.1 51.7 9.4 11.4

As shown in Table 3 above, it was confirmed that the effective sugars for forming the glycoprotein polymer according to Examples 1 to 3 were in the order of glucose > lactose = maltose. Guan et al. (2006) reported that the free amino group content of proteins decreases as glycoprotein polymerization progresses, and that glycoprotein polymerization may show differences in the degree of glycoprotein polymerization depending on the saccharides used in the reaction.

That is, it is judged that glucose, which is a monosaccharide, is the most effective in forming a glycoprotein polymer, and that disaccharide or polysaccharide is somewhat less effective.

On the other hand, it was confirmed that the glycoprotein polymers of Comparative Examples 1 and 2 were unsuitable for formation.

Test Example 7. Measurement of gelation properties

Foam capacity, foam stability, emulsion capacity and emulsion stability of the glycoprotein polymer prepared according to Examples 1 to 3 of the present invention and the heated protein prepared according to Comparative Example 1 (Emulsion stability) was measured.

division Example 1 Example 2 Example 3 Comparative Example 1 Comparative Example 2 Foaming force (%) 284 251 237 97 101 Emulsifying ability (%) 98.8 92.3 86.6 84.2 72.4

As shown in Table 4 above, the glycoprotein polymers prepared according to Examples 1 to 3 of the present invention have foam capacity and emulsion capacity compared to the heated proteins prepared according to Comparative Examples 1 and 2 ) was confirmed to be excellent.

In particular, it was confirmed that Example 1 was more excellent in foam capacity and emulsion capacity compared to Examples 2 and 3.

division Example 1 Example 2 Example 3 Comparative Example 1 Comparative Example 1 bubble stability 0 minutes 100 100 100 100 100 10 minutes 90.8 81.4 78.7 61.6 59.4 20 minutes 75.9 64.1 59.9 40.7 37.2 30 minutes 66.4 50.4 42.7 32.5 30.6 60 minutes 58.8 46.2 39.1 20.6 18.4 emulsion stability 0 minutes 100 100 100 100 100 20 minutes 98.7 96.8 95.4 90.4 81.5 60 minutes 95.1 91.1 89.8 83.4 78.7 90 minutes 92.8 84.2 82.0 68.7 62.9 120 minutes 90.6 80.4 78.2 60.4 54.7

As shown in Table 5 above, the glycoprotein polymers prepared according to Examples 1 to 3 of the present invention have foam stability and emulsion stability compared to the heated proteins prepared according to Comparative Examples 1 and 2 ) was confirmed to be excellent.

In particular, it was confirmed that Example 1 was superior to Examples 2 and 3 in foam stability and emulsion stability.

Hereinafter, a formulation example of a composition containing the extract of the present invention will be described, but the present invention is not intended to limit the present invention, but to describe it in detail.

Formulation Example 1: Preparation of feed composition

Example 1 glycoprotein polymer powder 0.1 kg, corn 25.5 kg, wheat 15.04 kg, wheat flour 8.15 kg, rice bran 7.4 kg, soybean meal 18 kg, okgurten 1 kg, chicken by-product 14 kg, animal fat 9 kg, processed salt 0.3 kg, An animal (dog, pet dog) feed composition was prepared by mixing 0.3 kg of tricalcium phosphate, 1 kg of limestone, 0.01 kg of choline chloride, 0.05 kg of vitamins, 0.05 kg of minerals and 0.1 kg of digestive enzymes.

Formulation Example 2: Lotion

Preparation examples of the lotion in the cosmetic containing the glycoprotein polymer of Example 1 are shown in Table 6 below.

ingredient content (wt%) Polymer of Example 1 5.0 glycerin 6.0 1,3-butylene glycol 3.0 PG 1500 1.0 allantoin 0.1 DL-Panthenol 0.3 EDTA-2Na 0.02 Benzophenone-9 0.04 Sodium Hyaluronate 5.0 ethanol 10.0 Polysorbate 20 0.2 preservatives, fragrances, colors a very small amount Distilled water remaining amount Sum 100

Formulation Example 3: Lotion

Preparation examples of lotions in the cosmetic containing the glycoprotein polymer of Example 1 are shown in Table 7 below.

ingredient content (wt%) Polymer of Example 1 3.0 propylene glycol 6.0 glycerin 4.0 triethanolamine 1.2 Tocopheryl Acetate 3.0 liquid paraffin 5.0 squalane 3.0 Macadamina Nut Oil 2.0 Polysorbate 60 1.5 Sorbitan sesquioleate 1.0 Carboxyvinyl Polymer 1.0 preservatives, fragrances, colors a very small amount Distilled water remaining amount Sum 100

Preparation Example 4: Nourishing Cream

Preparation examples of the nutritional cream in the cosmetic containing the glycoprotein polymer of Example 1 are shown in Table 8 below.

ingredient content (wt%) Polymer of Example 1 1.0 lipophilic monostearate glycerin 1.5 cetearyl alcohol 1.5 stearic acid 1.0 Polysorbate 60 1.5 Sorbitan Stearate 0.6 Isostearylisostearate 5.0 squalane 5.0 mineral oil 35.0 dimethicone 0.5 Hydroxyethyl Cellulose 0.12 glycerin 6.0 triethanolamine 0.7 preservatives, fragrances, colors a very small amount Distilled water remaining amount Sum 100

Formulation Example 5: Essence

Preparation examples of the essence in the cosmetic containing the glycoprotein polymer of Example 1 are shown in Table 9 below.

ingredient content (wt%) Polymer of Example 1 1.5 glycerin 10.0 betaine 5.0 PEG 1500 2.0 allantoin 0.1 DL-Panthenol 0.3 EDTA-2Na 0.02 Benzophenone-9 0.04 Hydroxyethyl Cellulose 0.1 Sodium Hyaluronate 8.0 Carboxyvinyl Polymer 0.2 triethanolamine 0.18 Octyldodecanol 0.3 Octyldodeces-16 0.4 ethanol 6.0 preservatives, fragrances, colors a very small amount Distilled water remaining amount Sum 100

Formulation Example 6: Emulsion for mask pack

Preparation examples of the emulsion for a mask pack among the cosmetics containing the glycoprotein polymer of Example 1 are shown in Table 10 below.

ingredient content (wt%) Polymer of Example 1 1.0 polyvinyl alcohol 15.0 Cellulose Gum 0.15 glycerin 3.0 PEG 1500 2.0 cyclodextrin 0.15 DL-Panthenol 0.4 allantoin 0.1 Monoammonium glycyrrhizinate 0.3 nicotinamide 0.5 ethanol 6.0 PEG 40 hydrogenated castor oil 0.3 preservatives, fragrances, colors a very small amount Distilled water remaining amount Sum 100

Claims (21)

(A) defatting insect fat;
(B) extracting salt-soluble protein from the defatted insect powder; and
(C) forming a glycoprotein polymer by grafting technology by reacting the salt-soluble protein with the sugar and then spray-drying for the Maillard reaction Way. The method of claim 1, further comprising, after step (C), (D) gelling the spray-dried glycoprotein polymer. The method according to claim 1, wherein the fat content of the insect powder defatted in step (A) is 0.1 to 10 g/100 g. The method according to claim 1, wherein the solvent used for degreasing the insects in step (A) is hexane, distilled water, anhydrous methanol or anhydrous ethanol. The method according to claim 1, wherein the salt-soluble protein in step (B) is mixed with an insect and a solvent in a weight ratio of 1: 2-5, homogenized at 10000 to 15000 rpm for 10 to 50 minutes, and then left for 30 to 120 minutes. Next, a method for producing an insect-derived food protein, characterized in that obtained by homogenizing at 10000 to 15000 rpm for 20 to 80 minutes. The method according to claim 5, wherein the solvent is at least one selected from the group consisting of saline solution, sodium hydroxide aqueous solution and sodium phosphate aqueous solution. The method according to claim 1, wherein the extraction in step (B) is performed under the conditions of pH 9-11. The method according to claim 1, wherein in step (C), salt-soluble protein and sugar are mixed in a weight ratio of 1: 2-10, homogenized at 10000 to 15000 rpm for 30 to 120 minutes, and then at 70 to 90 ° C. for 30 to 120 minutes. A method for producing an insect-derived food protein, characterized in that it is heated during a bath. 9. The insect according to claim 8, wherein the heated mixture is centrifuged at 1 to 5 °C at 2000 to 5000 xg for 5 to 50 minutes to recover the supernatant, and then the Maillard reaction is performed to obtain a glycoprotein polymer. Method for producing derived food protein. The method according to claim 1, wherein the saccharides are monosaccharides, disaccharides, oligosaccharides or polysaccharides. The method according to claim 1, wherein the saccharide is a monosaccharide. The preparation of insect-derived food protein according to claim 1, wherein, in step (C), the salt-soluble protein reacted with sugar is injected at a temperature of 140 to 170 ° C. for the Maillard reaction during spray drying. Way. The method according to claim 1, wherein the insect is at least one member selected from the group consisting of a grasshopper order, a coleopteran order, and a lepidopteran order. According to claim 1, wherein the insect is a mealworm larvae ( Tenebrio molitor , TM), long- lived beetle larvae (Allomyrina dichotoma , AD), white spot beetle larvae (Protaetia brevitarsis seulensis , PB), rice locust (Oxya chinensis sinuosa Mistshenko ), twin-star Crickets ( Gryllus bimaculatus ) and silkworm pupa ( Bombyx mori ) A method for producing an insect-derived food protein, characterized in that at least one selected from the group consisting of. The method according to claim 1, wherein the insect is at least one selected from the group consisting of mealworm larvae (Tenebrio molitor , TM), long- leaved beetle larvae (Allomyrina dichotoma , AD), and larvae of Protaetia brevitarsis seulensis (PB). A method for producing an insect-derived food protein. An insect-derived food protein prepared by the method of any one of claims 1 to 15. A gel product using the insect-derived food protein of claim 16 . A food composition using the gel product of claim 17 . The food composition according to claim 18, wherein the food composition is a fish cake composition, a sausage composition, a patty composition, a yokan composition, or a konjac composition. A feed composition comprising the gel product of claim 17 . A cosmetic composition comprising the gel product of claim 17 .

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