KR100606499B1 - Skin whitening materials isolated from starfishes by protease treatment - Google Patents

Abstract

The present invention relates to an extract for skin whitening derived from starfish by proteolytic enzyme treatment, and the present invention has an excellent effect of providing a bioactive material having a whitening effect of skin extracted from starfish.

Amr starfish, star starfish, bioactive substance, protease, whitening effect

Description

Skin whitening extract from starfish by protease treatment {Skin whitening materials isolated from starfishes by protease treatment}

1 is a graph comparing the amino nitrogen content with the enzyme-free group after the proteolytic enzyme treatment to the Amre starfish of the present invention.

Figure 2 is a graph comparing the amino nitrogen content with the enzyme-free group after treatment with protease to starfish according to the present invention.

Figure 3 illustrates a solvent sequence extraction process of the starfish of the present invention.

Figure 4 shows a flow chart for extracting the bioactive material derived from the starfish of the present invention using silica gel column chromatography.

The present invention relates to an extract for skin whitening derived from starfish by proteolytic enzyme treatment, and more particularly, the present invention, after sequential extraction of the starfish, isolating the active substance assayed for physiological activity through silica gel column chromatography. It relates to a novel bioactive substance having a skin whitening effect obtained by purification.

The starfish is a flat star, one of the most common marine animals on the coast. There are about 30 species of starfish on the coast of Korea, and the breeding rate is very high, spawning 2 million to 3 million birds per head. It is a marine invertebrate animal included in the sea lily, starfish, arachnid, and sea urchin rivers. 47 species of starfish have been reported in Korea, and northern species are richer than in the south. These species are widely distributed in the North Pacific, and are common to northern Japan and North America. They are Ctenodiscus , Small Starfish ( Leptychaster ), Black Starfish ( Luidia ), Pseudarchaster , Baby Starfish ( Henricia ), Sun Starfish ( Solaster ) and Cuboid Starfish ( Pteraster), octopus, starfish (Plazaster), Bulgari Accessories (Distolasterias Japan), ahpael starfish (Aphelasterias), and the country has been widely inhabit the southern coast one Amr starfish, which are said to give a damage to farms (Asterias), And so on.

Starfish are carnivorous, low-eating animals that respond very sensitive to food. Thus, shellfish, beetles, mollusks, such as oysters, small crustaceans, shellfish, sea urchins, and echinoderm such as spider starfish are fed. It also eats fish caught in a net, and many starfish can be damaged by oysters, mussels and shellfish farms. Although it is omnivorous in food habits, especially shellfish, the shellfish farm considers it as a pirate creature and fights it every year. However, starfish are not only able to move, but also slowly move cattle, which are not sacrificed by starfish. The number of starfish is declining as much as the rapidly decreasing water resources. Starfish are a part of the aquatic ecosystem, and as other resources decrease, the number naturally decreases, and humans do not need to define the starfish as an inhabitant pirate creature and control it artificially. The utility value of starfish can be said to be absent for now. Attempts have been made to use the collected starfish as a fertilizer substitute, but its effects have not yet been recognized, and starfish have been cooked to find out its useful value as food, but not as food. Recently, a large amount of zinc extracted from starfish has been introduced as a health food. In addition, research on making anticancer substances and functional livestock feed using starfish is being actively conducted.

In view of this, the present inventors have separated the material having physiological activity into a functional material of the marine new material as a part of increasing the useful value of the Amr starfish and the star starfish, which are known to greatly damage the shellfish farms of Korea. The present invention has been practiced to mass produce materials.

Accordingly, it is an object of the present invention to provide a bioactive material having a skin whitening effect from starfish.

The object of the present invention is to treat the starch after protease and solvent sequential extraction, assay the physiological activity of the solvent extract obtained from the above, the activity assayed fraction is purified by silica gel column chromatography, the fraction of the fraction Achieved by searching for whitening effects.

EMBODIMENT OF THE INVENTION Hereinafter, the structure of this invention is demonstrated concretely.

The present invention is the solvent sequence extraction step of the starfish sample; Physiological activity assay step of solvent extract; Performing silica gel column chromatography on the fractions of which activity is assayed; It consists of the whitening effect assay step of purified material.

The starfish used in the present invention were two starfish ( Asterina pectinifera ) and aster starfish ( Asterias amurensis ), which are known to greatly damage shellfish farms in Korea , and were sampled by removing water from the surface. blender Co., USA) was used as the sample for analysis.

Skin whitening extract derived from the starfish of the present invention was added to the starfish three times distilled water (w / v) and added to the protease and stirred for 3 to 5 hours at 50 ℃ and centrifuged to separate the supernatant and the residue Next, three times methanol (w / v) was added to the separated residue, followed by stirring at 4 ° C. for 5 to 6 hours, followed by vacuum filtration to recover the supernatant.

On the other hand, preferably, the starfish-derived extract for skin whitening is added after stirring with methanol, and three times acetone (w / v) is added to the residue separated by vacuum filtration and stirred at 4 ° C. for 5 to 6 hours. After that, the supernatant is recovered by vacuum filtration and the supernatant is recovered.

The extract for skin whitening derived from the starfish of the present invention is a silica gel column chromatography (200) using a CH ₃ Cl ₃ : MeOH solvent system in which the supernatant obtained in the solvent sequential extraction is mixed at a volume ratio (v / v) of 5: 5 or 7: 3. And a fraction obtained by carrying out the mesh).

For the analysis of the components of the starfish of the present invention, the general components were measured according to the AOAC method, moisture was atmospheric pressure drying method, crude protein was semi micro Kjeldahl method, fat was measured by Soxhlet method, ash was dry ash method, and amino nitrogen was measured by Formol method. Quantification Cannotinoids were analyzed by measuring absorbance at 445 nm after pretreatment, and constitutive amino acid analysis was performed using an amino acid autoanalyzer (Pharmacia Biochrom 20, US Pharmacia LKB Biotech).

The present invention was used to assay the antimicrobial activity of bioactive substances isolated from starfish. Experimental strains were Gram-positive bacteria among food-causing, pathogenic and food poisoning bacteria. Bacillus subtilis , Bacillus cereus , Staphylococcus aureus , Listeria monocytogenes ( Listeria) monocytogenes ), gram-negative bacteria, E. coli , Salmonnella typhimurium , 7 Enterobacter aerogenes , and Aspergillus flavus A total of nine species, including two Aspergillus nigers , were used.

Hereinafter, the specific method of the present invention will be described in detail with reference to Examples, but the scope of the present invention is not limited only to these Examples.

EXAMPLE

Example 1 Component Analysis of Starfish Samples

In order to isolate and purify the bioactive substance from the starfish of the present invention, Asterias amurensis and stars collected monthly by each month (May-July) in the seawater range of 5-10 m from the coast of Jinjin-myeon, Masan-si, Gyeongnam Starfish ( Asterina pectinifera ) is placed in an ice chest and transported to the laboratory within an hour to remove foreign substances on the surface of the sample, washed with water, and then partially removed surface moisture with a newspaper paper. It changed into and used as the sample for analysis.

According to the AOAC method, the moisture content was measured by atmospheric heating, the crude protein was measured by semi micro Kjeldahl method, the fat was measured by Soxhlet method, and the ash was dried ash method, and the amino nitrogen was determined by Formol method.

Cannotinoids were analyzed by measuring absorbance at 445 nm after pretreatment, and constitutive amino acid analysis was performed using an amino acid autoanalyzer (Pharmacia Biochrom 20, US Pharmacia LKB Biotech).

Bell Collection period moisture(%) Crude Protein (%) Crude Lipids (%) Ash content (%) Asterias amurensis In May 72.35 ± 0.32 10.32 ± 0.07 1.77 ± 0.13 13.39 ± 0.18 In July 72.80 ± 0.09 12.09 ± 0.04 2.55 ± 0.19 9.31 ± 0.30 Starfish ( Asterina pectinifera ) In May 65.18 ± 0.02 8.71 ± 0.11 0.44 ± 0.03 22.54 ± 0.25 In July 64.75 ± 0.03 10.03 ± 0.05 1.18 ± 0.16 22.37 ± 0.16

¹⁾ Mean ± SD (n = 3).

As shown in Table 1, the general composition of the Amr starfish and the star starfish was characterized by low moisture content and high ash content as compared to general fish and shellfish. The Amr starfish accounted for about 50% of the ash content of the total solids and over 60% for star stars. This was found to be very high compared to the ash content of 1 to 3% of general fish and shellfish. In addition, star starfish had lower moisture and protein contents than ash starfish, while ash content was higher. This shows that the composition of the general components is slightly different depending on the type of starfish. On the other hand, except for moisture and ash, both kinds had the highest protein content.

According to the above results, the constituent amino acids for the Amr starfish and the star starfish were analyzed.

ingredient Content (mg (%)) Asterias amurensis Starfish ( Asterina pectinifera ) O-phospho-L-serine 9.39 (0.15) ²⁾ 72.73 (0.85) Taurine 19.10 (0.18) 296.87 (3.49) O-phosphoethanolamine 79.50 (0.74) N / D Urea 25.13 (0.23) 75.92 (0.89) Aspartic acid1 298.23 (2.76) 508.48 (5.97) Hydroxy-l-proline 221.11 (2.05) 134.21 (1.58) L-Threonine 20.03 (0.19) 540.21 (6.35) Serine N / D ¹⁾ 409.19 (4.81) Asparagine        N / D          N / D Glutamic acid 2552.45 (23.65) 991.09 (11.64) Sarcosine 35.59 (0.33) 10.79 (0.13) α-Amino adipic acid 8.58 (0.08) 5.54 (0.07) Proline 3528.21 (32.71) 301.93 (3.55) Glycine 78.09 (0.72) 1025.11 (12.04) L-Alanine N / D 594.79 (6.99) L-Citrulline 33.92 (0.31) 73.79 (0.87) L-α-Amino-n-butyric acid 7.38 (0.07) 7.04 (0.08) L-Valine 23.47 (0.22) 389.32 (4.57) L-Cystine 54.11 (0.50) 66.79 (0.78) L-Methionine 12.16 (0.11) 8.29 (0.10) Cystathionine 934.82 (8.67) 12.88 (0.15) L-Isoleucine 3.96 (0.04) 300.53 (3.53) L-Leucine 11.99 (0.11) 482.43 (5.67) L-Tyrosine 22.28 (0.21) 220.76 (2.59) β-Alanine 766.83 (7.11) 8.73 (0.10) L-Phenylalanine            N / D 302.99 (3.57) D, L-β-Aminoisobutyric acid 69.26 (0.64) 12.29 (0.14) L-Homocystine 65.50 (0.61) 1.77 (0.02) γ-Amino-n-butyric acid 196.52 (1.82) 0.49 (0.01) Ethanolamine 46.47 (0.43) 58.79 (0.69) Aminonium chloride 292.10 (2.71) 268.26 (3.16) δ-Hydroxylysine 70.99 (0.66) 95.21 (1.12) L-Ornithine 30.46 (0.28) 31.20 (0.37) L-Lysine 494.96 (4.59) 464.03 (5.45) 1-Methyl-L-histidine 42.17 (0.39) 3.83 (0.04) L-Histidine 152.82 (1.42) 147.19 (1.73) 3-Methyl-L-histidine 0.88 (0.01) 6.06 (0.07) L-Anserine 24.91 (0.23) 8.49 (0.10) L-Carnosine 4.26 (0.04) 1.81 (0.02) L-Arginine 542.15 (5.03) 571.20 (6.71) Total 10786.98 (100) 8511.33 (100)

¹⁾ N / D: Not detected ²⁾ % total amino acid content

As shown in Table 2, the total content of the Amr starfish was 10,786.95 mg, more than 8,511.33 mg of the star starfish. In the case of Amr starfish, proline was the highest at 3,528.21 mg, which is 32.71% of the total amino acids, and glutamic acid was 2,552.45 mg (23.65%). Cystathionine was 8.37%, β-alanine was 7.11%, L-arginine was 5.03%, and L-lysine was 4.59%. In the case of starfish, glycine was 1,025.11 mg, accounting for 12.04% of the total amino acids. Next, glutamic acid (11.64%), L-arginine (6.71%), L-threonine (6.35%), aspartic acid (5.97%) and L-leucine (5.67%). From the above results, it can be seen that the types of amino acids are slightly different depending on the type of starfish. In particular, there are many amino acids, which are components of collagen, which makes them valuable as functional materials.

On the other hand, carotenoids act as a powerful scavenger against activated oxygen in the photosensitive oxidation reaction induced by light rays. In addition, it has been reported to play an important role in inhibiting the progress of the overall reaction by reacting with activated singlet oxygen formed during the photosensitive oxidation reaction and protecting cells and tissues from free radicals generated by the mechanism of inhibition of free radical reaction. have. In addition, it is known to exhibit useful functions such as inhibiting cancer and promoting strong breeding of animals, improving growth rate, suppressing disease occurrence, and improving color of fish meat. Therefore, the analysis of the content of these substances was considered to be of value in order to use them as basic data for the analysis of functional substances later, and the results are shown in Table 3.

Amr Starfish Star starfish Total carotenoids 0.62 mg 5.88 mg

As shown in Table 3, the total star carotenoid content of the Amr starfish was small at 0.62 mg, whereas the star starfish contained 5.88 mg. This suggests that the starfish contains more amounts than 1.1 mg of male and 3.2 mg of female. Therefore, if only stability is expected, it is expected to be of great value for use in pharmaceutical solvents, feed additives, and health foods.

Example 2: Sequential Solvent Extraction of Starfish of the Present Invention

As can be seen from the above, starfish had the highest protein content except water and ash. Therefore, the treatment of protein hydrolase may increase the extraction efficiency of bioactive substances. According to the extraction time of the protease Amr and rate of hydrolysis of the specific starfish was investigated by measuring the amount of amino nitrogen, Alcala azepin 0.6L ^TM (Alcalase 0.6L ^TM) and other pro-Mex as protease ^TM ( Protamex ^™ ) was used.

Also, the more I higher hydrolysis rate than the control group treated with a protease, in particular Alcala azepin 0.6L ^TM Pro other Mex the ^TM was a higher rate of hydrolysis than in the group treated with, as shown in 1 and 2. The decomposition rate continued to increase until 5 hours, and then decreased at 6 hours. However, in the case of the control group without addition of the enzyme, the hydrolysis rate was low during the 6 hours of extraction. In addition, as shown in Figure 2, in the control group not treated with proteolytic enzymes at 50 ° C the amino nitrogen content increased to 60.2 mg after 1 hour of reaction, and then up to 5 hours of reaction was almost constant range. Compared with the case of treatment with alkalase 0.6L ^{TM, it was found} that the starch has an autodigesting effect of starfish. At 4 ° C., the enzyme-free group showed little effect of autodigestion enzyme.

Figure 3 shows the extraction process of the bioactive material through sequential extraction of solvent from the starfish. According to the process of Figure 3, after the proteolytic enzyme treatment for 5 hours, the extraction was carried out for each solvent, and the yield and extraction efficiency for each solvent fraction, the yield for each extraction section is shown in Table 4.

menstruum Asterias amurensis Starfish ( Asterina pectinifera ) Control group ¹⁾ Protease Control Protease Alcala azepin 0.6L ^TM Protamex ^TM Alcala azepin 0.6L ^TM Protamex ^TM water 15.31 17.42 17.71 18.19 19.94 20.78 Methanol 9.27 11.13 12.16 5.50 7.70 7.93 Acetone 1.44 1.68 1.66 0.21 0.36 0.34 Ethyl ether 0.12 0.18 0.19 0.04 0.06 0.06 Ethyl acetate 0.05 0.09 0.09 0.02 0.05 0.04 gun 26.19 30.50 31.81 23.94 28.66 29.11 Residue 73.81 69.50 68.19 76.94 71.34 70.89

¹⁾ no treatment of protease; Unit: w / w%

As shown in Table 4, the extracts of each solvent showed the highest yields in water extracts of 15.31-20.78%, followed by methanol and acetone extracts, followed by ethyl ether and ethyl acetate extracts. Very low. And the addition of the enzyme compared to enzyme untreated group enzymes that were clearly darker the color of the extract to separate the protein degradation or coloring, even when high yield came out, Alcala azepin Pro other than 0.6L handle Mex ^{TM TM} It showed a higher yield, and the effect by Protamex ^TM was excellent.

Example 3: Biological Activity Assay of Starfish Solvent Extract of the Present Invention

In order to assay the physiological activity of the starfish solvent extract extracted by treating the protease in Example 2, the antimicrobial activity and the antioxidant activity were investigated.

Experimental Example 1: Antimicrobial Activity Assay

The antimicrobial activity of the solvent extract of the starfish of the present invention was searched according to the paper disk method. As experimental strains food Bacillus as decay, pathogenic and food poisoning caused fungus of the gram-positive subtilis (Bacillus subtilis), Bacillus cereus (Bacillus cereus), star field rocker coarse aureus (Staphylococcus aureus), L. monocytogenes jeneseu (Listeria monocytogenes ), gram-negative bacteria, E. coli , Salmonnella typhimurium , 7 Enterobacter aerogenes , and Aspergillus flavus A total of nine species, including two Aspergillus nigers , were used. The results are shown in Tables 5 and 6.

Antimicrobial Activity Assay after Proteolytic Enzyme Treatment of Solvent Extract of Amre Starfish menstruum ㎍ / disk Protamex ^TM Treatment Clear ring (mm) Gram-positive bacteria Gram-negative bacteria mold B.subtilis B.cereus S.aureus L.monocytogenes E.aerogenes E.coli S.typhimurium A.flavus A.niger water 500 ^-1) - - - - - - - - 1000 - - - - - - - - - 1500 - - - - - - - - - 2000 - - - - - - - - - Methanol 500 9 9 8.5 8.5 8.5 8.5 8.5 8.5 9 1000 11 10 10 9 10 11 9.5 9 9 1500 14 12 13 12 12 13 10 10 10 2000 15 15 16 13 13 15 13 12 12 Acetone 500 8.5 8.5 - 8.5 - - 9 - - 1000 10 9 9 9 8.5 9 9 9 9 1500 12 9.5 10 10 10 11 10 9 9 2000 13 10 11 12 11 12 11 11 10 Ethyl ether 500 8.5 - - - - 8.5 - - 8.5 1000 10 9 9 8.5 9 10 9 9 9 1500 10 11 10 10 9 10 10 10 9 2000 11 12 11 11 9 12 10 11 11 Ethyl acetate 500 - - - - - - - - - 1000 10 9 - 9 - 10 - - - 1500 11 10 9 10 - 11 - - 9 2000 12 10 9 10 - 11 9 - 10

¹⁾ -: Not detected

Antimicrobial Activity Assay after Proteolytic Enzyme Treatment of Solvent Extract of Starfish menstruum ㎍ / disk Protamex ^TM Treatment Clear ring (mm) Gram-positive bacteria Gram-negative bacteria mold B.subtilis B.cereus S.aureus L.monocytogenes E.aerogenes E.coli S.typhimurium A.flavus A.niger water 500 ^-1) - - - - - - - - 1000 - - - - - - - - - 1500 - - - - - - - - - 2000 - - - - - - - - - Methanol 500 9 9 9 9 9 8.5 8.5 8.5 8.5 1000 11.5 11 10 10 10 11 10 9 9 1500 14 13 14 13 12 13 12 11 12 2000 16 15 17 14 13 16 14 14 14 Acetone 500 8.5 9 10 9 8.5 8.5 8.5 - - 1000 10 10 11 9.5 9 9 9 - 10 1500 13 11 12 10 10 10 10 9 11 2000 14 13 13 12 11 12 10 11 12 Ethyl ether 500 8.5 8.5 8.5 8.5 - 8.5 - - - 1000 9 9 10 9 - 9 8.5 - 9 1500 10 10 11 10 9 10 9 - 10 2000 12 11 12 11 10 10 10 10 11 Ethyl acetate 500 - 8.5 - - - - - - - 1000 9 9 8.5 9 - - - - 9 1500 10 10 9 9 - 9 - - 9 2000 11 11 10 10 - 10 - - 10

¹⁾ -: Not detected

As shown in Tables 5 and 6, the antibacterial activity was stronger in the star starfish than the Amr starfish. In terms of the extraction solvents, methanol and acetone were the best, followed by ethyl ether and ethyl acetate, and the water extract showed no antimicrobial activity.

Experimental Example 2: Antioxidant Activity Assay

The antioxidant effect test of the starfish solvent extract of the present invention was measured by the electron donating ability (EDA%) for DPPH (2,2-diphenyl-1-picrylhydrazyl, Aldrich Co., USA) according to the method of Blios (1958). It was. In other words, add 3-4 mL of ethanol to 0.8 mL of the prepared DPPH solution, shake it vigorously for 10 seconds, adjust the amount of ethanol so that the absorbance value of the spectrophotometer is 0.95-0.99, and then take 0.2 g of the sample to adjust the appropriate amount of ethanol. And 0.8 mL of DPPH solution were added, vigorously shaken for 10 seconds, left for 10 minutes, and absorbance was measured at 525 nm.

Antioxidant Activity of Solvent Extracts from Starfish (EDA ₅₀ , mg / mL) menstruum Asterias amurensis Starfish ( Asterina pectinifera )   Control Protamex ^TM Control Protamex ^TM water ^-1) - 1.26 1.28 Methanol 1.91 1.67 0.72 0.68 Acetone - 3.80 3.13 3.91 Ethyl ether - 1.81 3.13 3.91 Ethyl acetate 1.93 6.17 2.06 2.93 gun 1.93 1.81 2.06 2.93 Residue 9.90 6.17 12.5 8.70

¹⁾ not detected; Unit: mg / mL

As shown in Table 7, star starfish were superior to Amr starfish, and both types showed the best antioxidant effect in methanol extract.

Example 4 Separation and Purification of the Invention Starfish Solvent Extract by Silica Gel Column Chromatography

In order to separate the physiologically active substance from the starfish solvent extract of the present invention assayed for its physiological activity in Example 3 in the order of methanol and acetone from the enzyme group treated with proteolytic enzyme (Protamex ^TM ) according to step 1 of FIG. After sequential extraction of solvent, silica gel column chromatography with CH ₃ Cl ₃ : MeOH (10: 0, 9: 1, 7: 3, 5: 5, 3: 7, 1: 9, 0:10) solvent system 50 mL of each fraction was taken using I (200 mesh)) and separated and purified into 7 fractions in total. Then, the whitening effect of the fraction was investigated.

The whitening effect of the methanol and acetone extracts of the starfish of the present invention isolated and purified according to Example 4 was modified by Chen and Kubo's method (2002), 0.1 mL of 5% (w / v) sample, 0.1 M sodium phosphate buffer. (pH6.8) After shaking a total of 4.5 mL of the reaction solution such as 1.8 mL, 0.1 mL (125 units) of tyrosinase of mushrooms and 1 mL of 2.5 mL L-DOPA, incubated for 10 minutes at 25 ° C Measured at 475 nm.

Fraction number Inhibitory activity (%) Starfish ( Asterina pectinifera ) Asterias amurensis Methanol Acetone Methanol Acetone One 2.52 ^-1) 3.98 - 2 8.92 2.49 2.08 5.10 3 43.28 94.96 33.12 3.58 4 2.05 - - 3.45 5 1.93 - - 3.39 6 - - - 3.08 7 - - - 1.59

¹⁾ -: Not detected

As shown in Table 8, as a result of measuring the whitening effect of each fraction obtained by silica gel column chromatography I (200 mesh), the star starfish showed a better effect than the Amur starfish, All three fractions showed a high effect. In particular, fraction 3 of the acetone extract of the starfish showed a very excellent effect of 94.96% inhibition.

As described through the above Examples and Experimental Examples, the present invention relates to an extract for skin whitening derived from starfish by protease treatment, and has an excellent effect of providing a novel bioactive substance having a skin whitening effect extracted from starfish. There is. Therefore, the present invention can manufacture functional products and cosmetics using the starfish-derived physiologically active substance as an active ingredient, which is a very useful invention in the functional product manufacturing industry.

Claims (5)

A composition for skin whitening comprising a starfish extract as an active ingredient. The method according to claim 1, wherein the starfish extract is added to the starfish by distilled water and the protease, a protease, stirred for 3 to 5 hours at 50 ℃ and centrifuged to remove the methanol three times (v) / w) is added and stirred at 4 ° C for 5-6 hours, the composition for skin whitening, characterized in that the supernatant obtained by vacuum filtration. The method according to claim 1, wherein the starfish extract is stirred by adding methanol and then added three times acetone (v / w) to the residue separated by vacuum filtration and stirred at 4 ℃ for 5-6 hours, and then vacuum A composition for skin whitening, characterized in that it is a supernatant obtained by filtration. The method of claim 2 or 3, wherein the supernatant is subjected to silica gel column chromatography (200 mesh) with a CH ₃ Cl ₃ : MeOH solvent system mixed at a volume ratio (v / v) of 5: 5 or 7: 3. Composition for skin whitening, characterized in that the obtained fraction. The composition of claim 1, wherein the starfish is Asterias amurensis or Asterina pectinifera .

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