KR100975278B1 - Extraction method of opuntia humifusa using enzyme hydrolysis - Google Patents

Abstract

PURPOSE: A method for manufacturing Pounitia humifusa extract is provided to effectively remove viscosity of Pounitia humifusa and to extract physiologically active material from Pounitia humifusa. CONSTITUTION: A method for manufacturing Pounitia humifusa extract comprises a step of adding rapidase or viscozyme or mixture of rapidase or viscozyme for 2-12 hours to Pounitia humifusa to remove viscosity of Pounitia humifusa. The ratio of rapidase or viscozyme is 2:2-1:3. A method for producing Pounitia humifusa extract comprises: a step of removing washing and removing Pounitia humifusa; a step of adding water and pulverizing; a step of treating with rapidase or viscozyme; a step of performing thermal treatment; a step of filtering or centrifuging the Pounitia humifusa extract; and a step of concentrating the Pounitia humifusa extract. A cosmetic composition or food composition contains the Pounitia humifusa extract.

Description

Extraction method of Opuntia humifusa using enzyme hydrolysis

The present invention relates to a cheonnyeoncho extraction method for removing the viscosity of cheonnyeoncho through enzymatic hydrolysis of cheonnyeoncho and cheonnyeoncho extract prepared by the method.

There are about 4,000 kinds of cactus known to date, and have been used for food as a source of carbohydrates, vitamins and minerals.

Cheonnyeoncho (Opuntia humifusa) is a native Korean cactus that grows in Chungnam and other regions. Opuntia ficus-indica var saboten, commonly known as palm cactus, grows to 1-2 m long and has many long and thick thorns. And grow to about 30 cm in size. Baeknyeoncho, which is grown on Jeju Island, is native to the tropics and is difficult to cultivate in the open field, while Korean native cactus, Chunnyeoncho, survives in the open field even in the dormant winter season, and is resistant to pests.

Although much research has not been conducted on the activity of cheonnyeoncho compared to naturalized baeknyeoncho, Park MK, Lee YJ, Kang ES (2005) Hepatoprotective effect of Cheonnyuncho (Opuntia humifusa) extract in rats treated carbon tetrachloride. J Korean Food Sic Technol 37: 822-826) reported the antioxidative activity of cheonnyeoncho stem extracts from hot water and the prevention of liver damage from carbon tetrachloride, including Lee KS, Kim MG, Lee KY (2004) Antimicrobial effect of the Extracts of cactus Chounnyouncho (Oputia humifusa) against food borne pathogens.J Korean Soc Food Sci Nutr 33: 1268-1272.). It was reported that the antimicrobial effect of Cactus extract was excellent against pathogenic food poisoning microorganisms. Kwon DK, Song YJ (2005) Effect of opuntia humifusa supplementation on endurance exercise performance in rats fed a high-fat diet.The Korean Journal of Exercise Nutrition 9: 183-188. It has been reported to have anti-arteriosclerosis effect by reducing hyperlipidemic concentration and increasing HDL-cholesterol concentration, and suggested the possibility of improving aerobic exercise ability.

However, while baeknyeoncho is actively used as food, such as chocolate, cheonnyeoncho is processed and commercialized in food, cosmetics, etc. despite the above-mentioned physiological activity. The main reason why processing and use of millennial plant is not active is due to the viscosity of pectin, which is the main ingredient of millennial plant, and in order to develop various products using millennial plant, it is necessary to improve the extraction method for materialization of millennial plant.

So far, attempts to remove the viscosity of cheonnyeoncho have been proposed for filtration, enzymatic hydrolysis, fermentation, and pulverization after freezing of cheonnyeoncho. For double enzyme enzymatic hydrolysis, cheonnyeoncho cannot be completely removed. There was a problem that it is inappropriate because it changes the (Korean Patent Registration No. 10-0763797).

Republic of Korea Patent No. 10-0763797

Park MK, Lee YJ, Kang ES (2005) Hepatoprotective effect of Cheonnyuncho (Opuntia humifusa) extract in rats treated carbon tetrachloride. J Korean Food Sic Technol 37: 822-826 Lee KS, Kim MG, Lee KY (2004) Antimicrobial effect of the extracts of cactus Chounnyouncho (Oputia humifusa) against food borne pathogens. J Korean Soc Food Sci Nutr 33: 1268-1272. Kwon DK, Song YJ (2005) Effect of Opuntia humifusa supplementation on endurance exercise performance in rats fed a high-fat diet. The Korean Journal of Exercise Nutrition 9: 183-188.

It is an object of the present invention to provide a cheonnyeoncho extraction method that effectively removes the viscosity of cheonnyeoncho.

It is an object of the present invention to provide a cheonnyeoncho extract and the composition comprising the cheonnyeoncho viscous effectively removed.

In order to achieve the above object, the present invention provides a method for preparing a cheonnyeoncho extract comprising the step of removing the viscosity of the cheonnyeoncho by adding an enzyme to the cheonnyeoncho ( Pounitia humifusa ).

In another aspect, the present invention provides a cheonnyeoncho extract prepared by the above production method.

In addition, the present invention provides a composition comprising the cheonnyeoncho extract prepared by the above method.

The preparation method of the cheonnyeoncho extract of the present invention effectively removes the viscosity of cheonnyeoncho, it is possible to extract the bioactive material of cheonnyeoncho to a high level. In addition, the yield of the cheonnyeoncho concentrate also has a higher effect than the conventional extraction method.

Figure 1 shows the viscosity of the hydrolyzate of cheonnyeoncho according to the hydrolase.
Figure 2 shows the total sugars and acidic sugar content of the millenium distillate enzymatic hydrolyzate according to the hydrolase.
Figure 3 is the polyphenol content of millennial enzyme hydrolyzate according to hydrolase.
Figure 4 shows the viscosity of the millennial hydrolyzate when mixed with Rapidase (viscozyme) (RV-0: control, RV-1 (rapidase: viscozyme = 4: 0), RV-2 (rapidase: viscozyme) = 3: 1), RV-3 (rapidase: viscozyme = 2: 2), RV-4 (rapidase: viscozyme = 1: 3), RV-5 (rapidase: viscozyme = 0: 4)).
Figure 5 shows the total sugar and acidic sugar content of millennial hydrolyzate when mixed with rapidase and biscozyme (RV-0: control, RV-1 (rapidase: viscozyme = 4: 0), RV-2 (rapidase: viscozyme =) 3: 1), RV-3 (rapidase: viscozyme = 2: 2), RV-4 (rapidase: viscozyme = 1: 3), RV-5 (rapidase: viscozyme = 0: 4)).
Figure 6 shows the polyphenol content of cheonnyeoncho extract upon treatment of rapidase and biscozyme mixed solution (RV-0: control, RV-1 (rapidase: viscozyme = 4: 0), RV-2 (rapidase: viscozyme = 3: 1), RV-3 (rapidase: viscozyme = 2: 2), RV-4 (rapidase: viscozyme = 1: 3), RV-5 (rapidase: viscozyme = 0: 4)).
Figure 7 shows the change in the viscosity of the millennial hydrolyzate according to the amount of millennial to the same amount of water compared to the amount of water (CW-1: 100g of millennial to 100 mL of water, CW-2: 80g of millennial to 100mL of water, CW-3: Add 40 g of cheonnyeoncho to 100 mL of water, CW-4: add 20 g of cheonnyeoncho to 100 mL of water).
Figure 8 shows the change in total sugar content of the cheonnyeoncho hydrolyzate according to the amount of cheonnyeoncho compared to the same amount of water (CW-1: added 100g cheonnyeoncho to 100mL of water, CW-2: added 80g of cheonnyeoncho to 100mL of water, CW-3 : 40 g of cheonnyeoncho in 100 mL of water, CW-4: 20 g of cheonnyeoncho in 100 mL of water).
9 shows the acidic sugar content of the millennial hydrolyzate according to the amount of millennial added (CW-1: 100 g of millennial to 100 mL of water, CW-2: 80 g of millennial to 100 mL of water, CW-3: 100 of water) add 40 g of cheonnyeoncho to mL, CW-4: add 20 g of cheonnyeoncho to 100 mL of water).
10 shows the change in the polyphenol content of the millennial hydrolyzate according to the amount of millennial to the same amount of water compared to the amount of water (CW-1: added 100 g of cheonnyeoncho to 100 mL of water, CW-2: added 80 g of cheonnyeoncho to 100 mL of water, CW- 3: add 40 g of cheonnyeoncho to 100 mL of water, CW-4: add 20 g of cheonnyeoncho to 100 mL of water).
FIG. 11 shows the viscosity change with hydrolysis time of Rapidis / Biscocozyme hydrolysates.
Figure 12 shows the change of total sugar and acid sugar content with hydrolysis time of Millipore rapidase / biscozyme hydrolyzate.
Figure 13 shows the change in polyphenol content with hydrolysis time of millennial Rapides / biscozyme hydrolyzate.
Figure 14 shows the physiological activity according to the manufacturing process of rapeseeds / biscozyme hydrolysis concentrate.
Figure 15 shows the DPPH radical scavenging ability according to the manufacturing process of the cheonnyeoncho rapidase / biscozyme hydrolyzate.
Figure 16 shows the ABTS radical scavenging ability according to the manufacturing process of cheonnyeoncho rapidase / biscozyme hydrolyzate.

The present invention provides a method of preparing a cheonnyeoncho extract comprising the step of removing the viscosity of the cheonnyeoncho by adding an enzyme to Poyeonia humifusa. In another aspect, the present invention provides a cheonnyeoncho extract prepared by the above method, and provides a composition comprising the cheonnyeoncho extract.

In addition, the present invention comprises the steps of: 1) washing and removing thorns of Pounitia humifusa ; 2) rinsing by adding water to rinsed and removed thorns; 3) treating and hydrolyzing enzymes in milled millennials; 4) heat treatment to remove enzyme activity; 5) filtering or centrifuging the millennial extract obtained in step 4); and 6) concentrating the filtered or centrifuged millennial extract obtained in step 5). It provides a method of producing a cheonnyeoncho extract comprising a. In another aspect, the present invention provides a cheonnyeoncho extract prepared by the above method, and provides a composition comprising the cheonnyeoncho extract.

In addition, the present invention comprises the steps of 1) washing and removing thorns of Pounitia humifusa ; 2) grinding and washing off thorns and thorns removed; and 3) extracting crushed millennials; 4) filtering and concentrating the extracted cheonnyeoncho extract; 5) treating and hydrolyzing the enzyme in the concentrated cheonnyeoncho extract; and 6) concentrating and filtering the hydrolyzed cheonnyeoncho extract. In another aspect, the present invention provides a cheonnyeoncho extract prepared by the above method, and provides a composition comprising the cheonnyeoncho extract.

In addition, the present invention 1) the step of removing the viscosity of the millennial by hydrolysis by adding the enzyme to Pounitia humifusa ; And 2) preparing a food composition, a pharmaceutical composition or a cosmetic composition using the cheonnyeoncho extract prepared in step 1).

The present invention comprises the steps of: 1) washing and removing thorns of Pounitia humifusa ; 2) rinsing by adding water to rinsed and removed thorns; 3) treating and hydrolyzing the enzymes in the milled millennials; 4) heat treatment to remove enzyme activity; 5) filtration or centrifugation of the cheonnyeoncho extract obtained in step 4); 6) concentrating the filtered or centrifuged cheonnyeoncho extract obtained in step 5); And 7) preparing a food composition, a pharmaceutical composition or a cosmetic composition using the cheonnyeoncho extract prepared in step 6).

In addition, the present invention comprises the steps of 1) washing and removing thorns of Pounitia humifusa ; 2) grinding and washing off thorns and thorns removed; and 3) extracting crushed millennials; 4) filtering and concentrating the extracted cheonnyeoncho extract; 5) treating and hydrolyzing the enzyme in the concentrated millennial extract; 6) concentrating and filtering the hydrolyzed cheonnyeoncho extract; And 7) preparing a food composition, a pharmaceutical composition or a cosmetic composition using the cheonnyeoncho extract prepared in step 6).

In another aspect, the present invention provides a food composition, a pharmaceutical composition or a cosmetic composition prepared through the method for producing a food composition, pharmaceutical composition or cosmetic composition.

Hereinafter, the present invention will be described in detail.

Pounitia humifusa of the present invention may be any part of cheonnyeoncho . A leaf, stem, root or fruit part of the cheonnyeoncho may be used, preferably the cheonnyeoncho stem or leaf, more preferably the cheonnyeoncho stem. Cheonnyeoncho may be used immediately after harvesting, but may be used at room temperature or refrigerated, or may be lyophilized or naturally dried. The cheonnyeoncho may be used in powder form by cutting or grinding to a suitable size for convenience.

According to the present invention, water is added to the millennial plant to prepare the millennial liquor, and the hydrolyzate may be hydrolyzed by treating the enzyme therewith. The mass of millennial plant treated per 100 mL of water is not limited. However, considering the viscosity removal of cheonnyeoncho and the content of physiologically active substance, it is preferable to add 20-100 g of cheonnyeoncho per 100 mL of water to enzymatic hydrolysis, more preferably 20 to 80 g of cheonnyeoncho per 100 mL of water. . The amount of millennial vinegar can be adjusted according to the use of cheonnyeoncho extract.In case of preparing food composition for immunostimulation, enzymatic hydrolysis by adding 20-80 g of cheonnyeoncho per 100mL of water will increase the acid sugar content. It can be advantageous, and in the case of preparing antioxidant food compositions, it is advantageous to add 40-80 g of cheonnyeoncho per 100 mL of water to increase the polyphenol content. In order to improve the texture when ingested, it is advantageous to add 20-40 g of millennial ethanol per 100 mL of water to completely remove the viscosity. Therefore, those skilled in the art can adjust the ratio of water and millennial during the preparation of millennial liquor for enzyme treatment in consideration of the use of the millennial enzymatic hydrolysis solution, the economics, the formulation of the product using the millennial hydrolysate, and the age of the consumer.

Enzymes of the present invention are cellulose, pectinase, hemicellulase, arabinase, beta-glucanase, xylanase, alpha- It may be one or a mixture thereof selected from the group consisting of alpha-amylase and glucoamylase. In addition, the enzyme of the present invention may be biscozyme, rapidides or mixtures thereof, preferably the enzyme of the present invention is a mixture of biscozyme and rapidides. The enzyme of the present invention is more preferably an enzyme mixture in which the ratio of rapidase: biscozyme is 3: 1 to 1: 3, still more preferably a mixture having 2: 2 to 1: 3, and most preferably 1: Mixture of three. However, the enzyme of the present invention is not limited to these enzymes, and it will be apparent to those skilled in the art that any enzyme capable of hydrolyzing cheonnyeoncho can be used in the present invention.

When treating enzymes to hydrolyze cheonnyeoncho, it is advantageous to maintain optimal pH conditions to increase enzyme activity. Therefore, in the preparation method of the cheonnyeoncho extract of the present invention, it will be apparent to those skilled in the art that the pH conditions in the step of hydrolysis by adding the enzyme to cheonnyeoncho may vary depending on the type of enzyme. For example, when adding Celulase Econitase CE, it is appropriate to maintain pH 4.0 to 5.5, and when using beta-glucanase (Ultraflo L, etc.) to maintain pH 5.5 to 6.5. When using Rapinase is maintained at pH 4.0 to 5.0, when using Viscozyme is maintained at pH 3.3 to 5.5, in the case of Rapinase / Viscozyme mixed enzyme liquid pH 4.0 to 5.0 conditions are preferred.

The hydrolysis of the present invention is preferably carried out for 2 hours or more after treating the enzyme. More preferably, the hydrolysis of the present invention is carried out for 2 to 16 hours, even more preferably for 2 to 12 hours. If hydrolysis is less than 2 hours, the viscosity of cheonnyeoncho is not completely removed.If the content of cheonnyeoncho for 100 mL of water exceeds 80 g, the viscosity of cheonnyeoncho is already removed when the hydrolysis time is 16 hours or more. As a result, there is less profit to perform hydrolysis anymore. However, when preparing the millennial liquor for the enzyme treatment, if the content of the millennial to 100 mL of water is more than 60 g to hydrolyze for 16 hours to completely remove the viscosity. However, continuous hydrolysis over 12 hours does not increase the bioactive components of millennial plants to significant levels. Therefore, those skilled in the art can adjust the hydrolysis time according to the use of the millennial extract and the amount of millennial to water in the millennial liquor prepared for the enzyme treatment. For example, in the preparation of millennial liquor for enzyme treatment, if the content of millennial to 100 mL of water is greater than 0 g and contains less than 60 g of cheonnyeoncho hydrolyzate as an active ingredient, to improve health functional food for immune improvement, hydrolysis It is preferable to make time into 2 to 12 hours, and when manufacturing antioxidant food containing a millennial hydrolyzate, it is preferable to hydrolyze for 2 to 8 hours. However, even if the hydrolysis for a time outside the hydrolysis time range is less effective than hydrolysis for 2 to 16 hours, the present invention is characterized by removing the viscosity of the millennial by hydrolysis by hydrolyzing the millennial By enzymatic treatment, hydrolysis in less than 2 hours or in excess of 16 hours is not within the scope of the present invention.

After hydrolysis by adding enzyme to the millennial liquor, the activity of the enzyme may be lost and / or sterilized and then concentrated by filtration or centrifugation. At this time, the enzyme activity loss / sterilization process may be performed using thermal sterilization, ultra-high temperature sterilization, high temperature short time sterilization, high temperature long time sterilization, radiation sterilization using gamma rays, ultrahigh pressure sterilization, ultraviolet sterilization, ultrasonic sterilization, electron sterilization or chemical sterilization. In addition, other methods may be used as long as the enzyme activity is lost / sterilized.

The present invention provides a cheonnyeoncho extract and a composition comprising the cheonnyeoncho extract prepared through a manufacturing method comprising the step of hydrolyzing by adding an enzyme to Pounitia humifusa to remove the viscosity of cheonnyeoncho . In addition, 1) washing and thorn removal of Pounitia humifusa ; 2) rinsing by adding water to rinse and thorn removal cheonnyeoncho ; 3) treating and hydrolyzing the enzyme in the crushed cheonnyeoncho ; 4) Heat-treating the enzyme activity; 5) filtering or centrifuging the millennial extract obtained in step 4); and 6) concentrating the filtered or centrifuged millennial extract obtained in step 5). It provides a cheonnyeoncho extract and a composition comprising the cheonnyeoncho extract prepared by the manufacturing method. In another aspect, the present invention is 1) washing and thorn removal of Pounitia humifusa ; 2) crushing the rinse and thorn removal cheonnyeoncho; 3) extracting the pulverized cheonnyeoncho; 4) filtering and concentrating the extracted cheonnyeoncho extract; 5) treating and hydrolyzing the enzyme in the concentrated cheonnyeoncho extract; and 6) a composition comprising the cheonnyeoncho extract and the cheonnyeoncho extract prepared using the manufacturing method comprising the step of concentrating and filtering the hydrolyzed cheonnyeoncho extract. To provide.

The compositions may be a pharmaceutical composition, a food composition or a cosmetic composition, the pharmaceutical composition may be a pharmaceutical composition for antioxidant or a pharmaceutical composition for enhancing immune function. In addition, the food composition may be a general food, health supplement food, functional food, may be a health functional food for improving liver function, antioxidant food or health supplement for immune function enhancement.

Health functional foods and pharmaceutical compositions for antioxidant of the present invention containing the extract of cheonnyeoncho by removing the active oxygen in the body, inhibits the reaction of fat, cholesterol and free radicals in the body to produce lipid peroxide, whereby the peroxide lipid clots It can prevent the embolism. In addition, antioxidant health functional foods and pharmaceutical compositions of the present invention by removing the active oxygen, blood circulation disorders due to thrombus, embolism, lipid peroxide associated with active oxygen, stroke, stroke, cerebral thrombosis, myocardial infarction, arteriosclerosis and By treating and inhibiting the degeneration of DNA, cells and tissues by free radicals, it prevents and treats atopy, allergy, tumor, arthritis, cataract and skin tumor, and inhibits the aging process.

Health functional foods and pharmaceutical compositions for improving immune function of the present invention, including the extract of cheonnyeoncho is allergic to cancer, multiple sclerosis, rheumatoid arthritis, Crohn's disease, autoimmune diseases such as diabetes or asthma, allergic rhinitis, conjunctivitis, atopic dermatitis Effective for iso-immune diseases. However, even if not a specific disease, health functional foods and pharmaceutical compositions for enhancing immune function of the present invention may lower the possibility of infection and onset of various bacterial and viral diseases by enhancing the overall immunity of the body.

The pharmaceutical composition for improving immune function, including the extract of the present invention cheonnyeoncho is Zadaxin (Thymosinalpha-1), EP-HBS (HBV Therapeutic Vaccine), HBV Core Antigen Vaccine, LPS, Con. It can be administered with an immunostimulant such as A or an immunomodulator such as Intron A (Interferon alpha-2b) or Pegasys (Peginterferon alfa-2a). In addition, the pharmaceutical composition for enhancing immune function of the present invention may further include the immunostimulant or immunomodulator.

The pharmaceutical composition for antioxidant and the immune function enhancing pharmaceutical composition of the present invention can be administered orally or parenterally and can be used in the form of general pharmaceutical preparations. Preferred pharmaceutical preparations include oral preparations such as tablets, hard or soft capsules, solutions, suspensions and the like, which can be used in the form of excipients in conventional pharmaceutically acceptable carriers such as oral preparations, Binders, disintegrants, lubricants, solubilizers, suspending agents, preservatives or extenders can be used.

The dosage of the antioxidant composition of the present invention and the pharmaceutical composition for enhancing immune function may be determined by a specialist according to various factors such as the patient's condition, age, sex, and complications, but is generally 0.1 per kg of adult. It may be administered at a dose of mg to 10 g, preferably 10 mg to 1 g. In addition, it is intended to contain a daily dose of the pharmaceutical composition or a dose of 1/2, 1/3 or 1/4 thereof per unit dosage form, and may be administered 1 to 6 times a day. However, in the case of long-term intake for health and hygiene or health control, the amount may be below the above range, and the active ingredient may be used in an amount above the above range because there is no problem in terms of safety.

The food composition of the present invention may be a general food, health supplement food, functional food, and may be antioxidant food or health supplement food for enhancing immune function. At this time, the food composition includes a food additive. The food composition comprising the cheonnyeoncho extract prepared through the manufacturing method of the present invention may be added as it is or used in conjunction with other food or food compositions, may be appropriately used in accordance with conventional methods. The mixed amount of the active ingredient may be suitably determined according to the purpose of use (prevention, health or therapeutic treatment). In general, the food composition of the present invention may be added in the amount of 5 to 70% by weight, preferably 7 to 60% by weight based on the total raw materials in the manufacture of food or beverage, but the flavor, consumer preferences, economics, etc. It is apparent to those skilled in the art that the amount can be increased or decreased accordingly.

There is no particular limitation on the kind of food. The food composition comprising the extract of the cheonnyeoncho as an active ingredient of the present invention can be used in the form of oral administration preparations such as tablets, hard or soft capsules, solutions, suspensions, etc., these preparations are acceptable conventional carriers, such as For example, in the case of oral preparations, excipients, binders, disintegrants, lubricants, solubilizers, suspending agents, preservatives or extenders may be used.

Examples of the food containing the extract of the present invention as an active ingredient, meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, Teas, drinks, alcoholic beverages, and vitamin complexes, but are not limited to these types of foods.

In another aspect, the present invention provides a cosmetic composition comprising the cheonnyeoncho extract prepared by the manufacturing method as an active ingredient, and provides a production method of the cosmetic composition comprising the manufacturing step of the cheonnyeoncho extract. In the manufacturing method of the cosmetic composition, the carrier that is acceptable for the manufacture of cosmetics may vary depending on the formulation of the cosmetic composition of the present invention. The cosmetic compositions of the present invention may be formulated into solutions, suspensions, emulsions, pastes, gels, creams, lotions, soaps, shampoos, surfactant-containing cleansing, oils, powder foundations, liquid foundations, cream foundations, sprays, and the like. The antioxidant cosmetic composition of the present invention may be more specifically a flexible lotion (skin, toner), astringent lotion, nutrition lotion (lotion), nutrition cream, massage cream, essence, gel, skin adhesive patch, powder, It may be prepared in the form of a conventional cosmetic such as skin external ointment, patch, suspension, emulsion spray, eye cream, cleansing cream, cleansing foam, cleansing butter, pack, hair essence or essence.

When the formulation of the cosmetic composition prepared by the method of the present invention is a paste, cream or gel, the carrier component is animal oil, vegetable oil, wax, paraffin, starch, trachant, cellulose derivative, polyethylene glycol, silicone, bentonite , Silica, talc or zinc oxide and the like are preferred.

When the formulation of the cosmetic composition comprising the cheonnyeoncho extract prepared by the preparation method of the present invention as an active ingredient is a solution or emulsion, a carrier, a solvating agent or an emulsifying agent may be used as a carrier component, and examples thereof include water, Preferred are ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzene alcohol, benzene benzoate, propylene glycol, 1,3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid ester of sorbitan.

In the case of the suspension of the cheonnyeoncho extract prepared by the production method of the present invention is a suspension solution, the carrier component is water, ethanol or liquid diluents such as propylene glycol, ethoxylated isostearyl alcohol, aluminum metahydroxy, bentonite, Agar or tracant and the like are preferable.

When the formulation of the cosmetic composition prepared by the preparation method of the present invention is a surfactant-containing cleansing, the carrier component may be aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, Methyltaurate, sarcosinate, fatty acid amide ether sulfate, alkylamidobetaine, fatty alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol. Fatty acid esters and the like are preferred.

Hereinafter, the present invention will be described in more detail by the following examples and experimental examples. However, the following Examples and Experimental Examples are only illustrative of the contents of the present invention and the scope of the invention is not limited by the Examples and Experimental Examples.

Example 1 Materials

Cheonnyeoncho used in this experiment was produced in Chungnam Yeongi-gun and received from Neo-Crema. For the decomposition of viscous substances of cactus, cellulase (Econitase CE), beta-glucanase (Ultralflo L), pectinase (Cytolase PCL 5), alpha-amylase (Spezyme Prime), glucoamylase (Optidex L- 400), Pectinex 5XL, Celluclast 1.5L, optidex L-400, Enzyme mixture Rapidase (pectinase, hemicellulose and cellulase) and Viscozyme (arabinase, cellulase, beta-glucanase, hemicellulase and xyl Raise), which were purchased from Vision Biocam (Table 1).

Enzyme


Main activity


Source

Optimum condition

pH

Temp (℃)
Econitase CE

Cellulase
Trichoderma sp.
4.0-5.5
55

Rapidase
pectinase, hemicellulase,
cellulase

Asp . niger & T. longibrachiatum

4.0-5.0

10-55


Viscozyme
arabinase, cellulase, β-glucanase,
hemicellulase, xylanase



Asp . Sp.


3.3-5.5


40-50
Ultraflo L
β-glucanase

Humicola insolens
6
40
Cytolase PCL 5
pectinase

Asp . Niger
2.5-5.0
10-55
Spezyme prime
α-amylase

Asp . niger
6.2-6.5
85-90
Optidex L-400
glucoamylase
Geobacillus
stearothermophil us

4.0-4.5
58-65
Celluclast 1.5L
cellulase

Trichderma reesei
4.5-6.0
50-60
Pectinex 5XL

polygalacturonase
Asp . niger
4.5
50

Characteristics of the enzyme

Example 2 Activity Analysis of Enzyme Hydrolysates

Experimental Example 1 Preparation of Enzyme Hydrolysates

After rinsing the stem of Cheonnyeoncho, the residual residue was removed, and 700 mL of water was added to 70 g of Cheonnyeoncho and crushed with a homeogenizer. Divide 70 mL each, add ultraflo L, Viscozyme, Spezyme prime, Cytolase PCL5, Celluclast 1.5L, Optidex L-400, Pectinex 5XL, Rapidase and Econase CE (addition amount of enzyme: 350 μL), and slowly stir at 40 ° C for 6 hours. Hydrolysis was carried out. Thereafter, the mixture was heated in boiling water for 5 minutes to remove enzyme activity, cooled, and filtered to use the supernatant as a millennial enzyme hydrolysis solution.

Experimental Example 2 Activity Analysis of Enzyme Hydrolysates

Viscosity

The viscosity of enzymatic hydrolysates was measured using a Brookfield viscometer (Model DV-II). The viscosity was measured under the conditions of 60 rpm at 30 ° C using 3 (Fig. 1).

Since the viscosity of cheonnyeoncho was mainly due to pectin, it was expected that the viscosity would be the lowest when treating pectinase (cytolase PCL5). However, the results of viscosity measurement showed that pectinase did not have a significantly higher viscosity removal effect than other enzymes (Cytoclase PCL5). Viscosity at treatment: 13.60 cP), starch degrading enzymes spezyme prime and optidex L-400 were found to increase slightly in viscosity rather than control (14.47 cP) (14.77 and 15.47 cP, respectively). In particular, Viscozyme treatment without pectinase significantly lowered the viscosity of the cheonnyeoncho extract (12.87 cP), and the viscosity of the Rapidase treatment group was 12.67 cP, the lowest among the enzyme hydrolysis groups. It showed a viscosity.

Total sugar and acid sugar content

The total and acidic sugar contents of the enzymatic hydrolysates of cheonnyeoncho were determined using the phenol-sulfuric acid method and Blumenkrantz and Asboe-Hansen methods (Blumenkrantz, N. and Asboe-Hansen, G. (1973) Anal. Biochem., 54, 484). Measured.

As a result, the total and acid sugar contents of the control group were 14.2 mg / mL and 461.0 mg / mL, respectively, while the hydrolyzate by 1.5L of celluclast was 35.7 mg / mL and 564.5 mg / mL, respectively. The highest content of biscozyme and rapidase hydrolyzate was 24.0, 16.9 mg / mL and 468.6 and 558.4 mg / mL, respectively. The acidic sugar was 638.4 mg / mL having the highest hydrolyzate by econase CE, but the lowest total sugar content was 15.0 mg / mL (FIG. 2).

Polyphenols

Millenium Enzyme Singer by Folin-Denis Method (Dewanto, V., Wu, X., & Liu, RH (2002b) .Processed sweet corn has higher antioxidant activity.Journal of Agricultural and Food Chemistry, 50, 4959-4964.) The polyphenol content of the degradation product was measured.

All millennial enzyme hydrolyzates contained 2.5-4 times higher polyphenols compared to the control (37.8 mg / mL) (FIG. 3). Among them, the polyphenol content of pectinase, Cytoclase PCL5 hydrolyzate, was the highest (154.0 mg / mL), Ultraflo L, Biscozyme, Spezime prime, Pectinex 5XL and Rapidases contained 120-140 mg / mL polyphenols.

Example 3 Activity Analysis of Bichonzyme / Rapidase Hydrolysates

As a result of Example 2, the contents of total sugar and acid sugar were relatively high, and the rapid viscosity and biscozyme having the lowest viscosity were selected, and hydrolyzate was prepared by treating their mixed enzyme solution.

Experimental Example 1 Preparation of Bichonzyme / Rapidase Hydrolysates

In order to measure the effect of the mixture of biscozyme and rapidase, which had the lowest viscosity, 500 ml of water was added to 100 g of cheonnyeoncho and crushed, followed by addition of enzyme (5 mL / 100 g cheonnyeoncho) for 9 hours at 40 ° C. Hydrolysis. At this time, the enzymes were added to Rapidase and biscozyme in the ratio of 4: 0, 3: 1, 2: 2, 1: 3 and 0: 4, respectively, to prepare a hydrolyzate.

RV-0 RV-1 RV-2 RV-3 RV-4 RV-5 Control  4: 0  3: 1 2: 2 1: 3 0: 4

RV-1 to RV-5: Rapides: Biscozyme ratio

RV-0: control (millennial hydrothermal extract)

Experimental Example 2 Activity Analysis of Biscozyme / Rapidase Hydrolysates

Viscosity

When the mixed enzyme solution of rapidase and biscozyme was added, it was confirmed that the viscosity was significantly reduced compared to the control (10.07 cP). The viscosity of Viscozyme alone (RV-5, 2.50 cP) was higher than that of Rapidase only (RV-1), but when they were mixed at a specific ratio, the viscosity was lower (RV-3 and RV-4). In particular, Rapids: Biscozyme 1: 3 was the lowest viscosity when mixed (RV-4.25CP), which can effectively remove the viscosity of the cheonnyeoncho extract during treatment by mixing Rapidis and Biscozyme in a specific ratio. It means that it can (Fig. 4).

Total sugar and acid sugar content

As a result of measuring the total sugar and acid sugar content in the mixed use (Fig. 5), the mixed enzyme solution of rapidase and biscozyme was found to have higher total and acid sugar content than the control group. The total sugar content was the highest at 18.5 mg / mL in RV-4 and the control group was 12.3 mg / mL. The uric acid content of the acidic sugar was 1.27 mg / mL in the control group, whereas RV-1 and RV-4 showed high contents of 1.94 mg / mL and 1.95 mg / mL, respectively. The total and acidic sugar contents of RV-4, which had the lowest viscosity, were the highest.

Polyphenols

The content of polyphenols in the control group was the lowest (34.62 mg / mL) and 98.31-106.81 mg / mL when rapidase and biscozyme were mixed, but there was no significant difference between the mixing ratios of enzymes (FIG. 6).

In summary, when the enzyme mixture mixed with Rapidis and biscozyme in a ratio of 1: 3 was added to the millennial plant, the viscosity of the millennial extract was most effectively removed, and the millennial plant such as total sugar, acid sugar, and polyphenol It can be seen that the content of the active ingredient is also high.

Example 4 Activity of Biscozyme / Rapidase Hydrolysates with Millennial Addition

During the preparation of the millennial hydrolyzate by treating the mixed solution with Rapidis and biscoza, the hydrolyzate, total sugar, acidic sugar and polyphenol content were measured to determine the activity of the millennial hydrolyzate according to the amount of millennial added.

Experimental Example 1 Preparation of Hydrolyzate

Crushed by adding 100 mL of water to 20, 40, 80, and 100 g of Cheonnyeoncho, and adding rapidase / biscozyme mixed enzyme solution (1: 3 mixture) (5 mL of enzyme solution per 100 g of millennial plant) for 12 hours. Digested.

CW-1
CW-2
CW-3
CW-4

Amount of millennial
(g / water 100 mL)

100
80
40
20

<Experimental Example 2> Activity analysis according to the amount of cheonnyeoncho added

Viscosity

Prior to hydrolysis, the higher the amount of millennial plants, the higher the viscosity. However, the viscosity rapidly decreased after treatment with Rapidase / biscozyme mixed enzyme solution (1: 3 mixture). In particular, 100 g of water and 20 g of Cheonnyeoncho (CW-4) completely removed the viscous material, and 40 g of Cheonnyeoncho (CW-3) almost eliminated the viscosity (FIG. 7).

Total sugar and acid sugar content

As a result of measuring the content of total sugar and acidic sugar during hydrolysis by varying the amount of millennial added to the same amount of water, it was confirmed that the content of total sugar and acidic sugar increased after hydrolysis compared to before hydrolysis. After hydrolysis, the total equivalent weight of 100g (CW-1) and the most added 20g (CW-4) were the lowest, but CW-2 and CW-3 had no significant difference in total equivalents (Fig. 8). On the other hand, the content of acidic sugars tended to increase in proportion to the amount of millennial added (Figure 9). Therefore, it may be desirable to determine the amount of millennial added according to the use of the millennial hydrolyzate. For example, in the case of the dietary supplement for immune enhancement comprising a cheonnyeoncho hydrolyzate as an active ingredient, the higher the content of acidic sugar is advantageous, it is preferable to add a lot of cheonnyeoncho compared to the amount of water to treat the enzyme.

Polyphenols

The content of polyphenol was increased when the amount of polyphenol was increased below 40 g before the hydrolysis, but the amount of polyphenol was about the same when the amount of polyphenol was added more than 40 g. On the other hand, the content of polyphenols after enzymatic hydrolysis showed a tendency to increase as the amount of millennial added increased (FIG. 10).

Taken together, the preparation of millennial liquor for enzymatic hydrolysis showed a tendency to increase the bioactive substance of hydrolyzate as the amount of millennial herb contained in the same amount of water increased. It is not proportional to. Therefore, it is preferable to hydrolyze the amount of millennial plant by adjusting the amount of millennial plant hydrolyzate according to the use of the hydrolyzate and the demand of the consumer (price, degree of effect, etc.).

Example 5 Activity of Millipore Rapids / Biscocozyme Hydrolysates with Hydrolysis Time

100 mL of water was added to 100 g of cheonnyeoncho and crushed, and then mixed with Rapids / Biscocozyme (mixed at a ratio of 1: 3) (5 mL of enzyme solution per 100g of cheonnyeoncho) and hydrolyzed at different temperatures at 40 ° C. It was.

Viscosity

As a result of measuring the change in viscosity according to the hydrolysis time (FIG. 11), the millennial plant showed a viscosity of 40.9 cP before hydrolysis, and the viscosity decreased rapidly to 1.9 cP after 2 hours of hydrolysis, after which the viscosity gradually decreased. After 11 hours of hydrolysis, the decomposition was so complete that no viscosity was measured. Therefore, hydrolysis for 2 hours or more is effectively judged to remove the viscosity of the cheonnyeoncho extract.

Total sugar and acid sugar content

As a result of measuring the content of total sugar and acidic sugar of millennial Rapidase / Viscozyme hydrolyzate according to hydrolysis time, total sugar and acidic sugar tended to increase with increasing hydrolysis time (FIG. 12). The total sugar content before hydrolysis was 5.05 mg / mL, but after 2 hours hydrolysis, it increased to 9.75 mg / mL, and then slightly increased and then decreased. After 8 hours, the total sugar content was slightly decreased. Steadily increased. Uronic acid, an acidic sugar, increased rapidly from 1088.34 mg / mL to 2685.51 mg / mL after 2 hours of hydrolysis, after which the hourly increase was not as large as the first 2 hours, but the uronic acid content increased to a significant level. It was. However, after 12 hours of hydrolysis, the increase in uronic acid content was insignificant over time. Therefore, if you want to prepare a health functional food for immune improvement containing the cheonnyeoncho hydrolyzate as an active ingredient, it would be preferable to set the hydrolysis time to 12 hours.

Polyphenols

When hydrolyzed cheonnyeoncho with Rapidase / Viscozyme mixed enzyme solution for 2 hours, the polyphenol content increased from 672.9 mg / mL to 820.13 mg / mL. Thereafter, up to 8 hours of hydrolysis, although the increase was not as great as the first 2 hours, it increased to a significant level (polyphenol content of 856.48 mg / mL at 8 hours of hydrolysis). However, the contents of 852.20 and 863.90 mg / mL were observed at 11 and 24 hours of hydrolysis, respectively, and the change in content was found to be insignificant as a result of periodic measurement until 24 hours of hydrolysis. Therefore, if the millennial hydrolyzate is used for antioxidant use, it is not necessary to hydrolyze more than 8 hours.

In consideration of the above results, it is preferable to adjust the hydrolysis time according to the use of the millennial hydrolyzate. For example, when preparing a health functional food for improving immunity comprising a cheonnyeoncho hydrolyzate as an active ingredient, it is advantageous to have a hydrolysis time of 12 hours, but when preparing an antioxidant food containing a cheonnyeoncho hydrolyzate, for 8 hours Only hydrolysis will achieve the desired effect.

Example 6 Antioxidant Activity and Immune Function Enhancement Activity of Millennial Concentrates by Millennial Rapids / Biscozyme Hydrolysis Process

Experimental Example 1 Preparation of Millennial Rapids / Biscocozyme Hydrolyzate and Millennial Hydrothermal Extraction / Concentrate

Process A

After milling 1.0 kg of cheonnyeoncho water and removing thorns, 2.5 kg of water was added thereto, followed by grinding, to prepare a millennial vinegar. 50 mL / kg of an enzyme mixture mixed with Rapidase / Biscocozyme 1: 3 was added to the millennial solution and hydrolyzed at 40 ° C. for 12 hours. And heat treatment at 95 ℃ for 15 minutes to remove the enzyme activity and sterilization process. The cheonnyeoncho hydrolysis solution from which the enzyme activity was removed was concentrated by filtration.

Process B

Wash 1.0 kg of cheonnyeoncho, remove the thorns and crush them

Alcohol was extracted. After that, the millennial extract was filtered and concentrated, and 50 mL / kg of an enzyme mixture mixed with Rapidase / Biscocozyme 1: 3 was added and hydrolyzed at 40 ° C. for 6 hours. And heat treatment at 95 ℃ for 15 minutes to remove the enzyme activity and sterilization process. The cheonnyeoncho extract from which the enzyme activity was removed was filtered and concentrated to prepare a concentrate.

Control

A millennial hot water extract / concentrate was prepared as a control. That is, 1.0 kg of cheonnyeoncho was washed with water and thorns were removed, and 2.5 kg of water was added thereto, followed by crushing to prepare a millennial vinegar. Thereafter, the millennial liquor was heated to 95 ° C. for 6 hours, followed by hot water extraction, followed by filtration and concentration.

Experimental Example 2 Physiological Activity of Rapids / Biscocozyme Hydrolysates

As a result of measuring the physiological activity of the cheonnyeoncho concentrate prepared in Experimental Example 1, the cheonnyeoncho rapidase / biscozyme hydrolyzate concentrate is a control of both the concentrate prepared by step A and the concentrate prepared by step B cheonnyeoncho hot water Compared with the extract / concentrate, the total sugar, acid sugar, and polyphenol content were all higher (FIG. 14 and Table 4). In addition, the efficiency of the concentrate was higher in the rapids / biscozyme hydrolyzate than the hydrothermal extract.

Total sugar (mg / mL) Acid Sugar (mg / mL) Polyphenol (mg / mL) Extraction and Concentration Yield (%) Millennial Concentrate Prepared by Process A 5,221.3 882.7 160.7 6.70 Millennial Concentrate Prepared by Process B 2,749.7 864.7 70.4 3.67 Cheonnyeoncho Hydrothermal Extraction / Concentrate (Control) 2,134.6 737.2 45.8 2.77

<Experiment 3> Antioxidant Activity of Rapids / Biscocozyme Hydrolyzate Concentrate

1.0 kg of cheonnyeoncho was washed with water and thorns were removed, and then 1.0 to 5 kg of water was added thereto, followed by grinding. The antioxidant activity of the millennial rapidase / biscozyme hydrolyzate and millennial hydrothermal extract / concentrate prepared by Process A and Process B were measured using the millennial liquors having different ratios of water and millennial. Antioxidant activity was measured by ABTS and DPPH radical scavenging ability.

ABTS radical scavenging activity was performed by the ABTS cation decolorization assay method (Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C. 1999. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radic Biol Med 26: 1231-1237.), ABTS 7.4 mM and 2.6 mM potassium persulphate were left in the dark for 1 day to form ABTS radicals, and then the solution was molar extinction coefficient (ε = 3.6 ×) at 734 nm to obtain an absorbance value of 1.0. 104 M-1 cm-1) was diluted with distilled water. 50 μL of the extract was added to 1 mL of diluted ABTS radical solution, and the change in absorbance was measured after exactly 90 minutes.

Antioxidant activity by the DPPH radical scavenging activity is Kim et al. (Kim DO, Lee KW, Lee HJ, Lee CY. 2002. Vitamin C equivalent antioxidant capacity (VCEAC) of phenolic phytochemicals. J Agric Food Chem 50: 3713-3717.) Was modified. 50 μL of the extract was added to 1 mL of 0.2 mM DPPH solution and the change in absorbance was measured after exactly 30 minutes at 520 nm.

ABTS radical scavenging activity and DPPH radical scavenging ability were calculated by the following equation.

<Equation 1>

ABTS radical scavenging ability (%) = (Ab-As) / Ab × 100

Ab: change in absorbance value when distilled water is added instead of concentrate

As: absorbance value when concentrate is added

<Equation 2>

DPPH radical scavenging activity (%) = (Ab-As) / Ab × 100

Ab: change in absorbance value when distilled water is added instead of concentrate

As: absorbance value when concentrate is added

As a result of the antioxidant activity measurement, the millennial rapidase / biscozyme hydrolyzate concentrate prepared by Process A had a higher DPPH radical scavenging activity than the millennial hydrothermal extract concentrate over the concentration range of the concentrate, and the millennial raffy prepared by Process B. Days / biscozyme hydrolyzate concentrates showed higher DPPH radical scavenging ability than hydrothermal extract / concentrate at low concentrations, but lower than hydrothermal extract / concentrate at high concentrations. However, there was not a significant difference in DPPH radical scavenging ability between both millipria rapheides / biscozyme hydrolyzate concentrates and controls (FIG. 15). On the other hand, for ABTS radical scavenging activity, the millennial rapidase / biscozyme hydrolyzate concentrate was significantly larger than the millennial hydrothermal extract concentrate over the concentration range of the concentrate, and the difference tended to increase with increasing concentration of the concentrate (FIG. 16). In particular, the millennial rapidase / biscozyme hydrolyzate concentrate prepared by Process A had significantly higher ABTS radical scavenging capacity than that prepared by Process B as well as the control.

Experimental Example 3 Immune Function Enhancing Activity of Rapidis / Biscocozyme Hydrolysates

In the millennial rapidase / biscozyme hydrolyzate concentrates prepared in Experimental Example 1 and the millennial hydrothermal extract / concentrate, the active polysaccharide content as a causative agent of anti-complement activity was measured. As a result, the millennial hydrothermal extract / concentrate contained 5.85 mg / mL of active polysaccharide, and the millennial rapidase / biscozyme hydrolyzate concentrates prepared by Process A and Process B were 9.07 mg / mL and 6.25 mg, respectively. It contained / mL of active polysaccharide. Therefore, when hydrolyzing Chun-na-cho with Rapidase / Biscocozyme 1: 3 mixed enzyme solution, the amount of active polysaccharide extracted from Cheon-nyeon was high.

The preparation method of the cheonnyeoncho extract of the present invention is not only effective in removing the viscosity of cheonnyeoncho, but also contains the active substance in the extract higher than the cheonnyeoncho extract by the conventional method. Therefore, the preparation method and the cheonnyeoncho extract using the cheonnyeoncho extract of the present invention is expected to facilitate the processing and use of cheonnyeoncho.

Claims (9)

Method of producing a cheonnyeoncho extract comprising the step of hydrolyzing by adding rapidase or biscozyme to Pounitia humifusa to remove the viscosity of cheonnyeoncho.
Method of producing a cheonnyeoncho extract comprising the step of hydrolysis by adding a mixture of rapidase and biscozyme to Pounitia humifusa to remove the viscosity of cheonnyeoncho.
The method of claim 2,
The mixture is a method for producing a cheonnyeoncho extract, characterized in that the rapids and biscozyme is mixed in a ratio of 2: 2 to 1: 3.
The method of claim 1,
Process for hydrolysis for 2 to 12 hours.
1) washing and thorn removal of Pounitia humifusa;
2) crushing by adding water to the washed and thorn-free cheonnyeoncho;
3) treating and hydrolyzing rapidase or biscozyme in the milled millennial;
4) heat treatment to remove enzyme activity;
5) filtering or centrifuging the cheonnyeoncho extract obtained in step 4); and
6) A method for producing a cheonnyeoncho extract comprising the step of concentrating the filtered or centrifuged cheonnyeoncho extract obtained in step 5).
A cheonnyeoncho extract prepared by the method of any one of claims 1 to 5.
Cosmetic composition comprising the cheonnyeoncho extract of claim 6.
Food composition comprising the cheonnyeoncho extract of claim 6.
The method of claim 8,
The food composition is a food composition, characterized in that the antioxidant food composition or health supplement food composition for immune promotion.

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