JPH05339299A - Production of protein-polysaccharide complex compound - Google Patents

Abstract

PURPOSE:To efficiently mass-produce a protein-polysaccharide complex compound bonded by aminocarbonyl reaction at a low cost while suppressing the discoloration (browning) of the product. CONSTITUTION:A mixture of a protein and a branched polysaccharide is treated under high pressure (1,000-10,000kg/cm<2>) to obtain the objective protein- polysaccharide complex compound bonded by aminocarbonyl reaction.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a method for producing a protein-polysaccharide complex useful for foods, cosmetics, pharmaceuticals and the like. More specifically, it relates to a method for producing a protein-polysaccharide complex bound by an aminocarbonyl reaction, which comprises treating a mixture of a protein and a branched polysaccharide at high pressure.

[0002]

2. Description of the Related Art Conventionally, proteins have been chemically modified (J. Agric.
Chem., 33,125,1985) and enzyme modification (Agric.Biol.Chem., 5
0,3025,1986) and denaturing the protein by heating etc. (Agric.Biol.Chem., 45,2775,1981) have been carried out to improve the functionality of the protein. It was However, since conventional chemical modification, that is, acylation, alkylation, amidation, deamidation, esterification, etc. of proteins and the process of binding a carbohydrate or a fatty acid to a protein, a drug is used, these compounds are used in terms of safety. There is a problem in using the protein obtained by the chemical modification of the above as a food material. In general, proteins have an emulsifying activity, but they become insoluble due to denaturation due to treatment such as heating, and when used as emulsifiers, quality defects occur.

Therefore, a protein in which a branched polysaccharide and a protein are bound by an extremely spontaneous and safe reaction called an aminocarbonyl reaction without activation treatment with a drug which has recently been problematic in safety- A polysaccharide complex has been proposed (JP-A-3-215498), and this protein-
The polysaccharide complex is soluble in water, exhibits higher emulsification activity than the complex obtained by activation treatment with a drug, has a narrow molecular weight distribution, and has favorable properties for quality control. Compared to emulsifiers, it has excellent acid resistance and salt resistance,
It also has the advantage that the emulsification properties are improved in an alkaline solution and by heating.

[0004] Especially, when lysozyme is used as a protein alone, it has no antibacterial activity against Gram-negative bacteria, but it has a new property that antibacterial activity against Gram-negative bacteria is exhibited by forming a complex with a polysaccharide. It is proposed to join. Conventionally, as a method for producing a protein-polysaccharide complex bound by an aminocarbonyl reaction, a protein and a branched polysaccharide are mixed at an appropriate ratio to prepare an aqueous solution, and after freeze-drying, the obtained powder is generally prepared. 50 ~
A method of reacting for 2 to 6 weeks under the conditions of 80 ° C. and relative humidity of 60 to 80% has been proposed. However, according to this method, as the protein-polysaccharide complex is formed by the aminocarbonyl reaction, a browning substance that is repeatedly dehydrated, decomposed, rearranged, polymerized, etc., is easily produced, and thus the obtained protein-
The polysaccharide complex is highly colored (browning), and when it is applied to foods, cosmetics, pharmaceuticals, etc. from the aspect of aroma and taste,
Uses and addition amounts are limited. In addition, this method has low production efficiency, difficult process control, and requires enormous equipment for mass production. Therefore, the product price of the protein-polysaccharide complex is considerably high, and its practicality is extremely poor.

[0005]

DISCLOSURE OF THE INVENTION The present invention provides a protein-polysaccharide complex bound by an aminocarbonyl reaction,
It is an object of the present invention to provide a method for efficiently producing a large amount at low cost with little coloring (browning).

[0006]

Means for Solving the Problems As a result of intensive studies to solve the above problems, the present inventors have found that amino acids can be efficiently treated by treating a mixture of a protein and a branched polysaccharide at high pressure. It was found that the carbonyl reaction proceeds to form a protein-polysaccharide complex with little coloring (browning), and based on this, the present invention has been completed. That is, the present invention provides a method for producing a protein-polysaccharide complex bound by an aminocarbonyl reaction, which comprises treating a mixture of a protein and a branched polysaccharide at high pressure.

The present invention will be described in detail below. The protein referred to in the present invention may be of animal origin or plant origin, and examples thereof include whole egg white, ovalbumin, lysozyme, milk protein, fish meat protein, soybean protein and wheat gluten. Examples of the branched polysaccharides include dextran, dextrin, pullulan, xanthan gum, guar gum, carrageenan, chondroitin sulfate, and partial decomposition products thereof. These branched polysaccharides need not be subjected to an activation treatment using a chemical such as cyanogen bromide, sodium periodate, or cyanuric chloride as an activator, and are subjected to an aminocarbonyl reaction as they are to form a complex with a protein. Can be formed.

In order to carry out the present invention, first, a protein and a branched polysaccharide are mixed and hydrolyzed at an appropriate ratio to prepare a mixture, which may be further freeze-dried or spray-dried. The water content of the mixture of protein and branched polysaccharide is not particularly limited, but is usually about 5 to 90% by weight. The pressure applied to the mixture of protein and branched polysaccharide is 1,000-10,000kg / cm
² and the pressure is less than 1,000 kg / cm ² , the aminocarbonyl reaction does not proceed efficiently and 10,000 kg / cm ²
If it exceeds, it is practically disadvantageous in terms of the apparatus cost. It is also possible to use heating at the same time as or after the pressurization. The high-pressure treatment time is appropriately selected depending on the type of protein and branched polysaccharide used, pressure, amount of water added, temperature, size, morphology, etc., but is usually about 5 minutes to 5 hours.

As a pressurizing device, a hydrostatic pressure of 1,000 to 10,0
Any device can be used as long as a pressure of 00 kg / cm ² can be applied. When a large amount of pressure is applied, a steel plunger can be hermetically inserted into a steel cylinder. Etc. are used. The pressure medium used in the pressure device is not limited, but the use of city water is preferable.
In this way, by treating the mixture with high pressure, the aminocarbonyl reaction proceeds efficiently, and a protein-polysaccharide complex with less coloring (browning) is formed. The protein-polysaccharide complex of the present invention can be widely applied as a raw material for foods, cosmetics, pharmaceuticals, etc., such as emulsifiers and antibacterial agents. Examples of the present invention will be shown below, but the present invention is not limited thereto.

[0010]

【Example】

Example 1 A mixture of ovalbumin and dextran (average molecular weight 75,000) (weight ratio 1: 5) was dissolved in 9 times amount of water and treated with a pressure device at 10,000 kg / cm ² , 40 ° C. for 5 hours. ,
Spray drying was performed to obtain a protein-polysaccharide complex. The obtained complex was subjected to gel filtration using Sephacryl S-300 (70 × 3 cm ² ) (eluent: 50 mM containing 50 mM NaCl).
mM acetate buffer (pH 5.0), flow rate: 0.36 ml / min).
As a result, the elution position of ovalbumin in the complex of the present invention moved to the polymer side from the elution position of ovalbumin alone, and that position almost coincided with the elution position of dextran and the elution pattern of both of them. It was confirmed that the ovalbumin-dextran complex was formed from the similarity of the proteins. The obtained complex was treated with SDS-
Polyacrylamide gel electrophoresis (0.1% SD
This was carried out according to the method of Laemmli (Nature, 227, 680, 1970) using a 10% acrylamide separation gel containing S and a 3% fixed gel). As a result, in the complex of the present invention, a single band of protein and carbohydrate staining appeared near the boundary between the fixed gel and the separation gel, which revealed that ovalbumin was covalently bound to dextran. ..

The average molecular weight of the obtained complex was determined using a low-angle light scattering technique coupled with high performance liquid chromatography (J. Agric. Food Chem., 36, 421, 1988). As a result, the average molecular weight of the complex of the present invention is about 200,000,
This value is in good agreement with the estimated value obtained from the binding ratio of ovalbumin to dextran in the present complex of 1: 3 and the molar ratio of 1: 1.6 to 2.2. Furthermore, the emulsification property of the obtained complex was evaluated by the method of Pearce et al. (J. Agric. Food Che
m., 26, 716, 1978), the absorbance (turbidity) immediately after emulsification, which indicates emulsifying activity, is about 4 times that of ovalbumin alone, and the half-life of the absorbance after emulsification, which indicates emulsion stability, is Ovalbumin alone was about 30 seconds, whereas that of the complex of the present invention was about 10 minutes, and the emulsion stability was excellent.

Example 2 Mixture of casein and guar gum enzymatic degradation product (weight ratio 1:
1) is dissolved in 10 times amount of water, freeze-dried,
It was treated at 000 kg / cm ² , 60 ° C. for 1 hour to obtain a protein-polysaccharide complex. The obtained complex was subjected to the gel filtration of Example 1, the SDS-polyacrylamide gel electrophoresis and the high-performance liquid chromatography coupled low angle light scattering technique. As a result, the casein-guar gum enzymatic degradation product complex was found to be It was revealed that it was formed. Further, as a result of measuring the emulsification property of Example 1 for the obtained composite, the emulsification activity and the emulsion stability were improved as compared with casein alone.

Example 3 A mixture of lysozyme and dextrin (weight ratio 1: 3) was dispersed in an equal amount of water, and 1,000 kg / cm ² , with a pressure device,
After treating at 90 ° C. for 20 minutes, it was freeze-dried to obtain a protein-polysaccharide complex. The resulting complex was subjected to gel filtration of Example 1, SDS-polyacrylamide gel electrophoresis and high-performance liquid chromatography coupled low angle light scattering technique, resulting in the formation of a lysozyme-dextrin complex. It became clear. Furthermore, as a result of measuring the antibacterial activity of the obtained complex by an antibacterial spectrum, A.
A strong antibacterial action was observed by adding 100 ppm in agar medium to Gram-negative bacteria of hydrophila, V. parahaemolyticus and E. coli.

Comparative Example 1 A mixture of ovalbumin and dextran (average molecular weight 75,000) (weight ratio 1: 5) was dissolved in 1 g of ovalbumin at a ratio of 100 ml of water, stirred for 30 minutes and then freeze-dried. The obtained powder was placed in a glass dish and allowed to react by being kept at 60 ° C. for 3 weeks in a desiccator adjusted to about 65% humidity with a saturated potassium iodide solution.

Test Example 1 Each of the composites obtained in Example 1 and Comparative Example 1 was dissolved in distilled water at a certain concentration and the absorbance at 470 nm was measured to compare the degree of browning. As a result, the absorbance of the complex of Comparative Example 1 was 0.035, whereas that of the complex of the present invention of Example 1 was 0.007, and browning was significantly reduced. The scent and taste of the composite of the present invention of Example 1 were better than those of the composite of Comparative Example 1.

[0016]

As described above in detail, according to the present invention, a protein-polysaccharide complex bound by an aminocarbonyl reaction can be efficiently produced in large quantities at low cost with little coloring (browning). The protein-polysaccharide complex can be suitably used for producing foods, cosmetics, pharmaceuticals and the like. Therefore, the industrial significance of the present invention is very great.

Front page continuation (72) Inventor Masayoshi Nishiyama 9-5 Akahori Shinmachi, Yokkaichi-shi, Mie Taiyo Kagaku Co., Ltd.

Claims (2)

[Claims] 1. A method for producing a protein-polysaccharide complex bound by an aminocarbonyl reaction, which comprises treating a mixture of a protein and a branched polysaccharide at high pressure. 2. The pressure for high-pressure treatment is 1,000 to 10,000 kg /
The manufacturing method according to claim 1, which is cm ^2.