CN109369819A - A kind of extracting method of Hijiki polysaccharide - Google Patents
A kind of extracting method of Hijiki polysaccharide Download PDFInfo
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- CN109369819A CN109369819A CN201811230177.5A CN201811230177A CN109369819A CN 109369819 A CN109369819 A CN 109369819A CN 201811230177 A CN201811230177 A CN 201811230177A CN 109369819 A CN109369819 A CN 109369819A
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- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 40
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 40
- 238000000034 method Methods 0.000 title claims abstract description 28
- 150000004676 glycans Chemical class 0.000 title claims abstract 6
- 108090000790 Enzymes Proteins 0.000 claims abstract description 41
- 102000004190 Enzymes Human genes 0.000 claims abstract description 41
- 230000007071 enzymatic hydrolysis Effects 0.000 claims abstract description 22
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 claims abstract description 22
- 244000005700 microbiome Species 0.000 claims abstract description 9
- 230000015556 catabolic process Effects 0.000 claims abstract description 7
- 238000006731 degradation reaction Methods 0.000 claims abstract description 7
- 238000012869 ethanol precipitation Methods 0.000 claims abstract description 7
- 229940088598 enzyme Drugs 0.000 claims description 40
- 241000195474 Sargassum Species 0.000 claims description 16
- 239000002002 slurry Substances 0.000 claims description 16
- 239000007788 liquid Substances 0.000 claims description 12
- 239000000463 material Substances 0.000 claims description 10
- 239000000654 additive Substances 0.000 claims description 9
- 230000001965 increasing effect Effects 0.000 claims description 9
- 230000000996 additive effect Effects 0.000 claims description 8
- 230000009849 deactivation Effects 0.000 claims description 8
- 239000000706 filtrate Substances 0.000 claims description 8
- 238000001914 filtration Methods 0.000 claims description 8
- 239000013049 sediment Substances 0.000 claims description 8
- 239000012535 impurity Substances 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 240000006439 Aspergillus oryzae Species 0.000 claims description 5
- 235000002247 Aspergillus oryzae Nutrition 0.000 claims description 5
- 241000186000 Bifidobacterium Species 0.000 claims description 5
- 239000002994 raw material Substances 0.000 claims description 5
- 108010059892 Cellulase Proteins 0.000 claims description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 4
- 229940106157 cellulase Drugs 0.000 claims description 4
- 229960000935 dehydrated alcohol Drugs 0.000 claims description 4
- 230000000694 effects Effects 0.000 claims description 4
- 238000005360 mashing Methods 0.000 claims description 4
- 238000003756 stirring Methods 0.000 claims description 4
- 238000001291 vacuum drying Methods 0.000 claims description 4
- 238000009777 vacuum freeze-drying Methods 0.000 claims description 4
- 238000010792 warming Methods 0.000 claims description 4
- 241000193830 Bacillus <bacterium> Species 0.000 claims 1
- 235000013399 edible fruits Nutrition 0.000 claims 1
- 239000003292 glue Substances 0.000 claims 1
- 238000000605 extraction Methods 0.000 abstract description 9
- 229930013930 alkaloid Natural products 0.000 abstract description 2
- 150000003797 alkaloid derivatives Chemical class 0.000 abstract description 2
- 150000002016 disaccharides Chemical class 0.000 abstract description 2
- 150000002772 monosaccharides Chemical class 0.000 abstract description 2
- 229920001542 oligosaccharide Polymers 0.000 abstract description 2
- 150000002482 oligosaccharides Chemical class 0.000 abstract description 2
- 150000004804 polysaccharides Chemical class 0.000 description 33
- 244000063299 Bacillus subtilis Species 0.000 description 4
- 235000014469 Bacillus subtilis Nutrition 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 238000011084 recovery Methods 0.000 description 4
- MYKOKMFESWKQRX-UHFFFAOYSA-N 10h-anthracen-9-one;sulfuric acid Chemical compound OS(O)(=O)=O.C1=CC=C2C(=O)C3=CC=CC=C3CC2=C1 MYKOKMFESWKQRX-UHFFFAOYSA-N 0.000 description 3
- 238000003556 assay Methods 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 239000000284 extract Substances 0.000 description 3
- 229920001543 Laminarin Polymers 0.000 description 2
- 241001494479 Pecora Species 0.000 description 2
- 229920000615 alginic acid Polymers 0.000 description 2
- 235000010443 alginic acid Nutrition 0.000 description 2
- 230000003712 anti-aging effect Effects 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 230000036039 immunity Effects 0.000 description 2
- DBTMGCOVALSLOR-VPNXCSTESA-N laminarin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)OC1O[C@@H]1[C@@H](O)C(O[C@H]2[C@@H]([C@@H](CO)OC(O)[C@@H]2O)O)O[C@H](CO)[C@H]1O DBTMGCOVALSLOR-VPNXCSTESA-N 0.000 description 2
- 150000002632 lipids Chemical class 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000003809 water extraction Methods 0.000 description 2
- 244000178993 Brassica juncea Species 0.000 description 1
- 235000011332 Brassica juncea Nutrition 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- PNNNRSAQSRJVSB-SLPGGIOYSA-N Fucose Natural products C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)C=O PNNNRSAQSRJVSB-SLPGGIOYSA-N 0.000 description 1
- 240000005979 Hordeum vulgare Species 0.000 description 1
- 235000007340 Hordeum vulgare Nutrition 0.000 description 1
- SHZGCJCMOBCMKK-DHVFOXMCSA-N L-fucopyranose Chemical compound C[C@@H]1OC(O)[C@@H](O)[C@H](O)[C@@H]1O SHZGCJCMOBCMKK-DHVFOXMCSA-N 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 241000264279 Sargassum fusiforme Species 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- 229960001126 alginic acid Drugs 0.000 description 1
- 239000000783 alginic acid Substances 0.000 description 1
- 150000004781 alginic acids Chemical class 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 235000020251 goat milk Nutrition 0.000 description 1
- 235000021384 green leafy vegetables Nutrition 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000002218 hypoglycaemic effect Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000001151 other effect Effects 0.000 description 1
- 239000000825 pharmaceutical preparation Substances 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- -1 polysaccharide sulfate Chemical class 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000013094 purity test Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Sustainable Development (AREA)
- Biochemistry (AREA)
- Materials Engineering (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Organic Chemistry (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a kind of extracting methods of Hijiki polysaccharide, including the following steps: (1) is beaten；(2) preliminary enzymatic hydrolysis；(3) specified microorganisms fermentative degradation；(4) secondary enzymolysis；(5) it filters；(6) ethanol precipitation；(7) it is dried in vacuo.The present invention is improved on existing extraction method of polysaccharides, and first using the method tentatively digested, monosaccharide and disaccharide, oligosaccharide, alkaloid etc. are removed.Then it is degraded, polysaccharide is sufficiently released, and optimize complex enzyme zymohydrolysis condition using specified microorganisms.
Description
Technical field
The present invention relates to a kind of extracting methods of Hijiki polysaccharide, belong to technical field of polysaccharide extraction.
Background technique
Sargassum fusifome (Sargassumfusiforme (Harv.) Setch) is Phaeophyta Sargassum plant, alias goat milk
Son, extra large barley etc..Widely distributed, Japan and Korea bank are all distributed, and in Coastal Area in Zhejiang Province, distribution is more.In Japan, sheep
Dish of dwelling is referred to as " longevity greens/mustard green "；In China, sargassum fusifome, which is used as medicine, history remote, Shennong's Herbal, Compendium of Material Medica etc.
Record its edible value and medical value.Sargassum fusifome nutriment rich in itself, wherein carbohydrate occupies very big
Ratio, imparting it has more biological function and higher Development volue.Sargassum fusifome is widely used in reducing blood lipid, anti-blood
Bolt, antitumor and elimination brain fag in terms of promoting the dietotherapies such as immunity of organisms, anti-aging, treat rheumatism, diabetes
Chemical industry associated additives and the raw material of adhesive etc. are also applied simultaneously in terms of the development of equal pharmaceutical preparations.
Hijiki polysaccharide (SPF) is a kind of water-soluble polysaccharide extracted from full algae sargassum fusifome, sargassum fusifome frond
Main component is mainly made of algin and algal polysaccharide sulfate.It is anti-that modern pharmacology research shows that Hijiki polysaccharide has
Tumour, hypoglycemic, reducing blood lipid, enhancing immunity of organisms, anti-aging and other effects.The a variety of pharmacological activity and its polysaccharide of sargassum fusifome
Structure is closely related.SPF is the similar acidic polysaccharose mixture of a kind of composed structure, and good water solubility, viscosity is high, mainly includes brown
Alginic acid (AcidicPolysaccharidesofAlgae, APA), fucose (Fucoidanpolysaccharidesulfate,
FPS) and laminaran (Laminaran), the content analysis of polysaccharide is to sargassum fusifome drug efficacy study and development and utilization in sargassum fusifome
Important evidence.
In recent years, the extraction of polysaccharide and purification process have rapid development, and new method also continues to bring out, including HPLC
Method, enzyme process, membrane separation process etc..But due to various aspects, these methods are not widely used.In mostly using at present
Property Hot water extraction or acidleach formulation.Various nutritional ingredients based on Hijiki polysaccharide are largely water soluble substances, because
This method that can use hot water extraction is extracted；This extracting method simple possible, it is easy to operate, not to equipment requirement
Height, but its production time is long, and polysaccharide yield is not high, and waste is serious.
Summary of the invention
The technical problems to be solved by the present invention are: providing a kind of raising Hijiki polysaccharide recovery rate, and impurity content
Few extraction method of polysaccharides.
In order to solve the above technical problems, the extracting method of Hijiki polysaccharide provided by the invention, including the following steps:
(1) it is beaten.
(2) preliminary enzymatic hydrolysis.
(3) specified microorganisms fermentative degradation.
(4) secondary enzymolysis.
(5) it filters.
(6) ethanol precipitation.
(7) it is dried in vacuo.
Step (1) mashing concrete operations are: sargassum fusifome being cleaned, the pure water of 2-3 times of weight, beater or homogeneous is added
It is beaten 10~20 minutes in machine.
Step (2) tentatively enzymatic hydrolysis concrete operations are: adjusting the pH to 4.2-5.2 of slurries；Complex enzyme liquid is added in slurries,
Temperature after enzymatic hydrolysis, is increased enzyme deactivation by enzyme additive amount 0.5-2.3mKat/g, enzymolysis time 1-3h.
Step (3) specified microorganisms fermentative degradation concrete operations are: the raw material after enzymatic hydrolysis being warming up to 37-40 DEG C, is adjusted
The pH to 7.9-8.3 of material；Then aspergillus oryzae, Bifidobacterium, 0.05-1.5 parts of Bacillus subtilis strain is added, sufficiently mixes
Even, ferment 5-10h.
The aspergillus oryzae that is added in this step, Bifidobacterium, Bacillus subtilis strain quality dwell according to sheep in step (1)
The quality of dish is standard, i.e., the weight of sargassum fusifome is 1 part.
Step (4) secondary enzymolysis concrete operations are: adjusting the pH to 5.3-6.1 of slurries；Complex enzyme liquid is added in slurries,
Temperature after enzymatic hydrolysis, is increased enzyme deactivation by enzyme additive amount 0.5-1.5mKat/g, enzymolysis time 2-3h.
Step (5) filtering concrete operations are: the material filtering that above-mentioned steps are obtained removes 100 mesh impurity below, from
The isolated filtrate of the heart.
Step (6) ethanol precipitation concrete operations are: dehydrated alcohol added into filtrate, stands 0.5-3h after stirring 15min,
It is centrifugated after being cooled to 10-15 DEG C, collects sediment.
Step (7) vacuum drying concrete operations are: step (6) being collected sediment vacuum freeze drying, obtain sargassum fusifome
Polysaccharide.
In step (2) and (4), complex enzyme liquid is cellulase, pectase and zytase, the ratio of their enzyme activity
For 3:1-2:1.
Polysaccharide yield (%)=extract quality/material quality.
The purity testing of Hijiki polysaccharide: assay is carried out using polysaccharide of the Anthrone-sulfuricacid method to preparation.
Beneficial effects of the present invention:
The present invention is improved on existing extraction method of polysaccharides, first using the method that tentatively digests, by monosaccharide and disaccharide,
The removal such as oligosaccharide, alkaloid.Then it is degraded, polysaccharide is sufficiently released, and optimize complex enzyme using specified microorganisms
Enzymatic hydrolysis condition (complex enzyme ratio, hydrolysis temperature, enzymolysis time).
The present invention extracts Hijiki polysaccharide recovery rate height, can achieve 41.5-50.3%；Extract the purity of Hijiki polysaccharide
It is high.Traditional extracting mode, Hijiki polysaccharide effective component mutability, color is deeper, and the polysaccharide extracted contains albumen
Matter impurity needs the step of additionally increasing except deproteinized, provides extraction process using the present invention, and protein is during the extraction process
It is removed, polysaccharide finished product purity is high.
Specific embodiment
Embodiment 1
Present embodiments provide a kind of extracting method of Hijiki polysaccharide, including the following steps:
Mashing: sargassum fusifome is cleaned, the pure water of 2 times of weight is added, is beaten 10 minutes in beater or homogenizer.
Preliminary enzymatic hydrolysis: the pH to 4.2 of slurries is adjusted；Complex enzyme liquid, enzyme additive amount 0.5mKat/g, enzyme are added in slurries
Time 3h is solved, after enzymatic hydrolysis, temperature is increased into enzyme deactivation.
Specified microorganisms fermentative degradation: being warming up to 37 DEG C for the raw material after enzymatic hydrolysis, adjusts the pH to 7.9 of material；Then plus
Enter aspergillus oryzae, Bifidobacterium, 0.05 part of Bacillus subtilis strain, mix well, ferment 5-10h.
Secondary enzymolysis: the pH to 5.3 of slurries is adjusted；Complex enzyme liquid, enzyme additive amount 1.5mKat/g, enzyme are added in slurries
Time 2h is solved, after enzymatic hydrolysis, temperature is increased into enzyme deactivation.
Filtering: the material filtering that above-mentioned steps are obtained removes 100 mesh impurity below, is centrifugally separating to obtain filtrate.
Ethanol precipitation: adding dehydrated alcohol into filtrate, stands 0.5h after stirring 15min, centrifugation point after being cooled to 10 DEG C
From collection sediment.
Vacuum drying: step (6) are collected into sediment vacuum freeze drying, obtain Hijiki polysaccharide.
In preliminary enzymatic hydrolysis and secondary enzymolysis, complex enzyme liquid is cellulase, pectase and zytase, their enzyme activity
Ratio be 3:1:1.
Use the recovery rate of Hijiki polysaccharide manufactured in the present embodiment for 41.5%.
Assay is carried out using thick Hijiki polysaccharide of the Anthrone-sulfuricacid method to extraction, purity reaches 87.6%.
Embodiment 2
The extracting method of Hijiki polysaccharide provided in this embodiment, including the following steps:
Mashing: sargassum fusifome is cleaned, the pure water of 3 times of weight is added, is beaten 20 minutes in beater or homogenizer.
Preliminary enzymatic hydrolysis: the pH to 5.2 of slurries is adjusted；Complex enzyme liquid, enzyme additive amount 2.3mKat/g, enzyme are added in slurries
Time 1h is solved, after enzymatic hydrolysis, temperature is increased into enzyme deactivation.
Specified microorganisms fermentative degradation: being warming up to 40 DEG C for the raw material after enzymatic hydrolysis, adjusts the pH to 8.3 of material；Then plus
Enter aspergillus oryzae, Bifidobacterium, 1.5 parts of Bacillus subtilis strain, mix well, ferment 10h.
Secondary enzymolysis: the pH to 6.1 of slurries is adjusted；Complex enzyme liquid, enzyme additive amount 1.5mKat/g, enzyme are added in slurries
Time 2h is solved, after enzymatic hydrolysis, temperature is increased into enzyme deactivation.
Filtering: the material filtering that above-mentioned steps are obtained removes 100 mesh impurity below, is centrifugally separating to obtain filtrate.
Ethanol precipitation: adding dehydrated alcohol into filtrate, stands 3h after stirring 15min, is centrifugated after being cooled to 15 DEG C,
Collect sediment.
Vacuum drying: step (6) are collected into sediment vacuum freeze drying, obtain Hijiki polysaccharide.
In preliminary enzymatic hydrolysis and secondary enzymolysis, complex enzyme liquid is cellulase, pectase and zytase, their enzyme activity
Ratio be 3:1:1.
Use the recovery rate of Hijiki polysaccharide manufactured in the present embodiment for 50.3%.
Assay is carried out using thick Hijiki polysaccharide of the Anthrone-sulfuricacid method to extraction, purity reaches 85.2%.
Claims (9)
1. a kind of extracting method of Hijiki polysaccharide, characterized in that it comprises the following steps:
(1) it is beaten；
(2) preliminary enzymatic hydrolysis；
(3) specified microorganisms fermentative degradation；
(4) secondary enzymolysis；
(5) it filters；
(6) ethanol precipitation；
(7) it is dried in vacuo.
2. the method according to claim 1, wherein step (1) mashing concrete operations are: sargassum fusifome is cleaned,
The pure water of 2-3 times of weight is added, is beaten 10~20 minutes in beater or homogenizer.
3. the method according to claim 1, wherein step (2) tentatively enzymatic hydrolysis concrete operations are: adjusting slurries
PH to 4.2-5.2；It is added complex enzyme liquid in slurries, enzyme additive amount 0.5-2.3mKat/g, enzymolysis time 1-3h, enzymatic hydrolysis terminate
Afterwards, temperature is increased into enzyme deactivation.
4. the method according to claim 1, wherein step (3) specified microorganisms fermentative degradation concrete operations are:
Raw material after enzymatic hydrolysis is warming up to 37-40 DEG C, adjusts the pH to 7.9-8.3 of material；Then aspergillus oryzae, Bifidobacterium, withered is added
It careless Bacillus 0.05-1.5 parts, mixes well, ferment 5-10h.
5. the method according to claim 1, wherein step (4) secondary enzymolysis concrete operations are: adjusting slurries
PH to 5.3-6.1；It is added complex enzyme liquid in slurries, enzyme additive amount 0.5-1.5mKat/g, enzymolysis time 2-3h, enzymatic hydrolysis terminate
Afterwards, temperature is increased into enzyme deactivation.
6. the method according to claim 1, wherein step (5) filtering concrete operations are: above-mentioned steps are obtained
Material filtering, remove 100 mesh impurity below, be centrifugally separating to obtain filtrate.
7. the method according to claim 1, wherein step (6) ethanol precipitation concrete operations are: adding into filtrate
Add dehydrated alcohol, stand 0.5-3h after stirring 15min, be centrifugated after being cooled to 10-15 DEG C, collects sediment.
8. the method according to claim 1, wherein step (7) vacuum drying concrete operations are: by step (6)
Sediment vacuum freeze drying is collected, Hijiki polysaccharide is obtained.
9. the method according to claim 1, wherein complex enzyme liquid is cellulase, fruit in step (2) and (4)
Glue enzyme and zytase, the ratio of their enzyme activity are 3:1-2:1.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113088546A (en) * | 2021-05-08 | 2021-07-09 | 潍坊市检验检测中心 | Preparation method of sargassum fusiforme polysaccharide and oligosaccharide |
| CN114107410A (en) * | 2021-12-02 | 2022-03-01 | 安徽坤大生物工程技术有限公司 | Method for extracting and fermenting sargassum fusiforme polysaccharide |
| CN114621980A (en) * | 2022-03-10 | 2022-06-14 | 温州大学 | Method for improving hypoglycemic activity of sargassum fusiforme extract |
| CN115702909A (en) * | 2021-08-16 | 2023-02-17 | 青岛康迈臣生物科技有限责任公司 | Sargassum fusiforme fermentation mixture and preparation method and application thereof |
| WO2023205915A1 (en) * | 2022-04-24 | 2023-11-02 | 中国农业科学院烟草研究所 | Preparation process of sargassum fusiforme oligosaccharide and use of sargassum fusiforme oligosaccharide in crop planting |
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| CN114621980B (en) * | 2022-03-10 | 2023-04-25 | 温州大学 | Method for improving hypoglycemic activity of sargassum fusiforme extract |
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