JPH0638687A - Production of hydrolyzed product of protein - Google Patents

Abstract

PURPOSE:To obtain a hydrolyzed protein having excellent safety and palatability while suppressing the formation of mutagenic halogen compounds harmful to human body. CONSTITUTION:A protein raw material is treated with an acid protease in acidic state to solubilize the protein, the insoluble components are removed and the residue is hydrolyzed while keeping the acid concentration to about 7.5-15 W/V% by adding hydrochloric acid to obtain hydrolyzed product of protein. The formation of mutagenic halogen compound originated from oil and fat can be suppressed by the treatment with hydrochloric acid under mild condition after the treatment of the protein raw material with acid protease. A hydrolyzed protein having excellent safety and palatability can be produced by this process.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION The present invention relates to a mutagenic substance which is harmful to the human body when a hydrolyzate of a protein is produced by reacting oils and fats present in the protein raw material with hydrochloric acid. The present invention relates to a method for producing a protein hydrolyzate which is excellent in safety and taste by controlling the production of halogen compounds contained therein.

[0002]

2. Description of the Related Art Conventionally, protein hydrolysates are defatted soybeans, wheat gluten, gluten meal (maize protein), plant proteins such as yeast, and animal proteins such as fish meal, bone extract, gelatin and casein.
It is produced by hydrolysis with molar hydrochloric acid and is widely used in food processing as a vegetable protein hydrolyzate (HPP), an animal protein hydrolyzate (HAP) or an amino acid seasoning. However, recently, in the protein hydrolyzate produced by the hydrochloric acid decomposition method, glycerin from the fats and oils present in the raw material reacts with hydrochloric acid, and 1,3-dichloropropanol (1,3-dichloropropanol (1,3- Dichloropropanol), 2,3-
Dichloropropanol (2,3-Dichloropropanol), 3-
Monochloropropanediol (3-Monochloropropandio
l, hereinafter abbreviated as 3-MCP) or 2-monochloropropane diol (2-Monochloropropandiol) was discovered, and these compounds were detected from HPP and HAP. As a method for obtaining a protein hydrolyzate that does not contain these mutagenic compounds, there is a method for producing a protein hydrolyzate by a protease without using the hydrochloric acid decomposition method. However, when a protein is decomposed only with an enzyme, the amount of free amino acid produced is generally small, and the resulting hydrolyzate often has a bitter taste. Furthermore, in the case of glutamic acid or a protein containing glutamine, even amino acids are not easily decomposed, the free glutamic acid content of protein hydrolysates by the enzymatic method is significantly lower than that by the hydrochloric acid decomposition method, and its taste is weak, so it is practically used as a seasoning. It was difficult to convert. On the other hand, a method of adding wheat gluten to an aqueous solution having a neutral protease to hydrolyze it, adding an acid to the hydrolyzed soluble protein, heating the mixture, and substantially deamidating the hydrolyzate has been reported. (Japanese Patent Laid-Open No. 3-5
3850).

[0004]

Means for Solving the Problems In order to solve the above-mentioned problems, the present inventors have studied a method for producing a protein hydrolyzate having a low mutagenic compound content and a strong taste. By decomposing the protein raw material with acidic protease, extracting and separating the protein into peptides, and then decomposing the obtained peptides with low concentration hydrochloric acid for a long time at low temperature, almost no mutagenic compounds are contained, The present inventors have completed the present invention by knowing that a seasoning having an excellent taste can be produced.
That is, the present invention is to solubilize the protein by allowing acidic protease to act on the raw material protein to remove the insoluble matter,
Further, add hydrochloric acid to adjust the acid concentration to about 7.5 to 15 W / V.
It is a method for producing a protein hydrolyzate, which is characterized in that hydrolysis is performed while keeping the content at%.

The raw material protein of the present invention is a protein raw material usually used for the production of protein hydrolysates. For example, vegetable protein such as defatted soybean, wheat gluten, corn gluten, gluten meal, defatted rice bran, etc., protein of fungi such as yeast, protein of algae such as chlorella, spirulina or protein such as fish meal, bone extract, gelatin, casein, collagen Such as animal protein. Of these, defatted soybean, wheat gluten, corn gluten, yeast, gelatin and casein are preferable. More preferably, defatted soybean and yeast. These raw material proteins may be used alone or in combination of two or more kinds, and the mixing ratio thereof is appropriately selected. Examples of the acidic protease include, for example, acid protease of Hiirotake origin (Takeda Pharmaceutical Co., Ltd.), panprosin (Yakult Pharmaceutical Co., Ltd.), Protease M (Amano Pharmaceutical Co., Ltd.), Newase F (Amano Pharmaceutical Co., Ltd.). )), Morcin (Sorijin Pharmaceutical Co., Ltd.), Sumiteam AP (Shin Nippon Kagaku Co., Ltd.), Procutase (Meiji Seika Co., Ltd.), Denapsin (Nagase Sangyo Co., Ltd.) and the like. Acidic proteases of the Hiirotake origin, panprosin and Protease M are preferred. These 1 type (s) or 2 or more types are used.

A method for solubilizing a raw material protein by causing an acidic protease to act on the raw material protein will be described below. First, an aqueous suspension of the starting protein is prepared. The concentration of the raw material protein in the aqueous suspension may be such that it can be stirred so that the enzymatic reaction in the subsequent step can be carried out efficiently, and the concentration is about 5 to 35 W.
/ W%. It is preferably about 5 to 15 W / W%.
The proteolytic rate (yield of nitrogen) of the enzyme increases as the concentration decreases. The suspension may be sterilized if necessary. For example, it may be heated at about 120 ° C. for about 20 minutes. The above suspension is adjusted to acidic. Acids are used as regulators.
For example, mineral acids such as hydrochloric acid, sulfuric acid and phosphoric acid are used.
Hydrochloric acid is preferred. The acidic pH to be adjusted depends on the acidic protease used, and the optimum p
Adjust to H. Usually the pH is about 2-5. It is preferably about 2.5-4. The amount of acidic protease used is about 0.1 to 7 W / W% with respect to the raw material protein. It is preferably about 0.5 to 5 W / W%. Titer is raw protein 10
It is about 50,000 to 3.5 million units for 0 g. It is preferably about 250,000 to 2.5 million units. The titer was measured by the Hagiwara modified method (Shiro Akahori, Enzyme Research Method Vol. 2, 238,
Asakura Shoten, 1956). The higher the amount of acidic protease used, the lower the molecular weight of the starting protein, and the milder the conditions for hydrochloric acid decomposition in the subsequent step. The reaction time is about 8 to 72 hours. It is preferably about 20 to 50 hours. The reaction temperature is appropriately selected depending on the optimum temperature of the enzyme used. Usually, it is about 30 to 60 ° C. It is preferably about 40 to 50 ° C.

Next, the insoluble matter is removed, and the solubilized raw material protein and the insoluble matter are separated. The method for removing the insoluble matter is not particularly limited. Insoluble matter is removed by, for example, centrifugation, filtration or decantation. When removing the insoluble matter, the enzyme in the solubilized protein may be inactivated by heating or the like. The solubilized protein is concentrated if necessary. The solid matter concentration may be in a state that can be stirred in order to efficiently perform the hydrochloric acid decomposition reaction in the subsequent step. About 15-50W
It may be / V%. In order to keep the salt content of the protein hydrolyzate as the final product at 50 W / W% or less, the solid concentration is preferably about 25-40 W / V%. Further, hydrochloric acid is added for hydrolysis. The acid concentration during hydrolysis is about 7.
Keep at 5 to 15 W / V%. The HCl / total nitrogen molar ratio ranges from about 0.8 to 1.5. Preferably about 1.
It is 0 to 1.5. Here, the HCl / total nitrogen molar ratio is hydrochloric acid molar concentration / total nitrogen molar concentration. Within this range, the higher the hydrochloric acid concentration and the higher the reaction temperature, the shorter time the amino acid is decomposed. The reaction temperature is about 90-11
It is 0 ° C. It is preferably about 95 to 105 ° C. The reaction time is appropriately selected depending on the hydrochloric acid concentration, the HCl / total nitrogen molar ratio and the reaction temperature. It is usually about 11 to 72 hours.
About 15-48 hours is preferred. After hydrolysis of hydrochloric acid, neutralize with base. Examples of the base include sodium hydroxide,
Potassium hydroxide, potassium carbonate, etc. are used. Of these, sodium hydroxide is preferred. Adjust the pH to about 5-6.5. The resulting protein hydrolyzate is filtered, if necessary.
The protein hydrolyzate powder may be obtained by treatments such as decolorization, concentration and spray drying.

[0008]

INDUSTRIAL APPLICABILITY According to the present invention, acid protease is allowed to act on a raw material protein, and then hydrochloric acid is allowed to act under mild conditions to reduce the amount of mutagenic halogen compounds caused by fats and oils, and to improve safety and taste. A protein hydrolyzate having excellent properties can be produced.

[0009]

The present invention will be described in more detail with reference to the following examples. Example 1 To 2.0 kg of defatted soybeans, 18.0 liters of water was added and suspended, and then heated at 120 ° C. for 20 minutes. Then, concentrated hydrochloric acid (3
6 W / W%) 220 ml was added to adjust the pH to 2.8, and 80 g of acid protease of Hiirotake origin (Takeda Pharmaceutical Co., Ltd., 5 × 10 ⁵ units / g) was added to the enzyme at 45 ° C. for 24 hours. It was disassembled. After completion of the reaction, the enzyme decomposed solution 1 was prepared by centrifugation at 5000 rpm for 20 minutes in an acidic state.
3.0 liters (solid concentration 9.2 W / V%) were obtained. This enzymatic degradation solution was concentrated to 3.6 liters. Concentrated liquid has a solid content of 30.0 W / V% and total nitrogen of 3.5 W / V
%, The acid concentration was 2.2 W / V%, and the specific gravity (20 ° C.) was 1.14. This concentrated solution is divided into 4 by 900g, about 36W
/ W% hydrochloric acid (A) 240 g, (B) 160
g, (C) 40 g, and (D) 0 g each were added, and water was added to make the total amount 1 liter. Each was equipped with a reflux condenser and hydrolyzed at 105 ° C. for 24 hours.

At this time, the hydrochloric acid concentration of each sample and HCl /
The total nitrogen molar ratio (hereinafter abbreviated as molar ratio) is (A) hydrochloric acid concentration 10.38 W / V%, molar ratio 1.44, (B) hydrochloric acid concentration 7.50 W / V%, molar ratio 1.04, (C) Hydrochloric acid concentration 2.88 W / V%, molar ratio 0.40, (D) Hydrochloric acid concentration 1.
It was 74 W / V% and the molar ratio was 0.24. After the completion of hydrolysis, the mixture was neutralized to pH 5.5 with 45 W / W% sodium hydroxide, decolorized with decolorizing carbon (“Carboraffin” Takeda Pharmaceutical Co., Ltd.), and spray-dried to obtain powder samples A and B. , C and D were obtained. On the other hand, to 600 g of defatted soybeans, 300 ml of 36 W / W% hydrochloric acid and 300 ml of water were added, and a reflux condenser was attached to carry out hydrolysis at 105 ° C. for 24 hours. At this time, the hydrochloric acid concentration was 21.0 W / V% and the molar ratio was 1.
It was 0. After completion of hydrolysis, neutralization is performed to filter out humic substances, the filtrate is decolorized, and then spray-dried to obtain powder sample E.
Got For the above powder samples, salt, total nitrogen, sodium L-glutamate (hereinafter abbreviated as MSG), 3-M
Analysis of CP was performed. 3-MCP analysis by Roland
Whitman (Lebens Mitter Unter Zufn und Forschung, 193, 224, 1
991) reported by the GC / MS method.

The results are shown in Table 1.

[Table 1] As is apparent from the above, the samples A and B prepared by the method of the present invention have a 3-MCP content of 2% of the conventional hydrochloric acid hydrolysis method.
The content was below, and the content of MSG was large and the quality was good.

Example 2 Wheat gluten (2.0 kg) was suspended by adding 18.0 liters of water, and then heated at 120 ° C. for 20 minutes. Then 3
After adjusting the pH to 3.0 by adding 200 ml of 6 W / W% hydrochloric acid, 40 g of acid protease of Hiiroketake origin (Takeda Pharmaceutical Co., Ltd., 5 × 10 ⁵ units / g) was added at 45 ° C.
The enzyme was decomposed for 24 hours. Then, the pH was adjusted to 4.0 with 5 ml of 36 W / W% hydrochloric acid, 20 g of panprosin (36,600 units of Yakult Honsha Co., Ltd.) was added, and enzymatic decomposition was carried out at 40 ° C. for 12 hours. After completion of the reaction, the reaction mixture was centrifuged at 5000 rpm for 20 minutes in an acidic state, and 16.5 liters of enzyme decomposition solution (solid content concentration 9.8 W / V
%) Was obtained. This enzyme-decomposed solution was concentrated to 4.5 liters. The solid content concentration of the concentrate is 35.9 W / V%, total nitrogen is 4.7 W / V%, hydrochloric acid concentration is 1.9 W / V%, and specific gravity (2
(0 ° C.) was 1.14. This concentrated liquid was divided into 900 g in 5 portions (A to E), 240 g of hydrochloric acid was added to each, and water was added to make the total amount 1 liter. A reflux condenser is attached to each of them for a predetermined time (A: 8 hours, B: 16 hours, C: 24 hours, D: 36 hours, E: 48 hours), 1
Hydrolysis was performed at 05 ° C. At this time, the hydrochloric acid concentration of each sample was 10.14 W / V%, and the HCl / total nitrogen molar ratio was 1.05.
Met. After completion of the hydrolysis, the mixture was neutralized to pH 5.5 with 45 W / W% sodium hydroxide, decolorized with decolorizing carbon (“Carborafin” Takeda Pharmaceutical Co., Ltd.), filtered and spray-dried to obtain powder samples A and B, C, D and E were obtained.

On the other hand, 36 W / W per 500 g of wheat gluten
% 300 ml of hydrochloric acid and 300 m of water were added, and a reflux condenser was attached to carry out hydrolysis at the boiling point for 24 hours. At this time, the hydrochloric acid concentration was 21.0 W / V% and the molar ratio was 0.795. After the completion of hydrolysis, neutralized humic substances, decolorization, filtration and spray drying were performed to obtain powder sample F. The above powder samples were analyzed for common salt, total nitrogen, MSG and 3-MCP.
The analysis of 3-MCP was performed by the GC-MS method of Example 1.

The results are shown in Table 2.

[Table 2] As is apparent from this, the 3-MCP content of Samples A to E is reduced to 2% of the conventional hydrochloric acid hydrolysis method. The MSG content of sample A is low. Therefore, it was found that the samples B to E prepared by the method of the present invention had good quality.

Claims (4)

[Claims] 1. A raw material protein is hydrolyzed by allowing an acidic protease to act on the raw material protein to solubilize the protein and remove insoluble matter, and then add hydrochloric acid to maintain the acid concentration at about 7.5 to 15 W / V%. A method for producing a protein hydrolyzate, which comprises: 2. A process according to claim 1, wherein the HCl / total nitrogen molar ratio upon acid hydrolysis is about 0.8 to 1.5. 3. The method according to claim 1, wherein the raw material protein is defatted soybean. 4. The method according to claim 1, wherein the starting protein is yeast.