TW201345537A - Tremellafuciformis berk polysaccharide for protecting retinal cells and manufacturing method thereof - Google Patents

Abstract

The present invention discloses a Tremella fuciformis berk polysaccharide for protecting retinal cells and a manufacturing method thereof. The method of manufacturing the Tremella fuciformis berk polysaccharide can obtain natural and non-toxic Tremella fuciformis berk polysaccharide without using chemical reagents. Furthermore, after measuring, it is proved that the Tremella fuciformis berk polysaccharide comprises antioxidants, such as total polyphenol and flavonoid, and has an effect on protecting the retinal cells.

Description

Tremella polysaccharide for protecting retinal cells and preparation method thereof

The invention relates to a tremella polysaccharide, in particular to a tremella polysaccharide which does not add a chemical agent in the process and can protect the retinal cells from oxidative damage and a preparation method thereof.

The literature indicates that edible medicinal mushrooms can provide a source of antioxidants and reduce free radicals such as superoxide anion or hydrogen peroxide produced by active oxygen in the human body. The active ingredients available from mushroom fruit bodies or mycelium extracts include polysaccharides, triterpenoids, proteins, nucleic acids, chitosan. Wait. Among the important active ingredients are glucans and phenols and flavonoids, and β-D-glucans (β-D-glucans) and total phenols and flavonoids have strong antioxidant activity.

The most well-known function of polysaccharides is to activate macrophages, anti-oxidation, detoxification and cholesterol-lowering, etc., by regulating human immunity to alleviate various pathogenic factors and achieve disease resistance. In recent years, studies have found that polysaccharides produced by hyphae of beneficial mushroom mites such as Ganoderma lucidum, Schizophyllum, Cordyceps sinensis, and Hericium erinaceus have strong anti-cancer and disease-resistance functions.

Tremella is a kind of higher basidiomycetes, known as the king of bacteria. Tremella fuciformis Berk, also known as white jelly fungi, belongs to the fungal community, the dinuclear subfamily, the basidiomycetes, the toadstools, the white fungus, the Tremellales, the Tremaceae (Tremellaceae). , Tremella (Tremella). According to statistics, there are about 40 species of Tremella in the world, and its form is composed of two parts, namely the mycelium of vegetative organs and the fruiting bodies of reproductive organs. The fresh white fungus fruit body is pure white and translucent; while it is dry, it is horny, the texture is hard and brittle, and it is white or beige, and the volume shrinks by about 1/10.

In general, the edible part of Tremella is a fruiting body, which is rich in dietary fiber and protein 6.7%~10%, carbohydrate 65%~71.2%, fat 0.6%~12.8%, crude fiber 2.4%~2.75%, Inorganic salts 4% ~ 5.4%, water 15.2% ~ 18.7% and a small amount of vitamin B group. Tremella protein contains a variety of amino acids. The main component also contains an anti-tumor polysaccharide (β-glucan), and thus gradually becomes a high-value agricultural product.

The main chemical structure of Tremella fuciformis polysaccharide is α-1,3-mannan (mannan), with β-D-xylose and β-D-glucuronic acid on the side. The position is followed by β-(1,2)D-xylobiose, which is acidic due to glucuronic acid and belongs to acidic heteropolysaccharide. The Tremella polysaccharides contained in the Tremella mesophyll are mainly divided into five structures according to their composition ratios: (1) Acidic polysaccharide: composed of xylose, mannose and glucuronic acid, in addition to a small amount of glucose and fucose. Its molecule contains a typical ethyl fluorenyl structure. (2) Neutral heteropolysaccharide: its molecular weight is about 8000, mainly composed of xylose, mannose, galactose and glucose. (3) Acidic oligosaccharides: O-β-D-glucuronic acid-(1-2)-O-α-D-mannose-(1-3)-O-α-D-mannose- (1-3)-D-mannose O-(β-D-glucuronic acid)-(1-2)-O-α-D-mannose-(1-3)-D-mannose 2-O -(β-D-glucuronic acid)-D-mannose. (4) Cell wall polysaccharide: it can be isolated to obtain two kinds of cell wall polysaccharides, namely, acid polysaccharide composed of D-glucuronic acid, D-mannose and D-xylose, and D-glucose, D-glucuronic acid, An alkali-insoluble polysaccharide composed of D-mannose or D-xylose. (5) Extracellular polysaccharide: mainly composed of fucose, arabinose, xylose, mannose and glucuronic acid.

There are two sources in the acquisition of white fungus. One is to extract from the mycelium of liquid culture, or to extract the edible fruit body cultivated in space bag, but the related technology of extraction is not mature, and it is impossible to commercialize Production. The traditional method for extracting polysaccharides from the traditional fungus fruit body is to treat the fungus fruit body with an alkaline chemical agent, rinse or adjust the pH value to neutrality, and then extract with hot water or ethanol (alcohol), by this extraction method, Although it can reach 27~30% of the extraction rate of Tremella polysaccharides, it is possible to use the chemical extraction method to make the obtained Tremella polysaccharides residual chemical substances. Therefore, in order to improve the above problems, the technique of extracting Tremella polysaccharides needs to be further improved to obtain a natural Tremella polysaccharide free of chemical substances, and to increase the extraction rate of Tremella polysaccharides.

In view of the above problems in the prior art, the object of the present invention is to provide a Tremella polysaccharide for protecting retinal cells and a method for producing the same, which solves the problem of extracting Tremella polysaccharides by using a chemical substance by a conventional technique, and protecting the retina with the extracted Tremella polysaccharides. cell.

According to the object of the present invention, a method for manufacturing a Tremella polysaccharide for protecting retinal cells is provided, the method comprising the steps of: drying the white fungus; soaking the white fungus in water at a selected ratio; extracting the white fungus extract by using hot water at 55 ° C to 95 ° C, and The Tremella extract is cooled to room temperature; the Tremella extract is centrifuged for the first time; the Tremella extract is filtered to obtain the Tremella concentrate; the Tremella concentrate is separated and concentrated by the mushroom membrane tube separation concentrator to obtain a molecular weight of more than 80,000. Tremella polysaccharides; alcohol is mixed with Tremella polysaccharide for a second centrifugation; the Tremella polysaccharide is washed by alcohol; the Tremella polysaccharide is dried; and the Tremella polysaccharide is allowed to stand for 12 to 48 hours under the environmental conditions of 28 ° C to 55 ° C. To remove the protein in the Tremella polysaccharide.

Preferably, the ratio of the white fungus to the water is 1:40 to 1:100.

Preferably, the white fungus can be immersed in hot water for 2 hours to 6.5 hours.

Preferably, the first centrifugation can be carried out at 4 ° C for 5 minutes to 15 minutes at 4000 rpm to 6000 rpm.

Preferably, the filtering step can be carried out using a filter paper having a pore size of from 4 micrometers to 10 micrometers.

Preferably, the white fungus extract is mixed with alcohol for 12 hours to 48 hours.

Preferably, the second centrifugation is carried out at a speed of 6000 rpm to 15000 rpm for 5 minutes to 15 minutes.

Preferably, the drying time of the white fungus extract is 0.5 hours to 5 hours.

Preferably, the Tremella polysaccharide of the present invention protects retinal cells, which may comprise an excipient or an additive for medicinal and food. Excipients or additives are selected from the group consisting of flavoring agents, sweeteners, preservatives, antioxidants, chelating agents, penetrants, lubricants, lozenge adjuvants, colorants, moisturizers, binders, and pharmaceuticals. In the group consisting of carriers.

The invention determines that the polysaccharide of Tremella fuciformis has total polyphenols and flavonoids, thereby exhibiting physiological activity, can effectively prevent oxidation activity, and has potential for development as a food of physiological antioxidant capacity. In the process of administering oxidative stress to the in vitro retinal pigment epithelial cells, the Tremella extract containing the polysaccharide and the total polyphenols protects the retinal cells and protects the retinal cells from oxidation.

According to the above, the Tremella polysaccharides for protecting retinal cells of the present invention and the method for producing the same can obtain pure and non-toxic natural Tremella polysaccharide extract without using chemical agents, and can be directly used for human body to achieve the effect of protecting retinal cells. .

In order to give your reviewers a better understanding and understanding of the technical features of the creation and the efficiencies achieved, please refer to the preferred examples and the detailed descriptions below.

Example 1: Method for producing Tremella polysaccharides for protecting retinal cells of the present invention

Please refer to FIG. 1 , which is a flow chart of a method for producing a tremella polysaccharide for protecting retinal cells of the present invention. The method for producing a tremella polysaccharide for protecting retinal cells of the present invention comprises the step of drying the white fungus in step S20. Next, step S21 is to soak the white fungus by water at a ratio of 1:40 to 1:100. In step S22, the white fungus extract is extracted by hot water at 55 ° C to 95 ° C, and the white fungus extract is cooled to room temperature, wherein the white fungus is immersed in hot water for 2 hours to 6.5 hours. Next, under the condition of 4 ° C, step S23 is to carry out the first centrifugation of the white fungus extract, the centrifugation speed is 4000 rpm to 6000 rpm, and the centrifugation time can be 5 minutes to 15 minutes. The Tremella extract is filtered in step S24 with a filter paper having a pore size of 4 μm to 10 μm to obtain a white fungus concentrate. After that, in order to obtain a polysaccharide having different effects, step S25 is to separate and concentrate the tremella concentrate by a mushroom membrane tube separation concentrator to obtain a tremella polysaccharide having a molecular weight of more than 80,000. Next, in step S26, the second centrifugation is carried out by mixing the tremella polysaccharide with alcohol, and the standing time is 12 hours to 48 hours, and the centrifugation is performed at a speed of 6000 rpm to 15000 rpm for 5 minutes to 15 minutes, and then the step S27 is borrowed. The tremella polysaccharide is washed by alcohol. Then, step S28 is to dry the tremella polysaccharide for 0.5 hours to 5 hours. Finally, in step S29, the Tremella polysaccharide is allowed to stand for 12 to 48 hours at 28 ° C to 55 ° C to remove the protein in the Tremella polysaccharide.

Example 2: Analysis of β-glucan (β-Glucan)

(1) Determination of total sugar content

In this embodiment, after extracting the Tremella polysaccharides from the LT1 Tremella line and the LT6 Tremella line by using the manufacturing method described in Example 1, 0.1 g of the Tremella polysaccharides of each line of the scales were added, and 1.5 ml of hydrochloric acid (37% by volume) was added thereto. The water bath was taken at 30 ° C and vortexed once every 15 minutes for 45 minutes. Next, after adding 10 ml of deuterium depleted water (DDW) to vortex, the secondary water was heated to 95 ° C for 2 hours, and cooled to room temperature. Add 10 ml of 2N potassium hydroxide (KOH), pour the contents of each tremella tube into a 100 ml dosing bottle, rinse the tube wall with 200 mM pH 5.0 sodium acetate buffer solution, and quantify 100 ml to seal The membrane was sealed with a vial and inverted and mixed, poured into a 250 cc centrifuge bottle and centrifuged at 1500 rpm for 10 minutes. Take 0.1 ml of the supernatant and add exo-1,3-β-D-glucanase (20 U/ml) and β-glucosidase (20 μg/ml). β-Glucosidase) 200 mM acetate buffer (pH 5.0) was placed in a 15 ml tube, vortexed, and incubated at 40 ° C for 1 hour; and 3 ml glucose oxidase/peroxidase was added. The mixture was prepared into a standard group and a blank group, and after taking a water bath at 40 ° C for 20 minutes, 1000 μl of the absorbance at a wavelength of 510 nm was measured in a cuvette.

1. Blank group: 3 ml of exo-1,3-β-D-glucanase (exo-1,3-β-D-glucanase) (20 U/ml) and β-glucosidase (and β -Glucosidase) with a pH 5.0 sodium acetate buffer solution of +0.2 ml.

2. Standard group: 3 ml of exo-1,3-β-D-glucanase (20 μL/ml) and β-glucosidase (β-) Glucosidase) Add 0.1 ml of pH 5.0 sodium acetate buffer solution plus 0.1 ml of D-glucose standard solution.

(2) Determination of α-glucan (α-Glucan) content

Each weighing scale LT1 Tremella strain and 0.1 g Tremella polysaccharide of LT6 Tremella strains were placed in a 50 ml refrigerated centrifuge tube, 2 ml of 2M potassium hydroxide (KOH) was added, and the magnet was placed in an ice bath for 20 minutes, then added. 8 ml of pH 3.8 sodium acetate buffer solution 1.2 M, with 0.2 ml of amyloglucosidase (1630 U / ml) and transforming enzyme (500 U / ml), vortexed in an ice bath at 40 ° C for 30 minutes , the same weight of each tube scale, centrifuged at 3000 rpm for 10 minutes, then take 0.1 μl of the supernatant into a 15 ml tube and add 3 ml of exo-1,3-β-D-glucanase (exo- In the 1,3-β-D-glucanase) (20 U/ml) and β-glucosidase (β-Glucosidase), prepare a standard group and a blank group, and take a bath at 40 ° C for 20 minutes, and take 1000 μl of buffer solution. The absorbance at a wavelength of 510 nm was measured in a cuvette.

1. Blank group: 3 ml of exo-1,3-β-D-glucanase (20 μL/ml) and β-glucosidase (β) -Glucosidase) Add 0.2 ml of a pH 5.0 sodium acetate buffer solution.

2. Standard group: 3 ml of exo-1,3-β-D-glucanase (20 μL/ml) and β-glucosidase (β) -Glucosidase) Add 0.1 ml of a pH 5.0 sodium acetate buffer solution with 0.1 ml of D-glucose standard solution.

(3) The content of α-glucan (α-glucan) was subtracted from the total sugar content to obtain the content of β-glucan (β-Glucan) in different strains of Tremella, and the results are shown in Table 1.

Example 3: Determination of total polyphenol content

In the present embodiment, the LT1 Tremella line and the LT6 Tremella extract were separately subjected to the following steps by the production method described in Example 1, to determine the total polyphenol content of each Tremella line.

(1) Formulation reagents:
1. 60% aqueous methanol.
2. Reagent A: 0.3% hydrogen chloride + 60% aqueous methanol.
3. Reagent B: 2% sodium carbonate solution (Sodium Carbonate).
4. Reagent C: 50% Folin-ciocalteu's phenol reagent.

(2) Formulation standard group: Take 100 mg gallic acid, pour into a 20 ml sample vial, add 10 ml of reagent A, and dilute to a standard solution of concentration 10 mg/ml with reagent A.

(3) Determination method: Take 50 μl of each concentration standard and the sample to be tested, put into a 1.5 ml microcentrifuge tube, add 1 ml of reagent B, vortex and place for 2 minutes, add 50 μl of reagent C in the dark. Vortex, place at room temperature for 30 minutes, pipet 1 ml to the colorimetric tube, and fill the blank reagent tube with 1 ml of secondary water, and measure the absorbance at 750 nm.

Please refer to FIG. 2 and FIG. 3 together, which is a standard curve of the total polyphenol absorbance and a schematic diagram of the total polyphenol content of the Tremella polysaccharides of the present invention for protecting retinal cells. Figure 2 shows the absorbance curve of total polyphenols at a wavelength of 750 nm, and the absorbance is proportional to the concentration. Using this standard curve against Figure 3, we can see the total of the Tremella polysaccharides of LT1 and Tremella LT6. Polyphenol content.

Example 4: Determination of flavonoid content

In the present embodiment, the LT1 Tremella line and the LT6 Tremella extract were separately subjected to the following steps to determine the flavonoid content of each Tremella line by the manufacturing method described in Example 1.

(1) Formulation reagents:
1. 70% aqueous ethanol solution.
2. Reagent H: 10% aqueous solution of aluminum chloride.
3. Reagent I: 1 M potassium acetate aqueous solution.

(2) Preparation of standard: Take 10 mg of Quercetin dehydrate, pour into a 20 ml sample vial, and add 10 ml of 70% ethanol solution. It was diluted with a 70% aqueous solution of ethanol to a standard solution of the desired concentration of 1 mg/ml.

(3) Experimental method: Take 0.5 ml of each standard and the sample to be tested, put into a 15 ml centrifuge tube, and take 0.5 ml of secondary water as a blank reagent tube. Add 2.8 ml of secondary water and 1.5 ml of 99.9% methanol, then add 0.1 ml of reagent H and 0.1 ml of reagent I, vortex, place at room temperature for 30 minutes, pipet 1 ml to the colorimetric tube, and measure Absorbance at 415 nm illumination.

Please refer to FIG. 4 and FIG. 5 together, which is a standard curve of flavonoid absorbance and a schematic diagram of the flavonoid content of the Tremella polysaccharides of the present invention for protecting retinal cells. Figure 4 is a graph showing the absorbance of flavonoids at a wavelength of 415 nm. From the standard curve, Figure 5 shows the flavonoid content of Tremella fuciformis LT1 and Tremella fuciformis LT6.

Example 5: Cell viability assay (MTT assay)

Retinal pigment epithelial cells (RPE-19) were seeded in 96 well microplates at 100 μl of 5× ^{10 4} cell/well cell volume and placed in an incubator for 24 hours to remove old cultures. Wash the cells with 100 μl/well of Phosphate Buffer Saline (PBS) and remove them, then add 100 μl/well of cell culture medium and pre-tested samples, and culture 24, 48 After 72 hours, the solution was removed, and then 100 μl/well of 0.5 mg/ml MTT solution was added, and the solution was removed after incubating in a 37 ° C cell culture chamber (containing 5% CO ₂ ) for 4 hours, and then 100 μl was added. /well dimethyl sulfoxide (DMSO) dissolves formazan crystals, and adds the same volume of DMSO to the blank well as a blank group using an enzyme-linked immunosorbent assay (ELISA reader) The absorbance was detected at a wavelength of 540 nm.

Please refer to Fig. 6, which is a schematic diagram showing the survival rate of retinal pigment epithelial cells (ARPE-19) by the Tremella polysaccharide (LT1) for protecting retinal cells of the present invention. In the figure, the concentration of different Tremella polysaccharides was measured, and the cell viability of the pigment epithelial cells (ARPE-19) was cultured for 24 hours and 48 hours, respectively, as shown in Fig. 6, and cultured in various concentrations of Tremella polysaccharide for 48 hours. The survival rate of the pigmented epithelial cells (ARPE-19) was higher than that of the pigment epithelial cells (ARPE-19) cultured for 24 hours.

Example 6: Protection of ARPE-19 cells damaged by H ₂ O ₂ oxidation

Tremella extract (LT1) protects ARPE-19 cells damaged by hydrogen peroxide (H ₂ O ₂ ) oxidation. RPE-19 cells are seeded in 12 well microplates at a cell size of 1000 μl of 6× ^{10 4} cell/well. After culturing for 24 hours in an incubator, the old culture solution was removed, and the cell surface was washed with 500 μl/well of PBS buffer, and the old culture solution was removed, followed by 1000 μl/well cell culture. Liquid and pre-tested samples, and after 12 hours of incubation, remove the solution, wash the cell surface with 500 μl/well of PBS buffer and remove, then add 1000 μl/well cell culture medium and 100 μl/well. 500 μM hydrogen peroxide H ₂ O ₂ and cultured for 2 hours. After removing the solution, wash the cell surface with 500 μl/well PBS buffer and remove it. Add 1000 μl/well cell culture medium and incubate for 4 hours. After removing the solution, the cell surface was washed with 500 μl/well of PBS buffer and removed. After adding 3% of formalin for 15 minutes, the cells were removed, and the cell surface was washed with 500 μl/well of PBS buffer and removed. Add 1% crystal violet for 15 minutes and remove with 500 μl/well PBS buffer After washing and removal of the cell surface, the extract buffer was added 300 μl / well, the measured brightness of the points in the 590 nm after 20 minutes.

Please refer to Fig. 7, which is a schematic diagram of the H ₂ O ₂ oxidative damage of the pigmented epithelial cells (ARPE-19) of the Tremella polysaccharide (LT1) protecting retinal cells of the present invention. Among them, *p indicates that the experimental values are different; **p indicates that the experimental values are significantly different. In the figure, the spectrophotometry of the pigment epithelial cells (ARPE-19) containing various concentrations of Tremella polysaccharides was oxidized by hydrogen peroxide (H ₂ O ₂ ) and then irradiated at a wavelength of 590 nm. As shown in the figure, healthy pigment epithelial cells (ARPE-19) without the addition of Tremella polysaccharide showed pigmented epithelium damaged by oxidation compared with pigment epithelial cells (ARPE-19) damaged by hydrogen peroxide (H ₂ O ₂ ) oxidation. The absorbance of cells (ARPE-19) is greatly reduced. However, if the Tremella polysaccharide is added to the pigment epithelial cells (ARPE-19), the absorbance can be increased compared with the pigment epithelial cells (ARPE-19) without the addition of Tremella polysaccharides. Thus, it can be shown that the Tremella polysaccharide has a protective pigment epithelium. The effect of oxidative damage on cells (ARPE-19).

Example 7: Repair of RPE-19 cells damaged by H ₂ O ₂ oxidation

RPE-19 cells were seeded in 12 well microplates at 1000 μl of 6× ^{10 4} cell/well cells, and placed in an incubator for 24 hours, the old culture solution was removed, and then 500 After washing the cell surface with μl/well PBS buffer and removing the old culture solution, add 1000 μl/well cell culture medium and 100 μl/well 500 μM hydrogen peroxide (H ₂ O ₂ ) and incubate for 2 hours. The solution was washed with 500 μl/well of PBS buffer and removed, then 1000 μl/well of cell culture medium and pre-tested samples were added, and after 24 hours of incubation, the solution was removed and 500 μl/well was added. After washing the cell surface with PBS buffer and removing it, wash the cell surface with 500 μl/well PBS buffer and remove it. Add 3% of formalin for 15 minutes, remove it, and buffer with 500 μl/well PBS. After washing the cell surface and removing it, add 1% crystal violet for 15 minutes, remove it, wash the cell surface with 500 μl/well PBS buffer and remove it, add 300 μl/well of extract buffer to The 590 nm split value was measured after 20 minutes.

Please refer to FIG. 8 , which is a schematic diagram of H ₂ O ₂ oxidative damage of retinal pigment epithelial cells (RPE-19) repaired by Tremella polysaccharide (LT1) for protecting retinal cells of the present invention. Among them, *p indicates that the experimental values are different; **p indicates that the experimental values are significantly different. As shown in the figure, the absorbance was greatly reduced after being oxidized by hydrogen peroxide (H ₂ O ₂ ) as compared with healthy retinal pigment epithelial cells without Tremella polysaccharide. However, the retinal pigment epithelial cells (RPE-19) with different concentrations of Tremella polysaccharides, after being oxidized by hydrogen peroxide (H ₂ O ₂ ), absorbance and retinal pigment epithelial cells (RPE-19) with Tremella polysaccharides are not included. Compared with the retinal pigment epithelial cells (RPE-19) of Tremella fuciformis polysaccharides, the Tremella polysaccharides have the effect of repairing injured retinal pigment epithelial cells.

In summary, the Tremella polysaccharide (LT1) for protecting retinal cells of the present invention and the method for producing the same are not used in the manufacturing process, so that a natural non-toxic Tremella polysaccharide can be obtained, and after the above examples, a molecular weight of 80 can be obtained. KDa Tremella polysaccharide. It has been proved by experiments that Tremella polysaccharide has the function of protecting or repairing retinal pigment epithelial cells, and can be added to eye drops medical health care products, or adding excipients or other chemical substances as raw medicines.

The above is intended to be illustrative only and not limiting. Any equivalent modifications or alterations to the spirit and scope of the invention are intended to be included in the scope of the appended claims.

S20~S29. . . Step flow

Fig. 1 is a flow chart showing the method for producing Tremella polysaccharides for protecting retinal cells of the present invention.
Figure 2 is a standard curve of total polyphenol absorbance.
Fig. 3 is a schematic view showing the total polyphenol content of the Tremella polysaccharides of the present invention for protecting retinal cells.
Figure 4 is a standard curve of flavonoid absorbance.
Fig. 5 is a schematic view showing the content of flavonoids of Tremella polysaccharides for protecting retinal cells of the present invention.
Fig. 6 is a schematic diagram showing the survival rate of the pigment ear cells (ARPE-19) by the Tremella polysaccharide (LT1) for protecting retinal cells of the present invention.
Fig. 7 is a view showing the survival rate of the pigment epithelial cells (ARPE-19) which are protected from H ₂ O ₂ oxidation by the Tremella polysaccharide (LT1) which protects the retinal cells of the present invention.
Fig. 8 is a schematic diagram showing the H ₂ O ₂ oxidative damage of the retinal pigment epithelial cells (RPE-19) repaired by the Tremella polysaccharide (LT1) protecting the retinal cells of the present invention.

S20~S29. . . Step flow

Claims (10)

A method for producing a Tremella polysaccharide for protecting retinal cells, comprising the following steps:
Dry a white fungus;
Soaking the white fungus in water at a selected ratio;
The Tremella extract is extracted by hot water of one of 55 ° C to 95 ° C, and the Tremella extract is cooled to room temperature;
The white fungus extract is subjected to a first centrifugation;
Filtering the white fungus extract to obtain a white fungus concentrate;
Separating and concentrating the white fungus concentrate by a mushroom membrane tube separation and concentrating machine to obtain a tremella polysaccharide having a molecular weight of more than 80,000;
Mixing the tremella polysaccharide with alcohol for a second centrifugation;
Washing the Tremella polysaccharide by alcohol;
The Tremella polysaccharide is dried; and the Tremella polysaccharide is allowed to stand for 12 to 48 hours under the environmental conditions of 28 ° C to 55 ° C to remove the protein in the Tremella polysaccharide. The method for producing a tremella polysaccharide for protecting retinal cells according to claim 1, wherein the selected ratio of the white fungus to water is 1:40 to 1:100. The method for producing a tremella polysaccharide for protecting retinal cells according to claim 1, wherein the white fungus is immersed in the hot water for 2 hours to 6.5 hours. The method for producing a tremella polysaccharide for protecting retinal cells according to claim 1, wherein the first centrifugation is carried out at a temperature of 4000 rpm to 6000 rpm for 5 minutes to 15 minutes at 4 ° C. The method for producing Tremella polysaccharides for protecting retinal cells according to claim 1, wherein the method is to filter using a filter paper having a pore size of 4 μm to 10 μm. The method for producing a tremella polysaccharide for protecting retinal cells according to claim 1, wherein the white fungus extract is mixed with the alcohol for 12 hours to 48 hours. The method for producing a tremella polysaccharide for protecting retinal cells according to claim 1, wherein the second centrifugation is performed at a speed of 6000 rpm to 15000 rpm for 5 minutes to 15 minutes. The method for producing a tremella polysaccharide for protecting retinal cells according to claim 1, wherein the white fungus extract has a drying time of 0.5 hours to 5 hours. A tremella polysaccharide for protecting retinal cells, which is produced by using the method for producing a tremella polysaccharide for protecting retinal cells according to any one of claims 1 to 8, which comprises the endowment of medicine and food. a agent or an additive. The Tremella polysaccharide for protecting retinal cells according to claim 9, wherein the excipient or the additive is selected from the group consisting of a flavoring agent, a sweetener, a preservative, an antioxidant, a chelating agent, and the like. A group of lubricants, lozenge adjuvants, colorants, humectants, binders, and pharmaceutical compatible carriers.