KR101478615B1 - Method for manufacturing of traditional adhesives using enzyme - Google Patents

Abstract

The present invention relates to a method for producing an adhesive using an enzyme. The method of manufacturing a conventional adhesive according to the present invention can reduce the manufacturing process of a conventional adhesive requiring a lot of time and effort by manufacturing a conventional adhesive using an enzyme, It can contribute to the economy by reducing dependence on imports. In addition, the conventional adhesive manufactured by the production method of the present invention has an effect that it can be used as a cultural material preservation material having excellent durability and storage property.

Description

TECHNICAL FIELD [0001] The present invention relates to a method for manufacturing a conventional adhesive using an enzyme,

The present invention relates to a method for producing an adhesive using an enzyme.

In preserving and repairing cultural properties in the form of paper records, traditional glue such as gypsum grass is used in Korea, and Japanese glazed grass is used in Japan. This is because the use of a chemical paste or a paste with an antiseptic is caused by the chemical change of the paper, the coloring of the paper, or the paper, which shortens the life span and makes it difficult to preserve the cultural property. Traditional adhesives have lower viscosity and less adhesive strength than ordinary grass, which has the advantage of minimizing changes in paper records.

 The manufacturing method and the process of the traditional adhesive differ from that of the Japanese pastes and the Japanese pastes. In Korea, the process of making the traditional grass is a process in which wheat flour is immersed in water for a long time and then the wheat flour is plowed with the wheat flour. This is a change occurring in the process of soaking the flour into the water and is mainly caused by the microorganism producing amylase In the case of Japanese scallops, however, the method of fermenting the grass after boiling the grass is used. The new glue is made of wheat starch, and the old glue is a grass that has been made into a poplar in the summer and poured water into the poplar and then sealed it for about 10 years. The production of traditional adhesives takes a long time from several months to several years, and the cost and manpower required thereby necessitates the modern manufacturing technology of traditional adhesives.

When the modern manufacturing technology of traditional adhesive is developed, economic, social and technological achievements are expected to be achieved by reducing the manufacturing process of traditional adhesives, which requires a lot of time and effort, Not only can it improve efficiency, but it can also contribute to economic aspects by reducing dependence on imports of preserved materials for cultural properties preservation. In addition, through the activation of the research and development of preservation materials for cultural properties from the social aspect, it will be possible to raise awareness of the importance of cultural properties and motivate them to attract social attention and support. It may be worthy of use as a preservation material of cultural properties that is durable and storage-friendly.

However, there is no modern manufacturing technology for such traditional adhesives, and accordingly, it is urgent to develop a modern manufacturing technique for traditional adhesives.

As a result of continuing research on the modern manufacturing technology of traditional adhesives, the present inventors have conducted an enzymatic reaction using wheat starch and have succeeded in using the low molecular weight starch produced by the enzyme reaction to prepare a traditional adhesive The present invention has been completed.

It is an object of the present invention to provide a method for the modern manufacture of traditional adhesives using enzymes.

The present invention provides a method for producing a conventional adhesive using an enzyme.

The method of manufacturing a conventional adhesive according to the present invention can reduce the manufacturing process of a conventional adhesive requiring a lot of time and effort by manufacturing a conventional adhesive using an enzyme, It can contribute to the economy by reducing dependence on imports. In addition, the conventional adhesive manufactured by the production method of the present invention has an effect that it can be used as a cultural material preservation material having excellent durability and storage property.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a schematic diagram of a process for producing a starch adhesive using an enzyme of the present invention. FIG.
Fig. 2 is an electron micrograph of live meal after extracting wheat flour and live wheat flour.
FIG. 3 is a graph showing an enzyme reaction characteristic of? -Amylase. FIG.
4 is a graph showing changes in amylose content according to the enzyme treatment concentration of the present invention.
5 is a graph showing a change in size of starch particles according to the enzyme treatment concentration of the present invention.
6 is a graph showing the change in viscosity of the starch adhesive according to the enzyme treatment concentration of the present invention.
7 is a graph showing the peel strength of the starch adhesive according to the enzyme treatment concentration of the present invention.
Fig. 8 is a diagram showing the peel strength of a scarf and a commercial scarf imported from Japan. [A: Tokyo Metropolitan Univ., B: Utsunomiya Univ., C: Untitled Workshop, D: Kumamoto Univ.
9 is a graph showing bending characteristics according to an enzyme treatment concentration.
10 is a graph showing the viscosity of the starch sample according to the enzyme treatment time and hydrochloric acid treatment.
11 is a graph showing the peeling strength of the starch sample according to the treatment time of the enzyme and the hydrochloric acid treatment.
Fig. 12 is a graph showing the comparison of the peel strength and viscosity of starch samples according to enzyme treatment time. Fig.
13 is a graph showing the antifungal property of each drug against E. coli.
Fig. 14 is a graph showing the antibacterial property of a drug-added adhesive to E. coli. Fig.

The present invention

(a) kneading wheat flour with distilled water, washing the dough with distilled water, and extracting raw flour;

(b) reacting an enzyme with the live mass extracted in the step (a); And

(c) washing and drying the enzyme-treated starch in the step (b);

A method for producing a conventional adhesive using an enzyme.

The present invention also provides a conventional adhesive produced by the above-mentioned production method.

Hereinafter, the present invention will be described in detail.

The step (a) is a step of removing the protein present in the flour and extracting the live flour. Generally, the protein present in the wheat flour is consumed by the microorganism during the flour cutting process. Therefore, Protein is removed from wheat flour to separate live births.

The step (b) is a step of reacting the raw material extracted in the step (a) with an enzyme.

It is preferable that the enzyme uses? -Amylase.

The enzyme reaction is preferably performed for 3 hours to 5 hours since the reaction product by the enzyme reaction is hardly produced after 5 hours.

The enzyme reaction is preferably performed at 50 ° C. or less, preferably 30 to 50 ° C. Since the α-amylase enzyme must act on the live phase in the present invention, when the temperature is high, This is because the characteristics are different.

The amount of the enzyme is preferably 4 to 1600 units / mL based on the starch solution. As the amount of enzyme injected increases, the enzyme reaction time can be shortened.

The enzyme reaction can be stopped with a solution of 90 to 99% ethanol or a solution of 0.1 to 0.2 N hydrochloric acid (HCL) to which 1 to 2% (v / v) of acetic acid is added. This is because, as described above, as the amount of the enzyme is increased, the enzyme reaction time can be shortened and the enzyme reaction time can be shortened.

The step (c) is a step of washing and drying the enzyme-treated starch in the step (b).

The method of manufacturing a conventional adhesive of the present invention may further include the step of (d) adding at least one agent selected from the group consisting of white gum, alum, ice sheet, yucca, and glyoxylate can do.

In the conventional adhesive prepared by the above method, the viscosity of the starch paste is reduced, the peel strength and the bending property are excellent, and also the anti-bacterial property is obtained due to the addition of the drug.

As described above, the conventional adhesive manufacturing method can reduce the manufacturing process of the conventional adhesive requiring a lot of time and effort, thereby improving the process efficiency.

Hereinafter, preferred embodiments of the present invention will be described in order to facilitate understanding of the present invention. However, the following examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited by the examples.

< Example 1 > Manufacture of Traditional Adhesive by the Manufacturing Method of the Present Invention

The wheat starches used in this experiment were separated from wheat flour purchased from Kwangyoung-myeon, Guryong-gun, Jeonnam Province. Twenty milliliters of distilled water was added to 50 g of wheat flour and kneaded. Then, the dough was washed with 500 ml of distilled water to extract the starch, and the extracted starch was centrifuged to remove the supernatant and freeze-dried for 36 hours. And suspended in 0.1 M sodium phosphate buffer (pH 7.0) such that the concentration of the starch separated from the flour was 100 mg / mL. After warming at 130 rpm for 5 minutes at 37 ° C where starch gelification did not start, 800 units of α-amylase per 1 mL of the reaction solution were added in 0.1 M sodium phosphate buffer, and the reaction was started. After 4 hours of reaction at 50 ° C., 1.5% v / v) acetic acid was added to the reaction solution. Then, the conventional adhesive of the present invention was prepared by washing and drying.

A schematic diagram of a process for producing a conventional adhesive using the enzyme of the present invention is shown in Fig.

< Example 2 > Manufacture of traditional adhesives with antimicrobial agents

For the preparation of adhesives with chemical additives, 10 g of white ginseng, alum, ice, and oriental were weighed out into 500 mL of distilled water, boiled for about 30 minutes, and 1 g of glyoxylic acid was weighed and dissolved in 100 mL of distilled water , And then mixed with Luria-Bertani medium (LB medium). All medicines, chemicals, and media were sterilized at 120 ° C for 20 minutes before use. This was added to the conventional adhesive prepared in Example 1 to prepare a conventional adhesive agent.

< Experimental Example 1 > Live meal  Confirmation of production of sample

The removal of proteins from fresh wheat flour and wheat flour was confirmed by electron microscopy. The results are shown in Fig.

As shown in Fig. 2, wheat flour contained a large amount of protein, so that the elliptical structure, which is generally observed in the live part, is unclear, and it was observed that the protein is attached to the live part. Electron micrographs after removal of proteins from flour generally confirmed the elliptical structure observed in the live cells and confirmed that the protein was removed from the starch particles.

< Experimental Example 2 > Characterization of enzyme reaction of α-amylase

The reaction concentration was adjusted to 25 mg / ml to measure the reducing sugar content by enzyme reaction time. The mixture was heated for 5 minutes in a shaking water bath (JSSB-30T, JSR, Gongju, Korea) set at 110 rpm at 37 ° C where starch gelification did not start, and then washed with 0.1 M sodium phosphate buffer 7.0), 1 U of α-amylase was added per 1 ml of the reaction solution, and the reaction was started. After the reaction at 0 second, 10 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours and 24 hours, The reaction was stopped using NaOH. After stopping the reaction, the sample was centrifuged at 10,000 rpm for 3 minutes using a centrifuge (VS-15000N, Vision Co., Bucheon, Korea), and the amount of reducing sugar was measured using the supernatant. The reducing sugar reaction was measured by a copper-bicinchoninate method. The same amount of copper-nonchynoninate reagent was added to 200 μl of the enzyme reaction solution, followed by color development at 80 ° C for 35 minutes. The absorbance was measured at 540 nm using a Microsample plate reader (Model 680, Bio-Rad, Hercules, Calif., USA).

The copper-undecynoninate reagent was prepared as follows. Solution A was prepared by dissolving 97.1 mg of disodium 2,2'-bicinchoninate in 45 mL of a liquid containing 2.735 g of sodium carbonate anhydrous, 1.2 g of sodium bicarbonate, (No. D8248, Sigma-Aldrich Co., St. Louis, Mo., USA) was dissolved and the final volume was adjusted to 50 mL. Solution B was mixed with the same amount every time it was used and used for color development.

Enzyme reaction on wheat starch To determine the reducing sugar content by time, α-amylase The reaction characteristics of the enzyme over time are shown in Fig.

As shown in FIG. 3, the starch was decomposed with increasing time, and the amount of reducing sugar was increased. The progress curve of the general enzyme reaction was shown. The reaction products by enzyme reaction increased rapidly until 4 hours and then increased slowly.

< Experimental Example 3 > Depending on enzyme treatment Raw  Physicochemical characterization

The activity of the enzyme was measured by using 1% (w / v) soluble starch as a substrate in a 0.1 M phosphate buffer (pH 7.0), at 37 ° C in a water bath (circulating water bath, jeio tech, Daejeon, Korea) After reacting for 10 minutes, the reaction was stopped by adding a solution of DNS (3,5-dinitrosalicyclic acid) in three times the volume of the total reaction solution. After stopping the reaction, the absorbance was measured at 540 nm and quantified using maltose as a reference material. The amount of enzyme required to produce 1 μmole of maltose for 1 minute was defined as 1 unit of enzyme.

1. Analysis of Amylose Content by Enzyme Treatment

In order to measure the change of the amylose content according to the enzyme treatment concentration, the enzyme treatment time was fixed to 4 hours, and the amount of the enzyme to be treated in the starch was 8, 16, 32, 64 and 400 Unit / The change in the content is shown in Fig.

As shown in FIG. 4, the amylose content of the starch treated with 8, 16, and 32 unit / mL enzymes did not change significantly, and the amylose content tended to decrease gradually with 64, 100 Unit / mL enzyme treatment. The reduction of amylose content is related to the decomposition of starch amalose resulting in the formation of low molecular weight substances, which is considered to be correlated with the viscosity decrease of the starch paste.

2. Analysis of changes in starch particle size by enzyme treatment

The size of starch particles was measured using a laser light scattering apparatus, and 500 nm of polystyrene particles were used as a standard material. The particle size of the starch according to the amount of enzyme treatment using the light scattering apparatus is shown in FIG.

As shown in Fig. 5, the particle size change of the starch according to the concentration of enzyme treatment sharply decreased to 32 Unit / mL, and there was no significant difference thereafter.

< Experimental Example 4 > Enzyme-Treated Wheat starch  Characterization of adhesive

Starch adhesive was prepared by heating the starch treated with time and concentration to 16% concentration in distilled water and diluted with distilled water at a ratio of 1: 3.

1. Viscosity analysis of starch adhesive

The viscosity of the gluten - milled starch treated with enzyme was measured using Brookfield Viscometer under Spindle 3, 50 rpm. The viscosity change of the starch paste according to the enzyme treatment concentration is shown in Fig.

As shown in Fig. 6, the viscosity of the starch paste treated with 16, 32, and 64 units / mL of enzyme in Spindle 2 was the lowest at 24 cP. The viscosity of starch paste did not change at 16, 32 and 64 unit / mL, decreased at 800 unit / mL and decreased significantly at 1600 Unit / mL. It was confirmed that the viscosity of the starch paste decreased as the concentration of enzyme treatment increased and the starch became low molecular weight.

2. Starch adhesive Peel strength  analysis

In order to measure the adhesive strength, Hanji was purchased from Mungyeong Sangik branch in Gyeongbuk (Gyeongbuk Intangible Cultural Heritage 30, Mungyeong Traditional Hanji Hanji). Table 1 shows the basis weight, thickness and optical properties of Korean paper.

Basis weight (g / m2) Thickness (mm) L * a * b * H V / C 34 0.15 79.59 1.40 16.44 0.84 Y 7.82 / 2.41

The adhesion of starch paste was measured by peel strength (N / 5 ㎝) experiment (KS K 0533: 2008). The peel strength change of the starch paste according to the enzyme treatment concentration of the present invention is shown in Fig. 7, and the peel strength by the furrow and the commercial furrow collected in Japan is shown in Fig.

As shown in Fig. 7, the change of the adhesive force according to the enzyme concentration decreased gradually after 64 units / mL, and decreased greatly from 0.2 unit / mL to 0.2 unit / mL and from 0.3 unit / mL to 1600 Unit / mL. In particular, it was confirmed that the peel strength was lower at 800 and 1600 Unit / mL than the starch pool reacted at 400 unit / mL for 24 hours.

Also, as shown in FIG. 8, the peel strength of the crushed and commercially available crumbs collected in Japan showed a difference of about 1 N / 5 cm, though it varies depending on the workshop manufactured. In the case of the starch adhesive according to the enzyme treatment concentration described above, it was confirmed that the starch adhesive exhibited the best adhesive force for the adhesion of starch paste prepared at an enzyme treatment concentration of 400 U / mL or more.

3. Adhesive  Bending characteristic analysis

The measurement results of the bending properties of the adhesive according to the enzyme throughput are shown in Table 2 and FIG.

sample
(Unit / mL) W 8 16 32 64 400 800 1600 B 0.1060 0.1003 0.0153 0.0027 0.1061 0.0269 -0.0990 0.0430 2HB 0.1783 0.7818 0.8648 0.8377 0.7024 0.9475 0.5131 0.6860

W: Adhesion by water

As shown in Table 2 and FIG. 9, when the adhesive is paper, it tends to fold and fold due to the bending measurement, and accurate measurement is difficult. However, when only the hysteresis for the bending is compared, the hysteresis gradually decreases according to the throughput of the enzyme, It is thought to be more flexible in terms of less resistance.

< Experimental Example 5 > 0.1N HCl The viscosity change and adhesion measurement

The viscosity and the adhesiveness were measured by treating with 0.1N HCl instead of 95% ethanol added with 1.5% (v / v) acetic acid to obtain a final pH of 2.0.

1. Viscosity change analysis

Fig. 10 shows the result of measuring the viscosity of a starch sample treated with 0.1N HCl to obtain a final pH of 2.0.

As shown in FIG. 10, in the case of the conventional method, there was almost no change in viscosity depending on the enzyme treatment time. However, when 0.1 N HCl was added to the final pH of 2.0, And the decrease was observed rapidly.

2. Adhesion Analysis

The peel strength in the longitudinal direction and the width direction was measured under the same conditions as those described above in the case of the adhesive force, and the results are shown in FIG.

As shown in FIG. 11, when the reaction was stopped by the conventional method, there was almost no change in the adhesive force as in the case of the viscosity. However, when 0.1 N HCl was added to the final pH of 2.0, the enzyme reaction time was increased Therefore, the peel strength tended to decrease overall.

3. Peel strength and  Correlation analysis of viscosity

It was concluded that there is a correlation between the peel strength and the viscosity based on the tendency that the viscosity tends to decrease as the enzyme treatment time increases. Thus, it is considered that there is a correlation between the peel strength and viscosity of the starch sample The relationship is shown in one graph and shown in Fig.

As shown in FIG. 12, when two points are represented by the x and y axes, the relationship of the straight line is shown, and in the created graph except for the relatively outlier data, the value of R ² is 0.9 or more, . From these results, it can be expected that the peeling strength can be predicted more conveniently by measuring the viscosity in the case of the adhesive using the same kind.

< Experimental Example 6 > Additives of Traditional Adhesives Abrasion resistance  analysis

Fig. 13 shows the results of the antibacterial activity against the E. coli, and Fig. 14 shows the results of the antibacterial activity analysis of the conventional adhesive with the medicament prepared in Example 2.

As shown in FIG. 13, it was confirmed that the lean body was reduced by about 10 times from 1.7 × 10 ⁶ to 7.4 × 10 ⁵ after 1 hour, 3.5 × 10 ⁴ after 2 hours, and 4.5 × 10 ³ after 4 hours. In the case of alum, the rate of bacterial killing was slightly faster than that of white ginseng, but after 4 hours, the number of dilutions was reduced, but no colonies were observed. The ice sheet started at 1.8 × 10 ⁶ , but no colonies were observed after 2 hours. The brittleness to the following fractions was found to be the weakest among the five different test samples, and glyoxylate was found to have the highest bacteriostasis.

As shown in FIG. 14, when compared with the experiment in which the adhesive was not added, the time required for the bacteria to die was slightly slower, and the same fungicidal activity was also observed in the order of the oriental starch alum, the ice cold starch glyoxylate.

Claims (7)

(a) adding wheat flour to distilled water, kneading the flour, and washing the flour to extract the protein-free raw meal;
(b) injecting an enzyme of 4 to 1600 units / mL based on the raw solution into the raw blood extract obtained in step (a), and then allowing the enzyme reaction at 30 to 50 ° C for 3 to 5 hours to stop the reaction; And
(c) washing and drying the enzyme-treated starch in the step (b);
&Lt; / RTI &gt; by weight of the enzyme. The method of claim 1, wherein the enzyme in step (b) is? -Amylase. delete delete The method according to claim 1, wherein the enzymatic reaction in step (b) is performed by adding 90 to 99% ethanol solution or 1 to 2N hydrochloric acid (HCL ) To stop the reaction. The method of claim 1, further comprising: (d) adding at least one agent selected from the group consisting of white ginseng, alum, ice, frankincense, and glyoxylate; &Lt; / RTI &gt; further comprising the steps of: A traditional adhesive prepared by the process of any one of claims 1, 2, 5 or 6.

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