KR20130060954A - Process for preparation of higher yield extract from aloe gel using enzyme and its antioxidant activity - Google Patents

Process for preparation of higher yield extract from aloe gel using enzyme and its antioxidant activity Download PDF

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Publication number
KR20130060954A
KR20130060954A KR1020110127285A KR20110127285A KR20130060954A KR 20130060954 A KR20130060954 A KR 20130060954A KR 1020110127285 A KR1020110127285 A KR 1020110127285A KR 20110127285 A KR20110127285 A KR 20110127285A KR 20130060954 A KR20130060954 A KR 20130060954A
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South Korea
Prior art keywords
aloe
extract
sugar hydrolase
enzyme
aloe gel
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Application number
KR1020110127285A
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Korean (ko)
Inventor
김영선
전유진
강민철
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제주알로에영농조합법인
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Priority to KR1020110127285A priority Critical patent/KR20130060954A/en
Publication of KR20130060954A publication Critical patent/KR20130060954A/en

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    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A23B - A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105Plant extracts, their artificial duplicates or their derivatives
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/88Liliopsida (monocotyledons)
    • A61K36/886Aloeaceae (Aloe family), e.g. aloe vera
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K8/00Cosmetics or similar toilet preparations
    • A61K8/18Cosmetics or similar toilet preparations characterised by the composition
    • A61K8/96Cosmetics or similar toilet preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97Cosmetics or similar toilet preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783Angiosperms [Magnoliophyta]
    • A61K8/9794Liliopsida [monocotyledons]
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILET PREPARATIONS
    • A61Q19/00Preparations for care of the skin

Abstract

      The present invention relates to a method for preparing a high yield of water-soluble aloe gel extract using a sugar hydrolase from an aloe gel and an antioxidant composition comprising the same. By using the hydrolase, the yield of the extract can be twice as high as that of the conventional extract, and by hydrolysis, the aloe hydrolyzate having high water solubility of the lyophilized micropowder and high yield of antioxidant activity with excellent free radical scavenging ability Decomposed extracts can be obtained.

Description

Process for preparation of higher yield extract from Aloe gel using enzyme and its antioxidant activity}
The present invention relates to an optimized enzymatic extraction method for the preparation of water-soluble substances using aloe gel as a main raw material and an antioxidant composition containing the enzyme extract obtained therefrom as an active ingredient.
Lilies and Aloe (hereinafter referred to as Aloe) are a kind of succulent plant including Aloe barbadenisis Miller and Aloe arborescen Miller var.natalensis Berger. It is known empirically to have efficacy. Aloe has long been used as a Chinese medicine in China and other Asian countries. Today, however, various physiological activities such as antibacterial, anti-inflammatory, cell regeneration, and immunity enhancement are reported in various fields, and they are used in various parts of the world. It is recognized as a product.
Known aloe products include aloe gel, aloe gel concentrate, and aloe gel powder.Aloe gel is a gel obtained from the leaves of edible aloe varieties (vera, aboresense (kitachi), saponaria). It is defined as saying that it contains 0.5% or more of solid content as a gel component.
Aloe gel shows a very low solids content of about 1 to 2% solids, and there is a lack of research on high yield extraction methods and efforts to enhance physiological activity. In general, the method of commercializing the aloe relies on the method of freezing-drying the aloe leaves, wet-drying, hot-air-drying and processing into powders, capsules, and the like. However, this powder is a powder before the raw material is extracted, so it contains a large amount of water-insoluble material. Alternatively, it may be concentrated and sold in liquid form using only aloe gel components. However, efforts to increase the physiological activity by extracting only the gel component of aloe or increasing the content of water-soluble solids of the extract and the extraction conditions are still insufficient.
The present invention is to provide a method for producing a water-soluble substance in a high yield using aloe glycolase. The present invention also aims to find the optimum ratio of enzymes and enzymes, select hydrolyzates with excellent physiological activity, and finely powder them for use in various industrial fields.
The present invention also provides an antioxidant composition containing aloe gel extract with enhanced physiological activity.
In one embodiment of the present invention provides a method for producing an aloe sugar hydrolase extract comprising the step of treating the aloe gel using a sugar hydrolase.
In consideration of the improvement of the yield aspect in the production method according to an embodiment of the present invention, the step of treating the aloe gel using a hydrolase is a reaction temperature of 50 to 60 ℃ in a buffer of pH 4.5 to 7 and the reaction It may be preferable to perform the time 3 hours to 12 hours.
In consideration of the yield and physiologically active aspects of the production method according to an embodiment of the present invention, the step of treating the aloe gel with a glycolytic enzyme is a ratio of substrate to enzyme of 1: 100 to 1: 10,000. It may be desirable to carry out within the range.
In the preparation method according to an embodiment of the present invention, the glycolytic enzyme is biscozyme, celluclast, amyloglucosidase (AMG), termamyl and ultraflo ( Ultraflo) alone or a mixture thereof, and the most preferred sugar hydrolase in terms of enhancing the biological activity is the viscozyme (Viscozyme).
In the preparation method according to an embodiment of the present invention, the buffer may be a single or a mixture selected from distilled water, acetate buffer and phosphate buffer.
In the preparation method according to an embodiment of the present invention, after the step of treating the aloe gel using a sugar hydrolase, it may be prepared into an aloe gel powder including filtration and lyophilization step.
In one embodiment of the present invention provides an aloe sugar hydrolase extract prepared by the method according to the embodiment.
The present invention also provides an antioxidant composition containing aloe sugar hydrolase extract as an active ingredient.
In view of the yield of antioxidant active ingredients, glycolytic enzymes include Viscozyme, Celluclast, Amyloglucosidase (AMG), Termamyl and Ultraflo. It may be a single or a mixture thereof selected from the above, and considering the improved aspect of the yield and antioxidant activity of the antioxidant active ingredient, it may be preferable that the glycolytic enzyme is biscozyme.
Antioxidant composition of the present invention can be used as a medicine, health supplements, cosmetics, food or food additives, of course.
According to the preparation method according to one embodiment of the present invention, a water-soluble aloe gel powder of about 2 times as compared to the aloe extraction method, for example, an extraction method by water, can be obtained. In addition, it was confirmed that the antioxidant active ingredient also exhibits high antioxidant activity when a specific sugar hydrolase is used. Therefore, enzymatic hydrolysis using sugar hydrolase of aloe gel is an industrially and economically useful extraction method for producing cosmetic raw materials or health functional food.
1 is a flow chart of the extraction process using sugar hydrolase of aloe gel.
Figure 2 shows the extraction yield (when substrate to enzyme ratio 1: 100) according to each extraction method using the five sugar hydrolases of aloe gel.
Figure 3 shows the extraction yield according to the substrate-to-enzyme ratio in the extraction method using the biskozyme of aloe gel.
Figure 4 is a graph showing the DPPH radical scavenging activity of the aloe sugar hydrolase extract compared to the water extract.
Figure 5 is a graph showing the hydroxyl radical scavenging activity of the aloe sugar hydrolase extract compared to the water extract.
Figure 6 is a graph showing the hydrogen peroxide scavenging activity of the aloe saccharide hydrolase extract compared to the water extract.
Figure 7 is a graph showing the intracellular reactive oxygen scavenging activity of the aloe saccharide hydrolase extract compared to the water extract.
The present invention removes the viscosity of the aloe gel used in various fields industrially through the enzymatic extraction method to obtain a water-soluble extract in high yield, and also powdered aloe gel hydrolyzate having excellent physiological activity to be used in various industrial fields I would like to.
This invention will be described in detail with reference to the accompanying drawings.
The present invention relates to a method for preparing an aloe sugar hydrolase extract comprising treating a sugar hydrolase from an aloe gel.
When the aloe gel is extracted with water, the yield of the water-soluble extract is too low. In addition, there is a limit to improving the physiological activity of the obtained water-soluble extract.
Therefore, in one embodiment of the present invention, the yield of water-soluble extracts is remarkably improved through enzymatic extraction using sugar hydrolase.
Enzymatic extracts using the sugar hydrolase of aloe can be obtained through a series of processes shown in FIG. Figure 1 includes the lyophilization process after the enzymatic extraction step, which can of course be omitted when used as a gel form itself. Specifically, referring to Figure 1, (iii) the aloe stem is cut after removing the epidermis of the stem portion. Next, (ii) sugar hydrolase is added to the crushed aloe gel and maintained at an appropriate pH and temperature. (Iii) Extract at a suitable temperature and pH for 3 to 12 hours using an extractor. (Iii) The aloe gel extract is neutralized and filtered. (Iii) Lyophilize the aloe sugar hydrolyzate.
The sugar hydrolase is not limited as long as it is food-acceptable. For example, Viscozyme, Celluclast, Amyloglucosidase (AMG), and Termamyl And it may be a single or a mixture thereof selected from Ultraflo.
The use of sugar hydrolase can recover the amount of extract twice as high as that of conventional extraction, for example, extraction with water, and the high yield of the water-soluble lyophilized micropowder by hydrolysis and excellent scavenging ability of active oxygen. Aloe hydrolyzed extract having excellent antioxidant activity can be obtained. In terms of improving physiological and antioxidant activity, biscozyme may be preferable among sugar hydrolase. Hydrolysis using such a sugar hydrolase may be performed in a buffer. Hydrolysis using a sugar hydrolase may vary in optimum conditions for each enzyme, but may be preferably performed at 50 ° C. to 60 ° C. for 3 to 12 hours in a buffer having a pH of 4.5 to 7.
Although there is no limitation as a buffer, for example, distilled water, acetate buffer or phosphate buffer may be used alone or in combination.
In addition, the step of treating the aloe gel using a sugar hydrolase is preferably performed in the ratio of substrate to enzyme in the range of 1: 100 to 1: 10,000 when considering the improvement of yield.
Filtration and lyophilization of the aloe gel after treatment with the sugar hydrolase may be used in the form of aloe gel powder.
The aloe sugar hydrolase extract prepared by the method according to the present invention was confirmed to have an excellent antioxidant effect through the scavenging measurement experiments such as DPPH radicals, hydroxyl radicals, hydrogen peroxide radicals and intracellular reactive oxygen scavenging activity experiments, etc. Could. Therefore, the composition containing the aloe sugar hydrolase extract of the present invention is useful for medicines, health supplements, cosmetics, food, food additives, etc. for the suppression or treatment of aging and various diseases caused by the oxides produced by active oxygen It may be used to, but is not limited thereto.
Hereinafter, the present invention will be described in detail with reference to Examples, but the present invention is not limited by these Examples.
Example 1
(1) Preparation of materials
Used aloe vera was purchased from Jeju Aloe and stored frozen at -40 ℃ until pulverized.
(2) washing, stripping and grinding of aloe
The aloe was washed and the shell was removed, and the aloe gel was crushed using a grinder and used.
(3) Enzyme Extraction and Filtration of Aloe Gel
Enzyme extraction using enzyme hydrolysis was used. That is, 100 ml of distilled water was added to 3 g of the lyophilized sample, and the enzymes were mixed with the aloe gel for 12 hours at a substrate-to-enzyme ratio of 1: 100. Enzymatic hydrolysis was carried out at the optimum temperature and pH of each enzyme for 12 hours, and the hydrolyzed samples were powdered by yield measurement and freeze-drying using a filter (Watman filter paper). In addition, the biscozyme enzyme having a high yield was selected and hydrolyzed to a substrate-to-enzyme ratio of 1: 1,000 to 1: 10,000, and the yield was measured. As a result, optimum hydrolysis conditions were set. When the hydrolysis was completed, the reaction was terminated by heating at 100 ℃, the reaction solution was filtered and lyophilized to prepare a hydrolysis extract. The composition and reaction conditions of the enzyme used at this time are shown in Table 1 below.
enzyme pH (buffer) Temperature (℃)

Party
Hydrolysis
enzyme
Viscozyme 4.5 (acetate buffer) 50
Celluclast 4.5 (acetate buffer) 50
AMG 4.5 (acetate buffer) 60
Termamyl 6.0 (phosphate buffer) 60
Ultraflo 7.0 (phosphate buffer) 60
Experimental Example 1 Measurement of Yield of Hydrolase Extract per Aloe Gel
Measurement of the yield of the sugar hydrolase extract of the aloe gel was obtained from the aloe gel hydrolyzate according to the preparation method according to the method of calculating the amount by weight.
As shown in FIG. 2, the results of extracting the aloe gel for each enzyme at a substrate-to-enzyme ratio of 1: 100 for 12 hours showed that the biscozyme enzyme showed the highest yield of 95.5%. As a result of using the biscozyme enzyme through the results of Figure 2 was able to obtain a high yield of the hydrolyzate. In Figure 3 was added to the substrate and the enzyme ratio 1: 1000 ~ 1: 10,000 in order to economically use the extraction for 12 hours, it can be seen that the yield of 69.3% even when added in a 1: 1,000 ratio.
Experimental Example 2 DPPH Radical Scavenging Activity of Aloe Sugar Hydrolase Extract
Free radicals are a major cause of oxidation in vivo, and DPPH (1,1-diphenyl-2-picrylhydrazyl) is widely used to evaluate the free radical scavenging activity of natural antioxidants. In the present invention, DPPH (1,1-diphenyl-2-picrylhydride) using an electron spin resonator (ESR spectrometer, JEX-PX 2000-300, JEOL, Japan) to confirm the antioxidant effect of the aloe sugar hydrolase extract Lazil) radical scavenging ability was measured. DPPH is a dark purple itself, a nitrogen-centered radical, which exists in a stabilized state by delocalization of radical electrons.
Aloe was extracted at 12 mg for 12 hours with the addition of an enzyme at 1: 100. When the extract was extracted with water, it showed 65.6% of activity at 2 mg / ml, but treated with biscozyme hydrolyzate. It was confirmed that the DPPH radical scavenging activity as high as 85.45% at 2 mg / ml, it was confirmed that the high scavenging activity and concentration-dependent activity in the aloe biscozyme hydrolyzate was increased. This is illustrated in FIG. 4.
The aloe water extract used as a control in this experiment and the following experimental examples was prepared by the following method; 100 ml of distilled water was added to 3 g of the lyophilized sample, and the aloe gel was extracted for 12 hours. The extracted sample was measured by a filter (Wattman filter paper), and the yield was measured. The reaction was terminated by heating under a condition heated at 100 ° C. The reaction solution was filtered and lyophilized to be used as a control.
Experimental Example 3 Hydroxyl Radical Scavenging Activity of Aloe Glycolytic Enzyme Extract
In order to confirm the antioxidant effect of the aloe sugar hydrolase extract according to the present invention, the hydroxyl radical scavenging ability was measured using an electron spin resonator (ESR spectrometer, JEX-PX 2000-300, JEOL, Japan). The hydroxyl radical scavenging ability is to generate hydroxyl radicals by the Fenton reaction in which iron (Fe) ions and hydrogen peroxide react to measure the amount of DMPO (5,5-dimethyl-1-pyrroline N-oxide) -OH. The hydroxyl radical scavenging rate is measured.
The results of measuring the hydroxyl radical scavenging activity are shown in FIG. 5. As a result, it was shown that 65.6% of the hydroxyl radical scavenging activity was obtained at 2 mg / ml when extracted for 12 hours by adding an enzyme at 1: 100 using water, but it was 78.7 at 2 mg / ml when extracted with biscozyme. It was confirmed that the hydroxyl radical scavenging activity was higher than that of the water extract.
Experimental Example 4 Hydorgen Peroxide Scavenging Activity of Aloe Hydrolyzate Extract
Hydrogen peroxide scavenging ability was measured using an electron spin resonance (ESR spectrometer, JEX-PX 2000-300, JEOL, Japan) to confirm the antioxidant effect of the aloe sugar hydrolase extract according to the present invention.
Hydrogen peroxide is a powerful oxidant that reacts with superoxide and is known to cause serious damage to the DNA of each organ in the body, causing disease and aging. The hydrogen peroxide scavenging activity was evaluated and the results are shown in FIG. 6. Aloe gel water extract showed 62.5% Hydrogen peroxide scavenging activity at 2 mg / ml, but high activity at 7 mg / ml of aloe biscozyme hydrolyzate was 72.3%.
Experimental Example 5 Intracellular Oxygen Scavenging Activity of Aloe Hydrolyzed Extract
When hydrogen peroxide (H 2 O 2 ) was treated to Vero cells, the free radicals were measured by extracting five kinds of sugar hydrolase and found to be similar to DPPH, hydroxyl radical scavenging activity and hydrogen peroxide. The highest scavenging activity of 83.1% was confirmed in the extract 2 mg / ml.

Claims (12)

  1. A method of producing an aloe sugar hydrolase extract comprising the step of treating the aloe gel using a sugar hydrolase.
  2. The method of claim 1,
    The step of treating the aloe gel using a sugar hydrolase is carried out at a reaction temperature of 50 to 60 ℃ and a reaction time of 3 to 12 hours in a buffer of pH 4.5 to 7.
  3. The method of claim 1,
    Treating the aloe gel with a sugar hydrolase, wherein the ratio of substrate to enzyme is performed in the range of 1: 100 to 1: 10,000.
  4. The method of claim 1,
    Glycolytic enzyme is one or a mixture thereof selected from Viscozyme, Celluclast, Amyloglucosidase (AMG), Termamyl and Ultraflo. Manufacturing method.
  5. The method of claim 1,
    Sugar hydrolase is a manufacturing method of Biscozyme (Viscozyme).
  6. The method of claim 1,
    Wherein the buffer is a single or a mixture selected from distilled water, acetate buffer and phosphate buffer.
  7. The method of claim 1,
    After the step of treating the aloe gel using a sugar hydrolase, the method comprising the step of filtration and lyophilization.
  8. Aloe sugar hydrolase extract prepared by the method of any one of claims 1 to 7.
  9. Antioxidant composition containing aloe sugar hydrolase extract as an active ingredient.
  10. The method of claim 9,
    Glycolytic enzyme is one or a mixture thereof selected from Viscozyme, Celluclast, Amyloglucosidase (AMG), Termamyl and Ultraflo. Antioxidant composition.

  11. The method of claim 9,
    Antioxidant composition wherein the sugar hydrolase is biscozyme.
  12. The method of claim 9,
    Antioxidant composition used as medicine, health supplement, cosmetic, food or food additive.
KR1020110127285A 2011-11-30 2011-11-30 Process for preparation of higher yield extract from aloe gel using enzyme and its antioxidant activity KR20130060954A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101502219B1 (en) * 2013-07-03 2015-03-13 제주알로에영농조합법인 A antioxidant composition containing enzyme extract or fractions of aloe

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101502219B1 (en) * 2013-07-03 2015-03-13 제주알로에영농조합법인 A antioxidant composition containing enzyme extract or fractions of aloe

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