JP3383851B2 - Functional protein material and method for producing the same - Google Patents

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a functional protein material and a method for producing the same. More specifically, the present invention relates to an organic solvent-soluble functional protein material and a method for producing the same.

[0002]

2. Description of the Related Art Conventionally, attempts have been made to add a protein to synthetic fibers in order to improve the feel of the synthetic fibers, the inhalation and release of moisture, and the heat retention.

For example, Japanese Patent Application Laid-Open No. 1-293143 proposes that fine powder of gelatin and silk is dispersed in a synthetic resin. However, there are variations in the particle size of the fine powder, and the fine powder is Since it is not uniformly dispersed in the resin, the synthetic fibers produced from the synthetic resin are not satisfactory in terms of touch, balance between inhalation and release of moisture, and heat retention. Moreover, the fine pulverization of gelatin and silk has a problem that handling is very difficult due to the complexity of the pulverization operation and the scattering of pulverized products.

However, in order to improve the dispersibility of the protein in the resin, it is obvious that preferable results can be obtained if the protein can be dissolved in the organic solvent that dissolves the resin.

As a protein which can be dissolved in an organic solvent, for example, a simple protein, protamine, is known, which is 60 to 90% ethanol (that is, an aqueous organic solvent).
However, it is not soluble in 90% or more of higher-purity ethanol and other organic solvents and cannot be put to practical use.

Further, Japanese Patent Application Laid-Open No. 52-25800 discloses that
A modified protein obtained by hydrolyzing an addition reaction product of a protein and an isocyanate compound in the presence of an organic solvent or an aqueous organic solvent can be used in an organic solvent such as dimethyl sulfoxide (DMSO), dimethylacetamide (DMA), or tetrahydrofuran (THF). It is stated that it is molten. However, such a protein has an infrared absorption around 1740 cm ^-1 ,
Since the side chain having an amide bond or urethane bond near 1222 cm ^-1 is cleaved and only the side chain having a urea bond remains, it is a so-called low molecular weight protein denatured product, which may reduce the function as a protein. is there. Moreover, the solubility of the modified protein in an organic solvent is not sufficient.

[0007]

Means for Solving the Problems As a result of intensive studies to solve the above-mentioned problems of the prior art, the present inventor found that a protein polymerized by cross-linking, not a hydrolyzate,
Succeeded in obtaining a material that has the property of being soluble in organic solvents,
The present invention has been completed.

That is, the present invention relates to the following functional protein material and a method for producing the same.

A functional protein material having the following physicochemical properties, which was obtained by adding an acid to diisocyanate as a crosslinking agent during the production. The material is obtained, for example, in Example 1 described later.

(A) Amino acid composition: Substantially the same as the starting protein.

(B) Solubility: Soluble in dimethylformamide, dimethylsulfoxide and dimethylacetamide but not in water.

(C) Weight average molecular weight: 100,000 to 300
10,000, preferably 400,000 to 1.5 million (GPC / LALLS
By law).

(D) Infrared absorption: 3 compared to the starting protein
Weak absorption of 440cm ^-1 ~3420cm ^-1, 1603
shoulder around cm ^-1, having a bottom in the vicinity of 1353cm ^-1, a peak in the vicinity of 1222cm ^-1 is high.

(E) Crosslinked structure: Protein is crosslinked by the reaction of diisocyanate (SDS-electrophoresis).

A functional protein material having the following physicochemical properties, which was obtained by using a diisocyanate as a crosslinking agent and further adding an alkylating agent during the production. The material is obtained, for example, in Example 2 described later.

(A) Amino acid composition: Substantially the same as the starting protein.

(B) Solubility: Soluble in dimethylformamide, dimethylsulfoxide and dimethylacetamide but not in water.

(C) Weight average molecular weight: 100,000 to 300
10,000, preferably 400,000 to 1.5 million (GPC / LALLS
By law).

(D) Infrared absorption: 3 compared to the starting protein
Weak absorption of 440cm ^-1 ~3420cm ^-1, 1740
cm in the vicinity of ^-1 has a shoulder peak in the vicinity of 1222Cm ^-1 increases.

[0020] Incidentally, in the case of using a large excess of diisocyanate, in addition to the infrared absorption, the bottom is observed in the vicinity of the shoulder and 1353cm ^-1 in the vicinity of 1600 cm ^-1 (e.g. Examples below 17).

(V) Crosslinked structure: Protein is crosslinked by the reaction of diisocyanate (SDS-electrophoresis).

A functional protein material having the following physicochemical properties, which was obtained by using a diisocyanate as a cross-linking agent and further adding a Schiffing agent at the time of production. The material is obtained, for example, in Example 10 described later.

(A) Amino acid composition: Substantially the same as the starting protein.

(B) Solubility: Soluble in dimethylformamide, dimethylsulfoxide and dimethylacetamide but not in water.

(C) Weight average molecular weight: 100,000 to 3,000,000 (by GPC / LALLS method).

(D) Infrared absorption: 3 compared to the starting protein
Weak absorption of 440cm ^-1 ~3420cm ^-1, 1740
cm ^-1 and in the vicinity of 1600cm no shoulder in the vicinity of ^-1.

(V) Crosslinked structure: Protein is crosslinked by the reaction of diisocyanate (SDS-electrophoresis).

A method for producing a functional protein material, which comprises mixing an aqueous protein solution and a cross-linking agent, and then adding at least one selected from an alkylating agent, a Schiff's agent and an acid.

In order to produce this protein material (hereinafter referred to as "this material"), first, an aqueous protein solution and a crosslinking agent are mixed,
A protein and a crosslinking agent are subjected to addition polymerization to obtain a crosslinked product of the protein.

The protein used in the present invention is not particularly limited, but those having a free amino group are preferable, and examples thereof include egg white protein of eggs such as chicken, quail, duck and goose, whey protein, serum protein and casein. Can be mentioned. The concentration of the aqueous protein solution is not particularly limited, but it is usually preferably about 1 to 5% by weight.

The cross-linking agent that reacts with proteins can react with free amino groups and alcoholic hydroxyl groups between proteins to cross-link the proteins by chemical bonds such as urea bond, urethane bond and acid amide bond. There is no particular limitation as long as it can control the hydrophobicity of the protein, and examples thereof include diisocyanate compounds, dialdehyde compounds and diketone compounds. Among them, the diisocyanate compound is highly reactive and can be preferably used. As the diisocyanate compound, conventionally known compounds can be widely used, and examples thereof include toluene diisocyanate (TDI), diphenylmethane diisocyanate (MDI), hexamethylene diisocyanate (HDI), and isophorone diisocyanate (IPD).
I), naphthalene diisocyanate (NDI), etc. 1
Examples thereof include compounds having two or more isocyanate groups in the molecule. The amount of the cross-linking agent used is not particularly limited, but usually the total number of moles of amino groups and alcoholic hydroxyl groups is calculated from the primary structure of the protein to be reacted, and the amount of cross-linking agent used (the number of moles) is determined accordingly. Good.

The reaction between the protein and the crosslinking agent is usually carried out by mixing (preferably vigorously mixing) the aqueous protein solution and the crosslinking agent. The condition of this reaction is not particularly limited as long as it causes crosslinking between proteins, but it is usually carried out at room temperature for about 1 hour or more.

In the present invention, a solution prepared by dissolving a crosslinking agent in an organic solvent may be reacted with an aqueous protein solution. In that case, the aqueous protein solution and the organic solvent solution of the cross-linking agent are mixed, the mixed liquid is separated into an aqueous phase and an oil phase, and the aqueous phase containing the cross-linked protein is separated and subjected to the next step. . As the organic solvent, a known organic solvent that enables the interfacial addition polymerization of the protein and the crosslinking agent can be used, and examples thereof include chloroform, hexane, and toluene.

After mixing the aqueous protein solution and the cross-linking agent, a mixed solution (if the aqueous solution of the protein and the organic solvent solution of the cross-linking agent are mixed, a separated aqueous phase. The oil phase also contains the protein. It is possible that the present material is produced by adding at least one selected from an alkylating agent, a Schiffing agent and an acid.

First, the case of adding an alkylating agent will be described. It is presumed that the addition of the alkylating agent alkylates (modifies) the residual amino groups, the phenolic hydroxyl groups and the carboxyl groups in the crosslinked product of the protein to obtain the intended material of the present invention as a gel. A wide variety of known alkylating agents can be used, and examples thereof include dialkyl sulfates such as dimethyl sulfate, alkyl sulfates, alkyl halides, and alkyl sulfonates. The use of dialkylsulfate, alkylsulfate, etc., not only has an effect as an alkylating agent but also has an effect as a pH adjusting agent. The addition amount of the alkylating agent may be appropriately selected according to the degree of the above crosslinking reaction, etc., but it is usually assumed that all amino groups are crosslinked, and the phenolic hydroxyl group and the carboxyl group are extracted from the primary structure of the protein to be reacted. The total number of moles of the crosslinking agent may be calculated, and the amount of the crosslinking agent used (number of moles) may be determined accordingly. The gel-like product obtained can be used as it is as the main material, but after removing water by usual separation means such as centrifugation and filtration, washing with water as necessary, and then using a vacuum belt dryer or freeze-drying. It may be dried with a machine and used as a powder product.

Next, the case of adding a shift agent will be described. The Schiffing agent modifies the amino group by performing a Schiffing reaction with the residual amino group in the protein cross-linked product. By adding a Schiffing agent, the present material can be obtained as a gel-like material as in the case of the alkylating agent. Examples of the Schiffing agent include conventionally known ones such as aldehydes. The obtained gel-like material can be separated and dried as described above.

The case where an acid is further added will be described. Acid is added to the aqueous phase to adjust the pH of the phase below the isoelectric point of the starting protein. As a result, a substance that is soluble in the organic solvent and insoluble in water is obtained. The resulting precipitate can be pulverized by the same separating means and drying means as above. Any known acid can be used as the acid, for example, hydrochloric acid,
Examples thereof include citric acid, succinic acid, acetic acid, lactic acid, tartaric acid, fumaric acid, malic acid, adipic acid, gluconodeltalactone, gluconic acid, ascorbic acid, levulinic acid and phthalic acid. If the crosslinking is insufficient or the hydrophobicity is insufficient, a precipitate will be deposited even if the pH is lowered by adding an acid, but this material may not be produced. Therefore, when the acid is used alone, the amount of the cross-linking agent used in all the steps is about 2 times or more the usual amount, preferably about 2 to 3 times.

The above-mentioned alkylating agent, Schiffing agent and acid can be used alone, but two or three of them may be used in combination.

In the present invention, for example, a reaction with a crosslinking agent is carried out for the purpose of advancing a reaction such as a crosslinking reaction advantageously, accelerating the reaction rate, and improving the performance and yield of the material. Before that, the protein may be subjected to a pretreatment, or the pH may be adjusted after the reaction between the protein and the crosslinking agent and before the reaction with the alkylating agent or the Schiffing agent.

The pretreatment method of the above-mentioned protein is not particularly limited, but examples thereof include dilution, pH adjustment, centrifugation, electrodialysis, heating and the like, and more specifically, Japanese Patent Application No. 2-41.
The method described in Japanese Patent No. 8876 can be mentioned. That is, for example, in the case of egg white protein, when diluted with water, the dilution ratio is usually 2 times or more, preferably 2 to
It is preferably about 5 times, more preferably about 2 to 3 times. Examples of water used for dilution include deionized water,
Distilled water, pure water, tap water, etc. may be mentioned.

To adjust the pH, first, an acid is added to the protein or its diluted product to adjust its pH from an acidic range to a neutral range.
In the present invention, the pH range is not particularly limited as long as it is in the acidic to neutral range, but it is generally preferable that the pH range is about 3-8. The acid to be added is not particularly limited, but an acid that becomes an acidulant in the food additive is suitable. Specific examples of such an acid include citric acid, succinic acid, acetic acid, lactic acid, tartaric acid, fumaric acid, malic acid, adipic acid, gluconodeltalactone, gluconic acid, ascorbic acid, hydrochloric acid and the like.

When aggregates or precipitates are precipitated when an acid is added to the protein or its diluted product, it is preferable to remove these by a usual separation means. For example, aggregates and precipitates may be removed through a filter of about 42 mesh, and if necessary, centrifugation may be performed at about 3000 to 7000 rpm.

When the protein is further heat-treated, the heating conditions are not particularly limited, but usually about 75 ° C. or higher, about 3 ° C.
It should be around 0 minutes or more. If a precipitate is formed after heating, it may be filtered with a filter of about 200 mesh.

Depending on the above treatment, p of the aqueous protein solution is
Although H may be on the acidic side in some cases, it is preferable to readjust the pH of the aqueous protein solution to the normal pH, that is, the alkaline side, before reacting with the crosslinking agent.

The following is an example of a preferred embodiment of the method of the present invention.

(A) An egg white aqueous solution having a concentration of about 1 to 5% by weight is treated by the method described in Japanese Patent Application No. 2-418876. That is, the pH of the egg white or its water dilution is adjusted to an acidic or neutral range, the ionic strength is lowered, and the pH of the resulting liquid is adjusted to 9.
Adjust to 0 or more and heat.

(B) The pH of the treated egg white aqueous solution is adjusted to an alkaline range (for example, pH 10 to 12).

(C) The egg white aqueous solution after the pH adjustment and the organic solvent solution of the cross-linking agent are mixed and vigorously stirred and allowed to stand to separate into an aqueous phase and an oil phase.

(D) The aqueous phase (pH 6.5 to 9) is fractionated, and the pH is adjusted to an alkaline range (for example, pH about 12).

(E) An alkylating agent is further added with stirring,
The precipitated gel-like substance is separated by centrifugation, washed with water and freeze-dried.

The material thus obtained is confirmed to be a protein from the results of amino acid analysis. The greatest feature of this material is that it is soluble in an organic solvent such as dimethylformamide (DMF) but not in water, even though it is a protein.

[0052]

EFFECTS OF THE INVENTION According to the present invention, it is possible to provide a protein material having a higher solubility in an organic solvent than conventional ones.
When the raw material and the synthetic resin are dissolved and mixed in an organic solvent and the solvent is removed, the raw material disperses very uniformly in the resin, so that the protein is far superior to the case of mixing fine resin powder with the resin. Can perform the desired function.

Since this material is soluble in an organic solvent, it can be dispersed very uniformly in a plastic film, sheet, tube or the like, and a new function can be imparted to the plastic.

For example, when this material is applied to synthetic fibers for clothes, it gives a non-greasy feel like a natural material, as compared to fibers made only of plastic, balance of inhalation and release of moisture, heat retention, etc. The excellent performance that the film has a good property and is less likely to cause dew condensation is exhibited.

Further, cosmetic puffs are conventionally manufactured by coagulating and vulcanizing porous rubber such as latex or by molding urethane foam, but they are satisfactory in terms of surface moist feeling and skin feel. I haven't got what I want. Therefore, by adding an organic solvent solution of the present material when producing a urethane foam, a sponge (puff) having a moist feeling similar to silk and a smooth texture can be obtained. It should be noted that the production of the urethane foam may be carried out according to the conventional method, and the addition amount of the present material may be appropriately selected according to the intended use of the sponge to be obtained.

The present material can be used for various purposes because it has a property due to a functional group such as a carboxyl group in its molecule. For example, this material has a surface active effect and can be used as a surfactant, a fiber auxiliary agent, an antistatic agent, a dye fixing agent, a dye auxiliary agent, and the like. In addition, (the carboxyl group of this material) and esters such as ascorbic acid and pantothenic acid can be used as cosmetic raw materials. This material can also be used as a raw material for a chelating agent that traps metal ions.

In addition, this material is, for example,
Recording paper, artificial skin, artificial organs, artificial leather, immobilizing agents for analytical and physicochemical tests, coating agents such as pesticides and fertilizers,
It can be applied to applications such as biosensors for LB film formation, microcapsule materials, and drug delivery systems.

[0058]

EXAMPLES The present invention will be further clarified by the following examples.

Example 1 Frozen egg white was left to thaw in warm water at 35 ° C., thawed, passed through a 32 mesh filter, diluted 2.5 times with tap water, and stirred gently with 2 M citric acid. PH of 6.
Adjusted to 8. This adjusted egg white is passed through a 42 mesh filter and further centrifuged (7,000 rpm, 10 minutes).
Then, the precipitate was removed by adjusting the pH. The resulting supernatant was electrodialyzed (electrodialysis: CS-O type, trade name, manufactured by Asahi Glass Co., Ltd., flow rate: 250 liter / hr., Constant voltage: 1
4 V) and the electric conductivity was 950 μS / cm. The dialysate was adjusted to pH 10 by adding 1N and 6N sodium hydroxide. This was heated in boiling water for 30 minutes and cooled to room temperature to obtain an egg white liquid that passed through a 100-mesh filter.

The egg white liquor thus obtained was adjusted to pH 12 by adding 1N and 6N sodium hydroxide. 2 liters of this egg white liquor was heated to 50 ° C. On the other hand, 35 g of toluene diisocyanate was mixed with 400 ml of chloroform, and this was added to 2 liters of warmed egg white liquor, and stirring was continued for 2 hours, then, the mixture was allowed to stand at room temperature and separated into an aqueous phase and a chloroform phase. The aqueous phase was centrifuged (8000 rpm, 10 minutes).

After adjusting the pH of this supernatant to 4 with citric acid,
It was centrifuged again (10000 rpm, 15 minutes). The collected precipitate was easily soluble in DMF, and when this solution was added to tap water, precipitation of protein was observed. The powder obtained by freeze-drying this precipitate was also soluble in DMF, and when this solution was also added to tap water, precipitation of protein was observed.

The physicochemical properties of the above material were measured according to the following methods.

[Amino Acid Composition] This material is hydrolyzed (6
N-HCl, 105 ° C, 24 hours), amino acid analysis.
For analysis, a high performance liquid chromatograph LC-6A amino acid analysis system manufactured by Shimadzu Corporation was used, and the mobile phase was
A sodium type mobile phase kit for amino acid analysis manufactured by Shimadzu Corporation was used, and an amino acid analysis kit OPA reagent manufactured by Shimadzu Corporation was used as a reaction solution. Mobile phase flow rate 0.3 ml / min. Column temperature 55 ° C. Detector RF-535 ((manufactured by Shimadzu Corporation), the wavelength _{E x 348nm, E m 450nm)} . By this analysis, the weight% of each amino acid was obtained, and this was converted into the composition ratio. The results are shown in Fig. 1.

[Weight Average Molecular Weight] The measuring method is GPC / L.
According to the ALLS method. The measurement conditions are as follows. Column: Tosoh Corp., TSK gel, GMH _XL (7.8 m
ID × 30 cm × 2), eluent: DMSO (30 mM-
(Including LiBr), flow rate: 1.0 ml / min, sample: this material, sample injection amount: 200 μl (1.05 mg / ml),
Measurement temperature: 40 ° C., detector: RI, LALLS, line filter pore size: 2.0 μm, standard sample: pullulan, measuring instrument: Tosoh Corporation light scattering photometer LS
-8000, differential refractometer RI-801 manufactured by Tosoh Corporation
1 and a light scattering data processing system kit manufactured by Tosoh Corporation. As a result, the weight average molecular weight of the raw material obtained in Example 1 was 782,000.

[Infrared absorption] JIR manufactured by JEOL Ltd.
-RFX3200, FT-IR SPECTROPHO
The infrared absorption spectrum was measured using TOMETER. The results are shown in Figure 2.

[Ultraviolet absorption] The powder of this material has a concentration of 3%.
Dissolve in DMF at (W / V) and then add 100 with DMF.
The 0-fold diluted sample was used as a sample, and the maximum absorption was examined. DMF was used for the baseline. The results are shown in Fig. 3. The ultraviolet absorption varies depending on the type of diisocyanate used, but when a diisocyanate having a benzene nucleus in the molecule is used, it is generally 266 nm to 2 nm.
Maximum absorption is observed between 80 nm.

[Crosslinking Structure (SDS Electrophoresis)] PS-5200 [trade name, manufactured by Advantech] as an electrophoretic device using an aqueous phase collected after the crosslinking reaction described later as a sample.
Electrophoresis is performed for 100 minutes at a gel concentration of 13% and a current of 30 mA. A graph showing the SDS electrophoresis pattern of the above material is shown in FIG. As shown in the figure, a substantially single band was detected at the upper end, and albumin, which is an egg white protein, was not detected. From this fact, it is considered that cross-linking between proteins occurred due to the reaction of diisocyanate and the polymer was polymerized.

Example 2 Frozen egg white was thawed by leaving it in warm water at 35 ° C. for 8 hours to thaw it.
After passing through a 2-mesh filter, 2. with deionized water.
The mixture was diluted 5-fold, and 2M-citric acid was added under gentle stirring to adjust the pH to 6.5. This adjusted egg white liquor is passed through a 42 mesh filter and further centrifuged (7,000 rp).
m for 10 minutes) to remove the precipitate by adjusting the pH.

The obtained supernatant was electrodialyzed (electrodialyzer: CS-O type, trade name, manufactured by Asahi Glass Co., Ltd., flow rate: 25).
0 liter / hr. , Constant voltage: 18 V, 20 minutes), and the electrical conductivity was 660 μS / cm. The dialysate was centrifuged (7,000 rpm, 10 minutes) to remove the precipitate by dialysis.

The supernatant was adjusted to pH 10 by adding 1N and 6N sodium hydroxide. 30 at 90 ℃
The mixture was heated for 1 minute, cooled to room temperature, passed through a 100-mesh filter, and an egg white liquid having an electric conductivity of 1.1 mS / cm was obtained.

The egg white liquor thus obtained was adjusted to pH 12 by adding 1N and 6N sodium hydroxide. 1 liter of this egg white liquor was stirred to form 3.3 parts of isophorone diisocyanate.
% (V / V) chloroform solution to 300 ml,
After continuing stirring for an hour and then allowing to stand at room temperature, an aqueous phase and a chloroform phase were separated. The aqueous phase was separated and adjusted to pH 12 again, 8 ml of dimethylsulfate was added thereto with stirring to obtain a gelled product, and 3 times the amount of ionized water was added, followed by centrifugation (5
(000 rpm, 10 minutes). The collected precipitate is DM
It was easily soluble in F, and when this solution was added to ionized water, precipitation of protein was observed. The powder obtained by freeze-drying the precipitate was also soluble in DMF, and when a small amount of the powder was added to deionized water, protein precipitation was observed. The infrared absorption spectrum of this material is shown in FIG. 5, and the ultraviolet absorption spectrum is shown in FIG.

Example 3 The supernatant obtained by the same procedure as in Example 2 up to electrodialysis and subsequent centrifugation was treated with 1 N and 6 without heating.
The pH was adjusted to 12 by adding N sodium hydroxide. 0.5 liter of this solution was added to 150 ml of a 5% (V / V) chloroform solution of isophorone diisocyanate with stirring,
After continuously stirring for 24 hours, the mixture was allowed to stand at room temperature to separate into an aqueous phase and a chloroform phase. Thereafter, the same procedure as in Example 2 was carried out to obtain a precipitate. The precipitate was easily dissolved in DMF, and when this solution was put in ionized water, precipitation of protein was observed. The powder obtained by freeze-drying the precipitate was also soluble in DMF, and when a small amount of the powder was added to deionized water, protein precipitation was observed. The infrared absorption spectrum of the obtained material is shown in FIG.

Example 4 The procedure of Example 2 was repeated except that 5M-lactic acid was used in place of 2M-citric acid in the first pH adjustment, and a precipitate easily soluble in DMF was obtained. It was

Example 5 Dried egg white was dissolved in ionized water (protein concentration 3%) and centrifuged (7,000 rpm, 10 minutes) to remove precipitates.
Thereafter, the operations after electrodialysis were performed in the same manner as in Example 2.
The obtained precipitate and its freeze-dried powder were soluble in DMF and insoluble in water.

Example 6 In Example 2, isophorone diisocyanate (IP
DI) instead of diphenylmethane diisocyanate (M
The same procedure as in Example 2 was repeated except that DI) was used, and DMF was used.
A precipitate soluble in water and insoluble in water and its freeze-dried powder were obtained. The infrared absorption spectrum of the obtained material is shown in FIG.

Example 7 In Example 6, hydrochloric acid was used instead of dimethyl sulfuric acid (pH of the aqueous phase (pH 6.8) after the diisocyanate reaction was adjusted with hydrochloric acid to pH 4.0), and the amount of MDI added D was operated in the same manner as in Example 6 except that
A MF-soluble and water-insoluble precipitate and its lyophilized powder were obtained. The infrared absorption spectrum of the obtained material is shown in FIG.

Example 8 In Example 3, the raw egg white was changed to whey protein,
It was dissolved in deionized water to a concentration of 5% (W / V), and the procedure after electrodialysis was carried out in the same manner as in Example 3 using this. The lyophilized powder obtained was soluble in DMF.
The infrared absorption spectrum of the obtained material is shown in FIG.

Example 9 In Example 2, electrodialysis was not performed, and p after heating and cooling was used.
DMF was operated in the same manner as in Example 2 except that H was changed to 10.
A water-insoluble and water-insoluble precipitate and its freeze-dried powder were obtained.

Example 10 In Example 2, the concentration of IPDI was changed to 7.7 (V / V).
In the same manner as in Example 2 except that acetaldehyde was used instead of dimethyl sulfate, a DMF-soluble and water-insoluble precipitate and its freeze-dried powder were obtained. The infrared absorption spectrum of the obtained material is shown in FIG.

Example 11 The procedure of Example 2 was repeated, except that hexane was used instead of chloroform, and the concentration of IPDI was 7.7% (V / V), and it was soluble in DMF. A water-insoluble precipitate and its freeze-dried powder were obtained.

Example 12 The procedure of Example 7 was repeated except that toluene was used in place of chloroform and citric acid was used in place of hydrochloric acid in the second pH adjustment, and was soluble in DMF. A water-insoluble precipitate and its freeze-dried powder were obtained.

Example 13 The procedure of Example 2 was repeated except that 100 ml of IPDI was used instead of the chloroform solution of IPDI to obtain a DMF-soluble and water-insoluble precipitate.

Example 14 A powder of serum protein (BPP manufactured by Taiyo Kagaku Co., Ltd.) was added to 1 part of ionized water.
After 0% (W / W) dissolution, cellophane tube (2
After dialysis with 7/32 inch) of 20 times amount of deionized water, centrifugation (7,000 rpm, 10 minutes) was performed to remove precipitates. The obtained supernatant has an electric conductivity of 0.409 mS / cm,
pH 8.45, A ₂₈₀ absorbance (× 100 times) 0.538
Met. The supernatant was deionized with A ₂₈₀ absorbance (×
(100 times), diluted to 0.396, adjusted to pH 10 with 1N sodium hydroxide, heated in boiling water for 30 minutes, cooled to room temperature, and passed through a 100 mesh filter. The obtained serum protein was adjusted to pH 12 with 1N and 6N sodium hydroxide. 1 liter of this serum protein was heated to 45 ° C. On the other hand, after mixing 24 g of TDI with 200 ml of chloroform, the serum protein 1 was heated.
The mixture was placed in a liter and stirred for 150 minutes, and then left at room temperature to separate into an aqueous phase and a chloroform phase. The aqueous phase was centrifuged (10,000 rpm, 10 minutes). The supernatant was adjusted to pH 4 with citric acid and then centrifuged again (10000
rpm, 15 minutes). The collected precipitate was easily soluble in DMF, and when this solution was added to tap water, precipitation of protein was observed. The powder obtained by freeze-drying this precipitate was also DM
It was soluble in F, and when this solution was also added to tap water, protein precipitation was observed. The infrared absorption spectrum of the raw material protein (serum protein) is shown in FIG. 12, and the infrared absorption spectrum of this material (functional serum protein) is shown in FIG.

Example 15 The serum protein of Example 14 was changed to whey protein, dissolved in deionized water to 3% (W / V), and centrifuged (10000 rpm, 10 minutes). The supernatant was adjusted to pH 10 with 1N sodium hydroxide, heated in boiling water for 30 minutes, cooled to room temperature and passed through a 100 mesh filter to obtain 1 liter of adjusted whey protein. Then, the same procedure as in Example 14 was performed. The TDI is 17.
The pH was adjusted to 3.3 using 4 g. This functional whey protein was also soluble in DMF and insoluble in water. The infrared absorption spectrum of the raw material protein (whey protein) is shown in FIG.
The infrared absorption spectrum of this material (functional whey protein) is shown in FIG.

Example 16 The serum protein of Example 14 was changed to powdered casein, dissolved in deionized water at pH 10 so as to be 5% (W / V), and then dialyzed with cellophane tube to determine the electric conductivity. 1.1
It was dropped to 1 mS / cm. 1N in the obtained casein solution
And 6N sodium hydroxide were added to adjust the pH to 12.
1 liter of this casein solution was heated to 45 ° C. on the other hand,
After mixing 19 g of TDI with 200 ml of chloroform,
The mixture was added to 1 liter of casein solution under heating and the stirring was continued for 150 minutes. Then, the same procedure as in Example 14 was performed. The obtained functional casein protein was also soluble in DMF and insoluble in water. The infrared absorption spectrum of this material (functional casein protein) is shown in FIG.

Example 17 The serum protein of Example 14 was separated into an aqueous phase and a chloroform phase, and the procedure up to the point where the aqueous phase portion was centrifuged (10000 rpm, 10 minutes) was carried out in exactly the same manner as in Example 14. afterwards,
The pH of the centrifugally separated aqueous phase was adjusted to pH 12, dimethylsulfate was added, and the mixture was stirred to obtain a gel. This gel-like material was centrifuged (10,000 rpm, 10 minutes) to obtain this material as a precipitate. The material was soluble in DMF and insoluble in water.
The infrared absorption spectrum of this material is shown in FIG. Example 18 The serum protein powder of Example 14 was treated with 3.0% ionic water (W
/ W) dissolved. The conductivity at this time is 5.41 mS / c
It was m. This solution is not dialyzed and adjusted to pH 12 without heating.
Soluble in DMF by the same procedure as in Example 14 except that the solution was prepared.
A water-insoluble precipitate and its freeze-dried powder were obtained.

Reference Example 1 (Mixing with resin) The freeze-dried powder of Example 1 was dissolved in DMF at 3%. On the other hand, urethane pellets were dissolved in DMF in 20% (W / W). When these two solutions were mixed, the protein concentration was about 0.1.
Diluted to%. This mixed solution was attached to a polyester cloth, DMF was removed in deionized water, and a urethane film was manufactured. Then, it dye | stained by the coma brilliant blue R250. As a control, a film made of only urethane was prepared and the difference in staining was examined. The urethane film containing this material was evenly and beautifully dyed, but the control was hardly dyed.

Reference Example 2 20 g of polyurethane pellets were placed in 80 g of DMF and left to stand for several days to be dissolved to prepare a 20 W / V% polyurethane solution.

To this solution was added the freeze-dried powder of Example 1
1 g of DMF was added to 0.7 g of DMF, and the protein solution dissolved by stirring for 1 day was added and mixed uniformly.

To this, 350 g of calcium sulfate was added,
The mixture was well stirred with a mixer, and the resulting mixture was pressed into a sheet having a thickness of 30 mm. The molded sheet was immersed in water to wash out DMF. This operation was repeated 3 times to wash out DMF almost completely.

Thereafter, the sheet is dried at 90 ° C. for 30 minutes,
It was punched into an elliptical shape and the shape was adjusted to obtain a sponge-like material. This product had an excellent moist feeling and smooth texture.

[Brief description of drawings]

FIG. 1 is a drawing showing the amino acid composition of the present raw material and raw material protein obtained in Example 1.

2 is an infrared absorption spectrum diagram of the present material obtained in Example 1. FIG.

FIG. 3 is an ultraviolet absorption spectrum diagram of the present material obtained in Example 1.

FIG. 4 is a view showing an SDS-electrophoresis pattern of the present material obtained in Example 1.

5 is an infrared absorption spectrum diagram of the present material obtained in Example 2. FIG.

FIG. 6 is an ultraviolet absorption spectrum diagram of the present material obtained in Example 2.

FIG. 7 is an infrared absorption spectrum diagram of the present material obtained in Example 3.

8 is an infrared absorption spectrum diagram of the present material obtained in Example 6. FIG.

9 is an infrared absorption spectrum diagram of the present material obtained in Example 7. FIG.

10 is an infrared absorption spectrum diagram of the present material obtained in Example 8. FIG.

FIG. 11 is an infrared absorption spectrum diagram of the present material obtained in Example 10.

FIG. 12 is an infrared absorption spectrum diagram of a raw material protein (serum protein) used in Example 14.

FIG. 13 is an infrared absorption spectrum diagram of the present material obtained in Example 14.

FIG. 14 is an infrared absorption spectrum diagram of a raw material protein (whey protein) used in Example 15.

FIG. 15 is an infrared absorption spectrum diagram of the present material obtained in Example 15.

16 is an infrared absorption spectrum diagram of the present material obtained in Example 16. FIG.

17 is an infrared absorption spectrum diagram of the present material obtained in Example 17. FIG.

─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Kazuo Yamakoshi 463 Kagasuno, Kawauchi Town, Tokushima City, Tokushima Prefecture Otsuka Chemical Co., Ltd.In the Tokushima Factory (72) Tadashi Tsukiyama 463 Kagasuno, Kawauchi Town, Tokushima Prefecture Tokushima Factory Co., Ltd. (56) Reference JP-A-52-25800 (JP, A) JP-A-50-94000 (JP, A) (58) Fields investigated (Int.Cl. ⁷ , DB name) C07K 14 / 415-14/765 C07K 1/10 BIOSIS (DIALOG)

Claims (6)

(57) [Claims] 1. A functional protein material having the following physicochemical properties. (A) Egg white protein of chicken, quail, duck or goose egg
Quality, whey protein, serum albumin and casein
Not a hydrolyzate of at least one source protein
It is a protein polymerized by cross-linking. (B) Solubility: Soluble in dimethylformamide, dimethylsulfoxide and dimethylacetamide but not in water. (C) Weight average molecular weight: 100,000 to 3,000,000 (GPC / L
By the ALLS method). (D) Infrared absorption: Compared with at least one raw material protein selected from egg white protein of chicken, quail, duck or goose egg, whey protein, serum albumin and casein, absorption of 3440 cm ^{-1 to} 3420 cm ^-1 Weak 1
603cm ^-1 near the shoulder has a bottom in the vicinity of 1353cm ^-1, a peak in the vicinity of 1222cm ^-1 is high. (V) Crosslinked structure: Protein is crosslinked by the reaction of diisocyanate (SDS-electrophoresis). 2. A functional protein material having the following physicochemical properties. (A) Egg white protein of chicken, quail, duck or goose egg
Quality, whey protein, serum albumin and casein
Not a hydrolyzate of at least one source protein
It is a protein polymerized by cross-linking. (B) Solubility: Soluble in dimethylformamide, dimethylsulfoxide and dimethylacetamide but not in water. (C) Weight average molecular weight: 100,000 to 3,000,000 (GPC / L
By the ALLS method). (D) Infrared absorption: Compared with at least one raw material protein selected from egg white protein of chicken, quail, duck or goose egg, whey protein, serum albumin and casein, absorption of 3440 cm ^{-1 to} 3420 cm ^-1 Weak 1
Has a shoulder near 740 cm ^-1 and 1222 cm ^-1
The peak in the vicinity becomes high. (V) Crosslinked structure: Protein is crosslinked by the reaction of diisocyanate (SDS-electrophoresis). 3. In red external absorption, further 1600 m ^-1
The functional protein material according to claim 2, which has a shoulder in the vicinity and a bottom in the vicinity of 1353 m ^-1 . 4. A functional protein material having the following physicochemical properties. (A) Egg white protein of chicken, quail, duck or goose egg
Quality, whey protein, serum albumin and casein
Not a hydrolyzate of at least one source protein
It is a protein polymerized by cross-linking. (B) Solubility: Soluble in dimethylformamide, dimethylsulfoxide and dimethylacetamide but not in water. (C) Weight average molecular weight: 100,000 to 3,000,000 (GPC / L
By the ALLS method). (D) Infrared absorption: Compared with at least one raw material protein selected from egg white protein of chicken, quail, duck or goose egg, whey protein, serum albumin and casein, absorption of 3440 cm ^{-1 to} 3420 cm ^-1 Weak 1
740cm no shoulder in the vicinity of ^-1 and around 1600cm ^-1. (V) Crosslinked structure: Protein is crosslinked by the reaction of diisocyanate (SDS-electrophoresis). 5. A diisocyanate compound is mixed with an aqueous solution of at least one protein selected from egg white protein of chicken, quail, duck or goose egg, whey protein, serum albumin and casein, followed by alkylating agent and Schiffing. At least 1 sort (s) chosen from an agent and an acid is added, The manufacturing method of the functional protein raw material of any one of Claims 1-4 characterized by the above-mentioned. 6. A mixture of at least one protein aqueous solution selected from egg white protein of chicken, quail, duck or goose egg, whey protein, serum albumin and casein and a diisocyanate compound, and then adding an acid. The method for producing a functional protein material according to claim 1.