JPH08322547A - Protein separation from liquid material - Google Patents

Abstract

PURPOSE: To efficiently separate protein from a liquid material in preventing destruction of floc without losing active components by mixing a liquid material containing protein with silica sol, mixing the mixture with chitosan and coagulating the protein. CONSTITUTION: Protein is separated by adding silica sol and chitosan to a liquid material containing protein to coagulate the protein, or adding silica sol to the material to coagulate the protein and adding chitosan to the resultant protein, blending and filtering to separate the protein. Adding amount of the chitosan is preferably 10-100g to 1kL of the liquid food.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a method for separating a protein from a liquid containing the protein.

[0002]

2. Description of the Related Art As a method for separating proteins from liquid substances containing proteins such as sake, mirin, wines, soy sauce, etc., there is known a method of separating proteins by aggregating or aggregating and precipitating them. For example, a method using silica sol is known in the process of hanging down sake (Japanese Patent Publication No. 59-33351). In addition to silica sol,
It is known that the use of a protein such as gelatin or a water-soluble gelatin obtained by lowering the molecular weight of gelatin or a low molecular weight gelatin further promotes aggregation and precipitation.

[0003]

However, the flocs formed by coagulating and precipitating gelatin and silica sol, the flocs are destroyed and the proteins are dispersed again in the filtration step involving pressurization and contact with water, Since a part thereof passes through the filtration membrane, it has a drawback that sufficient protein separation may not be possible.

The object of the present invention is to provide a method for solving such conventional problems, preventing the destruction of flocs, and efficiently separating a protein from a liquid substance without loss of an active ingredient. Especially.

[0005]

Means for Solving the Problems As a result of intensive investigations by the present inventors, a method capable of preventing the destruction of flocs in such a filtration step and efficiently separating a protein from a liquid matter was obtained. By using chitosan, it was found that the floc was prevented from being destroyed and the protein could be separated very efficiently, and the present invention was completed.

[0006] That is, the present invention is a method of adding silica sol to a liquid containing a protein to separate the protein by aggregating or aggregating and precipitating the protein. Is added. Here, the "process until filtration" means that the filtration process itself is included.

The first embodiment according to the present invention is a method of adding silica sol to a liquid substance containing a protein to cause the protein to aggregate and settle and separate, and is characterized in that silica sol and chitosan are added to the liquid substance. .

A second embodiment according to the present invention is a method in which silica sol is added to a liquid containing a protein to cause the protein to aggregate and settle to separate, and the aggregate is filtered.
The method is characterized in that after the chitosan is added to and mixed with the agglomerate, that is, after chitosan is body-fed, the mixture is filtered.

A third embodiment according to the present invention is a method in which silica sol is added to a liquid substance containing a protein to separate the protein by aggregating or aggregating and precipitating the protein, and further filtering the agglomerate with a filter. Is characterized in that it is filtered using a filter precoated with.

In the present invention, the agglomerate containing chitosan according to the first or second embodiment may be filtered using a filter precoated with chitosan according to the third embodiment.

The chitosan used in the present invention is known as a deacetylated product of chitin and is not particularly limited, but the deacetylation degree is 70.
% Or more, more preferably 85% or more. The form is not particularly limited, but when used for body feed or precoat, fine powder is preferable.

In the present invention, the amount of chitosan added is appropriately adjusted depending on the type of liquid product and the protein content of the liquid product, but in general, it is preferably about 10 to 100 g per kiloliter of liquid food. Second according to the invention
In the embodiment, chitosan is added to and mixed with the aggregate, and in the third embodiment, filtration is performed using a filter precoated with chitosan. The amount of chitosan used at this time is
It can be determined by converting it to the liquid substance that is the object of treatment,
For example, it is preferable to use chitosan in the above-mentioned addition amount, that is, 10 to 100 g per 1 kiloliter of the liquid substance.

The silica sol used in the present invention is not particularly limited, and a wide range of general silica sol can be used. Usually, a silica sol having a silica content of about 15 to 45% by weight is preferable. As a commercially available product, COPOLOK 300 (manufactured by Otsuka Chemical Co., Ltd., silica content: 30% by weight) can be particularly preferably used. Regarding the amount of silica sol added, in the case of silica sol having a silica content of 30% by weight,
Generally 100 to 300 for 1 kiloliter of liquid food
About 0 ml, preferably about 300 to 2000 ml is preferable. Silica content 20% by weight
In the case of the silica sol described above, about 300 to 3000 ml is preferable.

In the first embodiment according to the present invention, silica sol and chitosan are added to the liquid material. The order of adding silica sol and chitosan is not particularly limited, and they can be added in any order. After the silica sol and chitosan are added to cause the protein to aggregate and settle, the aggregate can be filtered by one or more filtration steps if necessary. Pressure filtration may be employed for such filtration.

In the second and third embodiments according to the present invention, silica sol is added to aggregate and precipitate the protein, and the aggregate is separated and filtered by a filter. Second
In the embodiment, after the chitosan is added to and mixed with the agglomerate, that is, the body is fed and then filtered. Also the third
In one embodiment, the filtration is performed using a filter precoated with chitosan. As a method of pre-coating on a filter, a method of dispersing chitosan in a liquid such as water, supplying this to a filter and filtering it to form a uniform coat layer of chitosan on the filtration surface of the filter, etc. Can be adopted.

In the present invention, when chitosan is used for body feed or precoat, it is possible to mix chitosan with a filter aid such as diatomaceous earth or cellulose or an adsorbent such as activated carbon.

In the present invention, proteins such as gelatin and wheat flour, polysaccharides such as alginic acid and carrageenan, persimmon astringent, tannic acid and the like can be used in combination within a range that does not impair the effects of the present invention. By using these in combination, the aggregation and sedimentation effect can be further enhanced.

Further, activated carbon may be used in order to adsorb pigments, impurities and the like in the step of flocculation and sedimentation. The amount of such activated carbon used is, for example, 100 to 50.
Generally, about 00 g / kiloliter is used.

The liquid material to which the present invention is applied is not particularly limited as long as it contains protein. For example, sake, mirin, wines, soy sauce, vinegar, animal and plant protein hydrolyzate, sauce , Extract-based seasoning microorganism culture fluid,
A protein liquid such as an enzyme liquid may be used. In addition, the present invention can be applied at various stages in the production process of these liquids, from raw material adjustment, purification, to waste liquid treatment.

[0020]

In the present invention, by adding chitosan, the floc of the protein aggregate becomes a firm and firm floc as compared with the conventional one. For this reason, the flocs are not easily broken, and the flocs are not destroyed even in a filtration process or the like involving pressurization. Therefore, unlike conventional methods, flocs are not destroyed and proteins are not redispersed in the filtration step. In the present invention, chitosan may be added in the flocculation / sedimentation step, or may be added to and mixed with the flocculate or the precipitate. The chitosan in this case may be liquid or solid such as fine powder.
In both cases, good flocs are formed.
Further, such a good floc can also be formed by filtering using a filter precoated with chitosan. In this case, chitosan is preferably solid such as fine powder.

Further, the flocs of the protein aggregates formed by the present invention have a small bulk, so that the proteins can be removed extremely efficiently.

[0022]

Example 1 In 500 ml of sake before being lowered, 0.25 g of activated carbon (manufactured by Takeda Pharmaceutical Co., Ltd., "Special Egret Egret"), silica sol (manufactured by Otsuka Chemical Co., Ltd., trade name "COPOLOK 300", silica content 3)
0.2 wt%) and 30 mg of chitosan (trade name "SK-50" manufactured by Koyo Chemical Co., deacetylation degree of 85% or more) were added, and allowed to stand for one day for clarification, and then the supernatant liquid Was measured for turbidity. The results are shown in Table 1.

Next, the supernatant was removed, and 1 g of diatomaceous earth was body-fed to the remaining black stain to filter (filter: "ADVANTEC 5A" manufactured by Advantech Toyo Co., Ltd., filter paper: "ADVANTEC KS-90-UH"). Advantech Toyosha). 500 ml of pre-filtered sake was put into this filter and filtered. The turbidity of the liquid after filtration (the turbidity of sake after re-filtration) was measured. The results are shown in Table 1.

Next, in order to recover the alcohol remaining in the black stain, water is put into the filter in which the black stain remains and the mixture is filtered at a set pressure of 1.0 kgf / m ² to obtain the turbidity of the filtrate ( The turbidity of water after filtration) was measured. The results are shown in Table 1.

Comparative Example 1 Instead of chitosan, 30 mg of low molecular weight gelatin (manufactured by Tomiyama Co., Ltd., trade name “Purified gelatin”, which was obtained by decomposing gelatin with an enzyme to make it soluble in cold water) was used. Coagulation, sedimentation and filtration were performed in the same manner as in. Table 1 shows the turbidity of the supernatant after standing, the turbidity of sake after re-filtration, and the turbidity of water after filtration.

Comparative Example 2 30 mg of high-molecular gelatin (manufactured by Nitta Gelatin Co., Ltd., jelly strength: 200 bloom) was used in place of chitosan, and coagulation sedimentation and filtration were carried out in the same manner as in Example 1 except that. Table 1 shows the turbidity of the supernatant after standing, the turbidity of sake after re-filtration, and the turbidity of water after filtration.

[0027]

[Table 1]

As is clear from Table 1, in Comparative Example 1 using low molecular weight gelatin, the clarity of the supernatant is good, but the turbidity of sake after re-filtration and the turbidity of water after filtration. It turns out that the flock has been destroyed. In Comparative Example 2 using high-molecular gelatin, the turbidity of sake after re-filtration and the turbidity of water after filtration are low, but the turbidity of the supernatant is high, and therefore the supernatant is filtered. It takes a long time to do. On the other hand, in Example 1 in which chitosan was used according to the present invention, the clarity of the supernatant was good, and the turbidity of the filtrate did not increase even during filtration of the cage,
Cages or protein aggregates can be filtered without destroying the flocs.

Further, in order to compare the filtration rates, silica sol and chitosan or low molecular weight gelatin were added to the sake before hanging in Example 1 and Comparative Example 1 and clarified by leaving still for one day. It filtered using filter paper (5C), stirring, and measured the filtration rate. The filtration rate was determined by measuring the amount of filtrate after 5 minutes and 10 minutes. The amount of filtrate after 10 minutes and after 20 minutes is shown in Table 2.

[0030]

[Table 2]

As is clear from Table 2, when chitosan is added according to the present invention, the filtration rate is higher than that when conventional low molecular weight gelatin is added.

Example 2 and Comparative Example 3 In 500 ml of sake before hanging, 0.25 mg of activated carbon,
0.3 ml of silica sol and 15 mg of low molecular weight gelatin were added, and the mixture was allowed to stand for one day for clarification. The black portion was taken out, 1 g of diatomaceous earth and 5 mg of chitosan were fed as a body, and placed in a filter. 500 ml of pre-filtered sake was put into this filter and filtered under a pressure of 1.0 kgf / m ² , and the turbidity of the filtrate was measured. The results are shown in Table 3.

Next, in order to recover the alcohol remaining in the black stain, water was put into the filter in which the black stain remained.
The mixture was filtered under a pressure of 0 kgf / m ² and the turbidity of the filtrate was measured. The results are shown in Table 3.

As a comparison, the turbidity of sake after re-filtration and the water after filtration when the same procedure as in the above example was conducted except that 1 g of diatomaceous earth was added without adding chitosan. Table 3 also shows the turbidity.

[0035]

[Table 3]

As is clear from Table 3, when chitosan was added to the protein black aggregate, which is a protein aggregate, and filtered in accordance with the present invention, the turbidity of the sake after re-filtration was reduced, and the turbidity of the water after filtration was reduced. The degree is getting smaller. Therefore, it can be seen that the flocs of the aggregates are less likely to be destroyed.

Example 3 To 500 ml of undiluted sake, 0.25 g of activated carbon, 0.3 ml of silica sol, and 15 mg of low-molecular gelatin were added and allowed to stand for one day for clarification. Chitosan (50 mg) was dispersed in 200 ml of ion-exchanged water in advance, and filter paper was precoated by a conventional method. 1 g of diatomaceous earth was added to the black stain generated by the hanging, and the body was fed and placed in this filter. 500 ml of pre-filtered sake was placed in this filter and filtered. The turbidity of the filtrate was measured, and the results are shown in Table 4.

Next, in order to recover the alcohol contained in the black stain, water was put into the remaining filter of the black stain to obtain 1.0 kgf.
The mixture was filtered under a pressure of / m ² and the turbidity of the filtrate was measured.
The results are shown in Table 4.

[0039]

[Table 4]

As is clear from the results in Table 4, the amount of chitosan used is slightly increased, but the turbidity and water content of the sake after re-filtration can also be obtained by filtering the chitosan with a filter pretreated. It is possible to reduce the turbidity of the water after the filtration when the is filtered, and it is possible to make the flocs of the aggregate less likely to be broken.

[0041]

EFFECTS OF THE INVENTION According to the present invention, when the protein is aggregated and settled from the liquid material to be separated, the flocs of the aggregate can be made less fragile than the conventional flocs. Therefore, the flocs are prevented from being destroyed even in the filtration step involving pressurization, and the protein can be separated almost completely and efficiently.

Further, according to the present invention, since the filtration rate can be improved, the processing time can be shortened. Furthermore, since the bulk of the agglomerates can be made smaller than in the conventional case, it contributes to the reduction of the amount of waste and the reduction of the load on the environment.

Claims (4)

[Claims] 1. A method for separating a protein by aggregating proteins by adding silica sol to a liquid containing a protein, wherein chitosan is added in the steps up to filtration of the agglomerates. Method. 2. A method for separating a protein by adding silica sol to a protein-containing liquid material to aggregate and separate the protein, wherein silica sol and chitosan are added to the liquid material. 3. A method of adding silica sol to a liquid containing a protein to aggregate the protein and further filtering the aggregate, wherein chitosan is added to and mixed with the aggregate and then filtered. Method for separating proteins from substances. 4. A method in which silica sol is added to a protein-containing liquid material to agglomerate proteins, and the agglomerates are filtered by a filter, which is characterized in that a filter precoated with chitosan is used for filtration. A method for separating proteins from a liquid material.