KR102418317B1 - Mutant Porphyra yezoensis 500PP with increased content of polysaccharide with antioxidant activity and preparing method thereof - Google Patents

Abstract

The present invention relates to mutant radish laver 500PP having an increased content of polysaccharide having antioxidant activity and a manufacturing method thereof, and the mutant radish laver 500PP has increased antioxidant activity and can be used as a health functional food. It can be effectively used to develop high value-added products.

Description

Mutant Porphyra yezoensis 500PP with increased content of polysaccharide with antioxidant activity and preparing method thereof

The present invention relates to mutant Porphyra yezoensis 500PP having an increased content of polysaccharide having antioxidant activity and a method for preparing the same, and more particularly, to a mutant having an increased content of polysaccharide having antioxidant activity by treatment with ethyl methanesulfonate. It relates to a radiant pattern laver 500PP and a manufacturing method thereof.

Seaweed ( Porphyra ) is a kind of seaweed belonging to the red algae, and Porphyra of the red algae (Rhodophyta), the red algae (Rho-dophyceae), the primitive red algae (Bangiophyci-dae), ) is an edible seaweed belonging to the genus.

Seaweed is a red algae that is mainly farmed and used as food in Korea, Japan, and China. It grows naturally by attaching to rocks in the sea. The types of laver that are cultivated are mainly radish laver ( Porphyra yezoensis ) and sea laver ( Porphyra tenera ). About 50 species are grown, and about 10 species are distributed in Korea, including Porphyra suborbiculata , Porphyra pseudolinearis , and Porphyra seriata .

According to the 2011 fisheries and marine environment statistics, the total domestic aquaculture production and total production amount was reported to be about 1.35 million tons, or 1.8 trillion won. Among them, the production of seaweed aquaculture is 900,000 tons, which is equivalent to 380 billion won, and the proportion of the total aquaculture is 66.5% of production and 20.8% of production amount, accounting for a very large proportion.

About 98% of the produced seaweed is produced in the aquaculture industry, so the proportion of the aquaculture industry in the production of seaweed is very high. The production of aquacultured seaweed in 2010 increased by about two times compared to that of 2000, and although there is a slight difference in production since 2005, it has been maintained at a similar level. Although the seaweed aquaculture industry is generally considered to be in a period of stabilization, it is expected that there may be differences in production and production amount in the future as there are various variables such as the implementation of the aquatic plant variety protection system.

According to 2011 fisheries and marine environment statistics, the total production and production amount of seaweed in Korea is about 1 million tons, or 400 billion won. Seaweed, kelp, and laver were surveyed as major products in the total seaweed production, and seaweed was the third largest, producing about 320,000 tons, accounting for about 32% of the total seaweed production. Since the main consumption forms of kelp and seaweed are as feed for shellfish farming such as abalone, seaweed accounts for the highest proportion of seaweeds used for food.

In 2011, seaweed production amounted to KRW 220 billion, accounting for the highest proportion (58% of total seaweed production) among all seaweeds. In view of the amount of production and the scale of production, seaweed occupies a very important position as the main product of seaweed farmers. The laver farming industry occupies a very large portion regionally, economically and industrially, but the laver seed industry is at a very small level. The production of seaweed seedlings slightly increased from 2.84 million boxes of 7 billion won in 2009 to 3.67 million boxes and 9.2 billion won in 2012. In the total production of laver seedlings, regular laver (same laver, radish laver) accounted for about 40% of the total, while it-bodied laver and seed-patterned laver each accounted for about 30%.

The production value of laver seedlings is 7 to 9 billion won in total, which accounts for a very small proportion in the total laver aquaculture industry (220 billion won), but it occupies a very important position in the industrial position because the production of laver seedlings directly affects the laver culture industry. do. Recently, the government selected laver as one of the top 10 strategic export items, established a nurturing plan to preoccupy the global market, and proposed the international standard for seaweed to the Asian Food Standards Committee in 2010. In addition, at the 34th International Food Standards Commission (CODEX) General Assembly in 2011, it was decided to establish a standard standard for seaweed with the consent of 184 member countries around the world. Accordingly, Korea can take a leading role in the process of enacting international standards for laver in the future as a proposal country, and by inducing international standards for seaweed to be as consistent with domestic standards as possible, it is expected to greatly help in maintaining and pioneering the global export market for seaweed. It is expected.

However, at present, about 25 to 40% of the total seaweed production is foreign seeds (hybrids mixed with Japanese), which may cause problems such as payment of royalties. In order to solve this problem, the development of new seeds as an import substitute is too difficult for a small laver seedling producer to carry out, and the development of excellent import substitute seeds is urgently required at the national level.

However, there are very few studies on the breeding of seaweed seeds. In Korea, studies on the improvement of laver varieties include the characteristics of infancy aquaculture targeting large seaweed and large radish laver, an adaptation test for deriving suitable varieties for each sea area, and low-salt resistance cultivar improvement by tissue culture of radish laver. However, it did not lead to practical use or industrialization. The Mokpo Branch of the Namhae Fisheries Research Institute has carried out breed improvement targeting of the black laver and radish laver that have been cultivated for a long time in the southern seas since 1994, as well as the stone laver that is native to the coast of Korea. (Namhae Fisheries Research Institute, 2004, Development of Seaweed Culture Varieties and Technologies for Regional Characteristics of the South Coast, 2003 Business Report, pp 571-600).

In Japan, breeding studies for improving laver varieties have been conducted since the 1960s, and in particular, large seaweed laver was developed in 1962 by growers and large radish laver in 1967 as fast-growing strains (Miura, A., PF). Fu and JA Shin, 1992. lnterspecific cross experiments between Porphyra yezoensis Ueda and P. tenera Kjellman (Bangiales, Rhodophyta) by using pigmentation variants. J. Tokyo Univ. Fish., 79, 103-120.). In addition, protoplast cell fusion studies were conducted to develop new varieties, but they were not cultivated as practical varieties.

Seaweed contains a large amount of sugar. The main sugars are free sugars such as isofloridoside and floridoside, hemicellulose, an insoluble polysaccharide as a cell wall component, and a water-soluble acidic filling material between cells. polysaccharides such as porphyran.

Among them, porphyran is a sulfur-containing polysaccharide extracted from seaweed, and it has been reported to reduce the incidence of colorectal cancer by facilitating intestinal activity and diluting the toxicity of toxic components when ingested. Facts such as proliferation-promoting effects have been revealed. In addition, porphyran has a very high value as an undigested dietary fiber, and its high viscosity is increasing its potential as an excipient for physical properties of food and its use value.

Until now, studies on seaweed have mainly focused on physicochemical properties of seaweed, but recently, interest in using bioactive substances as functional foods is increasing. However, in order to overcome the backwardness and vulnerability of the domestic seaweed processing industry, high value-added products are required, but seaweed processing mainly relies on simple processing.

Accordingly, the present inventors treated ethyl methane sulfonate to fermented laver while researching the culture and breed improvement of P. yezoensis to obtain mutant radish laver 500PP ( P. yezoensis 500PP), and to obtain a normal radish laver. It was confirmed that the polysaccharide content and its antioxidant activity were superior to laver.

Accordingly, it is an object of the present invention to provide a mutant radioactive laver ( Porphyra yezoensis ) 500PP deposited with accession number KCTC13957BP with an increased content of polysaccharides having antioxidant activity by treatment with ethyl methanesulfonate.

Another object of the present invention is to provide a method for preparing mutant radioactive laver 500PP having an increased content of polysaccharides having antioxidant activity, comprising the following steps:

A culturing step of culturing by treatment with ethyl methanesulfonate in radish pattern; and

A screening step of selecting radiopatterns having an increased content of polysaccharides having an antioxidant activity than those not treated with ethyl methanesulfonate from among the cultured radiopatterns.

Another object of the present invention is to provide a health functional food composition having antioxidant activity comprising a mutant radioactive laver deposited with accession number KCTC13957BP ( Porphyra yezoensis ) 500PP extract.

Another object of the present invention relates to the use of polysaccharides having antioxidant activity of 500PP mutant radiopattern prepared by treatment with ethyl methanesulfonate.

The present invention relates to a mutant radioactive laver ( Porphyra yezoensis ) 500PP having an increased content of polysaccharide having antioxidant activity and a method for preparing the same. increased compared to seaweed.

The present inventors treated ethyl methanesulfonate to radish laver to obtain mutant radish laver 500PP ( P. yezoensis 500PP) with increased polysaccharide content, and confirmed that the antioxidant activity of polysaccharides was superior to that of normal radish laver.

Hereinafter, the present invention will be described in more detail.

One aspect of the present invention is a mutant radioactive laver ( Porphyra yezoensis ) 500PP deposited with accession number KCTC13957BP with an increased content of polysaccharides having antioxidant activity by treatment with ethyl methanesulfonate.

The ethyl methanesulfonate may be treated at a concentration of 1 to 5 mM, 2 to 5 mM, or 3 to 5 mM, for example, may be treated at a concentration of 4 to 5 mM, but is limited thereto not. When the ethyl methanesulfonate is treated with a concentration of 1 mM or less, the frequency of occurrence of mutations may not be sufficient, and when treated with a concentration of more than 5 mM, the survival rate of seaweed may be significantly reduced.

The mutant radiograss 500PP may be obtained as spores released from radiograss treated with ethyl methanesulfonate.

In one embodiment of the present invention, the content of polysaccharides having antioxidant activity and antioxidant activity was increased from 500PP of mutant radiopattern prepared by treatment with ethyl methanesulfonate. Accordingly, the antioxidant activity of 500PP mutant radish laver was higher than that of wild-type laver not treated with ethyl methanesulfonate.

The polysaccharide having antioxidant activity may be a water-soluble sulfur-containing polysaccharide, and the sulfur-containing polysaccharide extractable from laver is usually referred to as porphyran.

Another aspect of the present invention is a method for preparing mutant radioactive laver ( Porphyra yezoensis ) 500PP deposited with accession number KCTC13957BP with increased polysaccharide content having antioxidant activity, comprising the following steps:

A culturing step of culturing by treatment with ethyl methanesulfonate in radish pattern; and

A screening step of selecting radiopatterns having an increased content of polysaccharides having an antioxidant activity than those not treated with ethyl methanesulfonate from among the cultured radiopatterns.

The ethyl methanesulfonate may be treated at a concentration of 1 to 5 mM, 2 to 5 mM, or 3 to 5 mM, for example, may be treated at a concentration of 4 to 5 mM, but is limited thereto not. When the ethyl methanesulfonate is treated with a concentration of 1 mM or less, the frequency of occurrence of mutations may not be sufficient, and when treated with a concentration of more than 5 mM, the survival rate of seaweed may be significantly reduced.

The culturing step may be performed in MGM (Modified Grund Media) medium, for example, C ₁₀ H ₁₄ O ₆ N ₂ Na ₂ ·2H ₂ O (Na ₂ EDTA), FeSO ₄ ·7H ₂ O, Na ₂ HPO ₄ ·12H ₂ O, NaNO ₃ , MnCl ₂ ·7H ₂ O, and may be carried out in MGM medium containing vitamin B12 (Cyanocobalamin), but is not limited thereto.

The MGM medium can be used as a seawater medium in which MGM medium is added to general seawater. The seawater may have a favorable effect on the growth of radish laver to use a mixture of surface water and deep water in a 1:1 ratio.

The culturing step may be performed at a light intensity of 10 to 50, 10 to 40, 10 to 30, 10 to 25, 15 to 50, 15 to 40, 15 to 30 or 15 to 25 umol photon m ^-2 s ^-1 and, for example, may be cultured at a light intensity of 20 umol photon m ^-2 s ^-1 , but is not limited thereto. Cultivation in the above temperature range shows the most excellent leaf length and leaf width of radish laver.

The culturing step may be performed in a light:dark cycle of 10:14 to 14:10 or 11:13 to 13:11, for example, it may be performed in a light:dark cycle of 12:12, but is limited thereto. it's not going to be

The culturing step may be performed at a temperature of 10 to 25°C, 10 to 20°C, or 15 to 25°C, for example, 15 to 20°C, but is not limited thereto.

The culturing step may include the following steps:

A first culturing step of culturing the radioactive laver treated with ethyl methanesulfonate;

a second culturing step of selecting and culturing the surviving radiographic laver in the first culturing step; and

A third culturing step of culturing the spores released by the radiopatterns in the second culturing step.

The selection step is 30 to 70, 30 to 60, 30 to 55, 40 to 70, 40 to 60, 40 to 55 or 45 to 70 umol photon m ^-2 s ^-1 , for example, 45 to 60 umol photon m It may be carried out after inducing spore formation from radiation patterning at a luminosity of ^-2 s ^-1 , for example, inducing spore formation at a luminosity of 45 to 55 umol photon m ^-2 s ^-1 it could be

In one embodiment of the present invention, the first culturing step may be performed at 15° C. for 12 weeks, but is not limited thereto.

In one embodiment of the present invention, the second culturing step may be carried out at 10 to 15 ℃ for 12 weeks, for example, it may be carried out at 15 ℃ for 12 weeks, but is not limited thereto.

In one embodiment of the present invention, the third culturing step may be carried out at 10 to 15° C. for 12 weeks, for example, it may be performed at 15° C. for 12 weeks, but is not limited thereto.

The release of the spores may be induced by increasing the culture temperature of the radish laver to 18 to 20 °C, for example, the culture temperature may be increased to 20 °C, but is not limited thereto. At this temperature, the release of spores rather than the growth of the fronds may be effected.

In one embodiment of the present invention, the radiation pattern was treated with 1.0 to 5.0 mM of ethyl methanesulfonate to maintain a light intensity of 50 umol photon m ^-2 s ^-1 or higher at 12:12 light:dark cycle, and a temperature of 15° C. was maintained. After culturing for a week, the spores are released from the fronds, and the separated spores are cultured for 12 weeks at a light intensity of 20 umol photon m ^-2 s ^-1 , 12:12 light:dark cycle, and a temperature of 15°C to have antioxidant activity. Mutant radish laver 500PP was selected by selecting radiopatterns having an increased polysaccharide content.

Another aspect of the present invention is a health functional food composition having antioxidant activity comprising an extract of 500PP mutant radish laver deposited with accession number KCTC13957BP.

The extract includes a crude solvent extract, a specific solvent soluble extract (solvent fraction), and a solvent fraction of the crude solvent extract, and the extract may be in a solution, concentrate or powder state.

The radioactive laver extract may be a crude extract obtained by extracting radioactive laver with water and one or more solvents selected from the group consisting of linear or branched alcohols having 1 to 4 carbon atoms, for example, ethanol as a solvent and may be obtained, but is not limited thereto.

The extraction method used in the present invention may be any method commonly used, for example, cold extraction, hot water extraction, ultrasonic extraction, or reflux cooling extraction method, but is not limited thereto.

The health functional food composition may include the polysaccharide fraction of the 500PP mutant radish laver extract.

The polysaccharide fraction may be a water-soluble sulfur-containing polysaccharide separated from seaweed, but is not limited thereto.

When the health functional food composition of the present invention is used as a food additive, the health functional food composition may be added as it is or used together with other foods or food ingredients, and may be appropriately used according to a conventional method. In general, in the production of food or beverage, the health functional food composition of the present invention may be added in an amount of 15% by weight or less, preferably 10% by weight or less, based on the raw material.

There is no particular limitation on the type of the food. Examples of foods to which the above substances can be added include meat, sausage, bread, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, tea, drinks, There are alcoholic beverages, vitamin complexes, and the like, and includes all foods in a conventional sense.

The beverage may contain various flavoring agents or natural carbohydrates as additional ingredients. As the above-mentioned natural carbohydrates, monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and natural sweeteners such as dextrin and cyclodextrin, synthetic sweeteners such as saccharin and aspartame may be used. . The ratio of the natural carbohydrate may be appropriately determined by the selection of those skilled in the art.

In addition to the above, the health functional food composition of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin , alcohol, carbonation agent used in carbonated beverages, and the like. In addition, the health functional food composition of the present invention may contain fruit for the production of natural fruit juice, fruit juice beverage, and vegetable beverage. These components may be used independently or in combination. The proportion of these additives may also be appropriately selected by those skilled in the art.

The present invention relates to mutant radish laver 500PP having an increased content of polysaccharide having antioxidant activity and a manufacturing method thereof, and the mutant radish laver 500PP has increased antioxidant activity and can be used as a health functional food. It can be effectively used to develop high value-added products.

1 is a mutant laver 500PP ( P. yezoensis 500PP ) prepared by treatment with ethyl methanesulfonate according to the present invention and general radiograss (P. yezoensis) that is not treated with ethylmethanesulfonate, respectively. The figure shows the antioxidant activity of the polysaccharide fraction.
2 is a graph showing the content of polysaccharides contained in each polysaccharide fraction derived from mutant radiograss 500PP prepared by treatment with ethyl methanesulfonate according to the present invention and normal radiograss that has not been treated.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, these examples are only for illustrating the present invention, and the scope of the present invention is not limited by these examples.

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

Example 1: Mutant Radial laver 500PP ( P. yezoensis 500PP)

Radiated laver ( Porphyra yezoensis ; P. yezoensis ) was purchased from the Seaweed Research Center of the National Institute of Fisheries Science and used.

Radiated laver was put in MGM (Modified Grund Media) medium, and then put in a plant culture dish (100×40 mm) in a volume of 100 mL, luminosity 20 umol photon m ^-2 s ^-1 , 12:12 people: Dark (Light:Dark) cycle, maintained at a temperature of 15 ℃ was cultured for 12 weeks.

After treatment with ethyl methanesulfonate to a concentration of 1 mM, 2 mM, 3 mM, 4 mM, and 5 mM, respectively, radiopatterned laver being cultured in MGM medium, each treated radiopattern sample is washed and then again MGM medium After culturing for 12 weeks at a light intensity of 20 umol photon m ^-2 s ^-1 , 12:12 light:dark cycle, and maintaining a temperature of 15°C, the viability of seaweed was analyzed through cell counting using a microscope.

After treatment with ethyl methanesulfonate, the surviving radish laver was transferred back to MGM medium and incubated for 12 weeks at a light intensity of 20 umol photon m ^-2 s ^-1 , 12:12 light:dark cycle, and a temperature of 15°C. Release was induced by increasing the incubation temperature of seaweed to 20℃, and the obtained spores were maintained in fresh MGM medium with a light intensity of 20 umol photon m ^-2 s ^-1 , 12:12 light:dark cycle, and the temperature was maintained at 15℃ for 12 weeks. cultured.

The MGM (Modified Grund Media) medium is C ₁₀ H ₁₄ O ₆ N ₂ Na ₂ 2H ₂ O (Na ₂ EDTA) 3.72 g, FeSO ₄ 7H ₂ O 0.28 g, Na ₂ in 1 liter (l) of sterilized seawater HPO ₄ 12H ₂ O 10.74 g, NaNO ₃ 42.5 g, MnCl ₂ 7H ₂ O 0.019197 g, vitamin B ₁₂ (Cyanocobalamin) 1 mg was used.

In the above, from the radioactive laver obtained by culturing the surviving radioactive laver after treatment with ethyl methanesulfonate in MGM medium, the radioactive laver having an increased content of polysaccharides having antioxidant activity is selected than that of radioactive laver that has not been treated with ethyl methanesulfonate. Therefore, it was named as mutant radish laver 500PP ( P. yezoensis 500PP).

Test Example 1: Measurement of antioxidant activity of polysaccharide fraction and polysaccharide of mutant radish laver 500PP

The method for preparing the polysaccharide fraction from radioactive laver and measuring the antioxidant activity of the polysaccharide fraction is as follows. Fat and alcohol-soluble substances were removed by repeating extraction under reflux 3 times for 2 hours at 60°C with 150 mL of 95% ethanol of 10 g of dried laver sample. The powder residue was extracted twice with 1 L distilled water at 60° C. for 2 hours. After filtration, the supernatant was concentrated with a rotary evaporator at 50°C. Then, 95% ethanol was added to the concentrate at 4° C. for 12 hours, the mixture was centrifuged, and the precipitate was dried to recover polysaccharides. The polysaccharide content was expressed as a percentage of the polysaccharide precipitate relative to the dried laver sample.

The sulfation activity of the polysaccharide fraction was measured by DPPH radical scavenging ability by the following method. First, an aqueous solution was prepared by diluting polysaccharides derived from extracted laver. The dilution concentration was applied differently depending on the concentration of the extracted polysaccharide, and specifically, it was diluted so that the absorbance value was between 0.2 and 0.5.

A solution of DPPH radical (molecular weight 394.32 g/mol) was prepared in methanol at a concentration of 0.2 mM and stirred for about 10 minutes. 150 ul of 0.2 mM DPPH solution was added to each well. 150 ul of methanol was added to the negative control group (Blank).

50 ul each of 3 experimental groups (triplicate) and 3 control groups were added to each well of the prepared polysaccharide test sample by concentration. A positive control group containing only DPPH and solvent (standard) and a negative control group containing only methanol instead of DPPH solution (blank) were also tested. The 96-well plate was placed in a 25°C incubator for 30 minutes. The absorbance (absorbance, optical density) was measured at 492 nm with a microplate reader, and the antioxidant power was compared by calculating the radical scavenging power (%) using the average.

Table 1 shows the antioxidant activity of the polysaccharide fraction derived from mutant radish laver 500PP ( P. yezoensis 500PP) and the antioxidant activity of the polysaccharide fraction derived from untreated common radish laver.

Mutant Radial laver 500PP Untreated radiated laver Polysaccharide antioxidant activity (%, w/w) 42.0 31.0 Polysaccharide content (%, w/w) 17.6 14.7

As can be seen in FIG. 1, the polysaccharide fraction derived from mutant radish laver 500PP had an antioxidant activity of about 42.0% (w/w), and the polysaccharide fraction derived from untreated general radish laver had about 31.0% ( w/w) was confirmed to have antioxidant activity. As can be seen in FIG. 2, the content of the extracted polysaccharide fraction was obtained in a yield of about 17.6% (w/w) from 500PP of mutant radiopatterned seaweed, but untreated general Radial laver was extracted in a yield of 14.7% (w/w).

Therefore, the mutant radish laver 500PP obtained in the present invention has an antioxidant activity of 30% or more higher in the extracted polysaccharide fraction, and the content of polysaccharide fraction having excellent antioxidant activity is higher by more than 20%, compared to normal radish laver that is not treated with ethyl methane sulfonate. had a value

Korea Institute of Bioscience and Biotechnology KCTC13957BP 20190919

Claims (12)

Mutant radioactive laver ( Porphyra yezoensis ) 500PP deposited with accession number KCTC13957BP with increased content of polysaccharides having antioxidant activity by treatment with ethyl methanesulfonate. According to claim 1, wherein the treatment amount of the ethyl methanesulfonate will be 1 to 5 mM, Mutant Radix 500PP. The mutant radiograss 500PP according to claim 1, wherein the mutant radiograss 500PP is obtained as spores released from ethyl methanesulfonate-treated radiograss. A method for preparing mutant radioactive laver ( Porphyra yezoensis ) 500PP deposited with accession number KCTC13957BP with increased polysaccharide content having antioxidant activity, comprising the following steps:
A culturing step of culturing by treatment with ethyl methanesulfonate in radish pattern; and
A screening step of selecting radiopatterns having an increased content of polysaccharides having an antioxidant activity than those not treated with ethyl methanesulfonate from among the cultured radiopatterns. The method of claim 4, wherein the treatment amount of the ethyl methanesulfonate is 1 to 5 mM. According to claim 4, wherein the culturing step is C ₁₀ H ₁₄ O ₆ N ₂ Na ₂ ·2H ₂ O (Na ₂ EDTA), FeSO ₄ ·7H ₂ O, Na ₂ HPO ₄ ·12H ₂ O, NaNO ₃ , MnCl ₂ ·7H ₂ O, vitamin B ₁₂ (Cyanocobalamin; cyanocobalamin) will be carried out in a medium containing, the method for producing a mutant radioactive laver 500PP. The method of claim 4, wherein the culturing step is performed at a light intensity of 10 to 50 umol/m ² s. The method of claim 4, wherein the culturing step is performed in a 12h:12h light:dark cycle. The method of claim 4, wherein the culturing step is performed at a temperature of 10 to 25°C. [Claim 5] The method of claim 4, wherein the culturing step comprises the following steps:
A first culturing step of culturing the radioactive laver treated with ethyl methanesulfonate;
a second culturing step of selecting and culturing the surviving radish in the first culturing step; and
A third culturing step of culturing the spores released by the radiopatterns in the second culturing step. A health functional food composition having antioxidant activity comprising a 500PP extract of mutant radioactive laver deposited with accession number KCTC13957BP. The health functional food composition according to claim 11, wherein the health functional food composition comprises the polysaccharide fraction of the 500PP mutant radish laver extract.

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