KR101778752B1 - Method for preparation of extracts of corn silk comprising maysin using Pectinex - Google Patents

Abstract

The present invention relates to a method for preparing a corn bean extract having an increased content of maysin comprising the step of treating a corn beard extract with a pectinex enzyme. The corn bean extract prepared by the method of the present invention not only increases the content of meicin but also has an increased antioxidative and whitening effect.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a method for preparing a corn beard extract comprising Mechine using Pectinex enzyme,

The present invention relates to a method for preparing a corn beard extract in which the content of meicin is increased by using Pectinex enzyme. Further, the present invention relates to a composition for the manufacture of maize, a cosmetic composition for whitening, and a cosmetic composition for antioxidation, which comprises a corn mustache extract and the above enzyme.

Corn is a one-year-old herb with Poipu, which is cultivated all over the world, including North America. It is native to the tropical Americas and grows in sizes of 1 to 3 meters. The corn beard, which is a flower of corn, is known as a jade bean with its generic name, and is sometimes called 'jade teddy bean' or 'jade'. The taste is sweet and refreshing. It has excellent pharmacological effect and is widely used for medicinal purposes. It is widely used as food, food additives, and fragrance.

In particular, corn beard has been reported to be effective for hypertension, hyperglycemia, hematochezia, hemostasis and the like, and has been reported to have diuretic effect, diabetic inhibitory effect, COX-2 inhibitory effect, liver protective effect and the like.

These corn beets are known to contain various flavonoids and anthocyanins. Recently, attention has been paid to the antioxidative effect of maysin, a kind of florobonoid derived from corn beard, (Korean Patent Publication No. 10-2015-0057084).

However, in the case of corn hot-water extraction for conventional macrocein extraction, there is a problem that the antioxidative effect is remarkably deteriorated due to deformation and destruction of mexin contained in the extract.

Under these circumstances, the present inventors have made intensive efforts to produce a corn extract having enhanced physiological activity and an increased content of meicin. As a result, it has been found that by treating Pectinex Ultra SP-L enzyme with cornstarch extract, It was confirmed that the content of meicin can be remarkably increased and the physiological activity can be enhanced, and the present invention has been completed.

It is an object of the present invention to provide a method for producing corn musth extract having an increased content of maysin comprising treating corn beard extract with Pectinex enzyme.

It is another object of the present invention to provide a composition for preparing maysin comprising corn musth extract and Pectinex Ultra SP-L enzyme.

It is another object of the present invention to provide a method for producing maysin comprising the step of treating the corn musth extract with Pectinex Ultra SP-L enzyme.

It is another object of the present invention to provide a whitening cosmetic composition comprising corn musth extract and Pectinex Ultra SP-L enzyme.

It is another object of the present invention to provide an antioxidant cosmetic composition comprising a corn musth extract and a Pectinex Ultra SP-L enzyme.

One aspect of the present invention provides a method for producing corn musth extract having an increased content of maysin comprising treating a corn beard extract with a Pectinex enzyme.

Specifically, the corn musth extract may be extracted with ethanol. More specifically, the extract may be extracted at 50 to 60 ° C for 2 to 3 hours at an ethanol concentration of 80 to 90%, but is not limited thereto.

The term " extract " of the present invention means an extract obtained from a corn using a solvent. In addition, in the present invention, the above-mentioned extract includes any of the extract obtained by the extraction treatment, the diluted or concentrated liquid of the extract, the dried product obtained by drying the extract, or any of these adjusted products or purified products. The solvent is not particularly limited, and any solvent known in the art can be used. Examples of the extraction solvent include water, distilled water, alcohol, or a mixed solvent thereof. When the alcohol is used as a solvent, it is more preferable to use a C ₁ to C ₄ alcohol such as methanol, ethanol, propanol , Butanol) can be used, but specifically ethanol may be used.

In addition, the extract may contain fractions.

The term of the present invention. &Quot; Fraction " means a result obtained by performing fractionation to separate a particular component or group of specific components from a mixture comprising various components. The fractionation method for obtaining the fraction in the present invention is not particularly limited and may be carried out according to a method commonly used in the art. As a non-limiting example of the above-mentioned fractionation method, there can be enumerated a method of treating the extract obtained by extracting corn with a predetermined solvent to obtain a fraction from the extract.

The kind of the fraction solvent used for obtaining the fraction in the present invention is not particularly limited and any solvent known in the art can be used. Non-limiting examples of the fraction solvent include polar solvents such as water, distilled water and alcohol; And non-polar solvents such as hexane, ethyl acetate, chloroform, and dichloromethane. These may be used alone or in combination of two or more. When alcohols are used in the fraction solvent, C ₁ to C ₄ alcohols can be preferably used.

In the above extract, corn can be used without limitation such as cultivated or commercially available.

In one embodiment of the present invention, corn musth extract was prepared under the conditions of an ethanol concentration of 83.1%, an extraction temperature of 52.7 캜, and an extraction time of 2.6 hours to confirm the effect of increasing the mechin content by pectinex enzyme treatment (Example 1) .

The term " Pectinex enzyme " of the present invention is an enzyme preparation comprising polygalacturonase, a pectinase from Aspergillus aculeatus .

Specifically, the pectinex enzyme may be Pectinex Ultra SP-L, but is not limited thereto.

The term " Pectinex ultra SP-L " of the present invention is a yellow liquid enzyme that simultaneously hydrolyzes pectin and also acts as a hydrolyzate of cellulose. However, the effect of increasing the content of maize bean, which is a type of flavonoid, which is not pectin or cellulose, by the treatment of the enzyme is not known. On the other hand, the corn extract prepared by the production method of the present invention has an effect of increasing the content of meicin and enhancing the physiological activity by the treatment with the Pectinex Ultra SP-L enzyme.

In one embodiment of the present invention, the corn extract treated with the Pectinex Ultra SP-L enzyme showed a maximum 234% increase in the total polyphenol content and a maximum 411% increase in the macron content, compared with the control without the enzyme treatment , A 132% improvement in DPPH radical scavenging ability, and a maximal inhibition effect of tyrosinase activity of 148% (Example 2).

The term " maysin " of the present invention is a type of flavonoid and is known to have an antioxidative effect. The corn extract prepared by the production method of the present invention not only increased the content of meicin but also had an effect of improving antioxidative and whitening effect by mexin.

In one embodiment of the present invention, optimized treatment conditions of the Pectinex Ultra SP-L were identified to produce a corn extract having an increased mexin content (Example 3).

Specifically, the step of treating the Pectinex enzyme with the corn musth extract may be treating Pectinex Ultra SP-L at a concentration of 1.00 to 2.00 mL / 100 L, and more specifically, the enzyme is treated with 1.00 To 2.00 mL / 100 L, at 20 to 40 < 0 > C for 5 to 8 hours.

In one embodiment of the present invention, it was confirmed that the optimum treatment conditions of the Pectinex Ultra SP-L were an enzyme concentration of 1.48 mL / 100 g, a reaction temperature of 21.52 ° C, and a reaction time of 7.78 hours through an experimental design using a central synthesis plan . The total polyphenol content, macroin content, DPPH radical scavenging activity and tyrosinase activity inhibitory effect of the corn extract prepared under the above conditions were 6956.41 mg / 100g, 4762.62 mg / 100g, 3208.73 mg / 100g, 88.43% and 74.86%, respectively Respectively. Overall desirability was 0.85 for each control (Example 4).

Another aspect of the present invention provides a composition for making maysin comprising a corn musth extract and a pectinex Ultra SP-L enzyme.

The term " corn musth extract ", " pectinex Ultra SP-L enzyme ", and " maysin "

The term " composition for the preparation of meicin " of the present invention includes corn beard extract and pectinex Ultra SP-L enzyme, and the enzyme can be treated with the extract to increase the content of meicin contained in corn musth extract. The composition may further comprise a composition capable of separating the macrocin, so that the macroin which is increased in content can be separated from the corn extract to produce the macrocin. The separation of meicase may be by a method that can be easily adapted by a person skilled in the art.

Another aspect of the present invention provides a method of preparing meiocin comprising treating corn musth extract with Pectinex Ultra SP-L enzyme.

The term " corn musth extract ", " pectinex Ultra SP-L enzyme ", and " maysin "

Specifically, the method for producing a macine of the present invention may further include a step of isolating macine by chromatography.

The term " separation of meisin " of the present invention can be any method that can be readily adopted by a person skilled in the art, but can be silica gel column chromatography, reverse phase column chromatography, high performance liquid chromatography and the like. The developing solvent for chromatography may be n-hexane, chloroform, methanol, water or a mixed solvent thereof, but is not limited thereto.

Another embodiment of the present invention provides a whitening cosmetic composition comprising a corn musth extract and a pectinex Ultra SP-L enzyme.

The term "corn musth extract" and "pectinex Ultra SP-L enzyme" of the present invention are the same as described above.

The term " whitening " of the present invention means any action that inhibits the synthesis of melanin to inhibit or prevent skin deposition of melanin. Specifically, it may inhibit the activity of tyrosinase involved in the production of melanin .

In one embodiment of the present invention, it was confirmed that the corn extract treated with Pectinex Ultra SP-L enzyme inhibited tyrosinase activity compared with the control without the enzyme.

The cosmetic composition of the present invention can be used as a cosmetic composition in the form of a solution, a topical ointment, a cream, a foam, a nutritional lotion, a softening lotion, a pack, a soft water, a latex, a makeup base, The formulation may be selected from the group consisting of liquids, pastes, gels, lotions, powders, soaps, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations, patches and sprays, no.

The cosmetic composition of the present invention may further comprise at least one cosmetically acceptable carrier to be incorporated in a cosmetic composition for general skin. Examples of the cosmetic composition include ordinary ingredients such as oil, water, a surfactant, a moisturizer, a lower alcohol, , A chelating agent, a coloring agent, a preservative, a perfume, and the like may be appropriately compounded, but the present invention is not limited thereto.

The cosmetically acceptable carrier contained in the cosmetic composition of the present invention varies depending on the formulation of the cosmetic composition.

When the formulations of the present invention are ointments, pastes, creams or gels, the carrier component may be an animal oil, a vegetable oil, a wax, a paraffin, a starch, a tracer, a cellulose derivative, polyethylene glycol, silicon, bentonite, silica, talc, zinc oxide May be used, but is not limited thereto. These may be used alone or in combination of two or more.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder and the like may be used as a carrier component. In particular, But are not limited to, propellants such as rocaborn, propane / butane or dimethyl ether. These may be used alone or in combination of two or more.

When the formulation of the present invention is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent may be used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, Propylene glycol, 1,3-butyl glycol oil and the like can be used, and particularly fatty acid esters of cottonseed oil, peanut oil, corn oil, olive oil, castor oil and sesame oil, glycerol aliphatic ester, polyethylene glycol or sorbitan May be used, but is not limited thereto. These may be used alone or in combination of two or more.

When the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspension such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Crystalline cellulose, aluminum metahydroxide, bentonite, agar or tracant, but are not limited thereto. These may be used alone or in combination of two or more.

When the formulation of the present invention is a soap, use is made of an alkali metal salt of a fatty acid, a fatty acid hemiester salt, a fatty acid protein hydrolizate, isethionate, a lanolin derivative, an aliphatic alcohol, a vegetable oil, glycerol, But is not limited thereto. These may be used alone or in combination of two or more.

Another aspect of the present invention provides an antioxidant cosmetic composition comprising a corn musth extract and a Pectinex Ultra SP-L enzyme.

The term " corn musth extract ", " pectinex Ultra SP-L enzyme ", and " maysin "

The term " antioxidant " of the present invention means, in a narrow range, free radicals produced in the body, hydrogen peroxide or peroxides generated from the free radicals, inhibition, reduction or control of the generation or reaction of hydroxy radicals produced from the hydrogen peroxide And a wide range means an action of inhibiting, reducing or controlling the generation of an oxidation reaction occurring in the natural world. Specifically, it means the antioxidative action of mexin contained in corn extract by DPPH radical scavenging ability.

In one embodiment of the present invention, the DDPH radical scavenging ability of the corn extract treated with the Pectinex Ultra SP-L enzyme was improved compared with the control without the enzyme.

The term " cosmetic composition " of the present invention is the same as described above.

 The present invention provides a corn extract having increased macine content by treating Pectinex Ultra SP-L enzyme with corn beard extract. The corn extract may have increased whitening and antioxidant effects by increasing the content of meicin.

Figure 1 shows the reaction surface contour of total phenol content (mg / 100g) by Pectinex Ultra SP-L enzyme treatment (reaction time, A: 2 hours, B: 5 hours, C: D: 20 DEG C, E: 40 DEG C, F: 60 DEG C, enzyme concentration, G: 0.5 mL / 100 L, H: 1 mL / 100 L, I: 1.5 mL / 100 L).
Figure 2 shows the reaction surface contour lines of the macroin content (mg / 100g) by Pectinex ultra SP-L enzyme treatment (reaction time, A: 2 hours, B: 5 hours, C: D: 20 DEG C, E: 40 DEG C, F: 60 DEG C, enzyme concentration, G: 0.5 mL / 100 L, H: 1 mL / 100 L, I: 1.5 mL / 100 L).
Figure 3 shows reaction surface contours of DPPH radical scavenging activity (%) by Pectinex Ultra SP-L enzyme treatment (reaction time, A: 2 hours, B: 5 hours, C: 8 hours, reaction temperature, D : 20 DEG C, E: 40 DEG C, F: 60 DEG C, enzyme concentration, G: 0.5 mL / 100 L, H: 1 mL / 100 L, I: 1.5 mL / 100 L).
Figure 4 shows reaction surface contours of the tyrosinase activity inhibition effect (%) by Pectinex Ultra SP-L enzyme treatment (reaction time, A: 2 hours, B: 5 hours, C: : 20 DEG C, E: 40 DEG C, F: 60 DEG C, enzyme concentration, G: 0.5 mL / 100 L, H: 1 mL / 100 L, I: 1.5 mL / 100 L).
Figure 5 shows the optimization of the multiply reactive surface of the Pectinex Ultra SP-L enzyme treatment of corn extract.

Hereinafter, the present invention will be described in more detail with reference to Examples. These embodiments are only for illustrating the present invention, and the scope of the present invention is not construed as being limited by these embodiments.

Example 1: Preparation of corn beard extract

The inventors of the present invention produced corn musth extract under conditions of an ethanol concentration of 83.1%, an extraction temperature of 52.7 ° C, and an extraction time of 2.6 hours. Using the Pectinex Ultra SP-L enzyme, Was developed.

The extract was concentrated under reduced pressure at 40 ° C using a vacuum rotary concentrator (BUCHI rotavapor R-124 and BUCHI water bath B-480, Flawil, Switzerland) to prepare an extract. Then, it was completely dissolved in lyophilized (Ilshin Biobase, KOREA) and 10% (w / v) ethanol at -50 ° C and used as a sample.

(9,500 pectinase unit / mL) of Novozym (Bagsvaerd, Denmark) and centrifuged at 4000 rpm for 10 minutes to obtain the supernatant. Then, it was filtered with filter paper (Whatman No. 2) and analyzed.

Example 2. Verification of Effect of Pectinex Enzyme Treatment

The enzyme concentration (0.5-1.5 mL / 100 L), the reaction temperature (20-60 ° C) and the reaction time (2 - 10 min) were examined to confirm the effect of the Pectinex Ultra SP- L of the corn bean extract prepared in Example 1, 8 hours) were analyzed for total polyphenol content, macine content, DPPH radical scavenging ability and tyrosinase inhibitory effect of corn beard extracts.

2-1. Total polyphenol content

The total polyphenol content of the sample was measured by modifying the Folin-Denis method. 3 mL of the Folin &Ciocalteau's phenol reagent diluted 1:10 with distilled water and 2.5 mL of 0.2% Na ₂ CO ₃ solution were added to 0.5 mL of the extracted sample solution, and the mixture was stirred at 25 ° C. for 30 minutes, The absorbance was measured.

The total phenol content of the corn bean extracts prepared in the above examples was expressed as the amount (㎍) of gallic acid corresponding to 1 g of corn beard by preparing a standard calibration curve.

As a result, the total polyphenol content in the enzyme-free treatment was 3120.49 mg / 100 g, and the content of the enzyme at the experimental point of interest was 6279.9-7308.9 mg / 100 g. From this, it was found that the treatment of Pectinex Ultra SP-L with corn beard extract was effective in increasing polyphenol content.

Especially, the increase of total polyphenol content by 234% was observed at the enzyme concentration of 1.0 mL / 100 L, reaction temperature of 20 ℃ and reaction time of 8 hours.

2-2. Maysin content

The macronutrient content of the sample was analyzed by HPLC and the retention time was compared using Meishin standard material (RDA). The value calculated by the peak area of the standard sample was calculated, Respectively. The HPLC analysis conditions for analyzing meiosis are shown in Table 1.

device YL9100 HPLC column Agilent 5 TC-C18 (2) 250 x 4.6 mm Column temperature 30 ℃ Mobile phase A - 10% ethanol + 0.1% phosphoricacid
B - 90% ethanol + 0.1% phosphoricacid Gradient conditions H ₂ O / ethanol linear gradient
(from 20% ethanol to 80% ethanol to 35minutes Flow rate 1.0 mL / min. Injection capacity 20 쨉 l Detector wavelength 340 nm

As a result, the content of meiocin in enzyme - free treatment was 800.67 mg / 100g, while the range of enzyme treatment in experimental area of interest was 2645.98-3288.05 mg / 100g. From these results, it was confirmed that the treatment of Pectinex Ultra SP-L with corn beard extract was effective in increasing the macro-nutritional content.

Especially, increase of macine content of 411% compared to the control was observed under the conditions of enzyme concentration 1.0 mL / 100 L, reaction temperature 20 ° C. and reaction time 8 hours

2-3. DPPH radical scavenging ability

The antioxidant activity of corn extracts was determined by the reduction of 2,2-diphenyl-1-picrylhydrazyl (DPPH) to determine the antioxidant properties and the ability of each sample Free radical scavenging ability.

0.2 mL of 0.2 mM DPPH ethanol solution was added to 0.2 mL of the corn horn extract sample solution, which was reacted at room temperature for 30 minutes, and the absorbance was measured at 517 nm. The control was performed in the same manner using distilled water instead of the sample, and the free radical scavenging ability (%) for each sample was calculated using the following equation.

Free radical scavenging ability (%) = (1-A / B) x 100

A: Absorbance of the sample, B: Absorbance of the control

As a result, it was confirmed that the scavenging ability in the enzyme - free treatment was 66.96%, while the range in the experimental point of the enzyme treatment was 87.63-88.61%.

From this, it was confirmed that the treatment of Pectinex Ultra SP-L with corn beard extract was effective in improving the free radical scavenging ability.

Especially, it was confirmed that the DPPH radical scavenging ability of 132% was increased at the enzyme concentration of 0.5 mL / 100 L, the reaction temperature of 40 ° C. and the reaction time of 2 hours.

2-4. Inhibition of tyrosinase activity

The inhibitory effect of tyrosinase, an enzyme involved in the production of melanin pigment, was examined to confirm the whitening effect of the enzyme - treated corn extract.

Specifically, 0.4 mL of a 2 mM L-tyrosine solution and 0.2 mL of a sample extract were mixed with 2.3 mL of potassium phosphate buffer (pH 6.5), and 0.1 mL of mushroom tyrosinase (220 unit / mL) was added After reacting in a 37 ° C water bath for 20 minutes, the absorbance was measured at 470 nm using a UV-visible spectrophotometer (A).

In order to measure the inhibitory effect of tyrosinase activity, 0.1 ml of distilled water was added instead of the enzyme solution, and the value (B) obtained by measuring the absorbance and 0.2 ml of distilled water instead of the extracted sample solution were used. .

Activity inhibition effect (%) = [1- (A-B) / C] 100

As a result, it was confirmed that the tyrosinase activity inhibition effect was 52.90% in the enzyme-free treatment, while the range in the experimental point of the enzyme treatment was 61.06-78.32%.

From this, it was confirmed that the treatment of Pectinex Ultra SP-L with corn beard extract was effective for inhibiting tyrosinase activity.

In particular, tyrosinase inhibitory activity was improved by 148% compared to the control at an enzyme concentration of 0.5 mL / 100 L, a reaction temperature of 40 ° C and a reaction time of 2 hours.

The results of the total polyphenol content, macrocyclic content, DDPH radical scavenging ability and tyrosinase activity inhibitory effect are shown in Table 2.

Experiment
number X1 ¹⁾ X2 ²⁾ X3 ³⁾ Total polyphenol content
(mg / 100g) Macine content
(mg / 100g) DPPH radical scavenging ability
(%) Inhibition of tyrosinase activity
(%) One 0 -One One 7308.9 ± 31.8 ^{4) a5)} 3288.05 88.45 0.07 ^ab 76.11 ± 2.50 ^ab 2 0 One -One 7052.4 ± 25.5 ^b 3043.21 88.28 0.03 ^abcd 72.57 ± 3.75 ^ab 3 0 0 0 6542.4 ± 17.0 ^cd 3122.40 87.99 + 0.17 ^bcde 72.12 + - 4.38 ^b 4 0 -One -One 6588.9 ± 31.8 ^c 3265.33 88.45 ± 0.20 ^ab 76.11 + 1.25 ^ab 5 -One 0 One 6492.9 + -53.0 ^def 2982.55 88.25 + - 0.14 ^abcd 72.57 ± 1.25 ^ab 6 -One -One 0 6483.9 ± 36.1 ^ef 3172.51 88.59 ± 0.27 ^a 72.12 ± 1.88 ^b 7 -One 0 -One 6588.9 + - 6.4 ^c 3090.80 88.61 + 0.03 ^a 78.32 ± 1.88 ^a 8 0 0 0 6503.4 ± 29.7 ^def 3069.58 88.49 + 0.14 ^a 72.57 ± 1.25 ^ab 9 One One 0 6479.4 ± 12.7 ^ef 2851.94 87.63 ± 0.07 ^e 72.57 ± 0.00 ^ab 10 0 0 0 6510.9 ± 6.4 ^de 3088.80 88.33 0.30 ^abc 71.68 ± 0.00 ^b 11 One 0 -One 6279.9 ± 2.1 ^h 3071.35 88.33 + 0.03 ^abc 73.45 ± 2.50 ^ab 12 One 0 One 6479.4 ± 17.0 ^ef 3010.99 88.21 ± 0.20 ^abcd 70.80 ± 1.25 ^b 13 One -One 0 6417.9 ± 14.8 ^g 3139.92 88.37 ± 0.44 ^abc 73.45 ± 5.01 ^ab 14 0 One One 6449.4 0.0 ^fg 2645.98 87.92 ± 0.20 ^cde 61.06 + - 2.50 ^c 15 -One One 0 6507.9 ± 10.6 ^de 2809.22 87.83 ± 0.14 ^de 65.49 ± 0.00 ^c

2-5. Statistical processing

Statistical analysis was performed using Duncan's multiple range test at p <0.05 after analysis of variance (ANOVA) using SAS 9.3 (SAS Institute Inc., Cary, NC, USA) .

Example 3 Experimental Design of Optimal Enzyme Treatment Process

Experimental designs for the optimal enzymatic treatment process used a central composite design and the optimal range of activity in the manual of Pectinex Ultra SP-L was determined as the interesting area. Three factors for the Pectinex Ultra SP-L were enzyme concentration (X ₁ ) 0.5-1.5 mL / 100 L, reaction temperature (X ₂ ) 20-60 ° C, reaction time (X ₃ ) 2-8 hours Interest areas are defined as shown in Table 3.

Factors Symbol Code value -One 0 One Enzyme concentration
(mL / 100 L) X1 0.5 One 1.5 Reaction temperature (캜) X2 20 40 60 Reaction time (hours) X3 2 5 8

Each independent variable was coded to have a code value in the range of -1 to 1 and was set to 15 intervals according to the central synthesis plan (Table 4).

Experiment
number X1 X2 X3 Enzyme concentration
(mL / 100 L) Reaction temperature
(° C) Reaction time
(time) One 0 -One One One 20 8 2 0 One -One One 60 2 3 0 0 0 One 40 5 4 0 -One -One One 20 2 5 -One 0 One 0.5 40 8 6 -One -One 0 0.5 20 5 7 -One 0 -One 0.5 40 2 8 0 0 0 One 40 5 9 One One 0 1.5 60 5 10 0 0 0 One 40 5 11 One 0 -One 1.5 40 2 12 One 0 One 1.5 40 8 13 One -One 0 1.5 20 5 14 0 One One One 60 8 15 -One One 0 0.5 60 5

The secondary polynomial regression curve for the reaction surface analysis in the present invention is as follows.

Here, X is an independent variable, X ₁ (enzyme concentration), X ₂ (reaction temperature) and X ₃ (reaction time), Y is a dependent variable (Y _n ), total polyphenol content (Y ₁ ), total flavonoid content (Y ₂ ), mechine content (Y ₃ ), DPPH radical scavenging ability (Y ₄ ), and tyrosinase inhibitory effect (Y ₅ ), and A i is a regression coefficient.

For the optimization of the enzyme treatment process, the response surface methodology was used. For the optimization of the ethanol extraction process according to the level of each factor, statistical methods such as analysis of variance, quadratic polynomial regression and response surface method were used. (SAS Institute Inc., Cary, NC, USA).

Example 4: Determination of optimal conditions for enzyme treatment

Derivation of a quadratic polynomial regression curve

The results of the analysis of the degree of the total polyphenol content, the macroin content, DPPH radical scavenging ability and tyrosinase inhibition effect of the corn beard extract according to the enzyme concentration, reaction temperature and reaction time in the interest zone set in Example 3 were analyzed Table 5 shows the results.

factor Total polyphenol content Total flavonoid content Macine content DPPH radical scavenging ability Inhibition of tyrosinase activity X1 ¹⁾ F value 26.93 2.74 6.36 0.91 0.83 Probe> F <.0001 0.0577 0.0018 0.4758 0.521 X2 ²⁾ F value 68.5 2.73 136.44 6.85 6.88 Probe> F <.0001 0.0581 <.0001 0.0012 0.0012 X3 ³⁾ F value 68.61 4.60 32.95 1.88 5.00 Probe> F <.0001 0.0085 <.0001 0.1536 0.0059 ¹⁾ Enzyme concentration (mL / 100 L)
²⁾ Reaction temperature (캜)
³⁾ Reaction time (time)

As can be seen from the above Table 5, the factors affecting the total polyphenol content were influenced by reaction time, reaction temperature, enzyme concentration, and all three factors showed significant difference within 5%.

The reaction temperature was the most influential factor for the macine content, and the reaction time and enzyme concentration were found to be in the order of. All three factors showed significant differences within 5%.

The DPPH radical scavenging activity was found to be affected by the reaction temperature, reaction time and enzyme concentration.

In addition, the effect of tyrosinase inhibition was confirmed in the order of reaction temperature, reaction time, and enzyme concentration. Significant differences were found in reaction temperature and reaction time within 5%.

Furthermore, the results of calculating the regression coefficients of the secondary polynomial regression curves to predict the total polyphenol content, macine content, DPPH radical scavenging ability, and tyrosinase inhibiting effect, which are dependent variables on the levels of the processing factors, are shown in Table 6 .

Parameter Estimate Total phenol content Total flavonoid content Macine content DPPH radical scavenging ability Inhibition of tyrosinase activity Intercept 6092.643 4599.794 2962.769 88.796 84.493 X1 ¹⁾ 1357,000 -94.270 377.225 -0.073 -17.049 X2 ²⁾ -9.616 0.395 5.242 0.017 0.168 X3 ³⁾ 3.215 -8.733 32.394 -0.152 -1.311 X1 * X1 -872.500 -34.718 -243.832 -0.173 4.576 X2 * X1 0.938 -1.960 1.883 0.001 0.144 X2 * X2 0.429 0.094 -0.098 0.000 -0.003 X3 * X1 49.250 53.624 7.982 0.040 0.517 X3 * X2 -5.513 -1,238 -1.750 -0.001 -0.048 X3 * X3 17.722 0.783 0.699 0.014 0.188
¹⁾ Enzyme concentration (mL / 100 L)
²⁾ Reaction temperature (캜)
³⁾ Reaction time (time)

The contribution of primary polynomial regression, quadratic regression, and cross product in the quadratic polynomial regression curves for total polyphenol content, macine content, DPPH radical scavenging ability, and tyrosinase inhibition Respectively.

As a result, as shown in Table 7, the contribution of the total polyphenol content was in the order of crossing regression, quadratic polynomial regression, and first order polynomial regression, and the contribution of meiocin content was in the order of primary polynomial regression, crossed regression, and secondary polynomial regression . The contribution of DPPH radical scavenging was in the order of primary polynomial regression, secondary polynomial regression, and cross regression. The contribution of tyrosinase inhibitory effect was in the order of primary polynomial regression, cross regression, and secondary polynomial regression. The total regression in the secondary polynomial regression curve fit test showed significant differences within 5% for all dependent variables.

Regressions Primary polynomial regression Secondary polynomial regression Crossrelation Total regress Total phenol content R ² 0.0421 0.4292 0.4856 0.9569 F value 6.52 66.44 75.18 49.38 Pr> F 0.003 <.0001 <.0001 <.0001 Total flavonoid content R ² 0.0202 0.0528 0.4471 0.5202 F value 0.28 0.73 6.21 2.41 Pr> F 0.8383 0.5439 0.0037 0.0487 Macine content

R ² 0.8074 0.0469 0.1148 0.9691 F value 174.25 10.13 24.78 69.72 Pr> F <.0001 0.0003 <.0001 <.0001 DPPH radical scavenging ability

R ² 0.5291 0.0885 0.0321 0.6496 F value 10.07 1.68 0.61 4.12 Pr> F 0.0003 0.2026 0.6155 0.004 Inhibition of tyrosinase activity

R ² 0.4563 0.0728 0.1479 0.677 F value 9.42 1.50 3.05 4.66 Pr> F 0.0004 0.2446 0.0522 0.002

4-2. Response surface analysis

The results of the analysis of the concentration of Pectinex Ultra SP-L, the reaction temperature, and the reaction time on the total polyphenol content in the corn beard extract were analyzed by the reaction surface analysis method as shown in FIG.

From the change in total polyphenol contents (A to C in FIG. 1) according to the enzyme concentration and the reaction temperature, it was confirmed that the reaction temperature had a greater effect on the total polyphenol content change than the enzyme concentration.

From the change of total polyphenol contents (D to F in FIG. 1) with enzyme concentration and reaction time, it was confirmed that the reaction time had a greater effect on the total polyphenol content change than the enzyme concentration.

Furthermore, from the change in total polyphenol content (G to I in FIG. 1) with the reaction temperature and the reaction time, it was confirmed that the reaction temperature and the reaction time had similar effects on the total polyphenol content change, The higher the total polyphenol content was.

The results of the analysis of the concentration of Pectinex Ultra SP-L, the reaction temperature, and the reaction time on the content of mexin contained in the corn mustache extract were analyzed by the reaction surface analysis method as shown in FIG.

From the changes in the macine content (A to C in FIG. 2) depending on the enzyme concentration and the reaction temperature, it was confirmed that the reaction temperature had a greater effect on the change of the macroin content than the enzyme concentration and the reaction temperature was low regardless of the reaction time And it was confirmed that the content of meiocin was higher.

In addition, it was confirmed that the reaction time had a greater effect on the change of the macine content than the enzyme concentration from the change of the macine content according to the enzyme concentration and the reaction time (FIG. 2 D to F) Was 0.10 mL / L, and the shorter the reaction time (60 min), the higher the macroin content.

Furthermore, it was confirmed that the reaction temperature had a greater effect on the change of the macro-chemical content than the reaction time, regardless of the enzyme concentration, according to the change of the macro-content according to the reaction temperature and the reaction time (G to I in FIG. 2). Regardless of the enzyme concentration, the lower the reaction temperature, the higher the macroin content.

The concentration, reaction temperature, and reaction time of Pectinex Ultra SP-L on the DPPH radical scavenging ability of the corn mustard extract were analyzed by the reaction surface analysis method as shown in FIG.

From the changes in DPPH radical scavenging activity according to the enzyme concentration and reaction temperature shown in FIGS. 3A to 3C, it was confirmed that the reaction temperature greatly affected DPPH radical scavenging ability compared to the enzyme concentration regardless of the reaction time, Regardless of enzyme concentration and reaction temperature, DPPH radical scavenging activity was higher.

From the changes of DPPH radical scavenging activity according to the enzyme concentration and the reaction time shown in D to F of FIG. 3, it was confirmed that the reaction time greatly affected the DPPH radical scavenging ability compared to the enzyme concentration regardless of the reaction temperature. The shorter the reaction time, the higher the DPPH radical scavenging activity.

From the changes in DPPH radical scavenging activity according to the reaction temperature and reaction time shown in G to I of FIG. 3, it was confirmed that the reaction temperature greatly affected the DPPH radical scavenging ability change, regardless of the enzyme concentration, The DPPH radical scavenging ability was higher.

The results of analyzing the concentration, reaction temperature, and reaction time of Pectinex Ultra SP-L on the inhibition effect of tyrosinase of corn mustache extract by the reaction surface analysis method are shown in FIG.

From the inhibitory effect of tyrosinase activity according to the enzyme concentration and reaction temperature shown in Figs. 4A to 4C, it was confirmed that the reaction temperature greatly affected the tyrosinase activity inhibition effect compared to the enzyme concentration regardless of the reaction time. Time reaction time (B and C in FIG. 4), it was confirmed that the tyrosinase activity inhibiting effect was high near the reaction temperature of 20 ° C.

From the inhibition effect of tyrosinase activity according to the enzyme concentration and reaction time shown in D to F of FIG. 4, it was confirmed that the reaction time greatly influences the tyrosinase activity inhibition effect compared to the enzyme concentration regardless of the reaction temperature. In addition, it was confirmed that the shorter the reaction time regardless of the reaction temperature, the higher the inhibitory effect of tyrosinase activity.

From the inhibition effect of tyrosinase activity according to the reaction temperature and the reaction time shown in G to I of FIG. 4, it was confirmed that regardless of the enzyme concentration, the reaction temperature had a significant effect on the tyrosinase inhibitory effect as compared with the reaction time.

4-3. Determination of optimal treatment conditions for Pectinex Ultra SP-L enzyme

The total polyphenol content, macine content, DPPH radical scavenging ability and tyrosinase inhibitory effect of corn beard extracts according to the enzyme concentration, reaction temperature, and reaction time of Pectinex Ultra SP-L were measured as satisfaction function the desirability function was used to determine the weight of the dependent variable to 1 (Figure 5).

As a result, optimum treatment conditions of the Pectinex Ultra SP-L enzyme were an enzyme concentration of 1.48 mL / 100 L, a reaction temperature of 21.52 ° C, and a reaction time of 7.78 hours. The total polyphenol content of the corn extract prepared under the above conditions, DPPH radical scavenging activity and tyrosinase activity inhibitory effect were found to be 6956.41 mg / 100g, 4762.62 mg / 100g, 3208.73 mg / 100g, 88.43% and 74.86%, respectively. Overall desirability was 0.85 for the control group.

From the above description, it will be understood by those skilled in the art that the present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. In this regard, it should be understood that the above-described embodiments are to be considered in all respects as illustrative and not restrictive. The scope of the present invention should be construed as being included in the scope of the present invention without departing from the scope of the present invention as defined by the appended claims.

Claims (11)

CLAIMS What is claimed is: 1. A method for producing corn musth extract having an increased content of maysin, comprising the step of treating corn mustache extract with Pectinex Ultra SP-L enzyme,
Wherein the corn musth extract is extracted at a concentration of 80 to 90% ethanol and at 50 to 60 DEG C for 2 to 3 hours.
delete delete delete 2. The method of claim 1, wherein the step of treating the enzyme comprises treating the enzyme with a concentration of 1.00 to 2.00 mL / 100 L.
6. The method according to claim 5, wherein the step of treating the enzyme is carried out at 20 to 40 DEG C for 5 to 8 hours.
A composition for making maysin comprising corn beard extract and Pectinex Ultra SP-L enzyme,
Wherein the corn musth extract is extracted at a concentration of 80 to 90% ethanol and at 50 to 60 DEG C for 2 to 3 hours.
CLAIMS What is claimed is: 1. A method for the production of maysin comprising the step of treating the corn mustache extract with Pectinex Ultra SP-L enzyme,
Wherein the corn musth extract is extracted at a concentration of 80 to 90% ethanol at 50 to 60 DEG C for 2 to 3 hours.
9. The method of claim 8, further comprising separating the meicin by chromatography.
delete delete

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