PREPARATION METHOD FOR TWO-SPOTTED CRICKET POWDER OR EXTRACT THEREOF, FOOD COMPOSITION COMPRISING SAME, AND USES OF SAME

Abstract

The present invention relates to: Two-spotted cricket powder and an extract thereof for food, drugs, and cosmetic materials; a preparation method of the Two-spotted cricket powder; a food composition or care food prepared using the powder; and uses of the food composition prepared using the Two-spotted cricket powder for the prevention and amelioration of sarcopenia and antioxidant effects. Sarcopenia-preventive functional food prepared using Two-spotted cricket is a highly value-added novel alternative protein-source food that can make a contribution in the future to the edible insects industry and across the industries related to functional food, inner beauty food materials, and food, and is also expected to contribute to diversifying food choices and improving nutrition among older populations that are increasing in an aging society.

Claims

1-22. (canceled)

23. A method of producing a Two-spotted cricket powder or extract, comprising the steps of: (a) fasting the Two-spotted cricket; (b) steaming the fasted Two-spotted cricket; and (c) pulverizing the steamed Two-spotted cricket with a pulverizer and thus obtaining powder.

24. The method of claim 23, further comprising hot air drying or freeze-drying the steamed Two-spotted cricket of the step (b).

25. The method of claim 24, wherein the hot air drying is performed at a temperature of 55° C. to 65° C. for 12 hours to 24 hours; or the freeze-drying is performed for 48 hours to 60 hours after rapidly freezing at a temperature of −60° C. to −80° C.

26. The method of claim 23, wherein the fasting of the step (a) is performed for 24 to 72 hours.

27. The method of claim 23, wherein the steaming of the step (b) is performed using 100° C. to 200° C. steam for 15 to 60 minutes.

28. The method of claim 23, wherein the pulverizing of the step (c) is performed by filling the pulverizer with nitrogen so that oxidation of powder is prevented.

29. The method of claim 23, wherein the Two-spotted cricket powder has antioxidant activity.

30. The method of claim 23, wherein the Two-spotted cricket powder contains one or more amino acids selected from the group consisting of histidine, threonine, arginine, valine, methionine, tryptophan, phenylalanine, isoleucine, leucine, lysine, aspartic acid, glutamic acid, asparagine, serine, glutamine, glycine, alanine, taurine, γ-aminobutyric acid (GABA), tyrosine, and proline.

31. The method of claim 23, further comprising the steps of: (d) degreasing the Two-spotted cricket powder; and (e) extracting protein from the degreased Two-spotted cricket powder and obtaining an extract containing protein.

32. The method of claim 31, wherein the degreasing of the step (d) is performed using one or more solvents selected from the group consisting of ethanol, hexane, and petroleum ether; or the degreasing of the step (d) is performed at a temperature of 35° C. to 45° C. for 40 minutes to 80 minutes.

33. The method of claim 31, wherein the extracting of protein of the step (e) is performed using one or more solvents selected from the group consisting of sodium hydroxide, hydrochloric acid, and a carboxylic acid; or the extracting of protein of the step (e) is performed at a temperature of 30° C. to 50° C. for 40 minutes to 80 minutes.

34. A composition for a food material, comprising: the Two-spotted cricket powder or extract produced by the method of claim 23.

35. The composition of claim 34, wherein the food material is a food material for an inner beauty food.

36. The composition of claim 34, wherein the composition has antioxidant activity.

37. The composition of claim 34, wherein the food material is for a Sunsik (roasted grain powder).

38. The composition of claim 37, wherein the Sunsik (roasted grain powder) further comprises one or more ingredients selected from the group consisting of brown rice powder, black sesame powder, almond powder, and broccoli powder.

39. The composition of claim 37, wherein the Sunsik (roasted grain powder) includes 1 part by weight to 3 parts by weight of brown rice powder, 8 parts by weight to 20 parts by weight of black sesame powder, 5 parts by weight to 18 parts by weight of almond powder, or 10 parts by weight to 40 parts by weight of broccoli powder based on 100 parts by weight of the Two-spotted cricket powder.

40. A method for ameliorating or treating sarcopenia, comprising: administering a composition comprising the Two-spotted cricket powder or extract produced by the method of claim 23 as an active ingredient, into a subject in need thereof.

41. The method of claim 40, wherein the powder or extract inhibits expression of a myostatin (MSTN) gene by suppressing activity of an MSTN promoter.

42. The method of claim 40, wherein the composition has antioxidant activity.

Description

DESCRIPTION OF DRAWINGS

[0063] FIG. 1 is a flowchart illustrating a method of producing a Two-spotted cricket extract according to the present invention.

[0064] FIGS. 2A and 2B illustrate the results of measuring the expression level of myostatin (MSTN) (FIG. 2A) and the activity of an MSTN promoter (FIG. 2B) in order to evaluate a muscle loss inhibitory effect of an extract (Cricket) and a protein extract (Cricket Protein) of a Two-spotted cricket powder of the present invention.

MODES OF THE INVENTION

[0065] Terms or words used in this specification and claims should not be interpreted as being limited to commonly used meanings or meanings in dictionaries, and, based on the principle that the inventors can appropriately define concepts of terms in order to describe their invention in the best way, the terms and words should be interpreted with meanings and concepts which are consistent with the technical spirit of the present invention.

[0066] Hereinafter, exemplary embodiments are presented to help the understanding of the present invention. However, the following exemplary embodiments are only provided to facilitate understanding of the present invention, and the contents of the present invention are not limited by the exemplary embodiments.

EXAMPLES

Example 1

Two-Spotted Cricket Selection Criteria for Food Materialization

[0067] Two-spotted cricket, which is the main ingredient of the present invention, was purchased from a cricket farm located in Saenggeuk-myeon, Chungcheongbuk-do (Chungcheongbuk-do, Republic of Korea). The Two-spotted cricket was fasted for two days to remove feces in the body and prevent the growth of microorganisms and thereby eliminate an unpleasant smell and improve taste.

Example 2

Determination of Optimal Pulverization Conditions for Food Materialization of Two-Spotted Cricket

[0068] After fasting for two days and washing, the Two-spotted cricket was pulverized through the following process. The washed Two-spotted cricket was blanched for about 10 minutes in 100° C. boiling water or steamed for about 30 minutes with 100° C. steam, and then hot air-dried at 60° C. for 18 hours or rapidly frozen at −70° C. and freeze-dried for 48 to 60 hours. The dried Two-spotted cricket was pulverized with a pulverizer to obtain a powder.

Example 3

Determination of Conditions for Producing Extract Having Highest Protein Content from Two-Spotted Cricket Powder

[0069] The Two-spotted cricket powder obtained by the method of Example 2 was degreased by solvent extraction with ethanol. To the obtained degreased Two-spotted cricket powder, 0.25 M sodium hydroxide (NaOH) was added in a weight ratio of 1:15 and mixed at 40° C. for 60 minutes using a hot plate (Vision Scientific Co., Ltd., Korea) and a magnetic stirrer. Subsequently, centrifugation was performed at 4° C. and 3,000 rpm for 10 minutes using a centrifuge (VS-24SMTi, Vision Science Co., Ltd., Korea), and only the supernatant was collected. After adjusting the pH of the obtained supernatant to a pH of 4.4, centrifugation was again performed at 4° C. and 3,500 rpm for 10 minutes, and thus a pellet was obtained. The pellet was put in a dialysis bag (12 kDa MWCO; Sigma-Aldrich Chemical Co., USA) and desalted for 12 hours. Subsequently, freeze-drying was performed for 36 hours, and thus a Two-spotted cricket protein extract powder was obtained. The obtained powder was maintained at a temperature of −24° C. or less for further experiments.

[0070] 10 mg of the Two-spotted cricket protein extract powder was input into 1 mL of DMSO, dissolved using an ultrasonic homogenizer (Sonicator Vibra cell, Sonics & Materials; amplitude control 40%), and then centrifuged at 13,000 rpm and 10° C. for 10 minutes, and only the supernatant was collected and transferred to a 1 mL Eppendorf tube (e-tube; micro centrifuge tube).

Example 4

Nutritional Composition Analysis of Extract and Protein Extract of Two-Spotted Cricket Powder

[0071] 4.1. Analysis of Three Major Nutrients (Carbohydrate, Fat, and Protein)

[0072] After pulverizing Two-spotted cricket, water content was analyzed by an atmospheric heating and drying method performed at 105° C., crude fat content was analyzed by a Soxhlet extraction method, and crude mineral content was analyzed by an ashing method performed at 550° C. Carbohydrate content was calculated as a value obtained by subtracting the content of water, crude protein, crude fat, and crude minerals from 100 g of sample. The crude protein content was analyzed with a nitrogen analyzer, and the analyzed nitrogen content was multiplied by a protein factor of 6.25 and expressed as the crude protein content.

[0073] The results of analyzing composition of the Two-spotted cricket powder are shown in Table 1 below. It was confirmed that the Two-spotted cricket powder contained 3.86% water, 4.41% minerals, 61.05% crude protein, 19.08% crude fat, and 11.60% carbohydrates, which shows a significantly higher protein content and lower fat content compared to other edible insects. Since the protein content is higher than the fat content, it is expected that the Two-spotted cricket powder has potential as a food material or food for preventing muscle-related diseases or muscle loss in the elderly and an increase in blood lipids in middle-aged and elderly people, and in particular, it is expected that Two-spotted cricket powder can be used as a beauty food material with potential as a weight loss material due to its low fat and carbohydrate content.

TABLE-US-00001 TABLE 1 Sample Crude Crude Crude Carbo- (%) Water minerals protein fat hydrates Two- 3.86 ± 0.23 4.41 ± 0.60 61.05 ± 1.06 19.08 ± 0.16 11.60 ± 0.74 spotted cricket powder

[0074] 4.2. Mineral Composition Analysis of Extract and Protein Extract of Two-Spotted Cricket Powder

[0075] To analyze mineral content, an extract and protein extract of the Two-spotted cricket powder were wet-decomposed, and the amounts of calcium (Ca), potassium (K), sodium (Na), phosphate (P), and zinc (Zn), which are minerals contained in large amounts, were measured using an optical emission spectrometer.

[0076] The results of analyzing the mineral content of the extract (cricket powder extract (CPE)) and the protein extract (cricket powder protein extract (CPPE)) of the Two-spotted cricket powder are shown in Table 2 below (units: mg/100 g). The mineral composition of the Two-spotted cricket powder extract was 110.8 mg/100 g of Ca, 724.7 mg/100 g of K, 289.3 mg/100 g of Na, 74.0 mg/100 g of P, and 22.1 mg/100 g of Zn, and the Two-spotted cricket powder protein extract contained 2.5 mg/100 g of Ca, 8.6 mg/100 g of K, 91.0 mg/100 g of Na, 98.0 mg/100 g of P, and 23.5 mg/100 g of Zn. From these results, it is expected that the Two-spotted cricket powder, which has a high Ca content, has potential for the prevention and amelioration of osteoporosis in the elderly and the promotion of health in growing children and adolescents. In addition, since K has a physiological function of lowering blood pressure, it is expected that the Two-spotted cricket powder extract will be useful as a functional material for preventing hypertension.

TABLE-US-00002 TABLE 2 Sample (mg/kg) Ca K Na P Zn CPE 110.8 724.7 289.3 74.0 22.1 CPPE 2.5 8.6 91.0 98.0 23.5

[0077] 4.3. Essential Amino Acid Composition Analysis of Extract and Protein Extract of Two-Spotted Cricket Powder

[0078] The amino acid content was measured using an amino acid analyzer. In one example of a measurement method, the measurement was performed by inputting 5 g of a sample and 40 mL of 6 N HCl into a round flask and mixing, and then performing hydrolysis by injecting nitrogen gas at 110° C. for 24 hours. After concentrating HCl under reduced pressure at 50° C., the concentrated sample was diluted by adding 50 mL of 0.2 N sodium citrate buffer (pH 2.2), and filtered with 0.45 μm filter paper. After filtering, the sample (30 μL) was analyzed using an automatic amino acid analyzer, an ion change column (4.6 mm×60 mm), and a visible detector. The analysis was performed at a buffer flow rate of 1 mL/min, a column temperature of 20 to 85° C., and a reaction temperature of 50 to 140° C.

[0079] The essential amino acid compositions of the extract (cricket powder extract (CPE)) and the protein extract (cricket powder protein extract (CPPE)) of the Two-spotted cricket powder are shown in Table 3 below. The Two-spotted cricket powder extract contained essential amino acids such as 10,380.00 mg/kg of histidine, 12,679.04 mg/kg of threonine, 32,189.42 mg/kg of arginine, 17,186.31 mg/kg of valine, 6,077.52 mg/kg of methionine, 340.03 mg/kg of tryptophan, 12,249.08 mg/kg of phenylalanine, 12,790.01 mg/kg of isoleucine, 25,382.52 mg/kg of leucine, and 12,050.90 mg/kg of lysine, and the powder protein extract contained 2,763.08 mg/kg of histidine, 710.63 mg/kg of threonine, 2,822.72 mg/kg of arginine, 1,147.28 mg/kg of valine, 679.55 mg/kg of methionine, 1,367.50 mg/kg of phenylalanine, 1,281.90 mg/kg of isoleucine, 2,753.34 mg/kg of leucine, and 900.17 mg/kg of lysine. The essential amino acids are amino acids which are not synthesized in the body and thus should be ingested with food, and the Two-spotted cricket powder extract contained a large amount of each of the 10 essential amino acids. Both the Two-spotted cricket powder and the powder protein extract also contained leucine, isoleucine, and valine which are branched chain amino acids (BCAAs), and since the BCAAs are known to be oxidized in skeletal muscle and used as an energy source for the muscle, have the ability to minimize a decrease in protein in muscle cells by inhibiting protein catabolism in the muscle cells, and are used as an energy source for muscles during long-term exercise, it is expected that Two-spotted cricket, which is rich in the BCAAs, will have high utility as a food that helps to prevent muscle loss in the elderly and patients by participating in the process of muscle strength enhancement and muscle building and have high utility as a material for a nutritional supplement or health functional food not only for growing children and adolescents, but also for the general public. In addition, since Two-spotted cricket is a suitable food substitute which meets the needs of people having the risk of increased blood lipids due to excessive meat consumption and modern-day people who value health, it is expected that Two-spotted cricket will be developed as a material for food or various functional foods, and as protein supplements for athletes, nutritional supplements for patients, and new raw materials for pharmaceuticals and functional cosmetics. Two-spotted cricket also contains a large amount of lysine, and since lysine is known to help maintain skin health and suppress skin aging by participating in collagen synthesis, it is expected that Two-spotted cricket will be useful as a skin beauty-related material or a beauty food.

TABLE-US-00003 TABLE 3 Sample (mg/kg) Amino acid CPE CPPE Histidine 10,380.00 2,763.08 Threonine 12,679.04 710.63 Arginine 32,189.42 2,822.72 Valine 17,186.31 1,147.28 Methionine 6,077.52 679.55 Tryptophan 340.03 NA Phenylalanine 12,249.08 1,367.50 Isoleucine 12,790.01 1,281.90 Leucine 25,382.52 2,753.34 Lysine 12,050.90 900.17

[0080] 4.4. Non-Essential Amino Acid Composition Analysis of Extract and Protein Extract of Two-Spotted Cricket Powder

[0081] The non-essential amino acid compositions of the extract (cricket powder extract (CPE)) and the protein extract (cricket powder protein extract (CPPE)) of the Two-spotted cricket powder extract are shown in Table 4 below. The Two-spotted cricket powder extract contained non-essential amino acids such as 19,451.30 mg/kg of aspartic acid, 36,085.54 mg/kg of glutamic acid, 18,447.13 mg/kg of serine, 639.89 mg/kg of glutamine, 21,944.58 mg/kg of glycine, 31,947.01 mg/kg of alanine, 3,309.13 mg/kg of taurine, 17,053.70 mg/kg of tyrosine, and 2,834.81 mg/kg of proline, and the Two-spotted cricket powder protein extract contained 1,571.11 mg/kg of aspartic acid, 3,266.08 mg/kg of glutamic acid, 730.02 mg/kg of serine, 2,617.74 mg/kg of glycine, 1,965.78 mg/kg of alanine, 83.44 mg/kg of taurine, and 1,351.96 mg/kg of tyrosine. Among the non-essential amino acids, proline is involved in collagen synthesis together with lysine, so it is expected that the Two-spotted cricket powder extract or the Two-spotted cricket powder protein extract will be useful as a skin beauty-related material or a beauty food material in the future.

TABLE-US-00004 TABLE 4 Sample (mg/kg) Amino acid CPE CPPE Aspartic acid 19,451.30 1,571.11 Glutamic acid 36,085.54 3,266.08 Asparagine 338.44 NA Serine 18,447.13 730.02 Glutamine 639.89 NA Glycine 21,944.58 2,617.74 Alanine 31,947.01 1,965.78 Taurine 3,309.13 83.44 γ-Aminobutyric acid (GABA) 104.35 NA Tyrosine 17,053.70 1,351.96 Proline 2,834.81 NA

Example 5

Confirmation of Antioxidant Effect of Extract and Protein Extract of Two-Spotted Cricket Powder

[0082] The antioxidant activity of the extract and the protein extract of the Two-spotted cricket powder was confirmed by measuring a total polyphenol content and DPPH radical scavenging ability. In one example of a measurement method, the total polyphenol content was measured according to a modification of the Folin-Denis method (Folin & Denis, 1912). After taking 1 mL of the diluted Two-spotted cricket powder extract and adding 1 mL of a sodium carbonate (Na.sub.2CO.sub.3; Sigma-Aldrich) solution thereto, the resultant was allowed to stand for three minutes and then reacted by adding 0.2 mL of a 50% Folin-Ciocalteu reagent (Sigma-Aldrich), and absorbance was measured at 750 nm using a microplate reader (BN 02514; Molecular Devices, USA). A total polyphenol content converted based on a standard curve obtained using tannic acid (Sigma-Aldrich) is shown. The DPPH radical scavenging ability of the Two-spotted cricket powder extract sample was measured according to a modification of the Blois method (1958). After adding a 1.5×10.sup.−4 M DPPH solution to 0.1 mL of the sample, the resultant was allowed to stand for 30 minutes in the dark at room temperature, and absorbance was measured at 517 nm using a microplate reader.

[0083] The results of measuring the total polyphenol content of the extract (cricket powder extract (CPE)) and the protein extract (cricket powder protein extract (CPPE)) of the Two-spotted cricket powder are shown in Table 5 below. The total polyphenol contents of the Two-spotted cricket powder extract and the powder protein extract were 6.61 mg GAE/g and 12.18 mg GAE/g, respectively, which shows that the total polyphenol content of the Two-spotted cricket powder protein extract was higher. Phenolic compounds are materials present in plants and can be used as indirect indicators of antioxidant activity, and they inhibit the generation of reactive oxygen species occurring in the body and prevent the oxidation of lipids and proteins in the body, thereby inhibiting cell damage and the abnormal growth and differentiation of cells, and thus are known as factors capable of preventing cancer by blocking the initiation stage of cancer development. Therefore, the above results suggest that the Two-spotted cricket powder protein extract having a higher total polyphenol content has better antioxidant ability and thus has potential as a material for preventing aging and various cancers.

[0084] DPPH radical scavenging ability is based on the principle that a DPPH radical, which is a stable free radical, undergoes a color change from purple (DPPH) to colorless (diphenyl-picrylhydrazine) and an absorbance change as it is reduced by sulfur-containing amino acids such as cysteine, glutathione, aromatic amines, and ascorbic acid, and is useful for measuring antioxidant ability, and the measurement results are shown in Table 5 below. The DPPH radical scavenging ability of the Two-spotted cricket powder extract and the Two-spotted cricket powder protein extract was very high, 68.25% and 80.37%, respectively, and the scavenging ability of the protein powder extract was higher. This is significantly higher than the reported DPPH radical scavenging ability of a brown mealworm ethanol extract which is 16.2%, so the Two-spotted cricket powder has a very high amino acid content and is expected to have a great value as an animal-based antioxidant food. Since antioxidants are known to suppress aging by inhibiting cellular oxidation in the body and scavenging active radicals, alleviate hypertension, and enhance various physiological functions such as immune function, it is expected that a production method of the present invention will be useful for a food material or food having excellent antioxidant activity, and furthermore, an inner beauty food.

TABLE-US-00005 TABLE 5 Sample CPE CPPE DPPH (%) 68.25 80.37 Total polyphenol (mg GAE/g) 6.61 12.18

[0085] When the results of Examples 4 and 5 are combined, it can be seen that the Two-spotted cricket powder extract and the powder protein extract contained a large amount of protein and a small amount of fat, had an excellent essential amino acid composition, and in particular, contained a large amount of valine, leucine, and isoleucine which are amino acids related to muscles. In addition, the Two-spotted cricket powder extract and the powder protein extract had high total polyphenol content and excellent DPPH radical scavenging ability. Therefore, it is expected that the Two-spotted cricket powder extract or the powder protein extract produced by a production method of the present invention will be useful as a high-quality protein food that can replace animal protein, a food material or food for promoting antioxidant activity or preventing muscle loss, and an inner beauty food.

Example 6

Confirmation of Sarcopenia Inhibitory Effect of Extract and Protein Extract of Two-Spotted Cricket Powder

[0086] To confirm the muscle loss inhibitory effect of the extract and the protein extract of the Two-spotted cricket powder, the expression level of MSTN, which is a marker gene indicating sarcopenia, and the activity of an MSTN promoter were comparatively measured after treating the extract and the protein extract of the Two-spotted cricket powder of the present invention. The treatment of 0.01 to 1 mg/mL of the Two-spotted cricket powder extract (labeled as “Cricket” in FIGS. 2A and 2B) or the Two-spotted cricket powder protein extract (labeled as “Cricket protein” in FIGS. 2A and 2B) or less did not show toxicity to mouse skeletal muscle cells (C2C12 cell line).

[0087] Upon the treatment of 10 μg/mL of the Two-spotted cricket powder extract or the Two-spotted cricket powder protein extract for 24 hours, the mRNA expression level of MSTN gene decreased to about 77% and 44%, respectively (FIG. 2A). In addition, as a result of measuring activity using an MSTN promoter luciferase vector, it was confirmed that the Two-spotted cricket powder extract or the Two-spotted cricket powder protein extract inhibited the promoter activity by about 35.9% and 13.6%, respectively (FIG. 2B). That is, the above results prove that the Two-spotted cricket powder extract or the Two-spotted cricket powder protein extract has the effect of inhibiting sarcopenia by suppressing MSTN mRNA expression by suppressing MSTN promoter activity.

Example 7

Preparation of Mixed Sunsik (Roasted Grain Powder) Using Two-Spotted Cricket Powder

[0088] To the Two-spotted cricket powder of the present invention, brown rice (contains carbohydrate source, vegetable protein, fiber source, various vitamins and minerals), black sesame (contains essential fatty acids and vegetable protein), almond (contains vegetable protein and essential fatty acids), broccoli (vitamins and minerals), perilla (essential fatty acids), and the like were added to prepare a Sunsik. Since a high-protein food is defined as a food containing 11 g or more of protein, the composition ratio of the Sunsik was adjusted so that the Sunsik contained 11 g or more of protein per serving.

[0089] More specifically, the Sunsik was prepared by mixing 26 g of the Two-spotted cricket powder with 0.5 g of brown rice powder, 4.0 g of black sesame powder, 3.0 g of almond powder, and 6.5 g of broccoli powder. Since the prepared Sunsik is rich in protein, it is expected to function as a high-quality source of protein and essential fatty acids.

[0090] The above description of the present invention is for illustration, and those of ordinary skill in the art to which the present invention pertains can understand that modifications into other specific forms are possible without changing the technical spirit or essential features of the present invention. Therefore, it should be understood that the exemplary embodiments described above are illustrative in all respects and not restrictive.

INDUSTRIAL APPLICABILITY

[0091] The present invention relates to a method of producing a Two-spotted cricket powder or an extract thereof, and a food composition and the like produced using the same. The present invention can be variously used for specific target subjects (the elderly, athletes, growing and developing adolescents, people in need of weight control, patients, etc.), and specifically, can be highly effective for treating sarcopenia in the elderly, muscle loss due to weight loss, and sarcopenic obesity, promoting the health of children and adolescents, and improving the athletic performance of athletes. In addition, a food composition, care food, functional food, or Sunsik produced using a Two-spotted cricket powder of the present invention or an extract thereof contains a large amount of essential amino acids, amino acids that help prevent sarcopenia, and essential fatty acids that help reduce inflammation and thus is expected to greatly contribute to preventing geriatric diseases and promoting health. Furthermore, the Two-spotted cricket powder or extract is expected to have high industrial applicability due to its high utility in various fields such as the food field, functional food field, pharmaceutical field, and cosmetics field.