JPH10265496A - Production of acylated peptide - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method for producing a high-purity acylated peptide better in performance than those produced by conventional methods when used as a surfactant. SOLUTION: This method for producing the objective acylated peptide of the formula (R<1> is a hydrocarbon group; R<2> is an amino acrid side chain; M is a hydrogen atom metal atom or ammonium; (n) is a number of >=1) comprises the following consecutive processes: (1) a protein or protein hydrolyzate is subjected to hydrolysis treatment in the presence of an alkali, (2) nitrogen-contg. matter produced in the process (1) is removed under reduced pressures, and (3) the resultant protein hydrolyzate freed from the nitrogen-contg. matter is acylated.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a method for producing an acylated peptide, for example, an acylated peptide or a salt thereof.

[0002]

2. Description of the Related Art Acylated peptides are anionic surfactants that have been known for a long time. They are highly safe because they are derived from proteins derived from animals and plants, and have low skin irritation. Used in In particular, it is said that demand has been increasing due to recent natural orientation.

[0003] Methods for producing acylated peptides have also been studied for a long time. In the production of acylated peptides, means for hydrolyzing proteins are roughly classified into three types.
That is, a method using an acid, a method using an alkali, and a method using an enzyme. As a method using an enzyme, for example, there is JP-A-60-137299. This publication discloses a method for producing an acylated peptide in which a protein is hydrolyzed with a protease and acylated with a fatty acid halide in the presence of an alcohol.

A method using an alkali has been known for a long time. For example, Japanese Patent Publication No. 38-6455 discloses a method in which a soybean protein and a wheat protein are alkali-treated and then reacted with a fatty acid chloride. However, it is known that when an alkali is used for hydrolysis, the amide group in the side chain of glutamine or asparagine is decomposed to form a carboxyl group and to generate ammonia (Akahori et al., Protein Chemistry, pp. 1, 29). , 1951, Kyoritsu Shuppan).

[0005]

When an alkali is used for hydrolyzing a protein, it is inevitable that volatile base components such as ammonia are generated as described above. When the acylation is performed without removing the generated ammonia and the like, the acylating agent such as an acid chloride reacts with the ammonia to generate an acid amide, and the yield of the acylated peptide as the target product is reduced. Instead, when the acylated peptide is used as a surfactant, it causes a decrease in the surfactant activity. In addition, since ammonia and the like are volatile, they cause odor and the like.

Accordingly, an object of the present invention is to obtain acylated peptides having high purity and exhibiting better performance when used as a surfactant than acylated peptides produced by a conventional production method. It is to provide a new manufacturing method.

[0007]

The present inventors have conducted intensive studies and have developed a method for producing an acylated peptide which incorporates a step of removing the volatile base component. That is,
The present invention provides, as essential steps, a step of hydrolyzing a protein or a protein hydrolyzate in the presence of an alkali; a step of removing the nitrogen-containing substance produced in the step under reduced pressure; a hydrolyzed protein from which the nitrogen-containing substance has been removed Or the following general formula (1) obtained through a step of acylating a protein hydrolyzate:

Embedded image (Wherein, R ¹ represents a hydrocarbon group, R ² represents an amino acid side chain, M represents a hydrogen atom, a metal atom or ammonium, and n represents one or more numbers). This is a method for producing acylated peptides.

[0008]

BEST MODE FOR CARRYING OUT THE INVENTION In the acylated peptides represented by the above general formula (1), R ² is an amino acid side chain. Examples of amino acids include glycine, alanine, valine, leucine, isoleucine, serine, threonine, cysteine, cystine, methionine, asparagine, aspartic acid, glutamine, glutamic acid, lysine, arginine, phenylalanine, tyrosine, histidine, tryptophan, proline, oxyproline and the like. Is mentioned.

The protein which is a raw material of the production method of the present invention includes, for example, silk, bone, cartilage, tendon, fascia, skin, fish scale and the like derived from eyebrows, eyebrows, raw silk, raw silk waste and the like. Gelatin, animal hair, derived from
Keratin derived from hair, feathers, nails, horns, hooves, scales, etc., silk fibroin, milk, goat milk, casein derived from human milk, etc., albumin, globulin, plant proteins and the like,
A partial hydrolyzate of these proteins may be used.

The hydrolysis treatment of the above-mentioned proteins or hydrolysates thereof is carried out in the presence of water and alkali. Although the amount of water used is not particularly limited, it usually acts as a reaction solvent, and is preferably 50 to 50% based on the amount of the starting protein.
It is about 300% by weight. It is also possible to use water and an organic solvent such as a lower alcohol, a lower ketone and a lower ether in combination.

The alkali used for the hydrolysis treatment of the protein or its hydrolyzate is, for example, sodium hydroxide, potassium hydroxide, lithium hydroxide, calcium hydroxide, barium hydroxide, sodium carbonate, potassium carbonate, sodium hydrogen carbonate. And inorganic alkalis such as potassium hydrogen carbonate, and organic alkalis such as triethylamine and pyridine, and sodium hydroxide or potassium hydroxide is particularly preferable. The amount of the alkali used is not particularly limited, but it is preferably 1 to 50% by weight, more preferably 5 to 30% by weight, based on the protein of the raw material.

The temperature at which the protein or its hydrolyzate is hydrolyzed with an alkali is not particularly limited.
If the reaction is carried out at too low a temperature, the reaction rate is low and the reaction is not efficient.If the reaction is carried out at a too high temperature, degradation or denaturation of the raw material protein occurs, and amino acid decomposition occurs.
It is preferably performed at 0 ° C, more preferably at 20 to 90 ° C, and most preferably at 40 to 80 ° C. The time for performing the hydrolysis treatment with an alkali is not particularly limited, but is preferably about 30 minutes to 24 hours, more preferably about 1 to 18 hours, and still more preferably about 3 to 12 hours. Further, a hydrolysis treatment with an acid or an enzyme may be performed before and after the hydrolysis treatment with the alkali.

When the above-mentioned hydrolysis treatment is carried out, the protein used as a raw material or its hydrolyzate is decomposed into a low-molecular-weight polypeptide. The molecular weight of the polypeptide can be controlled by controlling the reaction conditions of the hydrolysis treatment, for example, the amount of alkali, the amount of water, the treatment time, and the like.
When adjusted to 2,000, acylated peptides useful as surfactants are obtained. The degree of polymerization n of the polypeptide is 1 (when completely hydrolyzed) or more.

The nitrogen-containing substance produced by the above-mentioned hydrolysis treatment is a substance produced by decomposing the protein, and is usually ammonia or lower amine. In order to remove these under reduced pressure, the pressure in the reaction system may be reduced by an aspirator, a vacuum pump or the like.
Hg or less, preferably 50 mmHg or less, more preferably 20 mmHg or less. Although the temperature at the time of removing under reduced pressure is not particularly limited, it is usually higher or lower than the temperature at the time of the hydrolysis treatment.
It is about 0 ° C. The time for the removal under reduced pressure is not particularly limited, but is preferably about 10 minutes to 8 hours, more preferably about 10 minutes to 6 hours, and still more preferably about 10 minutes to 3 hours.

It is preferable to use a carboxylic acid or a carboxylic acid halide as the acylating agent used for acylating the polypeptide that has been subjected to the above-mentioned removal under reduced pressure. As the carboxylic acid or its halide, for example, formic acid or its chloride, acetic acid or its chloride, propionic acid or its chloride, butyric acid or its chloride, valeric acid or its chloride, caproic acid or its chloride, caprylic acid or its chloride, capric acid Or its chloride, lauric acid or its chloride, myristic acid or its chloride,
Palmitic acid or its chloride, stearic acid or its chloride, oleic acid or its chloride, erucic acid or its chloride, linoleic acid or its chloride, linolenic acid or its chloride, and the like. Usually, a raw material is a single or mixed fatty acid or a chloride thereof having about 8 to 22 carbon atoms. Usually, fatty acids obtained from natural fats and oils or fatty acids obtained by chlorinating them are used. Examples of natural fats and oils include linseed oil, olive oil, cacao butter, sesame oil, rice bran oil, safflower oil, soybean oil, camellia oil, corn oil, rapeseed oil, palm oil, palm kernel oil, castor oil, sunflower oil, cottonseed oil, coconut oil Vegetable oils such as oils,
Animal fats and oils such as beef tallow, lard, milk fat, fish oil, and whale oil. The carboxylic acid used in the acylation reaction or a halide thereof produces an acylated peptide having an R
¹ is determined.

In order to promote the acylation reaction and improve the yield, it is preferable to add a solvent such as a lower alcohol, a lower ketone, or a lower ether to the reaction system during the acylation.
Examples of lower alcohols include methanol, ethanol, propanol, 2-propanol, butanol, ethylene glycol, and propylene glycol.Examples of lower ketones include acetone, methyl ethyl ketone and methyl isobutyl ketone, and lower ethers include dimethyl ether and Diethyl ether,
Ethyl methyl ether and the like can be mentioned.

The temperature at which the acylation reaction is carried out is not particularly limited. However, if the reaction is carried out at a too low temperature, the reaction rate is low and thus the reaction is not efficient. It is preferably carried out at 0 to 100 ° C. because it causes amino acid decomposition.
It is more preferably performed at a temperature of from 90 to 90 ° C, most preferably at a temperature of from 20 to 80 ° C. The time for performing the acylation reaction is not particularly limited, but is preferably about 10 minutes to 12 hours, more preferably about 20 minutes to 8 hours, and still more preferably about 30 minutes to 6 hours.

When carboxylic acid chloride is used in the acylation reaction, it is preferable to adjust the pH of the carboxylic acid chloride to alkaline with an alkali to promote the reaction, and to add the carboxylic acid chloride to the reaction system.

The acylated peptides can be obtained by the above reaction, but after completion of the acylation reaction, hydrochloric acid, sulfuric acid,
Adjusting the pH to acidic with an inorganic acid such as nitric acid and washing with water is preferable because by-products, impurities, unreacted raw materials, unreacted catalyst, and the like can be easily removed. When this washing treatment is performed, an acylated peptide in which M is a hydrogen atom is obtained.

Further, the obtained acylated peptide or the acylated peptide which has been subjected to the above-mentioned washing treatment can be treated with an alkali or an amine to obtain an acylated peptide salt. In this case, M is determined by the alkali or amine used. Examples of the alkali include those described above, and examples of the amine include ammonia, methylamine, dimethylamine, ethylamine, diethylamine, (iso) propylamine, di (iso) propylamine, monoethanolamine, diethanolamine, triethanolamine, and monopropanolamine. , Dipropanolamine, tripropanolamine, ethylenediaminetetraethoxylate, ethylenediaminetetrapropoxylate and the like.

The production method of the present invention has a specific step of removing a nitrogen-containing substance produced by hydrolyzing a protein or a protein hydrolyzate under reduced pressure, so that a nitrogen atom is added to a side chain of asparagine, glutamine or the like. This is a particularly useful production method when a protein containing many amino acids is used as a raw material. Proteins containing a large amount of amino acids such as asparagine and glutamine are often plant-derived proteins. Examples of plant-derived proteins include rice, rice bran, wheat,
Hato barley, rye, barley, oats, corn, soybeans, almonds, nuts, sesame, peanuts, buckwheat, seaweed, linseed, iwanori, kanori, matsuna and other proteins or proteins obtained from these processed products No.

The acylated peptides produced by the production method of the present invention can be used as an anionic surfactant, and can be mainly used by mixing in a detergent. Examples of the detergent include a hair shampoo, a hair rinse, a hair conditioner, a hair treatment, a body shampoo, a kitchen detergent, a dish detergent, and a clothes detergent. In addition, permanent wave agents, hair creams, hair foams, hair bleaches, hair lotions, hair liquids, hair tonics, lotions, shaving creams, after shaving lotions, pre-shaving lotions, face lotions, moisture creams, cleansing creams, cold creams, etc. In cosmetics.

[0023]

EXAMPLES The present invention will be described more specifically with reference to the following examples. In the following examples, parts and percentages are by weight unless otherwise specified. (Example 1) Edible gelatin 100 was placed in a reactor capable of reducing pressure.
Parts, 10 parts of sodium hydroxide and 190 parts of water,
Hydrolysis treatment was performed by stirring and mixing at 60 to 70 ° C. for 6 hours. After the reaction system was cooled to 50 ° C., degassing was performed under reduced pressure at 20 mmHg for 1 hour to obtain a solid content of 50% and an average molecular weight of about 50.
0, a polypeptide having a volatile base component of 0.023% was obtained. Next, 30 parts of ethanol was added to 100 parts of the polypeptide, and 20 parts of lauric chloride was added dropwise at 50 ° C. while adjusting the pH to 10 with sodium hydroxide. After completion of the dropwise addition, the mixture was aged at 40 to 50 ° C. for 1 hour. Then, hydrochloric acid was added to adjust the pH of the system to 1, followed by washing with water, and then neutralized with sodium hydroxide to obtain a collagen hydrolyzed peptide lauric acid condensate sodium salt having a solid content of 35% and a transparent appearance.

Example 2 In a reactor capable of reducing pressure, 100 parts of soybean separated protein, 10 parts of sodium hydroxide and 190 parts of water were used.
Then, the mixture was stirred and mixed at 60 to 70 ° C. for 6 hours to perform a hydrolysis treatment. After cooling the reaction system to 50 ° C, 20
Vacuum deaeration was performed at 1 mmHg for 1 hour to obtain a polypeptide having a solid content of 48%, an average molecular weight of about 420 and a volatile base component of 0.018%. Next, 30 parts of ethanol was added to 100 parts of the polypeptide, and the mixture was adjusted to pH 1 with sodium hydroxide.
While adjusting to 0, 24 parts of lauric chloride was added dropwise at 50 ° C. After completion of the dropwise addition, the mixture was aged at 40 to 50 ° C. for 1 hour. Then, hydrochloric acid was added to adjust the pH of the system to 1, followed by washing with water, and then neutralized with sodium hydroxide to obtain a soybean protein hydrolyzed peptide lauric acid condensate sodium salt having a solid content of 33% and a transparent appearance. .

(Comparative Example 1) In a reactor capable of reducing pressure, 100 parts of edible gelatin, 10 parts of sodium hydroxide and 190 parts of water were added.
The mixture was stirred and mixed at 60 to 70 ° C. for 6 hours to carry out a hydrolysis treatment to obtain a polypeptide having a solid content of 30%, an average molecular weight of about 560, and a volatile base component of 0.81%. Then, 30 parts of ethanol was added to 100 parts of the polypeptide, and 12 parts of lauric chloride was added dropwise at 50 ° C. while adjusting the pH to 10 with sodium hydroxide. After dripping 4
Aged at 0-50 ° C for 1 hour. Thereafter, hydrochloric acid was added to adjust the pH of the system to 1, followed by washing with water, and then neutralized with sodium hydroxide to obtain a collagen hydrolyzed peptide lauric acid condensate sodium salt having a solid content of 35% and an opaque appearance. .

(Comparative Example 2) 100 parts of soybean separated protein, 10 parts of sodium hydroxide and 190 parts of water were placed in a reactor capable of reducing pressure.
The resulting mixture was hydrolyzed by stirring and mixing at 60 to 70 ° C. for 6 hours to obtain a polypeptide having a solid content of 30%, an average molecular weight of about 440 and a volatile base component of 1.8%. Next, 30 parts of ethanol was added to 100 parts of the polypeptide, and 24 parts of lauric chloride was added dropwise at 50 ° C. while adjusting the pH to 10 with sodium hydroxide. 40 after dropping
Aged at ５０50 ° C. for 1 hour. Thereafter, hydrochloric acid was added to adjust the pH of the system to 1, followed by washing with water, and then neutralized with sodium hydroxide to obtain a soybean protein hydrolyzate peptide lauric acid condensate sodium salt having a solid content of 33% and an opaque appearance. Was.

The purity and impurities of the acylated peptide salt obtained in the above Examples and Comparative Examples were determined by GPC (column: sh
index. Analysis was carried out with Asahipak 310, solvent: water / acetonitrile = 1: 1). The unit is%.

[0028]

[Table 1]

When the acylated peptide salts obtained in the above Examples and Comparative Examples were dried and analyzed by an infrared absorption spectrum by the potassium iodide tablet method, Comparative Examples 1 and 2 showed strong absorption of fatty acid amide. Was done. Further, the water solubility of the acylated peptide salts obtained in the above Examples and Comparative Examples was visually measured. The artificial hard water used was 100 ppm and 300 ppm of calcium carbonate.

[0030]

[Table 2]

From the above test, it was found that the Examples produced transparent solutions at all concentrations and solvents, whereas the Comparative Examples were cloudy and had poor performance as surfactants. In addition, foaming of the acylated peptide salt obtained in the above Examples and Comparative Examples was measured by the Ross-Miles method. As measurement conditions, the height of the foam after 5 minutes at a concentration of acylated peptide of 0.25% and a water temperature of 40 ° C. was measured.

[0032]

[Table 3]

In all the solvents, the examples showed good foaming, whereas the comparative examples showed poor foaming and poor performance as a surfactant. Further, the skin irritation of the acylated peptide salts obtained in the above Examples and Comparative Examples was measured. A patch test was performed on each of the five subjects for a 48-hour occlusion of humans, and the average score determined by the following determination method was evaluated.

[0034]

[Table 4]

At all concentrations, the examples showed good hypoallergenicity, while the comparative examples showed slightly irritating results. When the nitrogen-containing substance was removed under reduced pressure as in Examples 1 and 2, an acylated peptide or a salt thereof having high purity and good performance when used as a surfactant was obtained. In the case where the removal under reduced pressure is not carried out as in (2) and (3), only those containing more impurities and having inferior performance when used as a surfactant can be produced as compared with the case where removal under reduced pressure is carried out.

[0036]

The effect of the present invention is to provide a novel method for producing an acylated peptide. According to the present invention,
An acylated peptide having high purity and showing better performance than an acylated peptide produced by a conventional production method when used as a surfactant can be obtained.

Claims (5)

[Claims] 1. An essential step is a step of hydrolyzing a protein or a protein hydrolyzate in the presence of an alkali; a step of removing a nitrogen-containing substance produced in the step under reduced pressure; The following general formula (1) obtained through a step of acylating a protein or a protein hydrolyzate: (Wherein, R ¹ represents a hydrocarbon group, R ² represents an amino acid side chain, M represents a hydrogen atom, a metal atom or ammonium, and n represents one or more numbers). A method for producing an acylated peptide. 2. The method according to claim 1, wherein the acylating agent used for the acylation is a fatty acid halide. 3. The method according to claim 1, wherein the protein or the protein hydrolyzate is a plant-derived protein or a plant-derived protein hydrolysate. 4. An acylated peptide produced by the production method according to any one of claims 1 to 3. 5. A surfactant produced by the production method according to claim 1.