KR20180055339A - Method of preparing enzyme treated silkworm extract having high bioactive substances, the silkworm extract obtained thereby, and use thereof - Google Patents

Abstract

The present invention relates to a method for producing a silkworm enzyme hydrolysis extract with an enhanced antioxidant activity, a silkworm enzyme hydrolysis extract obtained therefrom, and a use thereof. The method of the present invention comprises: a first step of producing silkworm powder by drying silkworm; a second step of adding water to the silkworm powder of the first step, suspending the silkworm powder, and adding an acidic solution or a basic solution to prepare a silkworm powder having suitable pH; and a third step of adding a protease composed of any one or more of pancreatin or protease to the silkworm powder suspension of the second step to perform hydrolysis, thereby obtaining a silkworm enzyme hydrolysis extract having an enhanced antioxidant activity compared to when another protease is applied. According to the present invention, provided are a method for producing a silkworm enzyme hydrolysis extract, which improves an antioxidant activity by optimizing treatment conditions for improving the yield of an enzyme hydrolysis extract of a silkworm, and a silkworm enzyme hydrolysis extract obtained therefrom to have an enhanced antioxidant activity.

Description

[0001] The present invention relates to a method for producing a silkworm enzyme hydrolysis extract having enhanced antioxidation ability, a silkworm enzyme hydrolysis extract obtained therefrom, and a use thereof,

The present invention relates to a method for producing a hydrolysing enzyme of silkworm silkworm increased hydrolysis activity of silkworm silkworm, and more particularly, to a method for producing silkworm silkworm silkworm silkworm silkworm hydrolysis, And a method for producing the hydrolyzed silkworm enzyme hydrolyzate. The present invention also relates to a hydrolyzed silkworm silkworm enzyme hydrolyzate obtained by the hydrolysis of silkworm silkworm noodles.

Silkworm, which is known as a high quality and high protein source, is rich in various amino acids and has a low content of saturated fatty acids. Treatment of such highly functional silkworms with enzymes will induce the low molecular weight of various kinds of physiologically active substances and will be used as environment - friendly new processing technology.

In other words, the silkworm is one of the highly interested materials, which is currently being sold as a capsule or a product in the form of a dry powder or a hydrothermal treatment, as the preventive and therapeutic effects of adult diseases are known.

Indeed, since 1998, interest in the food production of the by-products of mulberry has begun to accumulate, and evaluation data on its physiological activity have been accumulating in the order of silkworm, mulberry leaf, and oder. Conventional studies have reported that silkworms have many physiological activity effects such as blood glucose lowering effect, constipation improvement effect, and liver toxicity recovery effect.

According to Dongbu Bohak or Basho Gangmok, silkworm has a special effect on the management of diabetes and senile dementia and strengthening liver function. In addition to the caterpillar, silkworm, cocoons, and pupa, each has its own active ingredient, and it does not even throw away the excreta.

Various techniques have been developed for the utilization of silkworms by paying attention to the value of silkworms. However, conventional techniques are generally extraction by high temperature pressurization treatment, freeze-dried powder or ring product, and products obtained by extracting some hot water extracts are sold There is a need for a more effective treatment method for bioactivity.

Accordingly, related prior arts have been known as follows.

Korean Patent Registration No. 10-1115510 (registered on Feb. 6, 2012) is a method of decomposing and extracting enzymes from silkworms, mulberry leaves, mulberry leaves, mulberry roots, . Korean Patent Registration No. 10-721644 (registered on May 17, 2007) discloses a health functional food in which silkworm powder is extracted with ethanol and hot water to improve hangover resolution activity. Korean Patent Registration No. 10-361085 (registered on Nov. 1, 2002) discloses an anti-diabetic health supplement composition containing an ethanol extract of silkworm powder.

Thus far, a wide variety of methods have been disclosed, including a fermentation method using a silkworm, enzymatic decomposition, or solvent extraction. The physiologically active substances of cocoons obtained by these methods are also very diverse, ranging from hyperlipidemia and fatty liver, antioxidant efficacy, thrombolytic efficacy, hangover resolution and diabetes treatment.

Nevertheless, there is still a need in the art for a method of obtaining a silkworm hydrolysis extract having a high active ingredient content by eliminating the problems of the prior art as described above and eliminating unnecessary steps, Improvement is still needed.

It is an object of the present invention to provide a method for producing a silkworm enzyme hydrolysis extract having improved antioxidant ability by optimizing treatment conditions for improving the yield of the enzyme hydrolysis extract of silkworm silkworm. The silkworm enzyme hydrolysis extract obtained through the above- An antioxidant activity and tyrosinase activity, and to provide a health food and a functional cosmetic comprising the same.

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are not intended to limit the invention to the particular embodiments that are described. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are not restrictive of the invention, There will be.

In order to accomplish the above object, the present invention provides a method for producing an enzymatic hydrolysis extract of a silkworm enzyme having enhanced antioxidant activity, comprising the steps of: preparing a silkworm powder by drying silkworm; adding water to the silkworm powder of the first step, A second step of suspending powder and then adding an acidic solution or a basic solution to prepare a powder suspension in an eye having a pH adjusted; and a second step of preparing a suspension of the silkworm powder of the second step, either pancreatin or protease And a third step of obtaining a silkworm enzyme hydrolysis extract having an enhanced antioxidative activity against the application of another protease by adding one or more proteolytic enzymes to hydrolyze the enzyme.

The proteinase of the third step is added in an amount of 0.1 to 10% by weight based on the weight of the silkworm powder.

The hydrolysates of hydrolysates of the silkworm enzyme of the present invention enhanced by the hydrolysis of silkworm silkworm hydrolyzate of the present invention were prepared by the above-mentioned method and enhanced by the antioxidant activity of the other proteolytic enzymes. The silkworm hydrolysis extract of the silkworm silkworm was obtained by filtration or centrifugation after filtration It is characterized by being in the form of a liquid, or a dry powder, and has a whitening effect.

The health food or functional cosmetic having the enhanced antioxidative and whitening effect of the present invention is characterized by containing the hydrolyzed extract of the silkworm enzyme as an active ingredient.

According to the present invention, there is provided a method for producing a silkworm enzyme hydrolysis extract having enhanced antioxidant ability by optimizing treatment conditions for improving the yield of the enzyme hydrolysis extract of silkworm silkworm, wherein the silkworm enzyme hydrolysis extract obtained through the above- It is possible to provide healthy foods and functional cosmetics containing the antioxidant activity and tyrosinase activity, thereby contributing to the increase of income of the farmers and processing companies and the health promotion of the people.

Fig. 1 is a diagram showing the color change of the enzyme hydrolysis extract of silkworm by enzyme treatment.
FIG. 2 is a view showing the extraction yield of the silkworm enzyme hydrolysis extract according to the pH condition of the enzyme treatment. FIG.
: Collected silk powder on the vertical axis is the remaining material after enzyme extraction
Fig. 3 is a diagram showing the extraction yield of the silkworm enzyme hydrolysis extract according to the enzyme treatment temperature condition.
: Collected silk powder on the vertical axis is the remaining material after enzyme extraction
FIG. 4 is a graph showing the reducing sugar content of the silkworm enzyme hydrolysis extract according to the pH condition of the enzyme treatment. FIG.
FIG. 5 is a graph showing the reducing sugar content of the silkworm enzyme hydrolysis extract according to the enzyme treatment temperature condition. FIG.
FIG. 6 is a diagram showing the protein degradation pattern of the silkworm enzyme hydrolysis extract according to the pH condition of the enzyme treatment. FIG.
Fig. 7 is a diagram showing the protein degradation pattern of the silkworm enzyme hydrolysis extract according to the enzyme treatment temperature condition.
8 shows the antioxidative activity of the silkworm enzyme hydrolysis extract according to the pH condition of the enzyme treatment using the ABTS method.
9 shows the antioxidative activity of the silkworm enzyme hydrolysis extract according to the enzyme treatment temperature condition using the ABTS method.
FIG. 10 is a graph showing tyrosinase inhibitory activity of the silkworm enzyme hydrolysis extract according to the pH condition of the enzyme treatment. FIG.
FIG. 11 is a diagram showing the tyrosinase inhibitory activity of the silkworm enzyme hydrolysis extract according to the enzyme treatment temperature condition. FIG.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, the present invention will be described in detail, and a detailed description of known functions and configurations that may unnecessarily obscure the gist of the present invention will be omitted.

The present invention relates to a method for producing a hydrolysing enzyme of silkworm silkworm increased hydrolysis activity of silkworm silkworm, and more particularly, to a method for producing silkworm silkworm silkworm silkworm silkworm hydrolysis, To enhance the antioxidant ability and to produce a silkworm enzymatic hydrolysis extract having a whitening effect.

The protease to be used for this purpose is at least one of pancreatin and protease, and it can be used as lipase, protease, which is applied to the enzyme hydrolyzate using silkworm, The present invention relates to the production of a silkworm enzyme hydrolysis extract which can be used for the development of high value-added products and products with improved functionality, which exhibits more enhanced antioxidative and whitening effects than the application of cellulase and the like, Each step will be described in detail.

Step 1) Production of silkworm powder

In this step, the silkworm is dried to produce a silkworm powder. First, a silkworm silkworm containing 75% of two silk fibroins and the rest consisting of sericin is prepared.

At this time, the silkworm species Bombix mori Although all species considered silkworms capable of obtaining silk fibroin can be used, among them, the most excellent cell proliferation effect is used, and the 5th-3rd day silkworm . This means that it is a F1 (hybridized generation) made by mating 125 kinds of sleep and 140 kinds of sleep.

In addition, any of the above-mentioned drying methods may be applied, but it is preferable to apply the freeze-drying method in order to minimize changes in the component of the silkworm. Freeze-drying is a method in which a sample containing an aqueous solution or water is frozen and reduced in pressure to a water vapor pressure or lower at that temperature to sublimate the water. The temperature and time range may be selected from conventional freeze-drying conditions, Is dried in a freeze dryer at 60 ° C to 80 ° C for 60 to 80 hours.

Step 2) Production of silkworm powder suspension

In this step, water is added to the silkworm powder of the first step to suspend the silkworm powder, and then an acidic or basic solution is added thereto to prepare a powder suspension .

At this time, the water is preferably added in an amount of 5 to 20 ml per 1 g of the silkworm powder. If the amount of water added is less than 5 ml, the content of the water is too small to form a liquid state. In this case, the enzyme reaction is not properly performed. When the water is added in an amount exceeding 20 ml, There is a problem that the reaction is not performed well.

In addition, sodium hydroxide (NaOH) or hydrogen chloride (HCl) is used to adjust the pH of the silkworm suspension, the pH of the silkworm powder suspension is adjusted to 5 to 7, and the silkworm powdery enzyme extract It was confirmed that the composition is suitable for use as an active ingredient in health foods or functional cosmetics (confirmed in the following experimental examples).

Step 3) Production of enzyme hydrolysis extract of silkworm

In this step, proteolytic enzyme is added to the silkworm powder suspension prepared in the second step, and at least one protease of pancreatin or protease is added to hydrolyze the other proteinase Thereby obtaining a silkworm enzymatic hydrolysis extract having enhanced antioxidant ability as compared with the application.

At this time, the pancreatin is a mixture of lipase, protease and amylase. The pancreatin is mixed in a weight ratio of 1:10 to 15:10 to 15, If the weight ratio is exceeded, there is a possibility that the desired enhanced antioxidant ability of the present invention may not be obtained.

In addition, the protease is added in an amount of 0.1 to 10% by weight based on the weight of the silkworm powder. When the protease is added in an amount of less than 0.1% by weight, the enzyme reaction is not performed well and the desired enhanced antioxidant ability of the present invention is not obtained. When the protease is added in an amount exceeding 10% by weight, It may become difficult to apply it as a health functional food or a cosmetic material due to a degrading enzyme.

Further, the enzyme hydrolysis conditions are 50 to 70 DEG C, preferably 50 to 60 DEG C

For 1 to 2 days, and when the hydrolysis of the silkworm is completed, the temperature is preferably raised to 90 DEG C to inactivate the silkworm.

The silkworm enzymatic hydrolysis extract prepared by the method for producing hydrolyzed silkworm enzyme of the present invention may be used as it is, but may be filtered to remove unreacted solids. The filtration is preferably performed at least once using a 0.1 to 3 탆 filter. When the pore size of the filter is less than 0.1 탆, the filter is extremely precise and ultrahigh (ultrafiltration method) is not economical. In the case of exceeding 3 탆, the filter has no sterilizing power and fungi and bacteria are contained in the final extract, There is a problem that it is likely to become.

The hydrolyzate of the silkworm enzyme of the present invention obtained by such filtration can be prepared into a liquid form obtained by centrifugal separation and a dry powdered product. The centrifugation is preferably performed at 1,000 to 1,300 rpm for 10 to 20 minutes.

In addition, the present invention relates to a process for producing a hydrolyzate of a silkworm silkworm mucilage produced by the process of the present invention, wherein the hydrolysis product of the silkworm silkworm mungbean is not subjected to strong acid treatment or prolonged high temperature treatment, And can be used directly in health food. In addition, the present invention provides a health food or functional cosmetic containing the hydrolyzed extract, having found that the silkworm enzyme hydrolysis extract obtained as described above has a whitening effect.

Hereinafter, the present invention will be described in more detail with reference to examples, but these examples are for illustrative purposes only and are not intended to limit the scope of protection of the present invention.

< Example  One. Antioxidant ability  Enhanced silkworm enzymatic hydrolysis Extract 1  Manufacturing>

The silkworm, 5 days old and 3 days old, was lyophilized at 75 ℃ for 70 hours and powdered to make silkworm powder.

10 ml of first distilled water was added to 1 g of silkworm powder prepared above and mixed at room temperature for 30 minutes to suspend the silkworm powder. The silkworm powder was suspended in 80% hydrogen chloride (HCl) or 1N sodium hydroxide (NaOH) .

Pancreatin, lipase, protease and amylase are composed of 8 USP U / g, 100 USP U / g and 100 USP U / g, respectively, in 10 ml of the silkworm powder suspension prepared above. And then hydrolyzed at an appropriate temperature for 24 hours using a shaking incubator.

The hydrolyzed silkworm silkworm hydrolyzate obtained after hydrolysis was enzymatically inactivated at 90 ° C, filtered using a filter paper of 2 μm, centrifuged at 1,300 rpm for 10 minutes to obtain a supernatant, Hydrolyzed extract 1 was prepared.

< Example  2. Antioxidant ability  Enhanced silkworm enzymatic hydrolysis Extract 2  Manufacturing>

The proteolytic enzyme hydrolyzed extract 2 of the present invention was prepared by the same method as in Example 1 except that protease (Vision Biochem) was used instead of Pancreatin as a protease.

< Comparative Example  1. Enzymatic hydrolysis of silkworm Extract 1  Manufacturing>

(Lipase, Vision Biochem) was used instead of Pancreatin as a protease to prepare silkworm enzyme hydrolyzed extract 1.

< Comparative Example  2. Enzymatic hydrolysis of silkworm Extract 2  Manufacturing>

The silkworm enzyme hydrolyzed extract 2 was prepared by the same method as in Example 1 except that cellulase (Vision Biochem) was used instead of Pancreatin as a protease.

< Experimental Example  1. Observation of color change of enzyme hydrolysis extract of silkworm by enzyme treatment>

1. Experimental Method

After preparing the hydrolyzed extract of silkworm enzyme of Examples 1, 2 and Comparative Examples 1 and 2, the color was observed with naked eyes and photographed. To observe the enzyme treatment at various pH, silkworm powder was mixed with distilled water and titrated with sodium hydroxide (NaOH) or hydrogen chloride (HCl), followed by treatment with each enzyme to observe the color change.

Observation of the color change by pH is to observe the stability of the silkworm enzyme hydrolysis extract as a food additive.

2. Experimental results

As shown in FIG. 1, it was observed that before the enzyme treatment, the milky color appeared at pH 3 and pH 5, green at pH 7 and 9, and pale green at pH 11, And 5 showed no color change, but pH 7 was black, pH 9 was black green, pH 11 was dark green.

In other words, when the pH of the silkworm suspension was maintained at 5 to 7 before the enzyme treatment, it was confirmed that the color changed to that of the silkworm powder without enzyme treatment after the enzyme treatment. This means that it can be used as a food additive.

< Experimental Example  2. Extraction of silkworm enzyme hydrolysis extract by enzyme treatment Confirm yield >

1. Experimental Method

In order to confirm the extraction yield of the hydrolysates of the silkworm enzymes produced in Examples 1, 2 and Comparative Examples 1 and 2, the silkworm solids filtered after each enzyme treatment were dried in a dry oven for 24 hours and then weighed %.

At this time, measurements were performed for each pH treatment and each temperature applied for each enzyme treatment.

2. Experimental results

The results of the experiment are shown in Tables 1 and 2 and FIGS.

pH of reaction (%) pH 3 pH 5 pH 7 pH 9 pH 11 non-treated 50 ° C 97 92 88 78 73 Pancreatin- Example 1 95 86 78 73 68 Lipase-Comparative Example 1 96 89 83 76 72 non-treated 60 ° C 93 80 73 65 57 Protease-Example 2 94 85 80 76 66 Cellulase-Comparative Example 2 91 86 75 66 60

Temperature of reaction (%) 30 ℃ 40 ℃ 50 ℃ 60 ° C 70 ℃ non-treated 88 86 80 73 85 Pancreatin- Example 1 92 83 75 74 82 Lipase-Comparative Example 1 85 80 80 84 90 Protease-Example 2 91 82 69 80 89 Cellulase-Comparative Example 2 85 83 76 75 93

As shown in Table 1 and FIG. 2, although the overall recovery rate is high due to the nature of the silkworm powder, it is confirmed that the higher the pH is, the lower the recovery rate is. It is considered that the components of silkworm powder are denatured under alkaline conditions and it is considered that the extraction efficiency is high. However, since the decomposition ability of enzymes and the possibility of denaturation of physiologically active substances are high, the pH before enzyme treatment is preferably 5 to 7 Could know. This was judged based on the results of the following experimental examples.

Also, as shown in Table 2 and FIG. 3, it was confirmed that the extraction yield was the highest when the application temperature was 50 to 60 ° C during the enzyme treatment, and it is preferable that the enzyme treatment is performed at 50 to 60 ° C , Which was confirmed to be the same as the recommended enzyme reaction temperature as the edible enzyme of the applied enzyme.

< Experimental Example  3. Determination of total reducing sugar content of hydrolysates of silkworm enzymes by enzyme treatment>

1. Experimental Method

A) Measurement of total reducing sugar content by 3,5-dinitrosalicylic acid (DNS) method

Each silkworm hydrolysis extract (Examples 1, 2 and Comparative Example 1, 2) was diluted 10 times with water, and then added with a DNS reagent. After the reaction was carried out at 105 ° C for 10 minutes, the temperature was rapidly lowered and distilled water was added to measure the absorbance of the UV absorbance photometer at 540 nm , The sugar content was calculated by substituting the measured value into the standard curve.

① Measurement of change in total reducing sugar content of silkworm powder by enzyme-treated pH

Pancreatin and lipase were reacted at 50 ℃ and 50 and 50 ℃, respectively. Protease and cellulase were also reacted at 60 ℃, which is the recommended optimum temperature. Was used at 60 ° C for 3 replicates, and D-glucose was used to obtain the standard curve.

② Measurement of total reducing sugar content of silkworm powder by enzyme treatment temperature

The pH of each enzyme was fixed at 7 and the experiment was performed.

2. Experimental results

The results of the experiment are shown in Tables 3 and 4 and FIGS.

pH of reaction (mg / ml) pH 3 pH 5 pH 7 pH 9 pH 11 non-treated 50 ° C 0.183 + - 0.001 0.335 0.003 0.498 ± 0.007 0.621 + 0.022 0.281 ± 0.001 Pancreatin- Example 1 0.181 ± 0.001 0.989 + 0.019 0.923 + 0.025 0.772 0.005 0.363 + 0.004 Lipase-Comparative Example 1 0.319 ± 0.001 0.860 ± 0.007 0.793 + 0.027 0.680 ± 0.007 0.347 ± 0.002 non-treated 60 ° C 0.154 + - 0.001 0.215 ± 0.003 0.403 0.003 0.154 + 0.004 0.215 ± 0.003 Protease-Example 2 0.148 ± 0.001 0.221 ± 0.001 0.399 ± 0.006 0.148 + 0.011 0.221 ± 0.003 Cellulase-Comparative Example 2 0.150 0.003 0.211 ± 0.002 0.433 0.002 0.150 0.004 0.211 ± 0.002

Temperature of reaction (mg / ml) 30 ℃ 40 ℃ 50 ℃ 60 ° C 70 ℃ non-treated 0.231
± 0.001 0.280
± 0.003 0.235
± 0.001 0.291
± 0.004 0.236
± 0.001 Pancreatin- Example 1 0.325
± 0.001 0.487
± 0.007 0.425
± 0.002 0.366
± 0.001 0.332
± 0.003 Lipase-Comparative Example 1 0.543
± 0.024 1.020
± 0.022 0.737
± 0.016 0.627
± 0.009 0.583
± 0.012 Protease-Example 2 0.443
± 0.005 0.877
± 0.006 0.543
± 0.010 0.460
± 0.010 0.490
± 0.012 Cellulase-Comparative Example 2 0.272
± 0.002 0.750
± 0.004 0.350
± 0.008 0.354
± 0.007 0.290
± 0.003

As shown in Table 3 and FIG. 4, the reducing sugar content of the pancreatin-treated group was measured at pH 5 and 7, and it was confirmed that pancreatin contained a different type of carbohydrase, α-amylase as a mixed enzyme, It is considered that carbohydrate present in silkworm powder was decomposed and the amount of reducing sugar was measured to be high, and it was found that protease does not affect reducing sugar production ability.

In addition, as shown in Table 4 and FIG. 5, the reducing sugar content of the pancreatin-treated group was measured at a temperature lower than the relatively low temperature of 30 and 40 ° C, and the pancreatin-treated sample was very high It was confirmed that the reducing sugar amount was measured.

< Experimental Example  4. Identification of total protein pattern of enzyme hydrolysis extract of silkworm by enzyme treatment>

1. Experimental Method

75 μl of each silkworm hydrolysis extract (Examples 1 and 2 and Comparative Examples 1 and 2) was taken out and mixed with 25 μl of NuPAGE 4x LDS sample buffer (Cat. No. NP0008, manufactured by Thermo Fisher Scientific) Heated for a while, allowed to stand on ice for a while, and then centrifuged at 13,000 rpm for 10 minutes. 5 ul of the supernatant was subjected to 4-12% gradient SDS polyacrylamide gel electrophoresis. After electrophoresis, the gel was stained with a coomassie brillant blu to confirm the protein pattern.

2. Experimental results

As shown in FIGS. 6 and 7, pancreatin and protease treatment results in a decrease in the overall band. Particularly, the protein band at 150 kDa and the thick protein band at 75 kDa decrease in intensity, On the other hand, it was confirmed that there was no change in the protein pattern of the hydrolysates of hydrolysates of silkworm enzyme obtained by treating lipase and cellulase (Comparative Examples 1 and 2).

< Experimental Example  5. Identification of Antioxidative Activity of Enzymatic Hydrolysis Extract by Enzyme Treatment>

1. Experimental Method

① Measurement of antioxidative effect by 2,2'-Azinobis [3-ethylbenzothiazoline-6-sulfonic acid] -diammonium salt (ABTS) method

: ABTS and potassium persulphate were reacted with ABTS and potassium persulphate for 16 hours to form ABTS cations. The solution was then diluted with PBS to an absorbance value of 734 nm of 0.7 The diluted solution was reacted with a sample diluted 100 times with water by hydrolysis of each silkworm extract (Examples 1, 2 and Comparative Examples 1 and 2) for 30 minutes at room temperature, and the absorbance was measured at 734 nm using a UV absorption spectrophotometer , The antioxidant capacity was calculated by the following equation, and the total of 3 replicates was performed. L-ascorbic acid was used as a control.

      ABTS radical scavenging activity (%) = (1- (A / B)) 100

       A: Absorbance in the presence of distilled water instead of the sample solution

       B: Absorbance with addition of extraction sample

2. Experimental results

The results of the experiment are shown in Tables 5 and 6 and FIGS.

Antioxidative activity by pH using ABTS method (%) pH 3 pH 5 pH 7 pH 9 pH 11 non-treated 50 ° C 36.2 ± 0.2 32.5 ± 0.3 26.8 ± 0.3 32.2 ± 0.1 34.6 ± 0.1 Pancreatin- Example 1 35.3 ± 0.4 51.5 ± 0.5 55.3 ± 0.2 43.1 ± 0.2 39.6 ± 0.2 Lipase-Comparative Example 1 31.4 ± 1.3 30.3 ± 1.5 28.5 ± 0.5 34.1 ± 0.3 33.1 ± 0.2 non-treated 60 ° C 28.0 ± 0.2 23.5 ± 0.1 31.6 ± 0.1 28.0 ± 0.1 23.5 ± 0.3 Protease-Example 2 22.1 ± 0.2 34.6 ± 0.4 54.3 ± 0.3 22.1 ± 0.5 34.6 ± 0.3 Cellulase-Comparative Example 2 23.2 ± 0.3 24.1 ± 0.3 31.2 ± 0.3 23.2 ± 0.1 24.1 ± 0.2

Antioxidant activity by temperature using ABTS method (%) 30 ℃ 40 ℃ 50 ℃ 60 ° C 70 ℃ non-treated 33.8 ± 0.2 48.9 ± 0.4 34.2 ± 0.7 55.1 ± 0.3 31.3 ± 0.3 Pancreatin- Example 1 38.8 ± 0.4 60.0 ± 0.8 37.7 ± 0.4 58.7 ± 1.1 37.1 ± 0.4 Lipase-Comparative Example 1 32.5 ± 0.6 57.2 ± 0.1 30.1 ± 0.1 32.5 ± 0.6 57.2 ± 0.3 Protease-Example 2 28.2 ± 0.5 32.5 ± 0.6 24.3 ± 0.2 28.2 ± 0.9 32.5 ± 0.5 Cellulase-Comparative Example 2 23.5 ± 1.0 32.6 ± 0.5 21.6 ± 0.2 23.5 ± 0.6 32.6 ± 0.3

As shown in Table 5 and FIG. 8, the antioxidative activity of pancreatin and protease-treated groups was higher than that of pH 3, which is a highly acidic condition, The highest antioxidant activity was observed at pH 7, which is the optimum pH of the enzyme, compared to untreated silkworm powder. The lipid and cellulase treatment groups (Comparative Examples 1 and 2) Respectively. In addition, as shown in Table 6 and FIG. 9, the antioxidative activities of pancreatin and protease were higher than those of pancreatin and protease treatment, , And the antioxidative activity similar to that of silkworm powder not treated with enzyme was measured in the lipase and cellulase treated groups (Comparative Examples 1 and 2). As described above, it is known that pancretin and protease are most suitable among the proteolytic enzymes applied in the production of the silkworm enzyme hydrolysis extract of the present invention. As shown in the result of Experimental Example 4, It is considered that the material has been extracted.

< Experimental Example  6. Identification of whitening activity of enzyme hydrolysis extract of silkworm by enzyme treatment>

1. Experimental Method

① Test for whitening effect by tyrosinase inhibition method

mushroom tyrosinase (1000 units / ml) was added to each of the L-DOPA solution and each silkworm hydrolysis extract (Examples 1 and 2 and Comparative Examples 1 and 2) dissolved in phosphate buffer, After the reaction, the absorbance was measured at 475 nm using a UV absorption spectrophotometer, and the inhibition effect of tyrosinase was measured by the following equation. A total of 3 repetitions were performed, and L-ascorbic acid was used as a control.

Tyrosinase inhibition effects (%) = [(A-B) / A] 100

       A: The absorbance was measured by adding distilled water instead of the extracted sample solution

       B: Absorbance was measured by adding extraction sample

Here, tyrosinase inhibits tyrosinase as an enzyme for producing melanin, which means that the melanin production is inhibited and thus the whitening effect is excellent.

2. Experimental results

The results of the experiment are shown in Tables 7 and 8 and FIGS. 10 and 11.

pH of reaction (%) pH 3 pH 5 pH 7 pH 9 pH 11 non-treated 50 ° C 50.8 ± 0.1 51.7 ± 1.5 52.1 ± 0.1 51.1 ± 1.3 52.4 ± 0.5 Pancreatin- Example 1 48.1 ± 0.3 47.6 ± 0.4 48.3 ± 0.5 48.4 ± 0.2 46.9 ± 0.1 Lipase-Comparative Example 1 46.3 ± 0.8 46.7 ± 0.3 46.2 ± 0.5 45.8 ± 0.4 45.3 ± 0.5 non-treated 60 ° C 50.3 ± 1.0 46.9 ± 2.9 44.5 ± 7.2 51.6 ± 0.8 51.7 ± 1.0 Protease-Example 2 48.6 ± 0.2 47.3 ± 0.1 47.6 ± 0.2 46.5 ± 0.1 45.8 ± 1.1 Cellulase-Comparative Example 2 47.9 ± 0.2 47.2 ± 0.1 47.7 ± 0.1 47.8 ± 0.5 44.2 ± 0.9

Temperature of reaction (%) 30 ℃ 40 ℃ 50 ℃ 60 ° C 70 ℃ non-treated 70.7 ± 0.1 70.2 ± 1.1 69.1 ± 1.2 68.1 ± 1.1 66.8 ± 2.3 Pancreatin- Example 1 51.7 ± 1.7 55.6 ± 0.9 53.7 ± 2.2 53.9 ± 1.1 54.9 ± 2.2 Lipase-Comparative Example 1 55.1 ± 1.7 54.5 ± 0.5 54.9 ± 1.5 52.5 ± 0.1 55.1 ± 0.2 Protease-Example 2 71.8 ± 1.1 63.4 ± 4.8 60.8 ± 1.5 80.7 ± 8.5 81.9 ± 15.3 Cellulase-Comparative Example 2 60.0 ± 0.2 58.2 ± 0.5 60.5 ± 0.1 61.7 ± 1.9 46.8 ± 3.2

As shown in Tables 7, 8 and 10, 11, pancreatin treatment showed similar inhibition of tyrosinase inhibition compared to that before the treatment in the pH condition, and in the proteas treatment, Compared with the control group.

In the experiment by enzyme treatment temperature condition, protease treatment was significantly higher than that of enzyme treatment, whereas in pancretin treatment, it was slightly lower than before enzyme treatment. However, this treatment significantly damaged the whitening efficacy of enzyme hydrolysis extract It is shown that it is not given, and pancretin treatment and protease treat treatment all have whitening effect.

As described above, the present invention provides a method for producing a silkworm enzyme hydrolysis extract having enhanced antioxidative ability by optimizing treatment conditions for improving the yield of the enzyme hydrolysis extract of silkworm silkworm. The silkworm enzyme hydrolysis extract Finds the enhanced antioxidant activity and tyrosinase activity and thus can provide health foods and functional cosmetics containing them, thereby contributing to an increase in the income of the silkworm farmers and processing companies and the health promotion of the people.

It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit or scope of the invention as defined by the appended claims. It will be possible. The scope of the present invention is defined by the appended claims, and all differences within the scope of the claims are to be construed as being included in the present invention.

Claims (7)

A first step of drying a silkworm to produce silkworm powder,
A second step of suspending the silkworm powder by adding water to the silkworm powder of the first step and then adding an acidic solution or a basic solution to prepare a powder suspension in an eye having a pH adjusted;
The silkworm powder suspension of the second step is subjected to enzymatic hydrolysis by adding a proteolytic enzyme consisting of at least one of pancreatin or protease to the silkworm enzyme hydrolysis, And a third step of obtaining an extract,
A method for producing an enzyme hydrolysis extract of silkworm enzyme with enhanced antioxidant ability.
The method according to claim 1,
Wherein the proteolytic enzyme of the third step is added in an amount of 0.1 to 10% by weight based on the weight of the silkworm powder.
A method for producing an enzyme hydrolysis extract of silkworm enzyme with enhanced antioxidant ability.
A hydrolytic extract of a silkworm enzyme produced by the method of claim 1, wherein the hydrolysis activity of the enzyme is enhanced by the addition of another protease.
The method of claim 3,
Characterized in that the silkworm enzyme hydrolyzed extract is in the form of a liquid obtained by filtration or centrifugation after filtration,
Enzymatic hydrolysis of silkworm enzyme with enhanced antioxidant activity.
The method of claim 3,
The silkworm enzymatic hydrolysis extract has a whitening effect,
Enzymatic hydrolysis of silkworm enzyme with enhanced antioxidant activity.
A health food having an enhanced antioxidative ability and a whitening effect, which comprises the hydrolysates of hydrolysates of the silkworm noodles according to any one of claims 3 to 5 as an active ingredient.
7. A functional cosmetic comprising the hydrolyzed hydrolyzate of the silkworm noodle according to any one of claims 3 to 5 as an active ingredient and having an enhanced antioxidative and whitening effect.

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