JPH04189833A - Solution of keratin in organic solvent and its production - Google Patents

Abstract

PURPOSE:To obtain the subject solution in a solvent useful as keratinous polymer membranes, films, fibers, sponges, etc., without using a protein denaturing agent by reducing a keratin-containing substance in a specific organic solvent such as dodecyl alcohol having a specified value or below of moisture content. CONSTITUTION:A keratin-containing substance (e.g. human hair, animal hair, cattle horn or hoof) is reduced in at least one organic solvent with <=50wt.% moisture content selected from alkoxy alcohols (e.g. a 2-methoxy alcohol) expressed by the formula (R is 1-3C alkyl; n is 2-4) and halogen-substituted 2-7C aliphatic alcohols (e.g. 2-chloroethanol) to afford the objective solution in the solvent.

Description

DETAILED DESCRIPTION OF THE INVENTION Two industrial fields of application [The present invention relates to a keratin organic solvent solution that does not involve irreversible denaturation of nosulfite bonds and a method for producing the same. The organic solvent solution of the present invention can be used for keratin polymer membranes, films, fibers,
Used to manufacture various industrial products such as sponges.

[Prior Art] Keratin, which exists as a structural protein in animal tissues such as hair, animal hair, and feathers, has long been viewed as a promising raw material for industrial materials such as membranes and fibers.

However, keratin is insoluble or sparingly soluble in common solvents. Therefore, in order to utilize keratin in natural raw materials, it is necessary to significantly shorten the molecular weight by decomposition, reduce the nosulfide bonds of keratin, or irreversibly protect the generated thiol groups by chemical treatment. That is, a natural product containing keratin is treated with a concentrated acid or alkali to produce a phosphorus hydrolyzate, and a reducing agent and a protein denaturing agent such as a highly concentrated urea aqueous solution are used in combination to remove the nosulfide bonds of keratin. An aqueous solution of keratin that undergoes reductive cleavage to form a 1,000-ol group, and is chemically modified with monoiodoacetic acid, sodium sulfite/sodium tetrathionate, etc. to prevent recombination of the thiol groups of the keratin, resulting in a keratin derivative, or reductive cleavage and proteolytic decomposition. It is used in the form of an aqueous keratin solution after its molecular weight is reduced by enzymes.

[Problem to be solved by the invention 2 However, keratin restored from protein obtained by oxidation treatment requires complicated chemical treatment and the recovery yield is extremely low. Further, in the reduction treatment, it is necessary to use a large amount of various denaturing agents as solubilization aids, making the operation extremely complicated. For example, keratin is processed under neutral or alkaline conditions in water or a water-based solvent such as methanol, ethanol, or amide in the coexistence of a reducing agent such as thiol and a protein denaturing agent such as urea. After dissolution, it is necessary to add a surfactant or add iodoacetic acid to the oxidation-unstable sulfhydryl group to perform a carphoquinolmethylation reaction. In addition, it is necessary to remove insoluble substances and denaturing agents such as urea by dialysis or the like in the middle of the process. Such dialysis requires large amounts of water and takes a long time to operate.

Therefore, conventional methods for extracting keratin from keratin-containing materials (1) require large amounts of protein denaturing agents such as urea, solubilization aids with surfactant effects, and reducing agents, and require uneconomical dialysis operations; If it is not included, the process will be complicated and require a long processing time, which will increase the product cost. Also (2)
Pollution countermeasures will also be needed to deal with the large amount of contaminated water generated during the extraction process.

Furthermore, (3) the use of keratin has been limited because it is difficult to mix keratin aqueous solutions with materials soluble in the battering machine solvent.

1. Means for Solving the Problems Therefore, the present inventor conducted extensive research on a method for extracting keratin from natural products containing keratin without using a protein denaturant. As a result, we unexpectedly found that by treating keratin-containing substances in a specific organic solvent in the presence of a reducing agent, keratin can be easily extracted without using any protein denaturing agents. I have barely completed it.

That is, the present invention provides (1) a keratin-containing material with a moisture content of 5
0% by weight or less of the general formula % Formula %: [In the formula, R is an alkyl group having 1 to 3 carbon atoms, and n is an integer of 2 to 4] Alcoquine alcohol represented by the formula and carbon not substituted with halogen A method for producing a keratin battering machine solvent solution, characterized by reducing the solution in at least one organic solvent selected from aliphatic alcohols of numbers 2 to 7, and (2)
) Alcoquino alcohol with a water content of 50 double stars or less and a maximum of II] and a halogen-substituted carbon number of 2
This invention provides a keratin a solvent solution which is obtained by dissolving 10 to 30% by weight of keratin in at least one organic solvent selected from 7 to 7 aliphatic alcohols.

The keratin-containing substance used in the present invention may be any substance containing true keratin, such as human hair, sheep, horse and cow hair, chicken and bird feathers, cow horns and hooves, as well as keratin powder itself. It is preferably used.

In addition, keratin powder can be prepared by various known methods (for example, T,
T, Sun and H, Green, J, Biol.
, Chem,, 253゜2053-2060 (1
978)).

Among the organic solvents used in the present invention, a representative example of the alcoquino alcohol represented by the general formula 1 is 2-methoquinoalcohol. It is preferable that the fatty acid alcohol of ~7 is substituted with chlorine or fluorine,
Preferably, it has 2 to 5 carbon atoms. Typical examples include 2-chloroethanol, 2,2.3.3-tetrafluoropropanol, and 2.2.2-trifluoroethanol. These organic solvents can be used as they are or with water added in an amount not exceeding 50% by weight. Other organic solvents may be added to these solvents to the extent that their effects are not impaired. The amount of these organic solvents used is preferably 10 to 30 times the weight of the keratin-containing material.

In the present invention, a reducing agent capable of reducing the nosulfite bond of keratin in a keratin-containing material to a thiol group is usually used. Such reducing agents include 2-
Mercaptoethanol, thioglycolic acid, toluene-
Examples include thiols such as ω-thiol, notothiothreitol, and othioerythritol; trialkylphosphines such as tripropylphosphine and tributylphosphine; and inorganic reduction compounds such as sodium hydrogen sulfite. These reducing agents reduce keratin sulfide bonds in keratin-containing materials and promote conversion to thiol groups.

The amount of reducing agent used is 0 per 10g of keratin-containing material.
The amount is preferably 0.05 to 0.050 mol, or preferably 0.05 to 0.020 mol per 10 g of the keratin-containing material from the viewpoint of reaction efficiency and economy.

To obtain keratin from the above ingredients, a keratin-containing material is immersed in an organic solvent containing a reducing agent, and
Shake for ~24 hours while warming at °C. The reaction is carried out, for example, at room temperature for 24 to 48 hours and at 50°C for about 24 hours.
At 00°C, 1 to 2 hours is sufficient. The thus obtained reaction product is filtered and centrifuged to remove insoluble matter, yielding an organic solvent solution containing sawa keratin.

The mechanism of such solubilization of keratin is said to be based on reducing the nosulfide bonds of keratin and converting them into thiol groups. In fact, the keratin organic solvent solution may be stored at a low temperature under an inert gas such as nitrogen, after the nosulfite bonds are regenerated by an oxidizing agent such as oxygen in the air and the viscosity gradually increases.

In addition, when reducing keratin and promoting extraction, especially when extracting keratin from tissues such as hard hair and nails, these raw materials are crushed, and a surfactant is added as a solubilizing agent to the keratin-containing material at a concentration of 10 to 50%. It is preferable to add % by weight.

Such surfactants include alkyl sulfates (e.g. sodium dodenol sulfate), alkyl ml acid ester salts,
Anion activators such as fatty acid alcohol phosphate ester salts and sulfosuccinate ester salts.

"In the formula, 1 to 2 of Rl, R2, R3 and R4 are linear or branched alkyl groups having 8 to 20 carbon atoms or hydroquinoalkyl groups, and the rest are straight chain or branched alkyl groups having 8 to 20 carbon atoms.
~3 alkyl group, hydroquinoalkyl group, or henonol group, and X is a halogen atom or a carbon number of 1 to 3
An aromatic quaternary amine salt with two alkyl sulfate groups or an alkylpyrinium halide] A cationic surfactant represented by the following formula.

Hetaino-based amphoteric surfactants such as N-carboxylic methyl, N-sulfoalkylated and imidazolinosulfonic acids of fatty acid amines (hydrophobic groups mainly contain 12 to 14 carbon atoms)
(The alkyl group is an anol group, and the counter ion is an alkali metal) 1. Nonionic surfactants such as polyoquinoethylene alkyl ether type, fatty acid ester type, polyethylene imino type, polycrycerin ether type, and ester type. The hydrophobic group is mainly an alkyl group or anol group having 12 to 14 carbon atoms.

Nato is mentioned.

The keratin obtained in the above manner has 1 amino acid + 00 residues, 5 to 10 lastins, and 0.5 to 2 nostino residues.
It has a molecular weight of 10.000 to 13o, ooo.

The obtained solvent solution is molded into various polymer membranes, films, fibers, sponges, etc. by known film forming and molding methods.

EXAMPLES Next, the present invention will be explained in more detail based on examples.

Example 1 Chicken wings] Og and 2-methoquinoethanol 200 m0.
The mixture was degassed and replaced with nitrogen.

Next, IOg of 2-mercaptoethanol was added in a nitrogen stream, and the container was tightly capped. The mixture was then stirred at 55°C for 24 hours, and then the reaction solution was cooled to room temperature.

This reaction solution was filtered to remove insoluble matter, and 180 mL of the desired keratin organic solvent solution containing a small amount of 2-mercaptoethanol was obtained using 2-methoquinoethanol as a solvent.

To remove the solvent, put 10 g of the solution into a cellophane dialysis tube, and add 2-mercaptoethanol (0.3%) to the external solution.
Dialysis was performed using pure water containing

Next, freeze-dry the contents of the tube to obtain keratin powder 01.
5-023 g was obtained. Therefore, the keratin concentration in the organic solvent liquid is estimated to be 15 to 2.3%. Amino acid analysis of the above keratin powder revealed that amino acids +0
There were 55 nosteins and 13 nostins per 0 residues. Further, when examined using polyacrylamide electrophoresis, proteins with a molecular weight of 15,000 to 70,000 were found to be the main components.

Example 2 10g of chicken wings, 200mf of 2-chloroethanol! A mixture of 15 mC and water was degassed and replaced with nitrogen. Next, in a nitrogen stream, 2-mercaptoethanol log was added, the stopper was sealed, and after stirring at 55°C for 24 hours, the reaction solution was returned to room temperature. This reaction solution was filtered to remove insoluble materials to obtain the desired keratin organic solvent solution using 2-chloroethanol as a solvent and containing a small amount of 2-mercaptoethanol.

The solvent was removed and freeze-dried from the solution in the same manner as in Example I, yielding 0.23-027g of keratin powder1.Thus, the keratin concentration in the organic solvent solution was estimated to be 2.3-27%. . Further, when examined by polyacrylamide electrophoresis, the main component was protein with a molecular weight of 15.000 to 70.000.

Example 3 10 g of wool, 3 g of sodium dodenle sulfate and 2
- A mixture of 200 ml of chloroethanol was degassed and replaced with nitrogen. Then, in a nitrogen stream, 10g of 2-mercaptoethanol was added, the mixture was tightly stoppered, and the mixture was stirred at 100°C for 2 hours, and then the reaction solution was cooled to room temperature for 7 hours. This reaction solution was filtered to remove insoluble matter, and the desired organic solvent solution of keratin was obtained using 2-chloroethanol as a solvent and containing a small amount of 2-mercaptoethanol.

The solvent was removed from the solution in the same manner as in Example I and lyophilized to obtain 0.18 to 020 g of keratin powder.
The keratin concentration in the organic solution was estimated to be 18-20%. Amino acid analysis of keratin powder revealed that there were 8 nostins and 2 nstins per 100 amino acid residues. In addition, when examined by polyacrylamide electrophoresis, the molecular weight ranged from 110.000 to 70.1) 0
00) The amount of protein produced is 1.

Example 4 5 g of nickeratin powder obtained from wool by a known method was immersed in 100 ml of a mixture of 2-methquinoethanol and 2-chloroethanol (volume ratio 1:1), and degassing and nitrogen substitution were repeated several times. After that, 5 g of 2-mercaptoethanol was added.The mixture was heated to 10FI!j at 45°C.
IJ stirred. The resulting reaction solution was centrifuged to remove insoluble materials to obtain a supernatant keratin organic solvent solution.

The solution was dialyzed and freeze-dried in the same manner as above to obtain 019-021 g of keratin powder, so the keratin concentration in the organic solvent solution was estimated to be 19-21%.

Amino acid analysis of keratin powder revealed that there were 8 nostins and 2 nstins per amino acid +00 residues.

In addition, when examined using polyacrylamide electrophoresis,
The main component was protein with molecules @IO,000 to 70,000.

Example 5 A container containing 1 g of chicken wings, 20 m& of 2,2,3,3-tetrafluoropropanol, and 1.5 g of 2-mercaptoethanol was tightly stoppered and shaken at 50° C. for 20 hours. The reaction solution was cooled to room temperature, filtered to remove insoluble matter, and mixed with the desired keratin organic solvent solution (approximately 20 rr J
) was obtained.

5 mQ of the solution was put into a cellophane dialysis tube, the external solution was dialyzed as pure containing 0.3% 2-mercaptoethanol, and the contents of the tube were freeze-dried to obtain 0.13 g of keratin powder. The keratin concentration was estimated to be 2.6%. In addition, we conducted an amino acid analysis of the above keratin powder and found that for every 100 amino acid residues, there are 2 nosteins and 6 nstins. Further, when examined by polyacrylamide electrophoresis, proteins with a molecular weight of 15,000 to 70,000 were found to be the main components.

Comparative Example 1 The following Table 1 shows the results of attempts to extract keratin in the same manner as in Example 1, but using the various solvents listed below instead of 2-methoquinoethanol.

Table 1 Raw materials ('C) (hour)
Keratin yield methanol chicken feather 50 2
4 0% ethanol 50 2
4 traces (3 familiar τ) // human hair 60
24 0% Wool 60 24
/Chloroform Chicken feather 50 24
No/No Saikisan 〃50 24
Tetrahydrofuran” 50 24
No/henzeno 〃 50 24
// [Effects of the invention: - Unlike conventional methods, the production method of the present invention does not require the use of a protein denaturing agent, and therefore does not require dialysis after the reduction treatment. Further, according to the present invention, an organic solvent solution containing keratin at a high concentration of 1 to 3% can be obtained. Since the keratin organic solvent solution of the present invention contains keratin at a high concentration, it has a wide range of uses.

Patent applicant: Takeda Pharmaceutical Company Limited

Claims (2)

[Claims] (1) A keratin-containing substance with a water content of 50% by weight or less with the general formula: RO-(CH_2)nOH [I] [wherein, R is an alkyl group having 1 to 3 carbon atoms, and n is an integer of 2 to 4] ] A method for producing a keratin organic solvent solution, which comprises reducing the keratin in at least one organic solvent selected from alkoxy alcohols and halogen-substituted aliphatic alcohols having 2 to 7 carbon atoms. (2) General formula with water content of 50% by weight or less: RO-(CH_2)nOH [I] [wherein R is an alkyl group having 1 to 3 carbon atoms, and n is an integer of 2 to 4] A keratin organic solvent solution obtained by dissolving 1.0 to 3.0% by weight of keratin in at least one organic solvent selected from alkoxy alcohols and halogen-substituted aliphatic alcohols having 2 to 7 carbon atoms. .