KR100705975B1 - Bioavailable fucoidan and methods for preparing the same - Google Patents

Abstract

The present invention comprises (a) 20-40% sulfuric acid group content, (b) a weight average molecular weight of 1,000-30,000 Da, (c) a polydispersity index of 1-6 and (d) 0-200 centipoise The present invention relates to a bioavailable fucoidan and a method for producing the same. Since the bioavailability of the present invention shows high sulfate content, small average molecular weight of 30,000 Da or less, low polydispersity index and viscosity, and very high water solubility, the human body absorption rate is greatly improved, and thus bioavailability is greatly improved. Fucoidan's unique physiological activities (inhibiting cancer cell proliferation, antioxidants, suppressing blood glucose elevation, anticoagulant, antioxidant, triglyceride and cholesterol levels, and promoting gastric ulcer healing) can be sufficiently exhibited in the human body. In addition, the improved physical properties of the bioavailable fucoidan of the present invention can solve the problems caused by non-uniformity of conventional fucoidan product quality.

Fucoidan, bioavailability, pyruvic acid, citric acid, acetic acid, vinegar, viscosity, water soluble, sulfate

Description

      Bioavailable Fucoidan and its preparation method {Bioavailable Fucoidan and Methods for Preparing the Same}

1 shows the results of gel chromatography analysis of the molecular weight of bioavailable fucoidan prepared by the method of the present invention.

2 is a Fourier transform infrared spectrophotometer (FT-IR) analysis to confirm the structural characteristics of the bioavailable fucoidan of the present invention.

3 is a result of analyzing the sulfuric acid group content of the bioavailable fucoidan of the present invention.

4a and 4b are the results of water solubility and viscosity analysis of the bioavailable fucoidan of the present invention.

5 is a result of comparing and analyzing the proliferation inhibitory function of squamous cell carcinoma which is a tumor cell of the bioavailable fucoidan of the present invention with the polymer fucoidan.

The present invention relates to a bioavailable fucoidan and a method for producing the same.

The seaweeds present in the ocean include functional polysaccharides that are widely used in the food industry. The representative ones are agar, alginic acid, carrageenan, and fucoidan, which has recently been tested for physiological activity. Etc. Fucoidan is a complex sulfated polysaccharide containing L-Fucose having an esterified sulfuric acid group and a small amount of galactose, xylose and glucuronic acid as main components. .

Fucoidan is a viscous polymer with a weight average molecular weight of 40,000-2,000,000 Daltons, and is mainly contained in mucus of mollusks such as abalone and squid, or brown algae such as seaweed or kelp, especially fucoidan extracted from seaweed spores. It has a molecular weight of 170,000 Daltons. Fucoidan, an acidic polysaccharide derived from brown algae, is a functional food material that has been recently developed in Japan and is in the early stages of development in Korea. The fucoidan, which is currently sold in Korea, is mainly composed of highly viscous polymers extracted from brown algae. .

The function of brown algae-derived fucoidan has been studied in Japan since the 1990s. It has been found to promote and is used as a raw material for health functional food ( Scand. J. Urol. Nephrol. , 40 (1), 2006; Am. J. Hematol. , 78 (1), 2005; Journal of the Korean Chemical Society , 46 (2); Special Issue, November issue of Monthly Food Industry, 2005).

Despite the wide range of functionalities of the polymer fucoidan, problems such as lowering of the human absorption rate due to high molecular weight, viscosity, and low water solubility, restriction of commercialization due to non-uniformity of quality due to the diversity of raw materials and manufacturing methods, etc. have not been solved.

Therefore, the technology of low molecular weight fucoidan into fluid fucoidan is a key technology for industrialization that can maximize the added value and added value as it can secure product uniformity and improve physical disadvantages.

Enzymatic hydrolysis is mainly studied to reduce the molecular weight of fucoidan. In Japan, Takara Shuzo co. Ltd registered a patent (US Patent No. 6,054,577) for the enzyme reaction product produced by enzymatic treatment of fucoidan on April 25, 2000, and on August 21, 2001, the microorganism, enzyme and fucoidan decomposing enzyme-producing enzyme were encoded. The patent was obtained by securing a gene (US Pat. No. 6,277,616). Since then, research has been actively conducted to secure new enzymes for decomposing polymer fucoidan in Japan and Russia, but is still in the early stage of development.

Recently, the physiological activity of low molecular weight fucoidan has been reported ( The Journal of Pharmacology and Experimental Therapeutics , 305 (1), 2003; Carbohydrate Research , 289, 1996). The current situation is spurring to develop technology for manufacturing.

Although the production of fucooligosaccharide by enzymatic method has the advantage of hydrolyzing only specific bonds, it is difficult to secure enzyme, solve the problem of liquefaction of polymer polysaccharide for enzymatic reaction, and high production cost by using enzyme in industrial process. There is a limit to practical use.

Throughout this specification, many papers and patent documents are referenced and their citations are indicated. The disclosures of cited papers and patent documents are incorporated herein by reference in their entirety, and the level of the technical field to which the present invention belongs and the contents of the present invention are more clearly explained.

The present inventors have made efforts to develop a low molecular weight bioavailable fucoidan excellent in physical properties and physiological activities, and to provide such a bioavailable fucoidan as a novel method that can overcome the limitations and problems of the conventional enzyme method. As a result, when treating a suitable organic acid such as pyruvic acid, the fucoidan polysaccharide can be effectively lowered in molecular weight, and the molecular weight of the fucoidan polysaccharide can be controlled by controlling the type and concentration of organic acid. The present invention was completed by confirming that physiological activity (eg, cancer cell proliferation inhibiting ability) is significantly improved compared to the polymer fucoidan.

Accordingly, it is an object of the present invention to provide a bioavailable fucoidan having excellent physical and physiological activities.

Another object of the present invention is to provide a method for producing a bioavailable fucoidan.

Other objects and advantages of the present invention will become apparent from the following detailed description, claims and drawings.

According to one aspect of the invention, the invention provides (a) a sulfuric acid group content of 20-40%, (b) a weight average molecular weight of 1,000-30,000 Da, (c) a polydispersity index of 1-6 and (d) 0- It provides a bioavailable fucoidan characterized by having a viscosity of 200 centi-poise.

The present inventors have made efforts to develop a low molecular weight bioavailable fucoidan excellent in physical properties and physiological activities, and to provide such a bioavailable fucoidan as a novel method that can overcome the limitations and problems of the conventional enzyme method. As a result, when treating a suitable organic acid such as pyruvic acid, the fucoidan polysaccharide can be effectively lowered in molecular weight, and the molecular weight of the fucoidan polysaccharide can be controlled by controlling the type and concentration of organic acid. It has been confirmed that the physiological activity (eg, cancer cell proliferation inhibitory ability) is significantly improved compared to the polymer fucoidan.

The present invention relates to a bioavailable fucoidan excellent in physical properties and physiological activity. The term "bioavailability" as used herein in connection with fucoidan is used to express fucoidan of low molecular weight, which has excellent absorption rate in water, high water solubility, and higher physiological activity than natural fucoidan of natural-occurring. It is used.

The bioavailable fucoidan of the present invention has a very small molecular weight compared to the conventional high-molecular weight fucoidan derived from seaweed spores (generally 170,000 Da or more). According to a preferred embodiment of the present invention, the weight average molecular weight of the fucoidan hydrolysis product is 1,000-20,000 Da, more preferably 3,000-10,000 Da, most preferably 1,500-3,000 Da.

As used herein, the term "weight average molecular weight" refers to the weight average molecular weight of fucoidan molecules having various molecular weights that may be present in the bioavailable fucoidan of the present invention. For example, when the bioavailable fucoidan of the present invention is obtained by treating an organic acid with a high molecular weight fucoidan, there may be fucoidan molecules having various molecular weights in the treatment product, and the weight of fucoidan molecules present in the organic acid treatment product. The average molecular weight is the weight average molecular weight described in this specification. For example, when the weight average molecular weight of the bioavailable fucoidan of the present invention is 1,500-3,000 Da, preferably the weight average molecular weight 3000-5000 Da: 1250-1500 Da: 600-750 Da: 140-210 Da = 30- It may be a mixture of fucoidan having 60: 12-35: 5-20: 15-25.

According to a preferred embodiment of the present invention, the sulfuric acid group content of the low molecular weight fucoidan of the present invention is 20-35%, most preferably 20-30%. Sulfuric acid groups are an important part of fucoidan's physiological activity ( Biochem Pharmacol , 65, 2003). Thus, the physiological activity of the low molecular weight fucoidan of the present invention (e.g., inhibiting tumor cell proliferation), which exhibits a high sulfate content, is greatly improved.

The bioavailable fucoidan of the present invention has a polydispersity of 1-6, preferably 1-5, more preferably 1-4, and most preferably 1-2. This small polydispersity index shows that the size distribution of the bioavailable fucoidan of the present invention is very narrow. Therefore, the bioavailable fucoidan of the present invention is a low molecular weight fucoidan having a uniform physical or physiological properties, and when it is commercialized, a bioavailable fucoidan product of uniform quality can be obtained.

The bioavailable fucoidan of the present invention (more specifically, an aqueous solution of bioavailable fucoidan) exhibits very low viscosity. Preferably, 0-100 centipoise, most preferably 0-50 centipoise. The low viscosity of the bioavailable fucoidan means that the solubility of the bioavailable fucoidan in water is nearly 100.

According to a preferred embodiment of the present invention, the component ratio of bioavailable fucoidan is carbon: hydrogen: oxygen: nitrogen: sulfur = 20-30: 3-5: 45-60: 0.05-1.0: 5.7-20, more preferably. Preferably 20-25: 4-4.7: 50-60: 0.05-0.5: 5.7-20, most preferably carbon: hydrogen: oxygen: nitrogen: sulfur = 23-25: 4.2-4.7: 52-58: 0.07 -0.2: 7.0-10

According to a preferred embodiment of the present invention, the bioavailable fucoidan of the present invention is obtained by treating pyruvic acid, citric acid, acetic acid or mixtures thereof with high molecular weight fucoidan. By adjusting the type and concentration of the organic acid, bioavailable fucoidan having desired physical properties and physiological activity can be obtained. Methods of making bioavailable fucoidans are described in detail below.

According to another aspect of the present invention, the present invention comprises the steps of (a) hydrolyzing the polymer fucoidan by treating pyruvic acid, citric acid, acetic acid or a mixture thereof to the polymer fucoidan; And (b) obtaining a fucoidan hydrolyzate, thereby providing a method for producing the bioavailable fucoidan of the present invention having the above-mentioned characteristics.

As used herein, the term "high molecular weight fucoidan" means a molecular weight of at least 170,000 Da, preferably at least 150,000 Da, more preferably at least 100,000 Da, most preferably at least 40,000 Da. According to one aspect of the present invention, the high molecular weight fucoidan is a macromolecular fucoidan extracted from various marine organisms such as seaweeds such as seaweed, kelp, shellfish and rhubarb and abalone, squid and starfish.

The present invention is a process for producing a biocompatible fucooligosaccharide desired by hydrolyzing the high molecular weight fucoidan efficiently.

In the present invention, the acid used for hydrolysis of fucoidan is pyruvic acid, citric acid, acetic acid or mixtures thereof. Preferably step (a) is carried out using a mixture of pyruvic acid and acetic acid. More preferably, the pyruvic acid to acetic acid ratio of the mixture of pyruvic acid and acetic acid is 1: 2-1: 20 based on volume. The inventors experimented with various acids and found that pyruvic acid, citric acid, acetic acid or mixtures thereof used as edible organic acids were very efficient for hydrolyzing fucoidan polysaccharides. Although pyruvic acid and citric acid are not a problem when developing a hydrolyzate of fucoidan as a food, it acts on a high molecular weight fucoidan and efficiently hydrolyzes to provide a fucoidan of a desired molecular weight, which can control the molecular weight of such fucoidan. It was confirmed that the acid.

In addition, as acetic acid used for hydrolysis, the thing of the form of edible vinegar, such as vinegar, synthetic vinegar, spirit vinegar, and other vinegar, can be used. The vinegar is an edible vinegar, the main component of acetic acid, brewed vinegar includes citric acid, lactic acid and succinic acid in addition to acetic acid, the other vinegar is composed of acetic acid.

When pyruvic acid, citric acid or acetic acid alone is treated in the hydrolysis step of the present invention, typically 0.001-10 N, preferably 0.01-10 N, more preferably 0.1-10 N, even more preferably 1- High molecular weight fucoidan is treated at a concentration of 8 N, most preferably 2-7 N. In addition, when treated with vinegar, synthetic vinegar, alcohol vinegar or other vinegar alone, typically 0.1-2%, preferably 0.4-1.5%, more preferably 0.8-1.2%, most preferably about 1% A high molecular weight fucoidan solution was prepared to produce vinegar with a pH of 6% of at least 20% (v / v), preferably at least 60% (v / v), more preferably at least 80% (v / v), More preferably, a 10-fold concentrated solution of vinegar with a minimum of 100% (v / v) and most preferably 6% acidity is treated with 50% (v / v).

When treated with pyruvic acid or a mixture of citric acid and vinegar, 1: 1-1: 20 is preferable based on the volume of organic acid solution and vinegar solution prepared at the same concentration, more preferably 1: 1- 1:10, most preferably 1: 1-1: 5 mixed acid.

The high molecular weight fucoidan as the starting material used in the hydrolysis step of the present invention is preferably 0.1-20% (w / v), more preferably 0.1-10% (w / v), most preferably 0.1- 6% (w / v).

In the case of hydrolysis, the reaction temperature is preferably 40-120 ° C., and the reaction time is 1-10 hours.

In the process of the invention, the step of obtaining fucoidan hydrolysis products can be carried out according to various protocols. For example, the hydrolysis product is filtered with a membrane filter having a constant molecular weight cut-off value (eg, a cut-off value of 1,000-50,000 Da). Subsequently, the filtrate was concentrated under reduced pressure to evaporate water and organic acids, and concentrated to obtain a liquid form, or after decolorization through ethanol washing to obtain fucoidan hydrolyzed powder.

According to a preferred embodiment of the present invention, the weight average molecular weight of the fucoidan hydrolysis product is 1,000-30,000 Da, more preferably 1,000-20,000 Da, even more preferably 3,000-10,000 Da, most preferably 1,500 -3,000 Da.

The molecular weight of the bioavailable fucoidan produced by the present invention can be controlled by the type of acid used and the concentration of the acid, which is one of the features of the present invention. That is, the gist of the present invention is not to lower the molecular weight of fucoidan by treating the acid, but to provide fucoidan having a molecular weight value excellent in bioavailability using an appropriate organic acid.

In the present invention, the fucoidan hydrolysis product preferably has a sulfuric acid group content of 20-40%, more preferably 20-35%, most preferably 20-30%. According to the method of the present invention, the final bioavailable fucoidan has a higher sulfuric acid group content than the high molecular weight fucoidan used as a starting material. Preferably, the sulfuric acid group content in the fucoidan hydrolysis product is increased by 1-20%, more preferably 2-15%, and most preferably 5-10%, higher than the high molecular weight fucoidan.

Sulfuric acid groups are an important part of fucoidan's physiological activity ( Biochem Pharmacol , 65, 2003). Thus, the physiological activity of the bioavailable fucoidan of the present invention, which exhibits high sulfate content, (eg, inhibiting tumor cell proliferation) is greatly improved.

According to a preferred embodiment of the invention, the fucoidan hydrolyzate exhibits a polydispersity of 1-6, more preferably 1-4, most preferably 1-2. This small polydispersity index shows that the size distribution of the bioavailable fucoidan of the present invention is very narrow. Therefore, according to the method of the present invention, it can be seen that a bioavailable fucoidan having a substantially constant size (molecular weight) can be obtained, and finally, a bioavailable fucoidan having a uniform physical or physiological characteristic can be obtained and commercialized therein. The bioavailable fucoidan product of uniform quality can be obtained.

The bioavailable fucoidans produced by the present invention (more specifically, aqueous solutions of low molecular weight fucoidans) exhibit very low viscosity. That is, the method of the present invention greatly reduces the viscosity of the high molecular weight fucoidan as the starting material. Preferably, the bioavailable fucoidan is 0-300 centipoise, more preferably 0-200 centipoise, more preferably 0-100 centipoise, most preferably 0-50 centimeter Represents Poise. The low viscosity of the bioavailable fucoidan indicates that the solubility (ie, water solubility) of the bioavailable fucoidan in water is nearly 100. In this respect, the method of the present invention can be expressed as a 'method of controlling the water solubility of fucoidan'.

Bioavailable fucoidan prepared by the method of the present invention is not simply reduced in molecular weight as compared to high molecular weight fucoidan, which is a starting material, but the ratio of member elements is changed throughout the molecule. According to a preferred embodiment of the present invention, the component ratio of the fucoidan hydrolysis products of the present invention is carbon: hydrogen: oxygen: nitrogen: sulfur = 20-30: 3-5: 45-60: 0.05-1.0: 5.7-20 More preferably 20-25: 4-4.7: 50-60: 0.05-0.5: 5.7-20, and most preferably carbon: hydrogen: oxygen: nitrogen: sulfur = 23-25: 4.2-4.7: 52 -58: 0.07-0.2: 7.0-10.

According to the method of the present invention, the bioavailable fucoidan produced is improved in physiological activity compared to high molecular weight fucoidan as a starting material (Example 7).

According to the method of the present invention, bioavailable fucoidan can be prepared with improved efficiency, convenience and economical efficiency as compared to the method using a conventional enzyme, and the molecular weight of the bioavailable fucoidan can be controlled. In addition, the method of the present invention does not affect the basic structure of fucoidan (see Example 3), not only does not impair the intrinsic physiological activity of fucoidan, but is also expected to discriminate activity according to molecular weight size.

Polymer fucoidan has many different physiological activities. However, in spite of such a wide range of functionalities, there are problems such as lowering of human absorption due to high molecular weight, viscosity, and low water solubility, restriction of commercialization due to non-uniformity of quality due to diversity of raw materials and manufacturing methods.

According to the present invention, the above problems of the polymer fucoidan can be almost completely solved. Since the low molecular weight fucoidan of the present invention exhibits high sulfate content, small weight average molecular weight of less than 30,000 Da, low polydispersity index and viscosity, and very high water solubility, the body absorption rate is greatly improved, and thus bioavailability is greatly increased. It is possible to fully utilize fucoidan's unique physiological activity (inhibiting cancer cell proliferation, anti-oxidation, hypoglycemia, anticoagulant, antioxidant, triglyceride and cholesterol concentration, and promoting gastric ulcer healing). In addition, the improved physical properties of the bioavailable fucoidan of the present invention can solve the problems caused by non-uniformity of conventional fucoidan product quality.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention. .

Example

Example 1: Preparation of low molecular fucoidan by hydrolysis of organic acid of fucoidan

Pyruvic acid, citric acid and acetic acid at a concentration of 2 N, typical edible organic acids, 100% (v / v) brewed vinegar with 6% acidity, synthetic vinegar and alcoholic vinegar, and a mixture of 2N pyruvic acid and 100% vinegar (50 / 50%, v / v) was added 1% (w / v) of the polymer fucoidan having a weight average molecular weight of about 170,000 Da and hydrolyzed with heating at 100 ° C for 4 hours. The resulting mixture was concentrated under reduced pressure for 6 hours, filtered through a membrane filter having a molecular weight of 30,000 Da, followed by hot air drying to obtain a powder. The hydrolysis results of fucoidan are summarized in Table 1.

Organic acid used Weight average molecular weight (Da) Polydispersity index Production amount (g) yield(%) Brewed Vinegar 1,500-3,000 2.4 0.75 75 Synthetic Vinegar 1,500-3,000 3.1 0.65 65 Alcohol vinegar 1,200-3,000 3.2 0.42 42 Other vinegar 10,000-15,000 3.6 0.34 34 Pyruvic acid 10,000-15,000 1.8 0.6 60 Citric acid 3,000-4,500 4.1 0.52 52 Acetic acid 2,000-3,000 3.8 0.53 53 Brewed vinegar + pyruvic acid 3,000-15,000 4.9 0.34 34 Synthetic Vinegar + Pyruvic Acid 3,000-15,000 4.8 0.46 46

As shown in Table 1, when brewed vinegar was used among edible organic acids, 0.75 g of fucoidan powder was obtained, which was the highest compared to other acids, and the yield thereof was 75%. However, the yield of other organic acids was 60% for pyruvic acid, 52% for citric acid, 53% for acetic acid, 65% for synthetic vinegar, 42% for alcohol vinegar and 34% for other vinegars. In addition, when vinegar and pyruvic acid, synthetic vinegar and pyruvic acid were mixed in a volume ratio of 1: 1, the yield was slightly lower than that of a single treatment of 34% and 46%, respectively.

Example 2 Weight average molecular weight of low molecular fucoidan produced by hydrolysis of organic acid

The molecular weight of the organic acid treated hydrolyzate obtained in Example 1 was determined by column (PL aquagel, OH30 * 2, Waters Co. Ltd.), RI detector (waters 410 Differential Refractometer, Waters Co. Ltd.), pump (waters 515 HPLC pump, Waters Co. Ltd., and gel permeation chromatography (Waters Co. Ltd.) with an autosampler (waters 717 plus Autosampler, Waters Co. Ltd.), pH 7 buffer solution, at 35 ° C., per minute At a flow rate of ml, polyethylene glycol (PEG) was analyzed as a standard sample (see Fig. 1).

 As can be seen in Figure 1, before hydrolysis, fucoidan shows a molecular weight of about 170,000 Da, but when hydrolyzed using a brewed vinegar having an acidity of 6%, depending on the concentration ratio 3000-5000 Da: 1250-1500 Da: 600-750 Da: 140-210 Da = 30-60: 12-35: 5-20: 15-25 The average molecular weight of the mixture in the form of mixed molecular weights is about 1800-2,300 Da. When the pyruvic acid was treated with 2N, the molecular weight was about 15,000 Da, and when the pyruvic acid was treated with 10N, the molecular weight was about 3,500 Da.

Therefore, when hydrolyzing the fucoidan organic acid, not only the molecular weight is significantly reduced, but also the fucoidan of the weight average molecular weight can be produced according to the organic acid treatment concentration and type. As a result, it can be seen that the molecular weight of fucoidan can be controlled by adjusting the type and concentration of the organic acid to be treated.

Example 3: FT-IR Spectrum of Fucoidan Organic Hydrolyzed

Fucoidan-induced Fourier Transform Infrared Spectrophotometer (FT-IR, Spectrum GX, PerkinElmer, prepared by treating 10% concentrated vinegar with 10% acidity in 1% polymer fucoidan solution at 50/100 (v / v)) Absorption spectrum results analyzed by USA) are shown in FIG. As can be seen in Figure 2, by showing a similar FT-IR spectrum before the brewing vinegar treatment it can be seen that the structural modification of the fucoidan by the production method of the present invention.

Example 4 Sulfuric Acid Group Content of Fucoidan Organic Hydrolyzed

The sulfuric acid content of the fucoidan prepared by treating the concentrated solution of 10 times concentrated brewed vinegar with 6% acidity in 1% polymer fucoidan solution at 50/100 (v / v) was measured.

Sulfuric acid content is 0.5-2 N hydrochloric acid solution and hydrolyzed the bioavailable fucoidan of the present invention at 110 ° C. for 4 hours, and then 0.2 ml of hydrolyzate, 3.8 ml of 4% TCA solution and BaCl ₂ -gelatin reagent , USA) was added and allowed to stand at room temperature for 20 minutes and analyzed by measuring absorbance at 360 nm. K ₂ SO ₄ was used as the standard sample.

As can be seen in Figure 3, the bioavailable fucoidan of the present invention was found to have a sulfuric acid group content of 4.6% higher than the high molecular weight fucoidan before hydrolysis, which plays an important role in physiological activity. In addition, when 10 N pyruvic acid was treated, it was confirmed that the sulfuric acid group content was 9.8% higher than that of high molecular weight fucoidan.

Example 5 Analysis of Main Elements of Fucoidan Organic Hydrolyzed

         Carbon, Hydrogen, Oxygen, Nitrogen and Sulfur, which are the major elements of Fucoidan, prepared by treating 50% (v / v) of a concentrated solution of 10 times concentrated vinegar with 6% acidity in 1% high molecular fucoidan solution. The content was analyzed using an elemental analyzer (Elemental Analyzer, EA1108, Fisons instrument, Italy). The results are summarized in Table 2.

division % Of element before hydrolysis  % Of element after hydrolysis  Carbon (C) 26.7 24.4  Hydrogen (H) 4.9 4.6 Oxygen (O) 48.3 55.4 Nitrogen (N) 0.8 0.1 Sulfur (S) 5.6 7.6 Ash 13.7 7.9 system 100 100

As shown in Table 2, the low-molecular-weight fucoidan of the present invention showed a slightly different aspect of the main element content, and in particular, the content of oxygen and sulfur element was increased compared to before hydrolysis. This experimental result is consistent with the analysis result in which the content of sulfuric acid group of fucoidan of the present invention was increased in Example 4.

Example 6: Water Solubility and Viscosity of Fucoidan Organic Hydrolyzed

1-10% (w / v) of fucoidan powder prepared by treating concentrated concentrate of brine vinegar with 10% acidity in 50% (v / v) of 1% high molecular weight fucoidan solution using distilled water. After preparing to the concentration, the viscosity according to the concentration was analyzed using a viscometer (Viscometer, LVT, Brookfield, USA).

As can be seen in Figure 4a, it can be seen that the solution of the bioavailable fucoidan of the present invention is transparent, the water solubility is significantly increased, the solution of the polymer fucoidan is very high suspension.

In addition, as shown in Figure 4b, the fucoidan of the present invention showed a significantly lower viscosity than the polymer fucoidan, which shows that the fucoidan of the present invention is excellent in water solubility.

Example 7: Empty analysis for confirming tumor cell proliferation inhibitory function

Bioavailable fucoidan prepared by treating concentrated concentrate of brewed vinegar with 10% acidity in 50% (v / v) of 1% polymer fucoidan solution in 50% (v / v) according to the present invention has malignant tumor cell proliferation inhibitory function After verifying that, adding the bioavailable fucoidan prepared in Example 1 to the squamous cell carcinoma cell suspension of tumor cells at a concentration of 0.5-5 0 mg per L and incubated for 2 days, 100 µl of empty (MTT: 3- (4,5-di-methylthiazol-2-yl) -2,5-diphenyl-tetrazolium bromide, Sigma, USA) was added to each groove. After 4 hours of incubation, empty dilutions were carefully removed, and then 100 μl of DMSO (dimethylsulfoxide) was added to each groove and shaken for 15-20 minutes. Absorbance was measured at 540 nm. Analyzed.

As can be seen in Figure 5, the bioavailable fucoidan of the present invention inhibits the growth of tumor cells in a concentration-dependent manner, and further shows about twice the tumor cell proliferation inhibitory ability compared to the high molecular weight fucoidan without brewing vinegar treatment. . In addition, it was confirmed that the tumor cell proliferation inhibiting function is almost absent when only vinegar is treated.

According to the method of the present invention, bioavailable fucoidan can be produced with improved efficiency, convenience, and economical efficiency compared to the method using a conventional enzyme, and the molecular weight of the low molecular weight fucoidan can not be adjusted by the enzyme method. In addition, the method of the present invention does not affect the basic structure of fucoidan, and does not impair the intrinsic physiological activity of fucoidan, and is expected to discriminate activity according to molecular weight size.

In addition, the bioavailable fucoidan of the present invention exhibits a high sulfate content, a small weight average molecular weight of less than 30,000 Da, a low polydispersity index and viscosity, and very high water solubility, thereby greatly improving the absorption rate of the human body, resulting in bioavailability. This greatly improves the unique physiological activity of fucoidan in the body (cancer cancer cell proliferation, antioxidant, blood sugar suppression, anticoagulant, antioxidant, triglyceride and cholesterol lowering function and gastric ulcer healing function). In addition, the improved physical properties of the bioavailable fucoidan of the present invention can solve the problems caused by non-uniformity of conventional fucoidan product quality.

Having described the specific part of the present invention in detail, it is apparent to those skilled in the art that the specific technology is merely a preferred embodiment, and the scope of the present invention is not limited thereto. Thus, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

Claims (15)

(a) 20-40% sulfate content, (b) weight average molecular weight of 1,000-30,000 Da, (c) polydispersity index of 1-6, and (d) viscosity of 0-200 centipoise Bioavailable fucoidan characterized in that it has a. The bioavailable fucoidan according to claim 1, wherein the weight average molecular weight of the bioavailable fucoidan is 3,000-10,000 Da. 3. The bioavailable fucoidan according to claim 2, wherein the weight average molecular weight of the bioavailable fucoidan is 1,500-3,000 Da. The method of claim 3, wherein the bioavailable fucoidan has a weight average molecular weight of 3000-5000 Da: 1250-1500 Da: 600-750 Da: 140-210 Da = 30-60: 12-35: 5-20: 15-25 The bioavailable fucoidan, characterized in that the mixture is a mixture of fucoidan. The bioavailable fucoidan according to claim 1, wherein the sulfuric acid group content of the bioavailable fucoidan is 20-30%. The bioavailable fucoidan according to claim 1, wherein the polydispersity index of the bioavailable fucoidan is 1-4. The bioavailable fucoidan according to claim 1, wherein the viscosity of the bioavailable fucoidan is 0-50 centipoise. [Claim 2] The bioavailability of claim 1, wherein the component ratio of the bioavailable fucoidan is 20-30: 3-5: 45-60: 0.05-1.0: 5.7-20 of carbon: hydrogen: oxygen: nitrogen: sulfur. Fucoidan. 7. The bioavailability according to claim 6, wherein the ratio of constituents of the bioavailable fucoidan is carbon: hydrogen: oxygen: oxygen: nitrogen: sulfur is 23-25: 4.2-4.7: 52-58: 0.07-0.2: 7.0-10. Fucoidan. The bioavailable fucoidan according to claim 1, wherein the bioavailable fucoidan is obtained by treating pyruvic acid, citric acid, acetic acid or a mixture thereof with high molecular weight fucoidan. (a) hydrolyzing the polymer fucoidan by treating pyruvic acid, citric acid, acetic acid or a mixture thereof with the polymer fucoidan; And (b) obtaining a fucoidan hydrolysis product. The method of preparing the bioavailable fucoidan according to any one of claims 1 to 10. 12. The method of claim 11, wherein step (a) is performed using a mixture of pyruvic acid and acetic acid. 12. The process of claim 11, wherein step (a) is performed using 0.1-10 N pyruvic acid, citric acid, acetic acid or mixtures thereof. 14. The process of claim 13 wherein the pyruvic acid to acetic acid ratio of the mixture of pyruvic acid and acetic acid is 1: 2-1: 20 based on volume. 12. The method of claim 11, wherein the acetic acid is provided in the form of vinegar, synthetic vinegar or spirit vinegar.

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