JPH05339300A - 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 irradiated with microwave to obtain the objective protein-polysaccharide complex compound bonded by aminocarbonyl reaction. There is no particular restriction on the frequency, output and irradiation time of microwave, however, the frequency is usually about 2450MHz and the microwave is radiated for 0.5-30min at an output of several to several tens kilowatt. If necessary, the microwave irradiation treatment can be carried out in an evacuated atmosphere having a vacuum degree of 20-200 Torr.

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 subjecting a mixture of a protein and a branched polysaccharide to microwave irradiation.

[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 from the viewpoint 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 emulsifying activity, but insolubilization occurs 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 emulsifying 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, with the formation of the protein-polysaccharide complex by the aminocarbonyl reaction, a browning substance that is repeatedly dehydrated, decomposed, rearranged, polymerized, etc., is easily produced, and 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 is low in production efficiency, difficult in process control, and requires a huge amount of 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 a mixture of a protein and a branched polysaccharide is subjected to a microwave irradiation treatment to efficiently perform amino treatment. It was found that the carbonyl reaction proceeds to form a protein-polysaccharide complex with less coloring (browning),
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 subjecting a mixture of a protein and a branched polysaccharide to microwave irradiation.

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.

To carry out the present invention, first, a protein and a branched polysaccharide are mixed and watered at an appropriate ratio to prepare a mixture. The water content of the mixture of protein and branched polysaccharide is appropriately selected depending on the type of protein and polysaccharide used, irradiation conditions, size, morphology, etc., but is usually about 5 to 60% by weight. The frequency of the microwave used in the present invention, the output and the irradiation time are not particularly limited, but the frequency is usually 2450 MHz.
It only needs to have a high frequency, and its output is 0 to several tens of kw.
Irradiate for 5 to 30 minutes. If necessary, vacuum 20
It is also possible to perform microwave irradiation treatment in a reduced pressure atmosphere of ~ 200 torr. A continuous type or a batch type can be used as the microwave induction heater. In this way, by subjecting the mixture to microwave irradiation, the aminocarbonyl reaction proceeds efficiently and a protein-polysaccharide complex with less coloring (browning) is formed. Protein of the present invention-
The polysaccharide complex 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.

[0009]

【Example】

Example 1 Ovalbumin, dextran (average molecular weight 75,000) and water were mixed at a ratio of 2: 10: 1 and subjected to microwave treatment (24
50 MHz, 50 kw, 5 minutes) to obtain a protein-polysaccharide complex. The obtained complex was treated with Sephacryl S-300 (70 ×
Gel filtration was performed using 3 cm (eluent: 10 mMN
50 mM acetate buffer (pH 5.0) containing aCl, 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 the elution position of dextran almost coincided with 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 subjected to SDS-polyacrylamide gel electrophoresis (using a 10% acrylamide separation gel containing 0.1% SDS and a 3% fixed gel, Laemmli's method (Nature, 227, 68).
0,1970)). As a result, the complex of the present invention showed a single band of protein and carbohydrate staining 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 weight ratio of the binding ratio of ovalbumin to dextran in this 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 emulsification 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 Casein, a guar gum enzymatic hydrolyzate and water were mixed at a ratio of 1: 1: 2 and subjected to microwave treatment (2450 MHz, 5 kw, 30 minutes) to give a protein-polysaccharide complex. Got 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.
It was revealed that a guar-gum enzymatic degradation product complex was formed. Further, as a result of measuring the emulsification characteristics of the obtained complex in Example 1, the emulsification activity and the emulsion stability were improved as compared with casein alone.

Example 3 Lysozyme, dextrin, and water were mixed at a ratio of 5: 6: 1 and subjected to microwave treatment (2450 MHz, 5 kw, 100 torr,
For 30 minutes) 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, the antibacterial activity of the obtained complex was measured by an antibacterial spectrum. As a result, A.hydrophila, V.paraha
10 against gram-negative bacteria of emolyticus and E. coli in agar medium
A strong antibacterial action was observed when 0 ppm was added.

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, while that of the complex of the present invention of Example 1 was 0.007, which markedly reduced browning. 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.

[0015]

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.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Masayoshi Nishiyama 9-5 Akahori Shinmachi, Yokkaichi-shi, Mie Taiyo Kagaku Co., Ltd.

Claims (1)

[Claims] 1. A method for producing a protein-polysaccharide complex bound by an aminocarbonyl reaction, which comprises subjecting a mixture of a protein and a branched polysaccharide to microwave irradiation.