Method for producing protein derived from dried laver processing by-products with improved antioxidant activity

Abstract

The present invention relates to a method for producing a protein derived from a dried seaweed processing by-product with enhanced antioxidant activity. The protein derived from a dried seaweed processing by-product produced through the method according to the present invention exhibits excellent antioxidant activity and processing suitability. Therefore, the protein derived from a dried seaweed processing by-product can be usefully utilized in processed foods or as a composition for antioxidant health functional foods.

Description

Technology Field

[0001] The present invention relates to a method for producing a protein derived from a dried seaweed processing byproduct with enhanced antioxidant activity. Background Technology

[0003] Gim is widely cultivated in Korea, primarily in the Jeollanam-do region, as well as in coastal areas such as Chungcheongnam-do and Busan. Depending on the raw materials and processing methods, it is produced in categories such as Dolgim, Jaeraegim, and Gimbapgim. The main raw material used for Dolgim ​​is Ibbadi Dolgim, also known as Gopchanggim ( Porphyra dentata ) and patterned stone laver ( Pyropia seriata It mainly uses ), and as a raw material for traditional laver and gimbap laver, it is true laver ( Pyropia tenera ) and radial patterned seaweed( Porphyra yezoensis ) is mainly used.

[0004] Dried laver has shown a steady upward trend in the seafood export market over the past decade and is establishing itself as the most important seafood export item. From $520 million in 2018 to $640 million in 2022, the export value grew by more than 20% over the past five years, and this export performance is comparable to that of major food export items such as ramen and tuna.

[0005] However, contrary to this boom, by-products generated during the processing of dried seaweed are on a continuous upward trend. These processing by-products are produced during the cutting process for shaping; while their nutritional content and value are comparable to the original product, they are discarded as by-products due to their unclear intended use. According to an investigation of dried seaweed processing plants, it is estimated that approximately 30% of the raw materials are being thrown away as waste. Therefore, given that the production volume of dried seaweed in 2022 was 48,000 tons, it is estimated that at least 15,000 tons of by-products were generated during that period. Although these processing by-products are exported at low prices to overseas manufacturers of seasoned seaweed and seaweed snacks in countries such as the United States and Thailand, as previously mentioned, their nutritional content and functionality are no different from the original product. Originally, laver is known as an excellent food ingredient with the highest protein content among seaweeds, and even dried laver processing by-products are excellent ingredients with a protein content of over 30%, but their value is not recognized because their use has not been established domestically, and they are being exported abroad.

[0006] Accordingly, the inventors extracted a protein derived from dried seaweed processing by-products through hydration and physical extraction, and confirmed that this protein has an excellent antioxidant effect and can be used as a processed food that can replace meat, thereby completing the present invention. Prior art literature

[0008] Republic of Korea Registered Patent No. 10-0828701 The problem to be solved

[0009] The objective of the present invention is to provide a method for producing a protein derived from dried seaweed processing by-products with enhanced antioxidant activity.

[0010] In addition, another objective of the present invention is to provide a protein derived from a dried seaweed processing by-product produced by the above method.

[0011] In addition, another objective of the present invention is to provide a processed food containing a protein derived from the processing by-product of dried seaweed.

[0012] In addition, another objective of the present invention is to provide a health functional food composition for antioxidant purposes comprising the protein derived from the processing by-product of dried seaweed as an active ingredient. means of solving the problem

[0014] To achieve the above objectives, the present invention provides a method for producing a protein derived from a dried seaweed processing byproduct with enhanced antioxidant activity, comprising the steps of: adding distilled water to dried seaweed processing byproduct powder to hydrate it, and then ultrasonically treating it to produce a dried seaweed processing byproduct protein extract; and filtering and drying the dried seaweed processing byproduct protein extract.

[0015] Next, the present invention provides a protein derived from a dried seaweed processing by-product produced by the above method.

[0016] Furthermore, the present invention provides a processed food containing a protein derived from the processing by-product of dried seaweed.

[0017] Finally, the present invention provides a health functional food composition for antioxidant purposes comprising the protein derived from the processing by-product of dried seaweed as an active ingredient. Effects of the invention

[0019] The protein derived from a dried seaweed processing by-product, produced by the method for producing a protein derived from a dried seaweed processing by-product with enhanced antioxidant activity according to the present invention, exhibits excellent antioxidant activity and processing suitability. Therefore, it can be usefully utilized in processed foods or health functional food compositions for antioxidant purposes that include the protein derived from a dried seaweed processing by-product. Brief explanation of the drawing

[0021] FIG. 1 is a diagram showing a process for extracting protein derived from dried seaweed processing by-products using distilled water in one embodiment of the present invention. FIG. 2 is a diagram showing a process for extracting protein derived from dried seaweed processing by-products through alkali and acid hydrolysis in one embodiment of the present invention. FIG. 3 is a figure showing a process for extracting protein derived from dried seaweed processing by-products through optimal ultrasonic treatment conditions in one embodiment of the present invention. FIG. 4 is a figure showing a protein material derived from dried seaweed processing by-products in one embodiment of the present invention. FIG. 5 is a figure showing the results of analyzing the characteristics of a protein derived from a dried seaweed processing by-product through electrophoresis in one embodiment of the present invention. Figure 6 is a figure showing the results of analyzing the antioxidant activity of a protein derived from a dried seaweed processing by-product in one embodiment of the present invention. Figure 7 is a figure showing the FE-SEM imaging results of a meat substitute containing protein derived from dried seaweed processing by-products in one embodiment of the present invention. Specific details for implementing the invention

[0022] Hereinafter, the present invention will be described in detail with reference to the attached drawings and embodiments thereof. However, the following embodiments are presented as examples of the present invention, and if it is determined that a detailed description of a technology or configuration well known to those skilled in the art may unnecessarily obscure the essence of the present invention, such detailed description may be omitted, and the present invention is not limited thereby. The present invention is capable of various modifications and applications within the scope of the claims set forth below and the equivalent scope interpreted therefrom.

[0023] Furthermore, the terminology used in this specification is used to appropriately describe preferred embodiments of the present invention, and may vary depending on the intent of the user or operator, or the conventions of the field to which the present invention belongs. Accordingly, the definitions of these terms should be based on the content throughout this specification. Throughout the specification, when a part is described as “comprising” a certain component, unless specifically stated otherwise, this means that it does not exclude other components but may include additional components.

[0024] Throughout this specification, '%' used to indicate the concentration of a particular substance is (w / w) % for solid / solid, (w / v) % for solid / liquid, and (v / v) % for liquid / liquid, unless otherwise noted.

[0025] The present invention will be described in more detail below.

[0027] In one aspect, the present invention relates to a method for producing a protein derived from a dried seaweed processing byproduct with enhanced antioxidant activity, comprising the steps of: adding distilled water to dried seaweed processing byproduct powder to hydrate it, and then ultrasonically treating it to produce a dried seaweed processing byproduct protein extract; and filtering and drying the dried seaweed processing byproduct protein extract.

[0028] The extract according to the present invention may be obtained by extracting and separating from nature using extraction and separation methods known in the art, and the "extract" defined in the present invention is extracted from dried seaweed processing by-products using a suitable solvent, and includes, for example, crude extracts, extracts soluble in polar solvents, or extracts soluble in non-polar solvents. Any pharmaceutically acceptable organic solvent may be used as a suitable solvent for extracting the extract from the above-mentioned dried seaweed processing by-product, and water or organic solvents may be used, but are not limited thereto; for example, various solvents such as purified water, alcohols having 1 to 4 carbon atoms including methanol, ethanol, propanol, isopropanol, butanol, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, hexane, and cyclohexane may be used alone or in combination. As for the extraction method, any one of the following may be selected and used: hot water extraction, cold maceration extraction, reflux cooling extraction, solvent extraction, steam distillation, ultrasonic extraction, elution, or pressing. In addition, the desired extract may undergo a conventional fractionation process and may be purified using a conventional purification method.

[0029] There are no limitations on the method for preparing the extract of the present invention, and any known method may be used. For example, the extract included in the composition of the present invention may be prepared in a powder state by additional processes such as vacuum distillation, freeze-drying, or spray-drying, from a primary extract obtained by the aforementioned hot water extraction or solvent extraction method. Additionally, a further purified fraction may be obtained from the primary extract using various chromatographic methods such as silica gel column chromatography, thin layer chromatography, or high performance liquid chromatography. Accordingly, in the present invention, the term "extract" encompasses all extracts, fractions, and purified products obtained at each stage of extraction, fractionation, or purification, as well as their dilutions, concentrates, or dried products.

[0030] In one embodiment of the present invention, the distilled water may be added in an amount of 30 to 50 times the weight of the dried seaweed processing byproduct powder, preferably 35 to 45 times, and more preferably 40 times, but is not limited thereto.

[0031] In one embodiment of the present invention, the hydration may be performed for 20 to 28 hours, preferably 22 to 26 hours, more preferably 24 hours, but is not limited thereto.

[0032] In one embodiment of the present invention, the ultrasonic treatment may be performed in a range of 650 to 850 kHz, preferably in a range of 700 to 800 kHz, more preferably in a range of 750 kHz, but is not limited thereto.

[0033] In one embodiment of the present invention, the ultrasonic treatment may be performed for 2,000 to 3,000 seconds, preferably 2,250 to 2,750 seconds, more preferably 2,500 seconds, at an amplitude of 60 to 100%, preferably 100%, but is not limited thereto.

[0034] In one embodiment of the present invention, the method may increase the content of a protein related to antioxidant activity, but is not limited thereto.

[0035] In one embodiment of the present invention, the antioxidant activity-related protein may be one or more proteins selected from the group consisting of Ribulose-1,5-bisphosphate carboxylase / oxygenase large subunit, Ribulose-1,5-bisphosphate carboxylase, Ribulose bisphosphate carboxylase large chain, R-phycoerythrinalpha chain, Phycoerythrinalpha subunit, C-phycocyaninalpha chain, Allophycocyanin gamma subunit and Allophycocyaninalpha chain, but is not limited thereto.

[0037] In one aspect, the present invention relates to a protein derived from a dried seaweed processing by-product produced by the above method.

[0038] In one embodiment of the present invention, the protein derived from the dried processing byproduct is selected from the group consisting of superoxide dismutase, sulfate adenylyltransferase, R-phycoerythrinalpha chain, ribulose-1,5-bisphosphate carboxylase / oxygenase large subunit, ribulose-1,5-bisphosphate carboxylase, ribulose bisphosphate carboxylase large chain, phycoerythrinalpha subunit, multifunctional fusion protein, mitogen-activated protein kinase, hypothetical protein BU14_0400s0008, hypothetical protein BU14_0399s0008, hypothetical protein BU14_0121s0018, histidine--tRNA ligase, DNA topoisomerase 2, C-phycocyaninalpha chain, allophycocyanin gamma subunit, allophycocyaninalpha chain, and 40S ribosomal protein. It may contain one or more proteins, but is not limited thereto.

[0039] In one embodiment of the present invention, the protein derived from the dried processing byproduct may comprise 35 to 45%, preferably 37 to 43%, more preferably 39.67%, of a protein with a molecular weight of 30 to 70 kDa, and 55 to 65%, preferably 57 to 63%, more preferably 60.33%, of a protein with a molecular weight of 1 to 30 kDa, but is not limited thereto.

[0041] In one aspect, the present invention relates to a processed food containing a protein derived from the processing by-product of dried seaweed.

[0042] The processed food of the present invention may be meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, chewing gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, or vitamin complexes containing protein derived from dried seaweed processing by-products, and includes all processed foods in the conventional sense.

[0043] In one embodiment of the present invention, the protein derived from the dried seaweed processing by-product may be added at a concentration of 1 to 10 ppm, 3 to 8 ppm, or 5 ppm, but is not limited thereto.

[0044] In one embodiment of the present invention, the processed food may be capable of replacing meat, but is not limited thereto.

[0045] In one embodiment of the present invention, the processed food may have enhanced antioxidant activity, but is not limited thereto.

[0046] In one embodiment of the present invention, the processed food may have improved processing suitability, but is not limited thereto.

[0048] In one aspect, the present invention provides a health functional food composition for antioxidant purposes comprising a protein derived from the processing by-product of dried seaweed as an active ingredient.

[0049] In addition to containing protein derived from dried seaweed processing by-products as an active ingredient, the food composition of the present invention may contain various flavoring agents or natural carbohydrates, etc., as additional ingredients, as in conventional food compositions.

[0050] Examples of the natural carbohydrates described above include monosaccharides, e.g., glucose, fructose, etc.; disaccharides, e.g., maltose, sucrose, etc.; polysaccharides, e.g., dextrin, cyclodextrin, etc., and conventional sugars, and sugar alcohols such as xylitol, sorbitol, erythritol, etc. The flavoring agents described above may advantageously use natural flavoring agents (taumatin), stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.), and synthetic flavoring agents (saccharin, aspartame, etc.). The food composition of the present invention may be formulated in the same manner as the pharmaceutical composition described above and used as a functional food or added to various foods. Foods to which the composition of the present invention may be added include, for example, beverages, meat, chocolate, food products, confectionery, pizza, ramen, other noodles, chewing gum, candy, ice cream, alcoholic beverages, vitamin complexes, and health supplements.

[0051] In addition, the above food composition may contain, in addition to the active ingredient protein derived from dried seaweed processing by-products, various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic and natural flavoring agents, coloring agents and thickening agents (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. Furthermore, the food composition of the present invention may contain fruit pulp for the production of natural fruit juice, fruit juice beverages, and vegetable beverages.

[0052] The functional food composition of the present invention may be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc. In the present invention, the term "health functional food composition" refers to a food manufactured and processed using raw materials or ingredients having functional properties useful to the human body pursuant to Article 6727 of the Act on Health Functional Foods, and means consuming it for the purpose of obtaining effects useful for health uses, such as regulating nutrients or physiological actions regarding the structure and function of the human body. The health functional food of the present invention may include conventional food additives, and unless otherwise stipulated, suitability as a food additive is determined according to the specifications and standards for the relevant item in accordance with the general provisions and general test methods of the Food Additives Codex approved by the Ministry of Food and Drug Safety. Items listed in the aforementioned "Food Additives Codex" include, for example, chemically synthesized compounds such as ketones, glycine, calcium citrate, nicotinic acid, and cinnamon acid; and natural additives such as persimmon dye, licorice extract, crystalline cellulose, sorghum dye, and guar gum. Examples include mixed preparations such as L-sodium glutamate preparations, alkaline additives for noodles, preservative preparations, and tar dye preparations. For instance, a health functional food in tablet form may be produced by granulating a mixture of the active ingredient of the present invention with excipients, binders, disintegrants, and other additives using a conventional method, and then compression molding by adding a lubricant, etc., or by directly compression molding the mixture. Additionally, the health functional food in tablet form may contain a binder, etc., as necessary. Among health functional foods in capsule form, hard capsules may be manufactured by filling a conventional hard capsule with a mixture of the active ingredient of the present invention mixed with additives such as excipients, and soft capsules may be manufactured by filling a capsule base such as gelatin with a mixture of the active ingredient of the present invention mixed with additives such as excipients. The soft capsules may contain plasticizers such as glycerin or sorbitol, coloring agents, preservatives, etc., as necessary.A health functional food in the form of a pill can be prepared by molding a mixture of the active ingredient of the present invention, excipients, binders, disintegrants, etc., using a previously known method, and if necessary, it can be coated with sucrose or other coating agents, or the surface can be coated with a substance such as starch or talc. A health functional food in the form of a granule can be prepared by making a mixture of the active ingredient of the present invention, excipients, binders, disintegrants, etc., into a granular form using a previously known method, and may contain flavoring agents, stimulating agents, etc., if necessary.

[0054] Hereinafter, embodiments of the present invention will be described in more detail with reference to the attached drawings. However, the following embodiments are intended only to embody the content of the present invention and are not to limit the present invention.

[0056] <Example 1> Preparation of Protein Extract from Dried Seaweed Processing By-products

[0057] The dried seaweed processing by-product used in this invention was a by-product generated after processing dried seaweed at a processing plant located in Seocheon-gun, Chungcheongnam-do. The dried seaweed processing by-product was powdered using a food grinder, and then proteins were extracted through hydration and physical treatment processes. The hydration and physical treatment processes were carried out as shown in Table 1 below, and seven types of extracts were prepared according to the process. Ultrasonic treatment was performed using a probe-type ultrasonic processor (VCX-750, Deail tech, Korea) with a 750 kHz output, with an amplitude and duty cycle of 60% each, and a treatment time of 1,500 seconds.

[0059] Extraction conditions extraction process Ultrasonic treatment P1 Extracted in distilled water for 24 hours X P2 Extracted in distilled water for 24 hours O P3 Extract in distilled water for 30 minutes X P4 Extract in distilled water for 30 minutes O AK1 Extraction in 1N NaOH for 24 hours X AK2 After extraction in distilled water for 24 hours, adjust to pH 11 using 1N NaOH. X AC Extraction in 9% HCl for 4 hours X

[0061] The extraction process of P1, P2, P3, and P4 in Table 1 above is shown in Fig. 1, and the extraction process of AK1, AK2, and AC in Table 1 is shown in Fig. 2.

[0063] <Experimental Example 1> Selection of Optimal Extraction Conditions

[0064] 1-1. Selection of Optimal Extraction Time and Ultrasonic Treatment Conditions

[0065] The protein content and protein extraction efficiency of four extracts (P1, P2, P3, P4) prepared in Example 1 with different hydration times and ultrasonic treatment conditions were measured and are shown in Table 2 below. Protein content was measured at a wavelength of 540 nm using a UV spectrometer (μ2, microdigtal, Sejong, Korea), and a calibration curve for calculating protein content was constructed using Bovine serum albumin (99%, Sigma-Aldrich, St. Louis, MO, USA).

[0067] Protein content (ppm) Protein extraction efficiency (%) P1 7,001.7 70.0 P2 8,984.7 89.8 P3 5,603.5 56.0 P4 6,124.4 61.2

[0069] As shown in Table 2 above, extraction efficiency was 56.0–61.2% when the hydration time was short (P3, P4), but 70.0–89.8% when the hydration time was long (P1, P2). The extraction efficiency according to ultrasonic treatment was 89.8% and 61.2% for the ultrasonically treated extracts (P2, P4), which increased by 19.8% and 5.2% respectively compared to the extracts not treated with ultrasonic treatment (P1, P3).

[0070] Through this, it was confirmed that tissue softening due to prolonged hydration during physical extraction is effective during ultrasonic extraction, and that ultrasonic extraction has a significant effect on increasing extraction efficiency.

[0072] 1-2. Selection of Optimal Extraction Solvent

[0073] The protein content and protein extraction efficiency of four extracts (P2, AK1, AK2, AC) prepared in Example 1 above with different extraction solvents and hydration times were measured and are shown in Table 3 below. The protein content was measured in the same way as in Experimental Example 1-1 above.

[0075] Protein content (ppm) Protein extraction efficiency (%) P2 8,984.7 89.8 AK1 9,208.3 92.8 AK2 8,723.5 87.2 AC 9,169.7 91.6

[0077] As shown in Table 3 above, the acid hydrolysis extract (AC) showed a yield 1.8% higher than the physical ultrasonic extract (PK2), and the base extract (AK1) showed a yield 3.0% higher than the physical ultrasonic extract (PK2).

[0078] Through this, it was confirmed that there was no significant difference between the extraction methods, with physical extracts at levels of 98.0% compared to the acid digestion method and 96.7% compared to the base extraction method. In addition, physical extraction is considered a suitable technology for materialization because there are no perception concerns regarding the use of acid or base, and it is easy to apply to the process using a portable probe-type ultrasonic processor.

[0079] Therefore, it was confirmed that the most optimal extraction condition is to extract dried seaweed powder by hydrating it in distilled water for an extended period followed by ultrasonic treatment, and the following experiment was conducted under these extraction conditions.

[0081] <Experimental Example 2> Selection of Optimal Ultrasonic Conditions

[0082] After hydrating dried seaweed powder, which was the optimal extraction condition selected in Experimental Example 1 above, in distilled water for 24 hours, a protein extract was prepared with amplitude, duty cycle, and treatment time as independent variables to select the optimal ultrasonic conditions for ultrasonic treatment. The reference frequency intensity for amplitude was 750 kHz, and the unit time for duty cycle was 20 seconds, and the conditions for each independent variable are shown in Table 4 below.

[0084] 1 2 3 4 5 Amplitude (%) 20 40 60 80 100 Duty cycle, %) 20 40 60 80 100 Processing time (Time, seconds) 500 1,000 1,500 2,000 2,500

[0086] When using the portable probe-type ultrasonic processor used in the present invention, the amplitude serves to control the intensity of the maximum output frequency; the frequency is fixed at 750 kHz, but the magnitude of the amplitude is adjusted proportionally to determine the amount of work applied during processing. To prevent heat generation during ultrasonic processing, the ultrasonic cycle is repeated, and the ratio of the cycle to the cycle is called the duty cycle. A duty cycle of 20% for a unit time of 20 seconds repeats the cycle of 4 seconds of operation followed by 16 seconds of suspension, and this is repeated until the total extraction method is achieved.

[0087] The protein content and protein extraction efficiency of the extracts prepared under the treatment conditions of Table 4 above were measured and are shown in Table 5 below. The protein content was measured in the same manner as in Experimental Example 1-1 above. It was determined that there was a significant change in efficiency if the increase in efficiency at each step was 5% or more.

[0089] Processing conditions Amplitude Duty cycle Processing Time Protein content (ppm) Growth rate (%) Protein content (ppm) Growth rate (%) Protein content (ppm) Growth rate (%) 1 8,496.2 - 8,043.5 - 8,141.0 - 2 8,675.5 2.11 8,825.7 9.72 7,883.8 -3.16 3 9,558.5 10.18 9,101.0 3.12 9,700.8 23.05 4 10,397.1 8.77 9,453.5 3.87 10,225.7 5.41 5 12,735.0 22.49 9,923.0 4.97 12,605.2 23.27

[0091] As shown in Table 5 above, regarding Amplitude, extraction efficiency continued to increase as the intensity increased; however, the rate of increase decreased after 60% processing, showing a very large increase of 22.49% at 100% processing. In the case of Duty Cycle, a significant increase was observed when increasing from 20% to 40%, but thereafter, the increase in extraction efficiency was at the 3–4% level, indicating a low increase efficiency relative to energy. Regarding Time, high growth rates of 23.05% and 23.27% were observed at 1,500 seconds and 2,500 seconds, respectively.

[0092] Through this, it was confirmed that extraction efficiency increased as the amplitude increased and the processing time increased, while an increase in duty cycle did not show a significant difference.

[0093] Therefore, the optimal ultrasonic treatment conditions for the preparation of dried seaweed powder extract were selected as an amplitude of 100%, a duty cycle of 40%, and a time of 2,500 seconds, and the following experiment was conducted under these ultrasonic treatment conditions.

[0095] <Experimental Example 3> Analysis of Protein Components and Characteristics Derived from Dried Seaweed Processing By-products

[0096] 3-1. Preparation of Protein Derived from Dried Seaweed Processing By-products

[0097] Protein extracts from dried laver processing by-products were prepared under the optimal conditions selected in Experimental Examples 1 and 2 above (see Fig. 3). Processing by-products generated after processing dried laver at a processing plant located in Seocheon-gun, Chungcheongnam-do were crushed into powder and hydrated for 24 hours in distilled water at a ratio of 40 times the weight of the processing by-product powder. Subsequently, proteins were extracted by ultrasonic treatment under conditions of Amplitude 100%, Duty cycle 40%, and Time 2,500 seconds. After protein extraction, primary filtration was performed using a 40-mesh sieve to remove insoluble residues, followed by secondary filtration using diatomaceous earth. To prevent denaturation of functional proteins, the mixture was treated by freeze-drying.

[0099] 3-2. Analysis of Crude Protein Content and Total Amino Acid Composition of Protein Derived from Dried Seaweed Processing By-products

[0100] The protein prepared in Experimental Example 3-1 above (see Fig. 4) has a deep purple color and, as a water-soluble material, has a scent characteristic of seaweed. To confirm the crude protein content and total amino acid composition of this protein, the crude protein content was measured using the semimicro Kjeldahl method, and the total amino acid composition was determined using a quantitative method with an automatic amino acid analyzer.

[0102] Compounds Unit Contein standard deviation Crude protein g / 100g 48.49 0.21 Aspartic acid mg / 100g 4,682.87 137.27 Threonine 1,811.13 164.60 Serine 1,849.09 74.28 Glutamic acid 6,967.47 664.88 Proline 2,608.39 10.38 Glycine 4,218.25 97.69 Alanine 4,580.45 72.55 Cysteine 186.93 0.73 Valine 1,386.18 113.16 Methionine 925.93 135.15 Isoleucine 1,320.15 12.09 Leucine 3,154.29 19.13 Tyrosine 1,365.27 60.64 Phenylalanine 1,680.42 27.65 Histidine 983.32 17.45 Lysine 3,955.31 173.78 Ammonia 0.00 0.00 Arginine 3,815.26 3.91 Total amino acid mg / 100g 45,490.71 1,785.34

[0104] As shown in Table 6 above, the crude protein content of the protein material derived from dried processing by-products is 48.49 g / 100g, which is similar to that of defatted soybean powder with a crude protein content of about 45%. In addition, the content of the seven essential amino acids (isoleucine, leucine, lysine, methionine, phenylalanine, threonine, and valine) among the total amino acid content was found to be 22.6%.

[0106] 3-3. Characterization of Proteins Derived from Dried Seaweed Processing By-products

[0107] To determine the characteristics and denaturation status of the proteins prepared in Experimental Example 3-1 above, SDS-gel electrophoresis was performed, and each protein was loaded in equal amounts of 150 μg. To cross-validate the determination of protein molecular weight and characteristics, 2DE-gel electrophoresis was performed simultaneously. After extracting protein spots from the 2DE-gel, the protein composition was determined by identifying the proteins at each location. The identification and differentiation of each protein were performed using the MASCOT program, which is shown in Figure 5. Figure 5A shows the results of SDS-gel electrophoresis, where sample 1 is the acid hydrolysis extract, sample 2 is the base hydrolysis extract, sample 3 is the ultrasonic extract, and sample 4 is the ultrasonic extracted and dried sample. In addition, Figure 5B shows the results of 2DE-gel electrophoresis, and the results of MALDI-TOF protein quantification analysis through 2DE-gel protein separation are shown in Table 7 below.

[0109] Protein Molecular weight (kDa) Area Ratio(%) Superoxide dismutase 27.46 3,794.84 0.38 Sulfate adenylyltransferase 48.20 2,302.81 0.23 R-phycoerythrinalpha chain 14.85 25,681.55 2.58 Ribulose-1,5-bisphosphate carboxylase / oxygenase large subunit 60.46 35,535.04 3.58 Ribulose-1,5-bisphosphate carboxylase 14.10 169,123.2 17.00 Ribulose bisphosphate carboxylase large chain 54.15 167,233.89 16.81 Phycoerythrinalpha subunit 13.99 26,366.775 2.65 Multifunctional fusion protein 13.81 246,612.85 24.80 Mitogen-activated protein kinase 52.71 34,449.65 3.46 Hypothetical protein BU14_0400s0008 53.53 10,850.91 1.09 Hypothetical protein BU14_0399s0008 51.89 58,879.17 5.92 Hypothetical protein BU14_0121s0018 52.88 51,977.77 5.23 Histidine-tRNA ligase 49.66 33,353.91 3.35 DNA topoisomerase 2 5.75 3,633.18 0.37 C-phycocyaninalpha chain 13.96 85,336.08 8.58 Allophycocyanin gamma subunit 27.87 18,686.06 1.88 Allophycocyaninalpha chain 13.76 17,415.245 1.75 40S ribosomal protein 28.75 3,370.46 0.34 Total protein - 994,603.39 100

[0111] As shown in Fig. 5A, the molecular weight of the dried seaweed protein is divided into a high molecular weight group of 40–50 kDa and 60 kDa and a low molecular weight group of 20 kDa, and the same results were observed in the 2DE-gel of Fig. 5B. In addition, the isoelectric point of the protein was found in acidic conditions around pH 2–7.

[0112] As shown in Fig. 5A, to confirm protein denaturation, samples extracted via acid digestion and those extracted using a basic solvent were simultaneously loaded and examined. In samples 1 and 2, a band of 40–50 kDa, representing a high molecular weight protein, was observed, but it appeared fainter compared to samples 3 and 4. Additionally, in samples 1 and 2, the band of 20 kDa, representing a low molecular weight protein, was faint and difficult to identify.

[0113] Through this, the effects of preventing protein denaturation and preserving protein due to physical extraction were confirmed.

[0114] As shown in Table 7 above, the protein identification results indicate that the protein with the highest content among the total proteins is Ribulose-1,5-bisphosphate carboxylase (hereinafter RubisCo) related protein (Ribulose-1,5-bisphosphate carboxylase / oxygenase large subunit, Ribulose-1,5-bisphosphate carboxylase, and Ribulose bisphosphate carboxylase large chain), accounting for 37.38% of the total protein composition. RubisCo is a peptide found in plants and algae that plays a key role in CO2 assimilation during photosynthesis. It is known to have antioxidant and antibacterial effects as an enzyme that prevents photooxidation and synthesizes energy through photocatalysis. The second most abundant protein is phytochemical-related protein (R-phycoerythrinalpha chain, phycoerythrinalpha subunit, C-phycocyaninalpha chain, allophycocyanin gamma subunit, and allophycocyaninalpha chain), which accounts for 17.44% of the total protein composition. Phytochemical-related proteins are associated with the pigment proteins of dried laver, and phycocyanin and phycoerythrin are known to have antioxidant effects as the blue and red pigments found in laver, respectively.

[0115] Through this, it was confirmed that the content of RubisCo-related proteins and phytochemical-related proteins, which are known to have antioxidant activity, is high in proteins derived from dried seaweed processing by-products.

[0116] Therefore, an experiment was conducted to measure the antioxidant activity of proteins derived from dried seaweed processing by-products.

[0118] 3-4. Analysis of Antioxidant Activity of Proteins Derived from Dried Seaweed Processing By-products

[0119] The total content of pigment protein and RubisCo protein contained in the protein derived from the dried seaweed processing by-product prepared in Experimental Example 3-1 was 54.82% (see Table 7), and the functionality was confirmed by measuring the antioxidant activity of the protein derived from the dried seaweed processing by-product accordingly.

[0120] The concentration of the DPPH reagent for confirming the antioxidant activity of the protein was prepared at 0.2 mM, and the protein sample was prepared in the form of a 0% aqueous solution and then diluted 20-fold in the DPPH reagent to confirm the activity. The measured DPPH antioxidant activity is shown in Figure 6 and Table 8.

[0122] Sample concentration (ppm) Average content (%) Standard deviation (%) control 0 0 0.625 14.06 4.73 1.25 45.86 3.36 2.5 71.97 0.85 5 84.42 0.33 10 80.62 1.04

[0124] As shown in Table 8 above, the DPPH activity of the protein material derived from dried seaweed processing by-products increased up to a protein concentration of 5 ppm, reaching a peak antioxidant activity of 84.42%, and then decreased to 80.62% at 10 ppm.

[0125] Through this, it was confirmed that the antioxidant activity of the protein material derived from dried seaweed processing by-products is best when the concentration is 5 ppm.

[0127] <Experimental Example 4> Preparation of a Meat Substitute Containing Protein Derived from Dried Seaweed Processing By-products and Confirmation of Characteristics

[0128] 4-1. Manufacture of meat substitutes containing protein derived from dried seaweed processing by-products

[0129] To produce a meat substitute containing protein prepared in Experimental Example 3-1 above, a meat substitute was prepared using the formulation ratios shown in Table 9 below. As a control, a meat substitute that did not contain protein derived from dried seaweed processing by-products was used. An extrusion machine was used to produce a low-moisture meat substitute using the formulation ratios shown in Table 9 below.

[0131] Defatted soybean powder Dried seaweed protein Wheat gluten corn starch Control group 1,500 g - 1,200 g 300 g 5% Protein Added 1,350 g 150 g 1,200 g 300 g

[0133] 4-2. Characteristic Analysis of Meat Substitutes

[0134] To analyze the protein mixing defects and fibrosis results of the meat substitute containing protein derived from dried seaweed processing by-products prepared in Experimental Example 4-1 above, FE-SEM (Field Emission-Scanning Electron Microscope) imaging was performed and is shown in Fig. 7.

[0135] As shown in Figure 7, the output results of the low-moisture meat substitute with the addition of a protein material derived from dried seaweed processing by-products showed a distinct purple color, but no morphological differences or changes in appearance due to poor mixing were observed. In addition, FE-SEM imaging results showed that the protein material derived from dried seaweed consists of irregular fragments and has a sharp shape; however, when the meat substitute is manufactured by adding this material, the shape resulting from these fragments is not observed, and the formation of a fibrous structure similar to the control group is observed.

[0136] Based on this, it is determined that poor mixing and impaired processing suitability do not occur when manufacturing meat substitutes containing protein derived from dried seaweed processing by-products.

[0138] 4-3. Analysis of Physicochemical Content of Meat Substitutes

[0139] To analyze the physicochemical components of the meat substitute containing protein derived from dried seaweed processing by-products prepared in Experimental Example 4-1 above, moisture, ash, crude protein, and crude lipid content were measured and are shown in Table 10 below.

[0141] Control group (g / 100g) Protein added from dried seaweed (g / 100g) moisture 52.2±0.0 57.6±0.1 Ash 0.015±0.0 0.020±0.0 crude protein 32.5±0.1 34.4±0.1 George 2.7±0.1 1.6±0.1

[0143] As shown in Table 10 above, the moisture content of the meat substitute containing protein derived from dried seaweed processing by-products increased by approximately 5%, while, in contrast, the crude fat content decreased. There was no significant change in the crude protein and ash content.

[0144] Through this, it was confirmed that the moisture content increased due to the increased water retention capacity upon addition, resulting from the characteristics of the protein material derived from dried seaweed processing by-products, which has high water solubility.

[0146] 4-4. Analysis of Antioxidant Activity of Meat Substitutes

[0147] To analyze the antioxidant activity of the meat substitute containing the protein derived from the dried seaweed processing by-product prepared in Experimental Example 4-1 above, the DPPH activity expression concentration was determined and is shown in Table 11 below.

[0149] Sample concentration (ppm) Control group Dried seaweed-derived protein additive Average content (%) Standard deviation (%) Average content (%) Standard deviation (%) control 0.00 0.00 0.00 0.00 2.5 -0.73 0.29 6.50 0.85 5 1.59 0.57 10.33 0.90 10 -6.94 0.50 8.17 2.89

[0151] As shown in Table 11 above, the control group exhibited negative values ​​when the sample concentration was around 5 ppm, regardless of the concentration, so it was determined that there was no antioxidant function. On the other hand, the group with added dried seaweed protein showed the highest antioxidant activity at a sample concentration of 5 ppm.

[0152] Through this, it was confirmed that there are no issues such as thermal denaturation or loss of functionality when protein materials derived from dried seaweed processing by-products are added, and that functionality is transferred.

[0154] Synthesizing these results, it was confirmed that a meat substitute produced using a protein derived from dried seaweed processing by-products, prepared by hydrating the dried seaweed processing by-products with distilled water for 24 hours and then ultrasonically treating them (amplitude 100%, duty cycle 40%, time 2,500 seconds), exhibited excellent antioxidant activity and processing suitability, thereby confirming that a meat substitute with enhanced functionality can be produced using a protein derived from dried seaweed processing by-products.

[0156] As described above, specific embodiments of the present invention have been described in detail; however, those skilled in the art who understand the spirit of the present invention will be able to easily propose other inventions that are inferior or other embodiments included within the scope of the spirit of the present invention by adding, changing, or deleting other components within the same spirit. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive. The scope of the present invention is defined by the claims set forth below rather than by the detailed description above, and all modifications or variations derived from the meaning and scope of the claims and equivalent concepts should be interpreted as being included within the scope of the present invention.

Claims

Claim 1 A method for producing a protein derived from a dried seaweed processing byproduct with enhanced antioxidant activity, comprising the steps of: adding distilled water to dried seaweed processing byproduct powder to hydrate it, and then ultrasonically treating it to produce a dried seaweed processing byproduct protein extract; and filtering and drying the dried seaweed processing byproduct protein extract, wherein the hydration is performed for 20 to 28 hours, the ultrasonically treatment is performed at 20 kHz in the range of 650 to 850 W, and the ultrasonically treatment is performed for 2,000 to 3,000 seconds with an amplitude of 60 to 100%, and the method is characterized by increasing the content of proteins related to antioxidant activity. Claim 2 delete Claim 3 delete Claim 4 delete Claim 5 delete Claim 6 A method for producing a protein derived from a dried seaweed processing by-product with enhanced antioxidant activity, wherein, in claim 1, the antioxidant activity-related protein is one or more proteins selected from the group consisting of Ribulose-1,5-bisphosphate carboxylase / oxygenase large subunit, Ribulose-1,5-bisphosphate carboxylase, Ribulose bisphosphate carboxylase large chain, R-phycoerythrinalpha chain, Phycoerythrinalpha subunit, C-phycocyaninalpha chain, Allophycocyanin gamma subunit, and Allophycocyaninalpha chain. Claim 7 Protein derived from dried seaweed processing by-products produced by the method of claim 1. Claim 8 In claim 7, the protein derived from the dried processing by-product is one selected from the group consisting of superoxide dismutase, sulfate adenylyltransferase, R-phycoerythrinalpha chain, ribulose-1,5-bisphosphate carboxylase / oxygenase large subunit, ribulose-1,5-bisphosphate carboxylase, ribulose bisphosphate carboxylase large chain, phycoerythrinalpha subunit, multifunctional fusion protein, mitogen-activated protein kinase, hypothetical protein BU14_0400s0008, hypothetical protein BU14_0399s0008, hypothetical protein BU14_0121s0018, histidine--tRNA ligase, DNA topoisomerase 2, C-phycocyaninalpha chain, allophycocyanin gamma subunit, allophycocyaninalpha chain, and 40S ribosomal protein. Protein derived from dried seaweed processing by-products, characterized by containing the above proteins. Claim 9 A processed food containing protein derived from the processing by-product of dried seaweed of Paragraph 7. Claim 10 A processed food according to claim 9, characterized in that the protein derived from the dried seaweed processing by-product is added at a concentration of 1 to 10 ppm. Claim 11 In claim 9, the processed food is characterized by being able to replace meat. Claim 12 In claim 9, the above processed food is characterized by having enhanced antioxidant activity. Claim 13 In claim 9, the above processed food is characterized by having improved processing suitability. Claim 14 An antioxidant health functional food composition comprising a protein derived from the processing by-product of dried seaweed of claim 7 as an active ingredient.

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