KR100827422B1 - Manufacturing method for extract of chlorella - Google Patents

Abstract

The present invention relates to a method for producing a chlorella extract, and more particularly, to obtain a chlorella extract containing chlorella growth factor (CGF). It is about.

The enzyme may be any one or more selected from cell wall degrading enzyme, protease.

Description

Manufacturing method for extract of chlorella}

The present invention relates to a method for producing a chlorella extract, and more particularly, to obtain a chlorella extract containing chlorella growth factor (CGF). It is about.

Chlorella is a kind of freshwater algae belonging to the green algae Chlorellaaceae. It is a spherical or elliptical single cell with a diameter of 10 μm or less commonly found in melted water and wetlands. There is no flagella, no movement, and the cell has one nucleus and one cup-shaped chloroplast. Pyrenoids are often absent. Grown cells proliferate through division, usually four daughter cells form endogenous spores (內 生 胞子). Currently about 10 species are known. They are commonly used in plant photosynthesis research because of their excellent photosynthetic capacity and easy cultivation.

Chlorella contains over 40 nutrients, including 5 major nutrients, and contains large amounts of minerals and minerals that are lacking in our modern people and children, which can solve nutritional imbalances, enhance immunity, release heavy metals, and suppress dioxin absorption As it is known to be effective against, it has become one of the well-being foods recently, and its intake is gradually increasing. Chlorella is an alkaline food with high alkalinity. Consuming chlorella steadily changes its acidity from alkaline to alkaline.

Various studies on the use of chlorella have been made, and among these, there are various methods for obtaining an extract of chlorella.

Conventionally, the method of preparing chlorella extract uses a method of simply extracting hot water by heating a chlorella suspension. This method has the advantage that the extraction process is simple, but the yield of the extract is low.

The present invention is to prepare a chlorella extract by the enzyme treatment and hydrothermal treatment in the chlorella suspension in order to improve the content of the chlorella growth factor and the yield of the extract in the production of chlorella extract.

Meanwhile, as a related art of the present invention, a method for preparing hot water extract from freshwater chlorella containing high iodine obtained by culturing freshwater chlorella in a culture solution containing water-soluble iodide or iodide in Korean Patent Publication No. 1989-0011604 and There is content about use.

However, the present invention is different from the method of producing a high yield of chlorella extract containing chlorella growth factor (CGF) by treating an chlorella suspension with an enzyme such as cell wall enzyme protease.

The present invention is to prepare a chlorella extract that can improve the content of the chlorella growth factor and the yield of the extract during the production of chlorella extract.

An object of the present invention is to provide a method for producing chlorella extract by extracting the chlorella suspension with enzyme treatment and hot water.

Chlorella extract of the present invention can be extracted with an enzyme treatment and hot water and then filtered to obtain a filtrate, can be stored by freeze drying.

On the other hand, the chloral extract of the present invention can be extracted with an enzyme treatment and hot water and then filtered to obtain a filtrate, and the filtrate can be lyophilized to obtain a dried product.

Method for producing a chlorella extract of the present invention for achieving the above-mentioned object is extracted after the reaction by adding an enzyme to the chlorella suspension.

In preparing the chlorella extract in the present invention, the pH of the chlorella suspension is preferably adjusted to 6.0 to 8.0.

In the present invention, the addition of an enzyme to the chlorella suspension may improve the yield of the chlorella extract by extracting the chlorella suspension by adding one or more enzymes selected from cell wall degrading enzyme and protease.

In the present invention, the addition of the enzyme to the chlorella suspension may be performed by adding the cell wall degrading enzyme to the chlorella suspension, followed by the addition of the protease, followed by extraction, thereby improving the yield of the chlorella extract.

In the present invention, the addition of an enzyme to the chlorella suspension may improve the yield of the chlorella extract by extracting the chlorella suspension by simultaneously adding the cell wall degrading enzyme and the protease.

The cell wall degrading enzyme may be any one or more selected from β-1,3-glucanase, beta cellulase, xylanase and pectinase. Can be.

When the cell wall lyase is added to the chlorella suspension in the above reaction, the cell wall lyase may be added at 0.5 to 2.5% of the chlorella solids of the chlorella suspension and reacted for 1 to 3 hours at the optimum temperature and pH of each enzyme.

The protease may be one or more commercial enzymes, and as the commercial enzymes, for example, papain, kojizyme, alkalase, esperase, nutrase, nutrase, Any one or more selected from flavozymes may be used.

The proteolytic enzyme is added to the chlorella suspension as described above, so that the proteolytic enzyme may be added to the chlorella solids in the chlorella suspension at a concentration of 0.5 to 2.5%, and reacted for 1 to 3 hours at the optimum temperature and the optimum pH of each enzyme.

In the above case, the cell wall degrading enzyme and the protease are added to the chlorella suspension at the same time, and the cell wall degrading enzyme is added at the same time as 0.5 to 2.5% of the chlorella solids of the chlorella suspension and the protease is 0.5 to 2.5% of the chlorella solids of the chlorella suspension at the same time. In consideration of the optimum temperature and the optimum pH of the two enzymes can be enzyme reaction for 1 to 3 hours under the appropriate conditions.

In the preparation of the chlorella extract of the present invention, an enzyme is added to the chlorella suspension and reacted to obtain chlorella extract by hydrothermal extraction for 20 to 60 minutes at 90 to 100 ° C.

The chlorella extract of the present invention may be reacted by adding an enzyme to the chlorella suspension, followed by hot water extraction to obtain a chlorella extract, and filtered by filtration means to obtain a filtrate. In this case, any filtering means may be used as long as the filtering means can be commonly used in the art. For example, the chlorella extract may be filtered by filtration means such as filter paper, filter cloth, ultrafiltration, or the like.

Chlorella extract of the present invention can be obtained by freeze drying the chlorella extract filtrate filtered by the filtering means. In this case, the freeze-drying may be carried out under a drying condition that can be commonly used in the art. For example, the chlorella extract filtrate may be frozen at −20 ° C. to 0 ° C. and then dried under reduced pressure to obtain a dried product of the chlorella extract.

The content of CGF in the chlorella extract prepared in the present invention can be measured by absorbance at 260 nm after diluting the extract, and the yield of the chlorella extract can be confirmed by measuring the weight by lyophilizing the chlorella extract.

In preparing the chlorella extract of the present invention, the chlorella extract was prepared from the chlorella suspension under various conditions. In order to obtain the chlorella extract according to the object of the present invention, it is preferable to prepare the chlorella extract under the aforementioned conditions.

On the other hand, the present invention includes a dietary supplement containing the chlorella extract prepared above.

The health supplement food containing the chlorella extract of the present invention may be used in the form of a liquid or the chlorella extract prepared in the above form in powder form.

The health supplement food containing the chlorella extract of the present invention is added to the health supplement food manufacturing process by adding the chlorella extract prepared by the above-mentioned method in the form of liquid or powder, or after the preparation of the base food. It can be easily obtained by adding a step of adding the chlorella extract of the present invention. At this time, the health supplement food containing chlorella extract may add an additive that can be used in foods to improve the taste and odor as necessary.

The content of the chlorella extract in the dietary supplement containing the chlorella extract of the present invention varies depending on the type of food, the taste, and the like, but is used in products such as tablets, capsules, pills, powders, and the like. E. chlorella extract may contain 0.1 to 10% by weight in liquid or powder form.

Hereinafter, the content of the present invention will be described in detail through examples. However, these are intended to explain the present invention in more detail, and the scope of the present invention is not limited thereto.

<Example 1>

Tunicase (trade name of β-1,3-glucanase, manufactured by Daiwa Kasei, Japan) was added as chlorella suspension (2%, pH 8.0) as 0 to 2.5% of chlorella solids, respectively. After reacting for 3 hours at 40 ℃ and extracted with hot water for 30 minutes at 95 ℃ to prepare a chlorella extract.

The chlorella extract prepared above was diluted 50 times to measure CGF with absorbance of 260 nm, and the chlorella extract prepared above was filtered through filter paper, and then frozen at -20 ° C. and freeze-dried under reduced pressure to measure the weight of chlorella extract. The yield of was measured and the results are shown in Table 1 below.

Table 1. CGF content and yield of chlorella extract according to the amount of tunicaase

Enzyme amount CGF content (260 nm absorbance, 50 fold dilution) Extract yield (%) 0% (control) 0.617 12.8 0.5% 0.741 18.8 1.0% 0.746 24.0 2.0% 0.747 23.7 2.5% 0.741 23.8

As shown in the results of Table 1 above, the yield of the extract is the best when the tunicaase is used as 1% of the chlorella solids during the enzymatic reaction by adding tunicaase as a cell wall enzyme to the chlorella suspension, especially in terms of the yield of the extract compared to the control. 2 times the increase was found to be.

<Example 2>

A chlorella extract was prepared by extracting chlorella suspension (2%, pH 7.5) at 95 ° C. for 30 minutes.

Papain as a protease was added to 1.0% of chlorella solids in a chlorella suspension (2%, pH 6.0), reacted for 3 hours at 40 ° C, and hydrothermal extraction for 30 minutes at 95 ° C to prepare a chlorella extract.

A chlorella extract was prepared by adding kojizyme as a protease to 1.0% of chlorella solids in a chlorella suspension (2%, pH 6.0), reacting for 3 hours at 40 ° C, and hydrothermal extraction for 30 minutes at 95 ° C. .

Alkalase as a protease was added to 1.0% of chlorella solids in a chlorella suspension (2%, pH 7.5) and reacted for 3 hours at 40 ° C, followed by hydrothermal extraction for 30 minutes at 95 ° C to prepare chlorella extract. It was.

Chlorella extract was prepared by adding esperase (1.0%) of chlorella solids as a protease to chlorella suspension (2%, pH 8.0), reacting for 3 hours at 40 ° C, and hydrothermal extraction for 30 minutes at 95 ° C. .

Each chlorella extract prepared above was diluted 50 times to measure CGF with an absorbance of 260 nm, and the chlorella extract prepared above was filtered through a filter paper, frozen at -20 ° C, lyophilized, and freeze-dried to weigh. The yield of the chlorella extract was measured and the results are shown in Table 2 below.

Table 2. CGF content and yield of chlorella extracts according to proteolytic enzymes

enzyme pH * CGF content (260 nm absorbance, 50 fold dilution) Extract yield (%) Control 7.5 0.505 12.0 papain 6.0 0.472 14.5 Kojizyme 6.0 0.499 15.5 Alkalase 7.5 0.566 20.8 Esperase 8.0 0.569 30.5

PH of chlorella suspension

As shown in the results of Table 2, it was found that the yield of the hydrothermally extracted chlorella extract was excellent after the addition of the proteolytic enzyme and the reaction compared to the hydrolyzed chlorella extract without the addition of the proteolytic enzyme. In particular, it was found that the CGF content and the chlorella extract yield were excellent in the hydrolysis of the proteolytic enzymes added to the chlorella suspension after the addition of alkalase and esperase.

<Example 3>

Esperase (produced by Novo) as a protease was added to chlorella suspension (2%, pH 8.0) at 0-2.5% of chlorella solids, respectively, reacted at 50 ° C for 3 hours, and then at 95 ° C for 30 minutes. Hot water extraction to prepare a chlorella extract.

The chlorella extract prepared above was diluted 50 times to measure CGF with absorbance of 260 nm, and the chlorella extract prepared above was filtered through filter paper, and then frozen at -20 ° C. and freeze-dried under reduced pressure to measure the weight of chlorella extract. The yield of was measured and the results are shown in Table 3 below.

Table 3. CGF Content and Yield of Chlorella Extract According to Esperase Content

Enzyme amount CGF content (260 nm absorbance, 50 fold dilution) Extract yield (%) 0% (control) 0.565 12.5 0.5% 0.551 19.5 1.0% 0.569 31.0 2.0% 0.611 31.0 2.5% 0.521 31.5

As shown in the results of Table 1, when the enzyme was added to the chlorella suspension as a protein enzyme, the yield of the extract was increased about 2 times compared to the control when the enzyme was used as the chlorella solids 1% during the enzyme reaction. On the other hand, when using esperase more than 1% of chlorella solids, there is no difference in extract yield. Therefore, it is recommended to add esperase as a protein enzyme to chlorella suspension when preparing chlorella extract, and to use 1% of chlorella solids during enzyme reaction. good.

<Example 4>

Tonicase (made by Daiwa Kasei, Japan) was added to a chlorella suspension (2%, pH 8.0) as a cell wall degrading enzyme at 1.0% of chlorella solids and allowed to react at 40 ° C for 3 hours. Then, as a protease, esperase was added at 1.0% of chlorella solids and reacted at 50 ° C for 3 hours. After completing the enzymatic reaction of the cell wall enzyme and protease, hydrochloric acid was extracted for 30 minutes at 95 ° C. to prepare chlorella extract (Experiment 1).

Tonicase (made by Daiwa Kasei, Japan) was added to chlorella suspension (2%, pH 8.0) as 1.0% of chlorella solids as a cell wall degrading enzyme, and at the same time, esperase was added as 1.0% of chlorella solids as protease. The enzyme was added and reacted at 50 ° C. for 3 hours. After the enzymatic reaction was terminated by hot water extraction for 30 minutes at 95 ℃ to prepare a chlorella extract (Experiment 2).

Chlorella suspension (2%, pH 8.0) was extracted with hot water at 95 ° C. for 30 minutes to prepare chlorella extract without enzymatic reaction (control).

50 mg of each of the chlorella extracts of the control group, experimental group 1, and experimental group 2 prepared above were measured by absorbance at 260 nm, and the chlorella extract prepared above was filtered with filter paper and then filtered at -20 ° C. Freeze, depressurize and freeze-dried to determine the weight of the chlorella extract yields and the results are shown in Table 4 below.

Table 4. CGF content and yield of chlorella extract according to enzyme treatment

Enzyme treatment CGF content (260 nm absorbance, 50 fold dilution) Extract yield (%) No treatment (control) 0.614 13.3 Sequential Processing (Experiment 1) 0.675 38.8 Concurrent Processing (Experiment 2) 0.748 43.3

As shown in the results of Table 4, the chlorella extracts of Experiment 1 and Experiment 2 subjected to enzymatic reaction and hydrothermal extraction of chlorella suspension were compared to the control chlorella extract of hydrochloric acid without chlorella suspension when the chlorella extract of the present invention was prepared. It was found that a large amount of CGF, the extract yield was also excellent.

On the other hand, in the chlorella extracts of Experimental Zone 1 and Experimental Zone 2, which were extracted after the enzymatic reaction to the chlorella suspension, cell wallase and proteolytic enzyme were simultaneously treated compared with Experimental Zone 1, which sequentially treated cell wall and protease. Chlorella extract of Experiment 2 was found to have better CGF content and extract yield.

As described above, although described with reference to a preferred embodiment of the present invention, those skilled in the art will be variously modified and modified within the scope of the present invention without departing from the spirit and scope of the present invention described in the claims below. It will be appreciated that it can be changed.

As the result of the above example, the chlorella extract extracted after reacting any one or more enzymes selected from cell wall degrading enzyme and proteolytic enzyme to the chlorella suspension has more chlorella growth factor (CGF) content than the chlorella extract extracted without reacting the enzyme. In addition, it can be seen that the extract yield is more improved.

Claims (7)

In the preparation of chlorella extract, After reacting xylanase as a cell wall degrading enzyme in a chlorella suspension of pH 7.0-8.0 with 1.0-2.5% of the chlorella solids of the chlorella suspension, Chlorella, characterized in that hydrothermal extraction at 90-100 ℃ for 20-60 minutes after reacting by adding at least one proteolytic enzyme selected from papain, cozyme, alkalase 1.0-2.5% of the chlorella solids of the chlorella suspension Method of Preparation of Extracts. delete delete delete delete delete delete