KR20040066344A - Hericium erinaceus exo-biopolymer having a hypolipidemic effect, method for isolation and the use thereof - Google Patents

Abstract

The present invention relates to an extracellular polysaccharide derived from a roe deer fungus mushroom having an hypolipidemic effect, and the extracellular polysaccharide produced from the mycelium liquid culture of the roe deer fungus mushroom was orally administered to hyperlipidemic rats as compared to the control group. There is an excellent effect of increasing the hypolipidemic effect depending on the concentration of. In addition, the extracellular polysaccharide of the present invention has an excellent effect of preventing atherosclerosis by reducing the concentration of total cholesterol, triglycerides, phospholipids, arteriosclerosis index and LDL cholesterol of plasma and maintaining the concentration of HDL cholesterol high. Moreover, the extracellular polysaccharide of the present invention has an excellent effect of reducing the synthesis of cholesterol by inhibiting the activity of HMG-CoA reductase of the liver. The extracellular polysaccharide of the present invention having the hypolipidemic effect is a glycoprotein having a molecular weight of 40 kDa, and is composed of 91.2% sugar and 8.8% amino acid, and the major sugars of the extracellular polysaccharide component are fucose, galactose and mannose. And the main amino acids are serine, aspartic acid, glutamic acid, and glycine.

Description

Extracellular polysaccharide derived from roe deer fungus with hypolipidemic effect, isolation method and use {Hericium erinaceus exo-biopolymer having a hypolipidemic effect, method for isolation and the use}

The present invention relates to an extracellular polysaccharide derived from the roe deer fungus having an hypolipidemic effect. More specifically, the present invention relates to an extracellular polysaccharide produced by the mycelium liquid culture of the rhinocyst beetle mushroom having a hypolipidemic effect in hyperlipidemic rats and a method for isolating the same.

Edible mushrooms contain high fiber, protein, trace elements, and low fat, making them well known as an ideal material for dietary prevention against atherosclerosis. The hypocholesterolemic effect of some mushrooms has already been studied. Thirsty mushrooms, mulberry mushrooms, and ganoderma lucidum are reported to have lower serum cholesterol levels in hyperlipidemic rats. However, early reports on the hypolipidemic effect of mushrooms were mainly limited to components extracted from fruiting bodies or mycelium. The components secreted from the culture mycelium into the culture medium have been reported to have various bioactive properties, but efforts to test the potential hypolipidemic effect have been very poor. Recently, bioactive extracellular polysaccharides produced from mycelial liquid culture of mushrooms have been studied. This was possible because the materials could be produced in a relatively simple purification step in the culture medium. Roe deer mushrooms are well known in the East as edible and medicinal mushrooms. This mushroom is called Yamabush Take in Japan and Houtou in China. Roe deer mushrooms have been proven to contain many biologically active components that exhibit interesting physiological activities such as synergistic, cytotoxic, antimicrobial and anticancer activity of neuronal growth factors.

Accordingly, it is an object of the present invention to provide an extracellular polysaccharide having a hypolipidemic effect produced from mycelium liquid culture of roe deer mushroom and its isolation method.

The object of the present invention is to culture the mycelia of the scab, the extracellular polysaccharide from the culture medium and obtain the extracellular polysaccharide orally administered to hyperlipidemic rats to obtain blood from the mice and analyze the blood to analyze the total cholesterol, The triglyceride, HDL (high concentration lipoprotein) cholesterol, phospholipids, LDL (low concentration lipoprotein) cholesterol content and arteriosclerosis index were investigated to investigate the hypolipidemic effect of the extracellular polysaccharide derived from Rhizome eryngii and the inhibition of the extracellular polysaccharide. Achievement by gel filtration on Sepahedz CL-6B column saturated with 0.2 M NaCl to identify the activator of blood, and then confirming the components and molecular weight of the extracellular polysaccharide and analyzing the amino acid and sugar components of the extracellular polysaccharide. It was.

Hereinafter, the configuration of the present invention will be described by examples.

Figure 1 is a graph showing the components of the extracellular polysaccharide of the roe deer fungus using Ceuzers CL-6B chromatography.

Figure 2 is a graph showing the molecular weight of the extracellular polysaccharide of the roe deer mushroom using HPLC.

The present invention is the mycelium liquid culture and the separation of the extracellular polysaccharide in the culture medium of the roe deer mushroom; Investigation of the hypolipidemic effect of the extracellular polysaccharide in hyperlipidemic rats; And, it comprises the component analysis step of the extracellular polysaccharide.

More specifically, the extracellular polysaccharide derived from the Roestock fungus of the present invention is obtained from the following steps:

A) Inoculate the culture medium of the roe deer mushroom into potato / dextrose agar medium and subculture at 4 ° C. every three months,

B) Put the culture solution into potato / dextrose broth (pH 5.0) and incubate in a rotary shaker (120 rpm) at 25 ° C.,

10) After 10 days of the culture, the culture medium was pulverized in aseptic state, and then the culture medium prepared in advance (glucose 20 g / L, MgSO ₄ · 7H ₂ O 0.5 g / L, KH ₂ PO ₄ 0.46 g). / L, yeast extract 2 g / L, peptone 2 g / L; adjusted to pH 4 before sterilization) at 1% (v / v),

D) culture the mycelia liquid for 12 days in a rotary shaker at 120 rpm, 25 ℃ condition containing the pulverized,

E) the supernatant obtained by centrifuging the mycelium liquid culture solution at 10,447 x g for 20 minutes, followed by centrifugation by ethanol precipitation,

F) obtained by diluting and dialysis with water and lyophilization.

In order to investigate the hypoglycemic effect of the extracellular polysaccharide derived from the present invention Roe beetle mushroom obtained from the above separation method, the hyperlipidemic rats derived from the diet were orally administered to the extracellular polysaccharide, and then blood was obtained from the mice. Investigate the blood content of total cholesterol, triglycerides, HDL (high concentration lipoprotein) cholesterol, phospholipids, LDL (low concentration lipoprotein) cholesterol, and arteriosclerosis index to investigate the hypolipidemic effect of extracellular polysaccharides It was.

Furthermore, gel filtration was carried out on a Ceuzers CL-6B column saturated with 0.2 M NaCl to identify the hypolipidemic activity of the extracellular polysaccharide derived from the present invention of the present invention. The amino acid and sugar components of the extracellular polysaccharide were analyzed.

The pharmaceutical composition may be prepared by mixing a composition for treating hyperlipidemia, or a composition for preventing atherosclerosis, comprising an extracellular polysaccharide having an hypolipidemic effect derived from a deer rot fungus or a pharmaceutically acceptable carrier. The pharmaceutical composition of the present invention may further include conventionally used excipients, disintegrants, sweeteners, glidants, anti-esthetics, etc., unit administration such as oral or parenteral formulations by conventional methods It may be formulated in a pharmaceutical form or in multiple dosage forms. The composition may be parenterally or orally administered as desired, and the exogenous polymer derived from the locust fungus per kg body weight per day may be administered from 0.05 g to 0.2 g, preferably from 0.2 g to 1 g. It can be divided into several doses. Dosage levels for a particular patient may vary depending on the particular compound to be used, body weight, patient's age, sex, health condition, diet, time of administration, method of administration, rate of excretion, severity of disease, and the like.

Each data value shown in each example is shown in the table expressed as the average ± standard deviation. The mean of each group was compared by single variance analysis and Duncan's multiple-range test. Statistical difference considered p <0.05 as a significant value.

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 Isolation of Extracellular Polysaccharide from Mycelia Liquid Culture of Roebuck Mushroom

In order to isolate the extracellular polysaccharide of the roe deer fungus of the present invention, first, the mycelium of roe deer fungus was obtained from the Rural Development Administration. Roe deer mushrooms were placed on a potato / dextrose agar (PDA, Diffcosa) slope and subcultured at 4 ° C. every three months. The culture medium of the Roebuck mushroom was spawned in a 250 mL-flask containing 100 mL of potato / dextrose broth (pH 5.0) in a rotary shaker (120 rpm) at 25 ° C. After 10 days, 50 mL of the culture was aseptically crushed and cultured at 1% (v / v) (glucose 20 g / L, MgSO ₄ · 7H ₂ O 0.5 g / L, KH ₂ PO ₄ 0.46 g / L). , 2 g / L yeast extract, 2 g / L peptone; pH was adjusted to 4 before sterilization). Mycelial liquid culture was carried out in a 500 mL-flask containing 200 mL of medium for 12 days in a rotary shaker (120 rpm) at 25 ℃. The culture medium obtained above was centrifuged (10,447 xg / 20 minutes), and then the supernatant was treated with ethanol. The ethanol precipitate was dissolved in water, dialyzed, and lyophilized to obtain extracellular polysaccharide.

Example 2: Hypoglycemic Effect of Extracellular Polysaccharides Derived from Roebuck Mushroom

The extracellular polysaccharide derived from the present invention of the present invention, the extract of the present invention in Example 1, was orally administered to the diet-induced hyperlipidemic rats to examine the growth reaction and hypolipidemic effect of the rats. First, Sprague-Dalley rats (4 weeks old) obtained from the Korea Advanced Institute of Chemical Technology were placed in stainless steel cages, respectively, and maintained at a constant temperature (22 ± 2 ℃) and humidity (55 ± 5 ℃), and the contrast was adjusted every 12 hours. Breeding in the nursery. AIN-76 modified diets were fed to rats for 6 weeks. The feed consists of 55.5% carbohydrates per weight, 14% fat (30% of total energy), 20% protein, 5% fiber and 0.5% cholesterol, increasing fat content to increase hyperlipidemia in mice Induced. Feed and water were provided to breeding animals sufficiently. After breeding in the nursery for 2 weeks, each rat group was orally administered saline (control) and extracellular polysaccharides at a concentration of 50-200 mg / kg body weight daily for 8 weeks at 8 am to 9 am. Dietary intake and body weight were recorded on different days each day. After the last oral administration, the rats were fasted for 14 hours and lightly anesthetized between 8 and 12 am with ether anesthesia to incise the abdomen to obtain blood from the aorta, and the blood sample was placed in a heparinized tube and 1,100 xg, 10 minutes. Plasma was separated by centrifugation. Cold saline was sparged in the liver, dissected and frozen at -70 ° C. Total lipids of liver were extracted by treating chloroform / methanol (2: 1, v / v) mixture.

Plasma total cholesterol, triglycerides, HDL (high concentration lipoprotein) cholesterol, and phospholipid concentrations were evaluated by enzyme test kit (Asan Pharmaceutical Co., Ltd., Korea). LDL (low concentration lipoprotein) cholesterol and arteriosclerosis index were calculated according to the following equation:

LDL cholesterol = total cholesterol-HDL cholesterol-(triglycerides / 5)

Arteriosclerosis Index = (Total Cholesterol-HDL Cholesterol) / HDL Cholesterol

Liver total cholesterol and triglycerides were analyzed according to the same method as the method for measuring total cholesterol and triglycerides in plasma after treatment with Triton X-100.

To separate liver microsome proteins, liver microsomes were prepared with a slight modification of Hulcher and Oleson's method. To 1 g of liver tissue, add 3 mL of cold grinding buffer (50 mM KH ₂ PO ₄ , 0.1 M sucrose, 50 mM NaCl, 30 mM EDTA, 2 μM dithiothreitol, pH 7.2) and grinder (Glass Co., Ltd.). , USA). It was centrifuged twice at 10,000 xg for 15 minutes in a Ti-50 rotor at 4 ° C, and then centrifuged at 100,000 xg for 1 hour to separate microsomal fractions. The fractions were suspended in ground buffer and again centrifuged at 100,000 × g for 1 hour. After centrifugation, the microsomal pellets were triturated with grinding buffer containing no dithiothreitol and then stored at -70 ° C. Proteins were quantified according to the Lowry method.

In addition, Shapiro's method in the fresh liver microsomes prepared above was slightly modified to measure the activity of HMG-CoA reductase. A mixture (60 μl) of microsomes (200 μg) and 500 nM NADPH (diluted with a reaction solution containing 0.1 M triethanolamine and 10 mM EDTA) was incubated at 37 ° C. for 15 minutes in advance. Then, 50 nM [ ¹⁴ C] HMG-CoA (NENTM, Life Sciences Products, USA) was added and incubated at 37 ° C. for 15 minutes. 15 μl of 10 M HCl was added and the reaction mixture was again incubated at 37 ° C. for 30 minutes to produce mevalonolactone from mevalonate. The mixture was centrifuged for 10,000 xg, 15 minutes and the supernatant was added to silica gel 60F254 TLC plate (Merck, Germany) and melononolactone standards were added together. The plate was developed in acetone: benzene (1: 1, v / v) and air dried. The area containing mevalonate was scratched with a clean razor blade and its [ ¹⁴ C] radioactivity was measured on a scintillation counter (Packard Tricap 1600 TR, Packard, The Netherlands). The results are expressed in mg of mevalonate pmole / min / microsomal protein.

Effect on Growth Response of Extracellular Polysaccharides Derived from Roebuck Mushrooms in Hyperlipidemic Rats Group (extrapolysaccharide, g / kg / day) Body weight (g / day) Dietary Intake (g / day) Control group ¹⁾ 4.5 ± 0.2 17.1 ± 0.4 50 4.3 ± 0.2 17.1 ± 0.5 100 4.6 ± 0.2 17.3 ± 0.4 150 4.4 ± 0.2 18.6 ± 0.2 200 4.2 ± 0.1 16.8 ± 0.2

¹⁾ was administered as saline instead of the extracellular polysaccharide derived from roe deer mushroom of the present invention as a control. Each value is expressed as mean ± standard deviation (n = 8).

As shown in Table 1, as a result of administering to the rats by varying the dose of the extracellular polysaccharide derived from the fungus of the present invention, all rat experimental groups were healthy and maintained normal weight for 4 weeks. In the four experimental groups, body weight and dietary intake were not significantly different compared to the control group. In addition, oral administration of extracellular polysaccharides did not affect the overall behavior of rats and no animals died. Therefore, oral administration of the extracellular polysaccharide derived from the Roe beetle mushroom did not have a detrimental effect on rats.

Effects of Extracellular Polysaccharides Derived from Roebuck Mushrooms on Plasma Total and LDL Cholesterol in Hyperlipidemic Rats Group (extrapolysaccharide, g / kg / day) Total cholesterol (mg / dl) LDL Cholesterol (mg / dl) ¹⁾ Control 100.6 ± 2.8 74.5 ± 4.4 50 80.0 ± 3.3 56.8 ± 3.7 100 74.1 ± 3.8 47.9 ± 4.4 150 73.1 ± 2.7 46.8 ± 5.2 200 67.5 ± 4.1 40.7 ± 6.9

¹⁾ Oral Saline

²⁾ Total Cholesterol-HDL Cholesterol-(Triglycerides / 5)

As shown in Table 2, the effects of extracellular polysaccharides derived from the fungus on the total cholesterol and LDL cholesterol of plasma showed that the total cholesterol and LDL cholesterol concentrations of plasma were significantly decreased in all experimental groups. The increase was further reduced.

It is generally known that LDL cholesterol in plasma is more involved in atherosclerosis than total cholesterol. In this example, the administration of the extracellular polysaccharide at a concentration of 200 mg / kg body weight lowered the total cholesterol and LDL cholesterol concentrations to 32.9% and 45.4%, respectively, compared to the control group.

The effect of extracellular polysaccharides on the extracellular polysaccharides of Roe deer fungi is thought to be due to the adhesion of the extracellular polysaccharides. The hypolipidemic effect continued to increase as the concentration of extracellular polysaccharides increased. This is associated with an increase in viscosity with increasing dose of extracellular polysaccharide. Because of these characteristics, it is possible to reduce the absorption of triglycerides and cholesterol by inhibiting the formation of colloidal particles in the small intestine and changing the physical characteristics of the intestinal mucosa of rats. However, the structural effect of the extracellular polysaccharide derived from the locust fungus cannot be excluded in showing the hypolipidemic effect. Moreover, like total cholesterol, LDL cholesterol continued to decrease with increasing daily dose of extracellular polysaccharides. Thus, it can be argued that the main reason for the decrease in total cholesterol concentration in plasma is that extracellular polysaccharides lower the concentration of LDL cholesterol. In addition, significant reductions in LDL and total cholesterol in plasma may be due to the inhibition of HMG Co-A reductase in liver tissue, resulting in reduced cholesterol production and / or sufficient removal of LDL cholesterol without subsequent regeneration in various tissues. It is feed. LDL values in plasma modify VLDL metabolism, including the conversion of VLDL (ultra low concentration lipoprotein) to LDL (low concentration lipoprotein). Basically, the initial VLDL is converted to mature VLDL and to LDL as triglycerides are lost by the action of LPL.

Effect of Extracellular Polysaccharides Derived from Roebuck Mushrooms on Plasma HDL Cholesterol and Atherosclerosis in Hyperlipidemic Rats Group (extrapolysaccharide, g / kg / day) HDL cholesterol (mg / dl) Arteriosclerosis index ¹⁾ Control group ¹⁾ 16.7 ± 1.7 5.03 ± 0.69 50 18.1 ± 1.3 3.42 ± 0.23 100 19.6 ± 0.7 2.79 ± 0.21 150 21.4 ± 2.0 2.42 ± 0.32 200 21.9 ± 3.1 2.08 ± 0.28

¹⁾ Oral Saline

²⁾ (Total Cholesterol-HDL Cholesterol) / HDL Cholesterol

Table 3 shows the effects on the plasma HDL cholesterol and atherosclerosis index of the extracellular polysaccharide derived from the Roestock fungus. In comparison to the control, extracellular polysaccharides increased the concentration of HDL cholesterol in plasma to 31.1% and reduced the atherosclerotic index to 58.7% at a concentration of 200 mg per kg body weight.

HDL cholesterol in the plasma as an indicator of anti- atherosclerosis is involved in the delivery of excess cholesterol for liver regeneration. Some types of dietary fiber have been reported to increase plasma HDL cholesterol levels, and Kim reported that L. edodes fruiting bodies increase plasma HDL cholesterol levels in rats. Therefore, the increase in HDL in this embodiment is considered to be due to the slow conversion of HDL to LDL or by some other method.

In addition, the extracellular polysaccharide of the roe deer fungus mushroom of the present invention increases the concentration of HDL cholesterol and significantly decreases the arteriosclerosis index, which may be helpful in preventing atherosclerosis.

Effects of Extracellular Polysaccharides Derived from Roebuck Mushrooms on Plasma Triglycerides and Phospholipids in Hyperlipidemic Rats Group (extrapolysaccharide, g / kg / day) Triglycerides (mg / dl) Phospholipids (mg / dl) Control group ¹⁾ 37.3 ± 1.7 99.1 ± 1.8 50 25.5 ± 1.7 83.7 ± 1.7 100 25.4 ± 1.6 83.3 ± 2.8 150 24.7 ± 1.2 82.8 ± 4.3 200 24.5 ± 1.1 80.4 ± 4.9

¹⁾ Oral Saline

As shown in Table 4, the extracellular polysaccharide at 200 mg / kg body weight compared to the control group decreased the concentrations of plasma triglyceride and phospholipid to 34.3% and 18.9%, respectively.

Effects of Extracellular Polysaccharides Derived from Roebuck Mushrooms on Liver Weight, Total Cholesterol, and Triglycerides in Hyperlipidemic Rats Group (extrapolysaccharide, g / kg / day) Weight (g / 100g body weight) Total cholesterol (mg / g) Triglycerides (mg / g) Control group ¹⁾ 5.45 ± 0.22 6.95 ± 0.15 ^NS 19.8 ± 1.3 50 4.93 ± 0.09 6.77 ± 0.20 17.7 ± 1.0 100 4.83 ± 0.12 6.28 ± 0.24 16.7 ± 1.4 150 4.85 ± 0.20 6.23 ± 0.21 16.0 ± 0.8 200 4.82 ± 0.08 6.15 ± 0.37 13.1 ± 1.1

¹⁾ Oral Saline

NS: no significance

As shown in Table 5, in the group to which the extracellular polysaccharide was administered, the total cholesterol concentration of the liver was not significantly different from the control group, but the concentration tended to decrease in the experimental group. In addition, the extracellular polysaccharide at the concentration of 200 mg / kg body weight reduced the triglyceride concentration to 33.8% in the liver compared to the control.

Hepatic cholesterol levels are associated with the activity and esterification of hepatic cholesterol synthase and the expression of apo B / E receptors, a major determinant of plasma LDL levels. That is, the accumulation of cholesterol in the liver can reduce the increase and accumulation of esterification, increase the secretion of cholesterol in the liver lipoproteins and reduce the absorption of plasma cholesterol by the LDL receptor.

Effect of Extracellular Polysaccharides Derived from Roebuck Mushrooms on Hepatic HMG-CoA Reductase Activity in Hyperlipidemic Rats Group (extrapolysaccharide, g / kg / day) HMG-CoA reductase activity of the liver (pmoles / min / mg microsome protein) Control group ¹⁾ 1488 ± 58 50 1451 ± 80 100 1286 ± 87 150 1255 ± 71 200 1188 ± 57

¹⁾ Oral Saline

As shown in Table 6, the HMG-CoA reductase activity of the liver was orally administered at 50, 100, 150, and 200 mg / kg of the present invention of the present invention of the present invention of the extract of the present invention, the fungus of the present invention, the beetle fungus, 2.5, 13.6, 15.7, 20.2%. Their concentrations decreased with increasing daily dose and may be proportionally related to the total cholesterol concentration of plasma in the experimental group. This correlation coefficient (r) was 0.886 at p <0.05. These results suggest that the main reason for the cholesterol-lowering effect of the extracellular polysaccharide of scavenger worm fungi is the reduction of cholesterol synthesis by inhibiting the HMG-CoA reductase activity in diet-induced hyperlipidemic rats. Therefore, the reduction of HMG-CoA reductase activity may be considered as one of the main reasons for the decrease in plasma total cholesterol concentration of the extracellular polysaccharide of the rotifera, and the extracellular polysaccharide affects the function of other enzymes involved in lipid metabolism. Indicates that it can have.

Example 3: Component Analysis of Extracellular Polysaccharides Derived from the Roe Deer Mushroom

As described in Example 2, the extracellular polysaccharide derived from the present invention of the Roe locust mushroom has hypolipidemic effect in diet-induced hyperlipidemic rats. Therefore, in order to determine the active source of the extracellular polysaccharide, it was analyzed by analyzing its components including molecular weight.

First, in order to investigate the molecular weight of the extracellular polysaccharide, the extracellular polysaccharide obtained from the culture medium was diluted with 0.2 M NaCl, and the effluent rate of 5 mL per tube volume in the column of Separz CL-6B saturated with 0.2 M NaCl. Gel filtration was carried out. The molecular weight of the extracellular polysaccharide was measured on HPLC using Shodex GS520 + GS320 + GS220 Packard columns. As standard, standard pullulan (P1600, 800, 400, 200, 100, 50, 20, 10, 5) was used for molecular weight measurement.

In addition, the components of the sugars and amino acids of the extracellular polysaccharide derived from the roe deer fungus mushroom of the present invention were analyzed. The total equivalent of the extracellular polysaccharide was measured according to the phenol sulfonic acid method using a mannose and galactose mixture (1: 1) as a standard. Sugar composition was analyzed by GC 3600 gas chromatography (Variang) according to the hydrolysis and acetylation method of Jones and Albersheim.

Total protein content of extracellular polysaccharide was measured according to Lowry's method using bovine serum albumin as a standard. The amino acid composition was analyzed by a Biochrome 20 amino acid automatic analyzer (manufactured by Pharmacia Biotech Co., Ltd.) equipped with a Na-type column after hydrolysis of the protein.

As shown in Figs. 1 and 2, the extracellular polysaccharides produced from mycelial liquid cultures of the Roe beetle mushroom showed only one peak in the Ceuzers CL-6B column chromatography, and the molecular weight of the extracellular polysaccharide was about It was measured at 40 kDa.

At this time, the vertical axis Kav of Figure 2 means (Ve-Vo) / (Vt-Vo), Vo is the empty volume, Vt is the total volume, Ve means the elution volume.

The total sugar and protein content of the extracellular polysaccharide were 91.2% and 8.8%, respectively. Detailed chemical analysis of the extracellular polysaccharide is shown in Table 7.

Composition of Sugars and Amino Acids of Extracellular Polysaccharides Produced from Mycelium Liquid Culture of Roebuck Mushroom. Sugar composition (%) ¹⁾ Amino Acid Composition (%) ²⁾ Fucose 20.1 Ribose 0.2 Arabinos 1.5 Xylose 0.3 Mannose 35.4 Galactose 34.1 Glucose 8.4 Aspartic Acid 10.5 Threonine 8.7 Serine 16.6 Glutamic Acid 10.5 Proline 0.3 Glycine 10.0 Alanine 6.7 Cysteine 3.5 Valine 6.4 Methionine 2.8 Isoleucine 3.3 Leucine 3.8 Tyrosine 1.8 Phenylalanine 8.9 Histidine 1.1 Lysine 3.7 Arginine 1.4 Total sugar content 91.2 Total protein content 8.8

¹⁾ : ratio per total

²⁾ : ratio to total amino acids

As shown in Table 7, the 17 amino acids make up the protein moiety, and the main amino acids are aspartic acid (10.5%), serine (16.6%), glutamic acid (10.5%), and glycine (10.0%). . Fucose (20.1%), mannose (35.4%) and galactose (34.1%) are the major carbohydrates of sugar moieties. From the above results, the mannose and galactose content in the total sugar of the extracellular polysaccharide of the roe deer mushroom accounted for almost 70%. The extracellular polysaccharides of the roe deer mushroom were similar to galactomannan and contained similar amounts of galactose and mannose.

On the other hand, different molar ratios of galactose and mannose affect the hypocholesterolemic effect of galactomannan in rats, and it has been reported to be apparent in the fenugreek rubber group including galactomannan with the highest galactose ratio. Therefore, it was found that the hypolipidemic effect of the extracellular polysaccharide derived from the scorpion mushroom was affected by its chemical composition and structure.

As described above, the present invention, as described through the examples, oral administration of the extracellular polysaccharide produced from the mycelium liquid culture of the roe deer mushroom fungus in hyperlipidemic rats, depending on the concentration of the extracellular polysaccharide compared to the control group There is an excellent effect of increasing the effect of blood. In addition, the extracellular polysaccharide of the present invention has an excellent effect of preventing the atherosclerosis by reducing the concentration of total cholesterol, triglycerides, phospholipids, arteriosclerosis index and LDL cholesterol of plasma and maintaining the concentration of HDL cholesterol high. Moreover, the extracellular polysaccharide of the present invention has an excellent effect of reducing the synthesis of cholesterol by inhibiting the activity of HMG-CoA reductase of the liver. The extracellular polysaccharide of the present invention having the hypolipidemic effect is a glycoprotein having a molecular weight of 40 kDa, and is composed of 91.2% of sugar and 8.8% of amino acids, and the main sugars of the extracellular polysaccharide component are fucose, galactose and mannose. Major amino acids were serine, aspartic acid, glutamic acid, and glycine. Therefore, the present invention is an extracellular polysaccharide derived from scavengerae fungus, which has a hypoglycemic effect and can provide a composition for treating and preventing hyperlipidemia using the extracellular polysaccharide as an active ingredient.

Claims (4)

A) Inoculate the culture medium of the roe deer mushroom into potato / dextrose agar medium and subculture at 4 ° C. every three months, B) Put the culture solution into potato / dextrose broth (pH 5.0) and incubate in a rotary shaker (120 rpm) at 25 ° C., 10) After 10 days of the culture, the culture medium was pulverized in aseptic state, and then the culture medium prepared in advance (glucose 20 g / L, MgSO ₄ · 7H ₂ O 0.5 g / L, KH ₂ PO ₄ 0.46 g). / L, yeast extract 2 g / L, peptone 2 g / L; adjusted to pH 4 before sterilization) at 1% (v / v), D) culture the mycelia liquid for 12 days in a rotary shaker at 120 rpm, 25 ℃ condition containing the pulverized, E) the supernatant obtained by centrifuging the mycelium liquid culture solution at 10,447 x g for 20 minutes, followed by centrifugation by ethanol precipitation, F) Separation method of extracellular polysaccharide in mycelium liquid culture of snail mushroom, which is obtained by diluting and dialysis with water, and then lyophilizing. It is separated by the separation method of claim 1, containing 91.2% of total sugar and 8.8% of protein, molecular weight of 40 kDa, and the content ratio of mannose and galactose, the major constituents, of 35.1% and 34.1%, respectively. An extracellular polysaccharide derived from a roe deer fungus with hypolipidemic activity. A composition for the treatment of hyperlipidemia, comprising extracellular polysaccharides derived from roe deer mushroom having hypolipidemic activity as an active ingredient. A composition for preventing atherosclerosis, characterized in that it contains an extracellular polysaccharide derived from a locust fungus isolated by the separation method of claim 1 as an active ingredient.