KR101092329B1 - Method for extracting pectin by physical and enzymatic treatment - Google Patents

Abstract

The present invention relates to a pectin extraction method by a physical and enzymatic combined treatment method that does not use existing chemical solvents and / or acid treatment methods. The method of the present invention has been improved yield of about twice the conventional method, and because it does not use a chemical process is an environmentally friendly and human-friendly process that can reduce the environmental pollution caused by by-products and waste generation.

Physical, enzymatic, pectin, extraction

Description

Method for extracting pectin by physical and enzymatic treatment

The present invention relates to a pectin extraction method by a physical and enzymatic complex treatment method that does not use a conventional chemical solvent and acid treatment method.

In general, each organ of many plants contains a polysaccharide called pectin, which is particularly present in meristems and milk tissues. Pectin is a colloidal carbohydrate obtained in or from a plant, and contains a large amount of galacturonic acid groups, which are chain-linked. Its form is a form of the cell wall component of the plant, which is combined with cellulose, hemicellulose, lignin, protein, minerals, and the like, and is called insoluble protofectin in water. A substance that is water soluble in pectin and which gels with sugar and acid under appropriate conditions is called pectin.

The general preparation of pectin consists of (1) extraction under high temperature acidity from plant starting materials, (2) purification of liquid extracts, and (3) isolation of extract pectin from liquids. Among them, in the acid extraction step, the plant material is usually treated at a pH of 1 to 2 and a temperature of 80 ° C to 100 ° C using dilute acid such as nitric acid, sulfuric acid, hydrochloric acid or other inorganic or organic acid. As a plant starting material generally used, the peel of citrus fruits obtained from the preparation of juice, and the press dregs of the apple obtained from the production of apple juice and cider are used.

Pectin has various characteristics according to the degree of esterification and molecular weight, and is mostly used in the food field as a gelling agent, a thickener, a stabilizer, and the like. The degree of esterification (DE) indicating the degree of esterification is a numerical value (%) indicating the proportion of esterified galacturonic acid (carboxyl groups) in the total galacturonic acid (total carboxyl groups), and according to the height of the methyl esterification degree, High methoxyl pectin (HM pectin) and low methoxyl pectin (LM pectin). Usually, DE is higher than 50% is called HM pectin, and 50% or less is called LM pectin. In general, many of the pectins extracted under high temperature and acidic conditions from plant starting materials are HM pectins, and in order to obtain LM pectins, it is necessary to further demethylate with acid, alkali, enzyme or ammonia. When extracting the pectin from the peel of citrus fruits, there was a drawback in that the sugar chain was cleaved simultaneously with hydrolysis of the ester by high temperature acidic conditions or demethylation treatment at the time of extraction and the molecular weight decreased. As a result, high molecular weight pectin is expected to have various characteristics, but a method for stably producing such pectin without chemical treatment is unknown.

The present invention solves the above problems, and the object of the present invention is to establish a natural extraction process capable of producing pectin environmentally friendly, to extract the natural pectin applicable to a variety of health functional food To provide a way.

In order to achieve the above object, the present invention provides a pectin extraction method through a physical treatment method and an enzymatic treatment method without using a chemical solvent, an acid, or a base.

In one embodiment of the invention, the physical treatment method is preferably at least one treatment method selected from the group consisting of homogenization, and hydrothermal extraction and / or autoclave, the physical treatment method further comprises a reduced pressure filtration process More preferably included but is not limited thereto.

In one embodiment of the invention, the enzyme is from the group consisting of arabase (arabanase), cellulose (cellulase), beta-glucanase, hemicellulase and xylanase (xylanase) It is preferably but not limited to one or more enzymes selected.

In another aspect, the present invention comprises the steps of a) mixing the solvent in the sample and stirring; b) homogenizing or hydrothermally extracting the stirred sample; And c) enzymatically treating the result of the homogenization or hydrothermal extraction.

In one embodiment of the present invention, the method preferably further comprises a pressure reduction filtration process between steps a) and b), but is not limited thereto.

In one embodiment of the present invention, step b) is more preferably to replace the hot water extraction process with an autoclave process, but is not limited thereto.

In one embodiment of the invention, the enzyme is from the group consisting of arabase (arabanase), cellulose (cellulase), beta-glucanase, hemicellulase and xylanase (xylanase) It is preferably at least one enzyme selected, and the enzyme treatment concentration is more preferably, but not limited to, diluted from one hundredth to one thousandth of the total treatment volume.

Hereinafter, the present invention will be described.

In the present invention, the conventional pectin extraction method is an acid treatment method, but the physical / enzyme complex treatment method developed in the present invention does not use any chemical solvent and acid, but only a pure physical treatment method and an enzymatic treatment method pectin. The extraction process is much simpler than the conventional method, and the extraction time is also shortened.

The GalUA content was 42% when the pectin was extracted by the physical / enzyme complex treatment method developed by the method according to the present invention, which was lower than the conventional acid treatment method (65%) but showed a yield of 20%. It is twice as effective as%).

An approximate process of the invention is shown in FIG. 1.

The present invention establishes a pectin extraction process through natural methods (physical and enzymatic processing methods), rather than conventional chemical processing methods, through pure physical and enzymatic treatment in which chemical methods such as organic solvents or acid treatments are eliminated. A pectin extract with a galacturonic acid content of 100% was prepared, and the yield (about 20%) was twice as high as that of the conventional method (about 10%). It is an eco-friendly and human-friendly process that does not emit.

Hereinafter, the present invention will be described in more detail with reference to non-limiting examples.

However, since the following examples are described for the purpose of illustrating the present invention, the scope of the present invention is not to be construed as limited by the following examples.

Example 1: Particle size separation process

After the juice from the tangerine, the remaining by-products are put in a hot air dryer (80 ℃) and dried for 2 days. The dried sample was sieved using a grinder to separate the powder obtained by sieving, and then separated by a total of five particle sizes smaller than 30-50 mesh, 100-200 mesh, 200-325 mesh, and 325 mesh (FIG. 2).

As can be seen in Figure 2 by the galacturonic acid analysis of the content of the pectin content by particle size, the smaller the particle size, the higher the content of the extracted pectin. However, 200-325 mesh particles were selected because it is not easy to obtain a large amount of pectin of 325 mesh or more.

Example 2: Physical Treatment Process 1 (homogenization vs sonication)

5g of citrus powder with a particle size of 200 mesh is mixed with 50ml of distilled water to make a suspension. After stirring for 1 hour at 500rpm using a stirrer, mechanical / physical treatment is performed using a homogenizer and an ultrasonicator. 1, 3, 5 minutes for the homogenizer and 20, 40, 60 minutes for the ultra sonicator and then filtered and lyophilized filtrate to determine the pectin content (Fig. 3).

As can be seen in Figure 3, there was no significant difference between Homogenization and ultrasonication treatment, but the homogenization showed a slightly higher pectin content. In addition, since the process time is much shorter during the homogernization treatment, a process of treating homogenization for 3 minutes was selected (FIG. 4).

Example 3: Physical Treatment Process 2 (Hot Water Extraction)

Attempt to extract hot water as a second physical treatment process. After stirring at 120 rpm for 1 hour using a shaking water bath (60, 80, 100 ° C.), the obtained filtrate was lyophilized using reduced pressure filtration and the pectin content was measured (FIGS. 5 and 6).

As can be seen in Figures 5 and 6, in the hot water extraction process, the pectin content and yield increase as the extraction temperature increases, showing the highest yield and pectin content when extracted for 1 hour at 100 ℃.

Example 4: Impurity Removal Process

4-1

 50 g of citrus flour with a particle size of 200 mesh is mixed with 50 ml of distilled water to make a suspension. The mixture is stirred at 500 rpm for 1 hour using a stirrer, filtered under reduced pressure using a miracloth, separated into a residue and a filtrate, and lyophilized. Was measured (Fig. 7).

As can be seen in FIG. 7, the result of analyzing the residue obtained after the filtration and the content of the pectin contained in the filtrate revealed that most of the pectin was present in the filtered residue.

4-2: Pressure Reduction Filtration

The impurity removal process of 4-1 was connected to the previously established homogenization 3 min treatment and hot water extraction process at 100 ° C. to obtain the content and yield of pectin after the sample was treated (FIGS. 8 and 9).

 As can be seen in Figures 8 and 9, it was found that the pectin content significantly increased when the impurity removal process through vacuum filtration was added before the homogenization 3 minutes treatment and the hot water extraction process at 100 ° C.

Example 5: Complex enzyme treatment process

5-1. Selection of Complex Enzymes

To further increase the extraction content of pectin, an enzyme treatment process was added after physical treatment. The enzymes used were celluclast and viscozyme as complex enzymes that degrade the cell wall (see FIG. 10 for the characteristics of each enzyme). Two different enzyme concentrations (100-fold, 500-fold dilution) were added to the sample solution subjected to the above-mentioned physical treatment process, followed by reaction at the optimum temperature of each enzyme (celluclast-55 ° C, viscozyme-40 ° C) for 1 hour. After filtering, the filtrate obtained by filtration was lyophilized to measure the pectin content (FIG. 11).

As can be seen in Figures 11 and 12, regardless of the dilution factor of the enzyme when using the viscozyme enzyme showed a high yield as well as pectin content.

5-2. Determination of enzyme reaction time

After adding 500 times dilute viscozyme enzyme, the enzyme reaction was performed at 40 ° C. for 1, 3, and 5 h to determine the pectin content according to the enzyme reaction time.

As can be seen in Figures 13 and 14, as a result of measuring the pectin content and the yield according to the enzyme reaction time (1, 3, 5h), even if the enzyme reaction is increased for more than 1 hour can obtain a clear effect of improving the pectin content There was no. Therefore, in order to shorten the process time, the enzyme reaction time was set to 1 hour.

5-3. Determination of Enzyme Treatment Concentration

The viscozyme enzyme was added to the sample solution by concentration (100-fold 500-fold 1000-fold dilution) and reacted for 1 hour. The sample solution was filtered and the filtrate was lyophilized to measure the pectin content (FIG. 15).

As can be seen in Figures 15 and 16, when the enzyme was diluted 1000 times, the yield as well as the pectin content was most increased.

Example 6: Autoclave Treatment Process

Instead of hot water extraction for 1 hour at 100 ℃ during the physical treatment process, autoclave treatment is used for shortening the process time and convenience. After the homogenization treatment, the sample solution was autoclave (121 ° C., 10 minutes), and the obtained filtrate after filtration after viscozyme treatment was lyophilized to measure pectin content (FIGS. 16 and 17).

As can be seen in Figure 17 and Figure 18, there was no significant difference between the 100 ℃ hot water treatment and autoclave, the process time can be significantly reduced from 1 hour to 10 minutes when using the autoclave.

1 is a schematic overview of the method of the present invention.

2 is a diagram illustrating a separation process by particle size.

3 and 4 is a comparison of the homogenization and sonication process and the results of the physical treatment process.

5 and 6 is a diagram comparing the hot water extraction process and the results thereof in the physical treatment process.

Figure 7 is a view showing the impurity removal process and its result.

8 and 9 is a diagram showing the additional pressure filtration process and the result.

10 is a table showing enzyme treatment conditions and enzyme composition.

11 and 12 are diagrams showing the process and results for the enzyme selection and the like.

13 and 14 is a view showing a process for determining the enzyme reaction time and the result accordingly.

15 and 16 are diagrams showing a process for determining enzyme treatment concentrations and the results.

17 and 18 are diagrams showing the autoclave treatment process and the result thereof.

Claims (10)

a) mixing and stirring water in the sample; b) physically treating the stirred sample by one or more methods selected from the group consisting of homogenization and hot water extraction by autoclave; And c) enzymatically treating the physically treated result; Without using a chemical solvent and acid or base, characterized in that it is made through a complex treatment by a physical treatment method and an enzymatic treatment method Pectin extraction method. The method of claim 1, Further comprising the pressure reduction filtration process before step b) Pectin extraction method. The method of claim 1, wherein the enzyme is at least one selected from the group consisting of arabase, cellulase, beta-glucanase, hemicellulase, and xylanase. Characterized in that the enzyme Pectin extraction method. The method of claim 1, The enzyme treatment concentration is characterized in that diluted from one hundredth to one thousandth of the total treatment volume Pectin extraction method. delete delete delete delete delete delete

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