KR101307051B1 - Nature seasoning using micro algae protein hydrolysate and the manufacturing method thereof - Google Patents

Abstract

The present invention relates to a composition for lowering cholesterol and a method for preparing the same, comprising a protein hydrolyzate obtained by hydrolyzing a protein of microalgae.
The cholesterol-lowering composition according to the present invention has no side effects by utilizing a natural microalgae-derived protein hydrolyzate, and can increase production yield by using ultrasonic waves.

Description

Nature seasoning using micro algae protein hydrolysate and the manufacturing method

The present invention relates to a composition for lowering cholesterol and a method for preparing the same, comprising a protein hydrolyzate obtained by hydrolyzing a protein of microalgae.

Cholesterol is an essential component of cells and plays an important role in cell growth, differentiation and development. In addition, corticosteroids, sex hormones, digestive enzymes to make bile acid material, and plays an important role in maintaining the strengthening of blood vessels.

However, when the cholesterol is ingested excessively, cholesterol in the blood penetrates into the innermost membrane of the blood vessels, and as a result of the release of various substances, the inner lining of the arteries grows and thickens. In addition, cholesterol and cells that consumed it accumulate and protrude into the blood vessels. This is called atherosclerosis, and is a representative case of arteriosclerosis caused by cholesterol.

Circulatory diseases such as arteriosclerosis have been the main cause of death in Korea since the 1970s. In addition, cholesterol causes adult diseases such as obesity, and lipid peroxides derived from cholesterol cause in vivo regeneration processes, and are known to have negative effects on biological functions such as cancer, aging, changes and destruction of biological membranes. Thus, the control of these lipids in vivo and externally is very important for modern people with a westernized diet, ie, an animal diet.

In order to lower blood cholesterol, blood cholesterol lowering agents such as nicotinic acid, triparanol, and butyrophenone have been developed. However, since it can cause side effects such as acne, it is necessary to find blood cholesterol lowering agents in natural products without side effects. Attempts are being made.

Accordingly, the present inventors have developed a blood cholesterol-lowering composition using a natural microalgae-derived protein hydrolyzate.

The present invention is to provide a composition for lowering cholesterol, including a microalgae-derived protein hydrolyzate and a method for preparing the same.

The present invention relates to a composition for lowering cholesterol comprising protein hydrolyzate of microalgae.

In addition, the present invention comprises the steps of a) eluting the protein by sonicating the microalgae; And

b) preparing a protein hydrolyzate using a protease to the protein;

It relates to a method for producing a composition for lowering cholesterol comprising a.

In addition, the present invention relates to a pharmaceutical composition for lowering cholesterol and a food composition comprising 1 to 5% of the composition.

The cholesterol lowering composition according to the present invention is substantially free of side effects by utilizing a natural microalgae-derived protein hydrolyzate, and can increase production yield by using ultrasonic waves.

Figure 1 shows the change in blood LDL-cholesterol content of the diet for 4 weeks.

The present invention provides a composition for lowering cholesterol comprising protein hydrolyzate of microalgae.

In the specification of the present invention, "microalgae" is a floating organism inhabiting fresh water and seawater, and includes algae, diatoms, coarse algae, green algae, red algae, yellow-brown algae and silver algae, and about 35,000 species are reported worldwide. However, it is estimated that there are potentially more than 200,000 species. Microalgae, like other plants, are photosynthetic and biosynthesize sugars and proteins in the interior, and are therefore responsible for basic production in freshwater and the ocean, and are important producers of food chains.

In the present invention, the protein hydrolyzate of the microalgae refers to a hydrolyzate produced by ultrasonically treating the microalgae to elute the protein and using the protease to the eluted protein. In addition, the microalgae, in particular, can be eluted protein by treating the cell wall of the microalgae.

In the present invention, the microalgae may include green algae (Chlorophyte), brown algae (Phaeophyceae), red algae (Rhodophyte), algae (Cyanophyta), diatoms (Bacillariophycea), coarse algae (Dinophyta) or algae (Haptophyta) In addition, the coccyx imitation may be Tetrselmis suecica .

The present invention also provides a method for preparing a composition for lowering cholesterol comprising microalgal protein hydrolyzate. More specifically,

a) sonicating the microalgae to elute the protein; And

b) preparing a protein hydrolyzate using protease on the protein;

It provides a method for producing a composition for lowering cholesterol comprising a.

The microalgae cell wall can be sonicated to elute the protein.

The microalgae include, but are not limited to, green algae (Chlorophyte), brown algae (Phaeophyceae), red algae (Rhodophyte), cyanophyta, diatoms (Bacillariophycea), warpweed algae (Dinophyta), or flax algae (Haptophyta). Can be.

In addition, the coccyx algae may include Tetrselmis suecica .

In the present invention, it is possible to calculate the optimum conditions of extraction frequency, ultrasonic extraction time and extraction temperature of the protein eluting (or extraction) in step a), in an embodiment of the present invention (Response Surface Methodology, RSM) Can be calculated using That is, as a result of calculating the extraction frequency, the ultrasonic extraction time and the extraction temperature as independent variables, the ultrasonic treatment can be performed at 26 ~ 28 ℃ for 4-5 hours at a frequency of 90-100 kHz in the ultrasonic generator.

In addition, in the present invention, the optimal condition for hydrolysis in step b) may be calculated using Response Surface Methodology (RSM). That is, as a result of calculating the hydrolysis time and pH as independent variables, the hydrolysis time may be 50 to 60 minutes and the pH may be 6 to 7.

The proteolytic enzyme may include an alcalase, and may use 2 to 3 L of the alkalase, and may be hydrolyzed at 45 to 60 ° C.

The present invention also provides a cholesterol-lowering pharmaceutical composition comprising 1 to 5% of a cholesterol-lowering composition comprising the protein hydrolyzate of the microalgae. Including less than 1% or more than 5% of the composition for lowering cholesterol including protein hydrolyzate of microalgae, there is a problem that the hypoglycemic effect may be lowered.

In addition to the microalgae-derived protein hydrolyzate, the pharmaceutical composition may further comprise nutrients, vitamins, electrolytes, flavoring agents, coloring agents, neutralizing agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH regulators, and stabilizers. It may further contain an agent, a preservative, glycerin, alcohol, a carbonation agent used in the carbonated beverage, and the like. Carriers, excipients or diluents may also include lactose, dextrose, sucrose, sorbitol, mannitol, ziitol, erythritol, maltitol, starch, acycia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline In the group consisting of cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, dextrin, calcium carbonate, propylene glycol, liquid paraffin, saline It may be one or more selected, but is not limited to any conventional carrier, excipient or diluent can be used.

In addition, the pharmaceutical composition of the present invention may further include conventional fillers, extenders, binders, disintegrants, surfactants, anti-coagulants, lubricants, wetting agents, fragrances, emulsifiers or preservatives, and can be used orally or parenterally. have.

Specifically, solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid preparations include at least one excipient in the composition, for example, starch, calcium carbonate, sucrose, and the like. (sucrose), lactose (lactose), gelatin, etc. are mixed and prepared.

In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid preparations for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweetening agents, have.

In addition, the formulation of the pharmaceutical composition of the present invention may be in a desired form depending on the method of use, and employing methods known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to the mammal Can be formulated. Examples of specific formulations include warnings, granules, lotions, linings, limonades, powders, syrups, ointments, liquids, aerosols, EXTRACTS, elixirs, ointments, liquid extracts, emulsions, Suspensions, premises, acupuncture, eye drops, tablets, suppositories, injections, spirits, capsules, creams, pills, soft or hard gelatin capsules.

Further, the pharmaceutical compositions of the present invention may be formulated using any of the known methods known in the art or by methods disclosed in Remington ' s Pharmaceutical Science (recent edition), Mack Publishing Company, Easton PA) .

The dosage of the pharmaceutical composition of the present invention depends on the condition and weight, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. Based on the cholesterol-lowering composition comprising the protein hydrolyzate of the microalgae of the present invention is preferably 0.0001 to 12g / kg, 0.01 to 10g / kg per day. The administration may be carried out once a day or divided into several times. The dose is not intended to limit the scope of the invention in any way.

In addition, the present invention provides a cholesterol-lowering food composition comprising 1 to 5% of a cholesterol-lowering composition containing protein hydrolyzate of microalgae.

Including less than 1% or more than 5% of the cholesterol-lowering composition comprising the protein hydrolyzate of the microalgae, there is a problem that the hypoglycemic effect may be lowered.

In the present specification, the term "food" means a natural product or a processed product containing one or more nutrients, and means a state in which it can be directly eaten through a certain processing process. It is intended to include additives, health functional foods and beverages.

Examples of the food to which the composition for lowering cholesterol containing the protein hydrolyzate of the microalgae of the present invention can be added include various foods, beverages, gums, teas, vitamin complexes, and functional foods. In addition, the food in the present invention includes special nutritional products (e.g., crude oil, young, baby food, etc.), processed meat products, fish products, tofu, jelly, noodles (e.g. ramen, noodles, etc.), health supplements, seasonings ( For example, soy sauce, miso, red pepper paste, mixed soy sauce, sauces, confectionery (e.g. snacks), dairy products (e.g. fermented milk, cheese, etc.), other processed foods, kimchi, pickles (various kimchi, pickles, etc.), beverages ( Examples include, but are not limited to, fruits, vegetable drinks, soy milk, fermented beverages, and the like. The food, beverage or food additive may be prepared by a conventional production method.

Functional food in the present invention is the control of biological defense rhythm, disease prevention and recovery of food groups or food compositions that have added value to the food by using physical, biochemical, biotechnological techniques, etc. It means a food that is designed and processed to fully express the function of the gymnastics for the back. The functional food may include a food-acceptable food-aid additive, and may further comprise suitable carriers, excipients and diluents conventionally used in the production of functional foods.

In the present invention, beverage is a generic term for drinking or enjoying a taste, and is intended to include functional beverages. The beverage is not particularly limited in addition to including the composition as an essential ingredient in the indicated proportions, and may contain various flavors or natural carbohydrates, etc. as additional ingredients, as in general beverages. Examples of such natural carbohydrates include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the flavoring agent other than the above, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used.

In addition to the above, the composition for lowering cholesterol containing the protein hydrolyzate of the microalgae of the present invention is a flavoring agent such as various nutrients, vitamins, minerals (electrolytes), synthetic flavoring agents and natural flavoring agents, colorants and neutralizing agents (cheese, chocolate) Etc.), and the salts of pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks and the like. These components can be used independently or in combination. The ratio of such additives is not so important, but selected from the range of 0 to 20000 parts by weight, 10 to 2000 parts by weight, and 100 to 200 parts by weight per 100 parts by weight of the cholesterol lowering composition comprising the protein hydrolyzate of the microalgae of the present invention. Can be.

Functional beverage in the present invention is a biological defense rhythm control, disease prevention and the like having a beverage group or a beverage composition that has added value to the beverage by using physical, biochemical, biotechnological techniques, etc. to function and express the function of the beverage to a specific purpose It means a beverage that is designed and processed to fully express the body control function relating to recovery and the like to the living body.

The functional beverage is not particularly limited in addition to the cholesterol lowering composition comprising the protein hydrolyzate of the microalgae of the present invention as an essential ingredient in the indicated ratio, and various flavors or natural like ordinary drinks Carbohydrates and the like may be included as additional components. Examples of such natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. .

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail with reference to the following examples. However, the following examples are intended to illustrate the contents of the present invention, but the scope of the present invention is not limited to the following examples. Embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art.

< Example  1> Microalgal Proteins Hydrolyzate  Produce

Example  1-1. Preparation of Microalgal Solids

Tetraselmis , the microalgae used in the experiment suecica was used directly in marine biology research and education center (Gangneung, Gangwon-do). The microalgae purchased and cultured were centrifuged to remove the culture solution. More specifically, the microalgae purchased and cultured were centrifuged at 7,000 rpm for 15 minutes to separate the supernatant and solids, and the supernatant was removed, and only the solids were used. The obtained microalgae was stored at -40 ° C until the experiment. It was.

Example  1-2. Extraction and Hydrolysis of Microalgae-Derived Proteins

The obtained solids of the microalgae were sonicated to extract internal protein and hydrolyzed by the addition of protease.

More specifically, 900 mL of water was added to 100 g, extracted at a frequency of 97.78 kHz for 4.49 hr at a temperature of 26.22 ° C., followed by mixing the extract and the remaining solids after extraction for 10 minutes in a constant temperature bath at 52.5 ° C. I was. The pH was then adjusted to 6.01 and 2.91 g of alcalase 2.4L were added and then hydrolyzed for 58.53 minutes. After removing the solids by centrifugation for 15 minutes at 7,000 rpm and then lyophilized at -80 ℃ to prepare 14.2 g of Tetraselmis suecica protein hydrolyzate.

< Example  2> Breeding and sampling of experimental animals

2-1. Breeding of test animals Diet

Experimental animals were used 6 weeks old C57BL / 6J male mice were purchased from the center experimental animals. It was bred in SPF (specific pathogen free) -animal laboratory automatically set to a temperature 22 ± 2 ℃, humidity 50 ± 5%, contrast cycle 12 hours (08:00 ~ 20:00), the feed composition table is shown in Table 1.

(g / 100g diet) Normal diet High-cholesterol diet - - HTS-1% HTS-5% Starch 10 10 10 10 Casein 20 20 19.3 16.6 Cellulose 5 5 5 5 Sucrose 50 50 50 49 Vitamin mix One One One One Mineral mix 3.5 3.5 3.5 3.5 DL-methionine 0.3 0.3 0.3 0.3 Choline bitartrate 0.2 0.2 0.2 0.2 Corn oil 10 5 5 5 Lard - 5 5 5 Cholesterol - 1.25 1.25 1.25 Sodium cholate - 0.25 0.25 0.25 HTS - - One 5

In the first week, the diet was adjusted to receive a normal diet (AIN-76 based modified diet), and in the second week, the mice were weighed to separate the groups. Separation of the group proceeded as follows.

① normal diet group does not contain cholesterol,

② High cholesterol dietary control group containing cholesterol (cholesterol 1.25% + cholic acid 0.25%)

③ Tetraselmis in high cholesterol feed group containing 1% suecica hydrolyzate

(HTS-1%)

④ Tetraselmis in cholesterol feed group containing 5% suecica hydrolyzate

(HTS-5%)

Statistical processing of the experimental data was shown using GraphPad Prism (Version 4.0), and analyzed by unpaired t-test to verify the significance of each experimental group.

2-2. Treatment and Sampling of Test Animals

After ingestion of the experimental diet containing the test substance for 4 weeks and 8 weeks, the experimental animals of each group were placed in a metabolic cage two days before the end of the experiment, and the feces were collected. The collected feces were stored at 4 ° C. until analysis.

Fasting for 12 hours before dissection, anesthesia using an anesthetic and orbital blood collection. The collected blood was placed in a BD SST tube and allowed to stand at room temperature for 30 minutes, followed by centrifugation (3000g, 15 minutes) to separate serum from the supernatant and analyzed by an automatic biochemical analyzer. After orbital blood collection, animals were euthanized with CO2 gas and liver and kidneys were extracted. The harvested organs were washed with PBS, removed with blood from the filter paper, and weighed. Liver tissue was stored at −70 ° C. until analysis for lipid analysis.

< Example  3> weight, Dietary intake , Dietary efficiency  And negative volume measurements

The experimental diet was ingested for 4 and 8 weeks, and the food and water were fed freely during the test period, and the body weight, dietary intake and drinking water were measured at a certain time every week. The food efficiency ratio (FER) was calculated by dividing the weight gain by the food intake over the same period and the results are shown in Table 2.

Content Feeding
period Normal diet High-cholesterol diet HCD ^{2 )} HTS-1% ³⁾ HTS-5% ⁴⁾ Body weight
(g) 0 weeks 20.83 ±± 0.27 20.94 ±± 0.36 20.79 ±± 0.28 20.82 ±± 0.28 4 weeks 24.46 ±± 0.30 24.69 ±± 0.53 24.01 ±± 0.48 24.82 ±± 0.35 8 weeks 26.98 ±± 0.54 27.78 ±± 0.81 27.70 ±± 0.67 27.18 ±± 0.34 Food intake
(g / day) 4 weeks 2.17 ±± 0.04 2.41 ±± 0.05 ^* 2.48 ±± 0.12 ^* 2.59 ±± 0.03 ^*** , ^# 8 weeks 2.43 ±± 0.16 2.52 ±± 0.01 2.67 ±± 0.17 2.77 ±± 0.23 FER ^{1 )} 4 weeks 6.06 ±± 0.40 5.67 ±± 0.45 5.09 ±± 0.40 5.98 ±± 0.45 8 weeks 4.88 ±± 0.38 5.27 ±± 0.61 5.10 ±± 0.41 4.20 ±± 0.26 Water intake
(ml / day) 4 weeks 3.21 ±± 0.16 3.06 ±± 0.16 2.89 ±± 0.13 4.17 ±± 0.25 ^* , ^## 8 weeks 3.19 ±± 0.13 3.08 ±± 0.24 2.91 ±± 0.15 4.30 ±± 0.01 ^* , ^#

¹⁾ FER (Food efficiency ratio) = (Weight gain (g) ÷ food intake (g)) × 100

²⁾ HCD, High cholesterol diet control group

³⁾ HTS-1%, High cholesterol diet including 1% enzymatic Tetraselmis suecica hydrolysate diet group

⁴⁾ HTS-5%, High cholesterol diet including 5% enzymatic Tetraselmis suecica hydrolysate diet group

Data are presented as means ± SEM.

-*; significant vs Normal diet (*; p <0.05, **; p <0.01, ***; p <0.001)

-#; significant vs High-cholesterol diet (#; p <0.05, ##; p <0.01, ###; p <0.001)

There was no increase in body weight by the diet including high cholesterol and HTS, and there was no difference in the food efficiency ratio (FER) between the groups.

< Example  4> Liver lipid composition analysis

Liver lipids were extracted and analyzed using the Folch method. Three times 0.9% NaCl was added to 100 mg of liver tissue and homogenized with a homogenizer. Add three times the amount of Folch solution (Chloroform: methanol = 2: 1) to the homogenizer, repeat this three times, and then centrifuge (3000 rpm, 10 minutes) to remove the upper layer, which is the water layer. Moved to the tube. In order to remove chloroform, a certain amount of the tube was placed in a new tube, completely dried using nitrogen gas, and then dissolved in a 10% Triton X-100 / isopropanol solution. And take a sample of LabAssay ^TM It was measured using a ^{triglyceride kit, LabAssay TM Cholesterol kit (} Wako, Japan) Results are given in Table 3.

(mg / g) Indicator Feeding
period Normal diet High-cholesterol diet HCD ^{1 )} HTS-1% ²⁾ HTS-5% ³⁾ Liver Tri-
glyceride 4 weeks 21.78 ±± 1.58 22.86 ±± 1.35 25.73 ±± 1.27 25.98 ±± 1.30 8 weeks 20.19 ±± 1.53 25.12 ±± 2.28 25.32 ±± 2.16 22.29 ±± 1.41 Total
cholesterol 4 weeks 7.28 ±± 1.08 21.55 ±± 0.90 ^*** 20.01 ±± 1.57 ^*** 25.96 ±± 0.69 ^{***, ##} 8 weeks 7.56 ±± 0.67 22.02 ±± 0.76 ^*** 20.38 ±± 1.09 ^*** 22.37 ±± 0.59 ^***

¹⁾ HCD, High cholesterol diet control group

²⁾ HTS-1%, High cholesterol diet including 1% enzymatic Tetraselmis suecica hydrolysate diet group

³⁾ HTS-5%, High cholesterol diet including 5% enzymatic Tetraselmis suecica hydrolysate diet group

Data are presented as means ± SEM.

-*; significant vs Normal diet (*; p <0.05, **; p <0.01, ***; p <0.001)

-#; significant vs High-cholesterol diet (#; p <0.05, ##; p <0.01, ###; p <0.001)

Neutral lipid content in liver tissue was not significantly increased by high cholesterol diet and HTS intake. The total cholesterol content in liver tissue was significantly increased in the high cholesterol diet compared to the normal diet group ( p <0.001), and the total cholesterol content in the liver tissue group was higher than the control group in the high cholesterol diet for 4 weeks. Cholesterol content was significantly higher ( p <0.01).

< Example  5> Feces  Lipid Composition Analysis

Fecal lipids were extracted by the same method as the extraction method of liver lipid, and neutral lipids and total cholesterol content were measured, and the results are shown in Table 4.

(mg / g) Indicator Feeding
period Normal diet High-cholesterol diet HCD ^{1 )} HTS-1% ²⁾ HTS-5% ³⁾ Feces Triglyceride 4 weeks 1.45 ±± 0.33 2.72 ±± 0.51 2.23 ±± 0.61 3.31 ±± 0.21 ^** 8 weeks 0.79 ±± 0.04 3.42 ±± 0.73 2.02 ±± 0.67 4.67 ±± 0.77 ^** Total
cholesterol 4 weeks 5.30 ±± 1.42 93.35 ±± 3.39 ^*** 78.21 ±± 6.44 ^*** 93.07 ±± 2.89 ^*** 8 weeks 10.35 ±± 1.46 84.93 ±± 4.51 ^*** 79.52 ±± 5.97 ^*** 86.46 ±± 4.23 ^***

¹⁾ HCD, High cholesterol diet control group

²⁾ HTS-1%, High cholesterol diet including 1% enzymatic Tetraselmis suecica hydrolysate diet group

³⁾ HTS-5%, High cholesterol diet including 5% enzymatic Tetraselmis suecica hydrolysate diet group

Data are presented as means ± SEM.

-*; significant vs Normal diet (*; p <0.05, **; p <0.01, ***; p <0.001)

-#; significant vs High-cholesterol diet (#; p <0.05, ##; p <0.01, ###; p <0.001)

The content of triglyceride in feces tended to increase in the high cholesterol diet compared with the normal diet, but it was not significant. Intake of HTS-5% for 4 and 8 weeks showed a significant increase in triglyceride content compared to the normal diet ( p <0.01), but no significant difference was observed in the high cholesterol diet control group.

The total cholesterol content in the feces was significantly increased in the high cholesterol diet group compared to the normal diet group ( p <0.001). On the other hand, no significant difference was observed between the HTS intake group and the high cholesterol diet group.

One mechanism of hypercholesterolemia regulation is to promote the excretion of cholesterol, which is present in high concentrations in the body, and to lower the lipid content in serum and liver. However, as a result of this, it promotes cholesterol excretion by HTS. The action is considered inadequate.

< Example  6> Serum Lipid Composition Analysis

Serum lipid components were analyzed by lipid biochemistry (Automated chemistry analyzer, Konelab 20XT, Thermo), and analyzed lipid components (total cholesterol, triglyceride, HDL-cholesterol, LDL-cholesterol) and the results are shown in Table 5 and FIG. .

(mg / dL) Indicator Feeding
period Normal diet High-cholesterol diet HCD ^{1 )} HTS-1% ²⁾ HTS-5% ³⁾ Triglyceride 4 weeks 55.84 ±± 6.31 23.86 ±± 1.37 ^** 32.44 ±± 3.32 ^* , ^# 22.20 ±± 2.06 ^*** 8 weeks 64.08 ±± 4.90 32.08 ±± 3.04 ^*** 31.55 ±± 3.93 ^*** 23.21 ±± 2.06 ^*** , ^# Total
cholesterol 4 weeks 158.20 ±± 7.90 147.50 ±± 18.22 135.80 ±± 8.85 146.40 ±± 11.81 8 weeks 138.50 ±± 6.93 138.10 ±± 7.08 143.40 ±± 3.89 160.90 ±± 12.44 HDL
cholesterol 4 weeks 74.19 ±± 3.53 60.84 ±± 6.15 56.78 ±± 2.55 ^** 58.71 ±± 4.92 ^* 8 weeks 65.84 ±± 2.91 57.45 ±± 2.83 59.45 ±± 1.48 61.06 ±± 3.54 LDL
cholesterol 4 weeks 15.22 ±± 2.37 66.65 ±± 5.97 ^*** 44.74 ±± 4.50 ^*** , ^# 70.85 ±± 6.77 ^*** 8 weeks 9.78 ±± 1.20 52.88 ±± 1.74 ^*** 44.28 ±± 6.77 ^*** 59.71 ±± 2.08 ^*** , ^#

¹⁾ HCD, High cholesterol diet control group

²⁾ HTS-1%, High cholesterol diet including 1% enzymatic Tetraselmis suecica hydrolysate diet group

³⁾ HTS-5%, High cholesterol diet including 5% enzymatic Tetraselmis suecica hydrolysate diet group

Data are presented as means ± SEM.

-*; significant vs Normal diet (*; p <0.05, **; p <0.01, ***; p <0.001)

-#; significant vs High-cholesterol diet (#; p <0.05, ##; p <0.01, ###; p <0.001)

Serum triglyceride content was significantly decreased by high cholesterol diet (Week 4; p <0.01, Week 8; p <0.001), compared with the high cholesterol diet in the group receiving HTS-1% for 4 weeks. Neutral lipid content increased significantly ( p <0.05). There was no significant difference between the HTS-1% intake group and the high cholesterol diet control group for 8 weeks, and the neutral lipid content was significantly decreased in the HTS-5% intake group ( p <0.05).

Serum total cholesterol content was not significantly different in all groups who consumed a normal diet, a high cholesterol diet and a diet containing HTS.

The HDL-cholesterol content in serum tended to decrease with high cholesterol diet, but there was no significant difference, and it was significantly decreased in the group receiving HTS-1% and -5% for 4 weeks compared to the normal diet group ( p <0.05), cholesterol diet was not significantly different compared to the control group.

The LDL-cholesterol content was significantly increased in the high-cholesterol diet compared to the normal diet group, and showed a significant decrease in the high-cholesterol diet group intake of HTS-1% for 4 weeks compared with the control group ( p <0.05). The group ingested HTS-5% for 8 weeks showed a significant increase compared to the high cholesterol diet control group ( p <0.05).

Taken together, the microalgal protein hydrolyzate did not significantly affect the total cholesterol synthesis in the liver and increased excretion to the stool, but 4 weeks of HTS-1% intake of the normal high cholesterol diet Compared to the blood LDL-cholesterol significantly decreased, compared with the blood cholesterol reduction effect.

< Example  7> Microalgae Proteins Hydrolyzate  Contains cholesterol lowering composition Formulation Example

Hereinafter, the formulation examples of the composition will be described, but the present invention is not intended to be limited thereto but merely to be described in detail.

Formulation example  1. Preparation of tablets

Cholesterol lowering composition containing protein hydrolyzate of microalgae ..... 100 mg

Corn starch ............................... 68 mg

Lactose .................................. 90 mg

Microcrystalline Cellulose ............. 40 mg

Magnesium stearate ..................... 2 mg

According to the conventional preparation method of tablets, the above ingredients were added in the prescribed amounts, uniformly mixed, stirred, and then granulated. After drying, using a tableting machine, a desired tablet containing 100 mg of a composition for lowering cholesterol containing protein hydrolyzate of a single microalgae was prepared.

Formulation example  2. Preparation of Capsule

Cholesterol-lowering composition containing protein hydrolyzate of microalgae ......................................................... 100 mg

Corn starch ................................ 68 mg

Lactose ................................... 90 mg

Microcrystalline cellulose ........................ 40 mg

Magnesium stearate ...................... 2 mg

According to the conventional method for preparing capsules, gelatin capsules of a suitable size to include a composition for lowering cholesterol containing 100 mg of microalgae protein hydrolysate per capsule after adding the above ingredients in a given amount and mixing them uniformly. Filled in to prepare the desired capsule.

Formulation example  3. Manufacturing of beverage

Cholesterol lowering composition containing protein hydrolyzate of microalgae ........... 100 mg

Concentrated Juice ........... 2 g

Sucrose ......................................... 12 g

Sodium Citrate ... 100 mg

Fragrance ............... 70 mg

Water ............................................

According to a conventional method for preparing a beverage, the above ingredients are mixed in an appropriate amount of water in a given amount, heated to dissolve, cooled, and filled into a container to prepare 100 mg of microalgae protein per bottle of a 200 ml beverage. The beverage containing the composition for lowering cholesterol containing a degradation product was prepared.

Claims (11)

A pharmaceutical composition for lowering LDL-cholesterol comprising 1% protein hydrolyzate by alcalase of Tetraselmis suecica .
delete delete a) eluting the protein by sonicating Tetraselmis suecica ; And
b) preparing a protein hydrolyzate using an alcalase to the protein;
Method for producing a pharmaceutical composition for LDL-cholesterol lowering comprising a.
delete delete 5. The method of claim 4,
The ultrasonication of step a) is a method for producing a pharmaceutical composition for lowering LDL-cholesterol, characterized in that the treatment at 26 ~ 28 ℃ for 4-5 hours at a frequency of 90-100kHz in the ultrasonic generator.
5. The method of claim 4,
The hydrolyzate of step b) is produced by hydrolysis at pH 6-7, 50-60 minutes, 45 ~ 60 ° C characterized in that the production method of LDL-cholesterol lowering pharmaceutical composition.

delete delete delete

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