JP4649568B2 - Protein removal filter medium, protein removal method, and drooping method - Google Patents

Description

  The present invention relates to a protein removing filter medium for removing protein from a liquid, a protein removing method using the protein removing filter medium, and a drooping method.

  The evaluation items for brewing liquids, particularly sake, include so-called “sae” and “teri”, and sake with white turbidity cannot be rated highly because it is bad. The cause of this white turbidity is protein turbidity called white bokeh and fire caused by fire eradication bacteria. To remove such turbidity, filtration with diatomaceous earth or activated carbon is used. In this case, it is impossible to remove them, so an operation called burning and lowering is required. In addition, when mirin is used for cooking and processing, there may be a cloudiness phenomenon or a cracking phenomenon that reduces the value of the product with the occurrence of bubbles. As a causative substance of this bite, protein can be cited, so it is still necessary to perform a drooping operation.

  As a conventional method for lowering brewing liquids such as sake, tannin, gelatin, silica, silicon dioxide sol, etc. are mixed to forcibly form koji, and precipitate and remove it as a hard flock, protease There are known a method of cleaving the ends of a protein which is a turbid substance, binding the proteins together at the cut portion and aggregating them, a method of filtering through an ultrafiltration membrane, and the like. These methods are described in detail in “Enhanced and revised Sake Manufacturing Technology” published by the Japan Brewing Association.

  However, among the above conventional methods, in the method using tannin, gelatin, silica, silicon dioxide sol, etc., the amount of addition varies depending on the quality of the liquor. There was a problem that it took days or excessive amount of the suspending agent. In particular, gelatin has been avoided from recent BSE (bovine spongiform encephalopathy) problems and allergen labeling problems.

  Also. The method using protease has a problem that it takes a long time to process and it is difficult to mix between lots.

  In addition, the method using an ultrafiltration membrane reduces the filtration rate when the osmotic pressure increases, and also removes components other than the causative agent of the koji so that the taste of the sake changes and the taste is lost. was there.

  In order to solve such a problem, for example, a method (see Patent Document 1) for lowering the brewing liquid by bringing the raw sepiolite into contact with the baked porous body and the brewing liquid has been proposed. There was a problem that coloring may occur. In addition, a method in which soy sauce or the like is brought into contact with a porous body obtained by firing a raw zeolite ore at a temperature of 55 ° C. or higher (see Patent Document 1), or a method in which a brewing liquid is brought into contact with unfired sepiolite in a predetermined temperature range (Patent Document) 2) has also been proposed. However, since the treatment temperature is high, there is a risk that the coloring degree of the product may increase or the scent of the product may change, so that an additional activated carbon treatment step or the like has to be added.

For this reason, there is a need for a technique that can lower the protein by quickly removing the protein in the brewing liquid without impairing the quality. If such a technique can be obtained, it can be applied not only to lowering the brewing liquid, but also to removing proteins from extracts obtained from various animals and plants.
JP 2000-106863 A JP 2000-152765 A JP 2001-128657 A

  The present invention has been made in view of the above problems, and is a protein removing filter medium capable of quickly removing a protein in a target liquid without deteriorating the quality of the liquid, and for removing the protein. It is an object of the present invention to provide a method for removing a protein using a filter medium, and a method for lowering a brewing liquid using the method for removing a protein.

  As a result of diligent research, the present inventors have performed protein removal using a filter medium that has a protein removal performance required for lowering without heating and can quickly remove a protein without causing a change in quality. The technique has been found and the present invention has been completed.

  That is, the filter medium for removing protein according to claim 1 is characterized by comprising a porous body obtained by adding a strong acid to kaolin mineral, and subjecting it to hydrothermal treatment and firing treatment.

  The protein removing method according to claim 2 is characterized in that a strong acid is added to kaolin mineral, and a liquid to be removed is brought into contact with a filter medium made of a porous material obtained by hydrothermal treatment and baking treatment. .

  The invention according to claim 3 is characterized in that, in claim 1, the filter medium is packed in a column, and the liquid to be subjected to the protein removal is passed through the column.

  The invention according to claim 4 is characterized in that, in claim 2 or 3, the liquid to be subjected to protein removal is a liquid obtained from a plant or an animal.

  The dripping method according to claim 5 is characterized in that dripping of the brewing liquid is carried out using the method according to claim 2 or 3.

  According to the invention of claim 1, by bringing the protein removal filter medium into contact with the liquid to be removed, the protein in the liquid can be captured by the filter medium and quickly adsorbed and removed. The protein concentration in this liquid can be reduced quickly and efficiently by removing the protein of this liquid, and at this time, treatment such as heating is unnecessary, and the liquid quality change and the occurrence of coloring are suppressed. It is something that can be done. In addition, the filter medium can be easily regenerated and used repeatedly even if its capacity is reduced.

  According to the second aspect of the present invention, the protein in the liquid to be removed can be trapped by the filter medium and quickly adsorbed and removed, and the protein in the liquid can be removed quickly and efficiently. The protein concentration can be reduced, and at this time, treatment such as heating is unnecessary, and the change in the quality of the liquid and the occurrence of coloring can be suppressed. In addition, the filter medium can be easily regenerated and used repeatedly even if its capacity is reduced.

  According to the invention of claim 3, it is possible to remove the protein from the liquid by continuous treatment, and an operation for separating the liquid and the filter medium is unnecessary, and the protein can be removed more efficiently. Is something that can be done.

  Moreover, according to the invention which concerns on Claim 4, protein can be removed from the liquid obtained from animals and plants, for example, it prevents that the cloudiness etc. which originate in protein arise in the processed goods which used this liquid as a raw material. Can do.

  Moreover, according to the invention which concerns on Claim 5, while removing a protein from a brewing liquid, while being able to perform a drooping quickly and efficiently, generation | occurrence | production of the quality change of a brewing liquid can be prevented. .

  Hereinafter, the best mode for carrying out the present invention will be described.

  The filter medium used in the present invention is obtained using kaolin mineral as a raw material.

  Kaolinite used as a kaolin mineral is a hydrated aluminum silicate, the most typical kaolin mineral. It produces small pseudohexagonal flaky and scaly crystals belonging to the triclinic system. There are also lumps, dense earths, and clay lumps.

  In addition to the kaolinite, the kaolin mineral may be one or more selected from dickite, nacrite, and halloysite as main components, and may be either a natural product or a synthetic product. An example of such a kaolin mineral is Sasame clay.

  A porous material can be obtained by adding a strong acid to the kaolin mineral and subjecting it to a hydrothermal treatment and a baking treatment. This porous material can be used as a filter medium for removing protein (hereinafter referred to as filter medium).

  As the strong acid, one having a pH of 4.0 or less in a state where kaolin mineral is made into a 10% slurry can be used. This strong acid may be either an inorganic acid or an organic acid.

  The hydrothermal treatment is preferably performed at 100 ° C. or higher for 1 hour or longer. The treatment temperature is particularly preferably in the range of 100 to 250 ° C. If the hydrothermal treatment conditions are weak, the resulting slurry or porous body has a small amount of pores. Conversely, if the hydrothermal treatment conditions are too strong, the kaolin mineral will be decomposed and the fuel consumption will be excessive and uneconomical. The problem that there is.

  The slurry or powder after the hydrothermal treatment is granulated as necessary. As the granulation method, any of a number of granulation methods such as a tumbling granulation method, a powder fog drying granulation method, a stirring granulation method, a vacuum drying granulation method, and a fluidized bed granulation method may be used. Here, when the filter medium is packed in a column and used, the smaller the particle size, the larger the contact area with the liquid. Therefore, the filtration efficiency increases. However, if the particle size is too small, the pressure loss is reduced. The problem arises that the flow of liquid becomes worse due to the increase. For this reason, it is preferable that the average particle diameter of a filter medium is 10 micrometers or more, and it is preferable to set it as the range of 100 micrometers-2 mm practically according to the viscosity etc. of a liquid.

  Moreover, although a baking process can be performed on suitable conditions, the range of 350-1000 degreeC is preferable for baking temperature, and the range for 1-5 hours is preferable for baking time. Here, according to the differential thermal analysis data, the water contained in the form of OH in the kaolin mineral begins to dehydrate from around 400 ° C., dehydrates with a large endothermic peak around 600 ° C., and this dehydration is over. After that, it becomes metakaolin, and an exothermic peak due to crystallization of γ-alumina or mullite appears around 970 ° C. to 1000 ° C., and firing shrinkage starts. Therefore, when the firing temperature is 350 ° C. or lower, the water of crystallization is not sufficiently released, resulting in insufficient metakaolinization and a reduced amount of pores. As a result, the amount of pores is reduced. Accordingly, the firing temperature is preferably in the range of 350 to 1000 ° C. as described above.

  The filter medium obtained in this manner changes the crystal structure of the raw kaolinite ore as a raw material in the above production process, a peak of pore size distribution occurs in the range of 50 to 1000 nm and the specific surface area becomes extremely large. It becomes suitable for capturing proteins, and proteins with a molecular weight of about 3500 or more or about 5000 or more can be adsorbed and removed. Here, the protein includes polypeptides. At this time, depending on the filter medium, it is difficult to remove low molecular weight peptides, which are said to be indispensable for umami components, and it is possible to sufficiently reduce only proteins that cause wrinkles and the like.

  When such a filter medium is brought into contact with a liquid containing a protein, a protein having a molecular weight of 5000 or more in the liquid is trapped in the pores of the filter medium and quickly adsorbed and removed, and the adsorbed protein is eluted in the liquid again. Nothing will happen.

  In addition, when the protein trapping performance by such a filter medium is lowered, the filter medium has high chemical resistance and heat resistance, so that the filter medium is treated with various chemicals or subjected to heat treatment, etc. The protein can be easily removed or denatured from the filter medium without alteration, and the filter medium can be regenerated and reused by regenerating the protein capturing performance. Here, various methods can be cited as a method for regenerating the filter medium, for example, treatment with an aqueous solution such as treatment with an aqueous sodium hypochlorite solution, treatment with an aqueous sodium hydroxide solution, treatment with an aqueous urea solution, A baking treatment at a temperature of 500 to 600 ° C. can be mentioned.

  The protein removal performance as described above is suitable for lowering in various brewing liquids such as sake, mirin, fermented seasoning, beer, sparkling sake, soy sauce, white soy sauce, wine, etc. By bringing the brewing liquid into contact with the filter medium, it becomes possible to quickly and efficiently remove the protein that causes koji from the brewing liquid and perform the kneading. In addition, at this time, the filter medium can be processed without heat treatment, and the removal of components other than the causative agent of the koji is also suppressed, and the deterioration of the quality of the brewing liquid due to the koji lowering is suppressed. can do.

  In addition, the removal of protein by the filter medium can be applied not only to lowering the brewing liquid, but also to removing protein from liquids obtained from plants such as vegetables and fruits, and animals such as livestock and seafood. it can. Examples of such liquids include liquids that come out when animals and plants are collected, frozen, and thawed, extracts obtained by treating the animals and plants with water or an organic solvent (such as ethanol), and extracting the umami components of the animals and plants. Can be mentioned. When the protein is removed from the liquid using the filter medium, white turbidity caused by the protein can be prevented from occurring in a processed product using the liquid as a raw material. For example, by removing the protein from the flavor raw material solution extracted from boiled bonito, murojiji, bonito extract, kelp extract, etc. by boiling it in water or hot water, the flavor raw material solution is soy sauce, mirin, fermented seasoning. It is possible to prevent white turbidity from appearing in the soup stock such as udon and soba produced by mixing with ingredients.

  As a specific method for removing the protein from these liquids, for example, by putting a filter medium into the liquid to be removed, it is possible to bring them into contact with each other. At this time, in the process in which the filter medium quickly settles in the liquid, the protein is adsorbed and removed from the filter medium, and the protein is efficiently removed.

  In addition, a method in which a filter medium is packed in a column and a liquid to be removed of protein is passed through the column is also mentioned. In this case, continuous processing is possible by passing a liquid through the column, and an operation for separating the liquid and the filter medium is not required, and the protein can be removed more efficiently.

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

[1] Removal of protein from sake (filter material)
Hydrochloric acid was added to Sasame clay mainly composed of kaolinite produced in the Seto district of Aichi Prefecture to prepare a 10% slurry at pH 0.5. This slurry was sealed in a Teflon (registered trademark) interior type bomb and left at 220 ° C. for 18 hours by circulating hot air drying to perform hydrothermal treatment. The slurry was taken out, filtered, washed with water, dried, and sample The powder was obtained by grinding with a mill.

  This powder and the slurry filtered and washed with water were granulated using a spray dryer to obtain a granulated dried product.

  And the granulated dry body was fired at 700 ° C. for 2 hours in an electric furnace to obtain a granular filter medium having an average particle diameter of 170 μm.

FIG. 1 shows the results of measuring the cumulative pore volume (cm ³ / g) and the log differential pore volume (cm ³ / g) in the pore diameter range of 10 to 100 nm by the mercury intrusion method for this filter medium.

(Untreated sample 1)
Sake produced with a brewing maker in Aichi Prefecture and having a cloudiness that has not been lowered is designated as untreated sample 1.

(Processed sample 1)
3.1 g of the filter medium was packed in a column (manufactured by Biorat) having a diameter of 8 mm and a length of 170 mm.

  And the sample which supplied the said untreated sample 1 from the upper end in the state which has arrange | positioned the said column to the up-down direction, and let the column pass through from the lower end was derived | led-out, and the processed sample 1 was obtained. At this time, a sample of 120 to 140 ml was passed over about 1 to 2 minutes.

(Electrophoresis test)
Each of the untreated sample 1 and the treated sample 1 was dialyzed in distilled water using a dialysis tube (Spectrum. Medical Industries, spec / por3, molecular weight cut off 3500) to remove low molecular weight sugars. After that, freeze-dried and dissolved in 0.8 ml of distilled water, mixed with a sample buffer for SDS-polyacrylamide gel electrophoresis (SDS-PAGE) having the composition shown in Table 1 below at a ratio of 1: 1, An electrophoresis sample was prepared.

  Subsequently, the electrophoresis sample was subjected to SDS-PAGE using an electrophoresis apparatus (anatech Co., Ltd., 3072w type) and a 15% polyacrylamide gel having the composition shown in Table 2 below. In the test, 10 μl of each electrophoresis sample was added to the concentrated gel, and electrophoresed at a constant current of 5 mA until the sample entered the concentrated gel, 10 mA in the concentrated gel, and 15 mA in the separated gel.

  The gel after electrophoresis was stained with a staining solution having the composition shown in Table 3 below, and decolorization was performed with a decoloring solution having the composition shown in Table 4 below.

(Quality test)
The Baume degree, alcohol concentration, acidity, and amino acid degree of each sample were analyzed in accordance with the section of sake in the National Tax Agency's commentary method.

(Results and discussion)
FIG. 2 shows the results of the electrophoresis test for the untreated sample 1 and the treated sample 1. As a result, various proteins ranging from low molecules to high molecules exist in the untreated sample 1, and this is considered to be a causative substance of turbidity. On the other hand, in the treated sample 1, it was confirmed that the protein content was reduced in a wide molecular weight range, and it was confirmed that the removal of the protein by the filter medium was a very useful means for lowering.

  Further, according to the quality test result of each sample, the untreated sample 1 and the treated sample 1 both have a Baume degree of +5, an alcohol concentration of 17.4, an acidity of 19.0, and an amino acid degree of 1.20. No change was observed, and it was confirmed that the quality of sake was not changed by removing the protein with the filter medium, and the low molecular weight peptide was not removed.

[2] Regeneration of filter medium The same filter medium is used to repeatedly remove the protein from the sake, and the sample obtained at the 100th time is treated sample 2.

  Next, with respect to the filter medium after obtaining the treated sample 2, an aqueous solution of sodium hypochlorite having an effective chlorine concentration of 0.1% is circulated in the column for 5 hours, and then distilled water is passed through this column. The filter medium was subjected to a regeneration treatment by washing until the effective chlorine concentration of the effluent derived from No. 1 was 0.1 ppm or less.

  About the filter material after this reproduction | regeneration, the protein removal process was performed with respect to the untreated sample 1 similarly to the removal of the protein from the said sake, and the processed sample 3 was obtained.

  An electrophoretic test was performed on the treated samples 2 and 3. As a result, as shown in FIG. 2, in the processed sample 2 obtained using the repeatedly used filter medium, the protein was not sufficiently removed, whereas it was obtained using the regenerated filter medium. In the treated sample 3, it was confirmed that the protein content was reduced in a wide molecular weight range, and it was confirmed that the protein removal performance of the filter medium was recovered by the regeneration treatment.

3. Removal of protein from mirin (untreated sample 2)
Rice bran was prepared by kneading glutinous rice having a rice polishing ratio of 80% for 46 hours.

  Moreover, the glutinous rice was immersed at 15 ° C. for 6 hours, drained, steamed for 50 minutes, and then allowed to cool to 30 ° C.

  Next, charging was performed with the charging composition shown in Table 5 below, and after charging, a proteolytic enzyme (Sumiteam LP50) manufactured by Shin Nippon Chemical Co., Ltd. was added in an amount of 1 / 10,000 by weight with respect to the total rice, and the lid was covered. After sealing, saccharification and aging were performed in a constant temperature bath at 30 ° C. During this period, once in 7 days, the mash was mixed uniformly, and after 30 days from the preparation date, the mash was left in the upper tank, and left for 20 days. .

(Processed sample 4)
36.7 g of a filter medium (manufactured by Biorat Co., Ltd.) having a diameter of 23 mm and a length of 150 mm was packed with the same filter medium as in the above-mentioned “removal of protein from sake”.

  And the sample which supplied the said unprocessed sample 2 from the upper end in the state which has arrange | positioned the said column in the up-down direction, and let the column pass through from the lower end was derived | led-out, and the processed sample 4 was obtained. At this time, a 500 ml sample was passed over about 3 to 5 minutes.

(Heat-treated sample and comparative samples 1 to 10)
800 ml of the untreated sample was put into a 900 ml capacity bottle, boiled and reached 87 ° C., maintained for 3 minutes, further subjected to heat treatment of water cooling and rapid cooling to 30 ° C. to obtain a heat treated sample.

  To 300 ml of this heat-treated sample, persimmon astringent was added at a ratio shown in Table 7 and stirred for 1 minute. Next, the oridia diluted 50 times was added at the ratio shown in Table 7, stirred for 30 seconds, and allowed to stand for 24 hours to be lowered. Subsequently, the supernatant and the phlegm portion were separated to obtain supernatants as comparative samples 1 to 10.

(Comparative samples 11-18)
To 300 ml of the above-mentioned untreated sample, koji astringent was added at a rate shown in Table 8 without stirring, and stirred for 1 minute. Next, the oridia diluted 50 times was added at the ratio shown in Table 8, stirred for 30 seconds, and allowed to stand for 24 hours to be lowered. Next, the supernatant and the phlegm portion were separated to obtain supernatants as comparative samples 11-18.

(Quality test)
About untreated sample 2, heated sample, treated sample 4 and each comparative sample, Baume degree, alcohol concentration, extract content, pH, acidity, amino acid degree, total sugar content, total nitrogen content, chromaticity (absorbance at 430 nm) Was measured. Baume degree, alcohol concentration, extract content, acidity, amino acid degree are the terms of phosphorus in the analytical method prescribed by the National Tax Agency, the total sugar content is the same as the term of the sake mother and moromi, the total nitrogen content is also the term of the raw rice, pH, chromaticity (430 nm Was also measured according to the sake section. The protein content was derived as the total nitrogen content multiplied by 6.25.

(Nikiri measurement)
About each sample, the quantitative determination of Nikiri was performed by boiling, water addition, an addition reaction, and a TCA reaction. Boiling, hydration, and kneading reactions were each carried out by measurement using the Yamashita method. As for the TCA reaction, 2.5 ml of 50% TCA (trichloroacetic acid) was added to 2.5 ml of the sample and stirred, and then the absorbance at a wavelength of 660 nm was measured with respect to the sample that was allowed to stand for 10 minutes.

  In addition, 150 ml of a sample was put in a pan and the appearance of foaming when heated and boiled was observed. When the water does not aggregate like when boiling water (-), when the foam aggregates over the entire liquid surface (±), not only the bubbles are aggregated but also the foam rises further. The case was evaluated as (+), and the case where the bulges were remarkable was evaluated as (++).

(Test results and discussion)
Test results for each sample are shown in Tables 6-8. In addition, in the evaluation of the comparative samples in Tables 7 and 8, those for which no evaluation is described indicate that the supernatant liquid and the cocoon part could not be separated at the time of lowering and evaluation was impossible.

  As a result, in the processed sample 4, the protein content was greatly reduced. Moreover, it was confirmed that the difference between the boiling reaction, the hydrolysis reaction, the addition reaction, and the TCA reaction was small and the boiling bubbles were small, and the biting phenomenon was reduced. As a result, it was confirmed that in the treated sample 4, the protein causing the biting was removed with high efficiency. Further, in the untreated sample 2 and the treated sample 4, no change was observed in the degree of baume, alcohol concentration, extract content, pH, acidity, amino acid content, and total sugar content, and the quality change occurred due to the protein removal by the filter medium. It was confirmed that the low molecular weight peptide was not removed.

  On the other hand, in the heated sample, the biting phenomenon and the protein content are reduced to some extent, and when heat treatment is performed in combination with astringent astringent or ORIDIA after heat treatment or unheated as in each comparative sample, Although the protein was removed and the biting rate was reduced, it was not sufficient as compared with the treated sample 4, and coloration occurred when heated.

4). Removal of protein from soup stock (untreated sample 3)
9 g of bonito (shaved bonito) was poured into 200 ml of water to suspend it, heated to 100 ° C. over 11 minutes, then held for 5 minutes to boil. This was filtered through two Kimwipes (registered trademark) and the knots were filtered off and cooled to obtain 162 ml of untreated sample 3.

(Processed sample 5)
The same filter medium as in the above-mentioned “removal of protein from sake” was packed in 35.4 g in a column (manufactured by Biorat) having a diameter of 23 mm and a length of 150 mm.

  And the sample which supplied the said untreated sample from the upper end in the state which has arrange | positioned the said column to the up-down direction, and let the column pass through from the lower end was derived | led-out, and the processed sample 5 was obtained. At this time, a 100 ml sample was passed over about 3 to 4 minutes.

(Evaluation test)
About the said untreated sample 3 and the processed sample 5, 30 microliters of electrophoresis samples were prepared with the method similar to the case of said "removal of protein from sake", and the electrophoresis test was implemented.

  The result is shown in FIG.

  Further, the protein content in each sample was measured by a protein assay manufactured by Bio-Rad Co., Ltd. In this case, bovine serum albumin was used as a standard (standardization) as a protein.

  As a result, the measurement result of the protein content in the untreated sample was 0.30 mg / ml, whereas the measurement result in the treated sample 5 was 0.00 mg / ml.

  From the above results, it was confirmed that the protein was removed by the treatment with the filter medium.

It is a graph which shows the result of having measured the pore distribution of the filter medium in an Example. It is the photograph which image | photographed the electrophoretic test result about the processed samples 1-3 in the Example and the unprocessed sample 1. FIG. It is the photograph which image | photographed the electrophoresis test result about the processed sample 5 and the unprocessed sample 3 in an Example.

Claims (5)

  A filter medium for removing a protein, comprising a porous material obtained by adding a strong acid to kaolin mineral and subjecting it to a hydrothermal treatment and a baking treatment.   A protein removal method comprising contacting a liquid to be removed with a filter medium comprising a porous material obtained by adding a strong acid to kaolin mineral and subjecting it to hydrothermal treatment and baking treatment.   The protein removal method according to claim 2, wherein the filter medium is packed in a column, and a liquid to be removed of the protein is passed through the column.   The protein removal method according to claim 2 or 3, wherein the liquid to be subjected to protein removal is a liquid obtained from a plant or an animal.   A method for lowering a brewing liquid using the method according to claim 2 or 3.