JPS58174232A - Polypeptide emulsifier - Google Patents

Abstract

PURPOSE:To obtain an emulsifier having emulsification stability without spoling the waste of the emulsion product and without any drawback in nutrition by constituting the same of 5-50 pieces of amino acids obtd. by acting proteolytic enzyme on total casein. CONSTITUTION:A polypeptide emulsifier is constituted of 5-50 pieces of amino acids obtd. by acting proteolytic enzyme (e.g.; chymosin) on alphaS-casein and/or beta- casein. THe conditions for acting the proteolytic enzyme on the starting raw material vary largely with the kinds of the enzyme used; in general, in the case of the alphaS-casein, the enzyme is acted on the casein of 0.1-20wt% concn. for one minute- one week at 1/2-1/50,000 enzyme concn. based on the concn. of the casein, 2-12pH and 3-60 deg.C reaction temp. Techniques such as gel filtration, ion exchange chromatography, electrophoresis for sepn., or the like are applied for separating the intended polypeptide from the reaction mixture obtd. by the above-mentioned enzyme decomposition.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an emulsifier comprising a polypeptide obtained by decomposing whole Cain tract or α1-casein and/or l-casein with tII leukomatase.

Conventionally, fatty acid monoglyceride, sorbitan fatty acid ester, egg yolk, lecithin, alginic acid, gelatin, and other O proteins have been used as emulsifiers in the production of processed cheese. Scale salt) is also used as an emulsifier.

However, commonly used monoglyceride emulsifiers are not suitable for producing food emulsions from the viewpoint of flavor, and protein-based emulsifiers generally have weak emulsifying power and, in addition, have a poor flavor. The disadvantage is that it impairs the

In addition, it has been reported that a partial hydrolyzate of casein is used as an emulsifier (#Open 1854).
-95747, page 1), but in this report, the peptide itself having a specific number of a
There is no mention of using it as an emulsifier, nor does it suggest anything about the emulsifying properties of polypeptides.90 Report on the use of enzymatic decomposition products of soybean protein as an emulsifier (JP-A-Sho! $ No. 5-39725,
JP-A-56-26171 and JP-A-1856-4255
No. 5) is also seen, but enzymatic decomposition products of soybeans generally have a poor flavor and are not necessarily satisfactory in terms of nutritional value.In addition, soybean protein is relatively rigid and cannot be heated or treated with alcohol beforehand. It has the disadvantage that it is not susceptible to enzymatic action unless it is subjected to denaturation treatment 11.

The present invention has been developed in view of the above-mentioned current situation, and it does not impair the flavor of the manufactured carbon dioxide product, has no nutritional defects, and improves emulsion stability, especially emulsion stability during heating. The purpose is to provide an excellent emulsifier.

By the way, "1 air meter" in Komu means 1 m1 of oil in water.
(0/W), water-in-oil m1 (Wlo), which tastes each industrial mulch soda and double-process mulch bottle tit, 0/W
Examples of the emulsion can include cheese, tin, dressing, spread 40 food O#t crab shampoo, cosmetics, liquid nutritional supplements, etc., and examples of the t&W10 emulsion can include foods such as marcarin and putter.

Therefore, the emulsifier according to the present invention can be applied to the production of these zero-process mill meal products.

The present invention will be explained in detail below.

The emulsifier according to the present invention consists of a polypeptide composed of 5 to 50 amino acids obtained by treating whole casein or αS-casein and/or β-casein with a protease. % mold.

Casein, which is used as a starting material in the preparation of the emulsifier of the present invention, is different from soybean protein in that it has a relatively loose higher-order structure, so it can be used raw without pretreatment such as heating or alcohol treatment. It has the advantage that even one trench is not susceptible to enzymatic decomposition.

The above-mentioned whole casein as a starting material is whole milk, skim milk, or acid casein obtained by adjusting the OpH of skim milk to around 4.6, or lactic acid casein obtained by fermenting milk*gi. β-casein strain Whole casein fraction, for example, obtained by fractionation using the urea method [For αS-casein, see Zittle at ml, J, Dairy 8c.
i, 4G, 118B (1963), Hlpp @tal, J, Am, C for β-casein.
h@m, Soc, F 4.4822 (1952)].

Among these raw materials, from the viewpoint of separation and purification of polypeptides obtained by enzymatic degradation, it is preferable to use αS-casein and/or β-casein obtained by pre-fractionation from total casein.The present invention Then total casein or αS-casein and/or β
- Obtained by decomposing casein tII white matter fraction ps# element,
Although a polypeptide composed of 5 to 50 akyno acids is used as an emulsifier, the proteolytic element used in ζζ is not particularly limited and a wide variety of enzymes can be used. Examples include chymosin, papain, pepsin, plasmin, trypsin, α-xotrypsin, and peptidase derived from lactic acid 1.

In addition, in order to impart high emulsifying power to the peptide obtained by the above-mentioned enzymatic degradation, the balance between hydrophilicity and hydrophobicity is important. It is preferable to use an enzyme that is highly specific in terms of decomposition and acts as a proteolytic enzyme, rather than one that enzymatically decomposes the enzyme randomly as seen in other substances. Examples of such enzymes include chymosin and pepsin.

In the present invention, the conditions under which these proteolytic enzymes are allowed to act on the above-mentioned starting materials vary greatly depending on the type of enzyme used, but in general, in the case of α-casein, the casein 1/2 to 1150
Enzyme concentration of ρ00, pH 2-12, reaction temperature 3-60℃
Let it act for 1 minute to 1 week.

For example, when using chymosin (rennet) as the d1 enzyme and αS-casein as the starting material, the negative value of αHatake-casein is 1.6% by weight, the rennet concentration is 0.4% by weight, and the pH is 6, 4, and 30°C. This enzymatic decomposition produces a polypeptide having 23 to 24 amino acids L1T2 from the N-terminus, and the remainder is αs-I casein. The degree of decomposition when a proteolytic enzyme is applied to α-casein can be determined simply by quantifying the amount of nitrogen soluble in TCA (IJ dichloroic acid), but it is more accurate. To measure
)] to analyze the migration pattern of the enzymatic decomposition reaction mixture, and this measurement allows the fi] of αl-casein to be analyzed.
This makes it possible to know what kind of peptides ``Regret'' has been broken down into.

It is preferable to apply various techniques such as ion exchange chromatography, high performance liquid chromatography, and separation electrophoresis to the reaction mixture obtained by the wI elemental decomposition described above, or a combination of these techniques may be used.

The polypeptide used in the present invention is a separated polypeptide consisting of 5 to 50 amino acids, preferably 10 to 30 amino acids, as described above, and has a balance between hydrophilic groups and hydrophobic groups. Preferably harvested ones.

In other words, in order to increase the emulsifying power of a polypeptide, the balance between hydrophilicity and hydrophobicity within the polypeptide must be maintained. It is necessary to make the number between 50 and 50. In other words, the above balance is difficult to maintain even if the number of amino acids is less than 5 or more than 50.

In polypeptides with such hydrophilic and hydrophobic amphipathic properties, the hydrophobic groups bind to fat globules, while the hydrophilic groups exit into water to form stable emulsified carbon atoms, similar to the well-known emulsifiers. It becomes like this.

Next, the emulsifying properties of the polypeptide in the present invention will be described.

0/Wl! Cheddar cheese with a ripeness of 4 months, which exemplifies the emulsifying properties for cheese, which is a raw material.
A mixture of 25 grams of dry powder of a polypeptide containing amino lt2 (or 24@) prepared as shown in the example below dissolved in 180 grams of water was added to 19 milled in a Kupar [supplied to an emulsifying pot]. and 800 rp at 40℃
Emulsification is performed by stirring at a rotation speed of rn, and then heat sterilization (9
The emulsion stability of the Grossessners obtained by heating (at 0° C. for 30 minutes) was examined.

For comparison, processed cheese was obtained in the same manner by applying phosphate (molten salt) and soda casein, which have been conventionally used as emulsifiers, to the polypeptide, and their emulsion stability was also investigated.

As a result of step 7, when the polypeptide was added and stirred for about 3 minutes, the fat in the cheese was dispersed into the protein-aqueous phase and a stable emulsion system was obtained, but when the casein was added In Kameno, no matter how much stirring was continued, the fat separated due to heating.9 The fat did not disperse into the protein-water phase and did not emulsify.
When the 1J phosphate tube was added, an emulsified system was obtained by stirring for about 4 minutes.

Next, the oil-off value of cheese prepared by adding polypeptide and phosphate was measured by the following method, and the results were as follows.

Measurement of oil-off value Place the cheese sample on knee paper in a thermostat at 30℃ (humidity 901℃)
1 in a cheese sample held for 24 hours and then depressurized.
The value tm of IP1F is set to a fixed value.

The smaller value of 4 has better emulsion stability.

Cheese Added with Phosphate 2.3 As seen in the above results, the emulsion stability of the processed cheese prepared using the polypeptide according to the present invention as an emulsifier is significantly superior to that produced by the conventional method.

The emulsifier of the present invention can be applied to the emulsification of various oils and fats, such as the above-mentioned milk fat, such as coconut oil, safflower oil, castor oil, cottonseed oil, corn oil, palm oil, palm kernel oil, rapeseed oil, and vegetable oil. Examples of animal oils include bean oil, bean oil, sunflower oil, olive oil, cocoa butter, and animal oils such as lard, het, mutton tallow, and whale oil.

In order to use the polypeptide as an emulsifier of the present invention to emulsify these oils and fats, the polypeptide is dissolved in water and emulsified.
is added to oil and fat, and an emulsifying machine such as a high-speed emulsifying machine is applied to form an emulsion.

The amount of polypeptide added varies depending on the type of oil to be emulsified11, but it is usually 0.01 to 30 g/g6 for the oil/fat to be emulsified.
N, t [In the production of processed cheese, when the raw material cheese is semi-hard, 0.2
The range is preferably 1 to 10 parts by weight, preferably 1 to 10 parts by weight.

The emulsion prepared using the polypeptide according to the present invention as an emulsifier has excellent emulsion stability as described above, and has particularly good stability against heat, so it is suitable for use in sterilization processes when applied to food processing. This method is particularly advantageous for improving the quality and hygiene of industrial soda products because no destruction or phase conversion occurs, and high-temperature resin can be filled into the container immediately.

In addition, the polypeptide can be used in high amounts without adversely affecting the flavor of the product and without any nutritional drawbacks.
Rather, it has the advantage of being nutritionally enriched.

Examples are shown below.

Example Preparation of polypeptide: αS-casein 15JF was dissolved in 0.1 M acetate buffer (pH
64) Dissolved in 11 and heated to 30C. Add 0.7 μ of rennet to the resulting solution and react for 3 hours, then add 8
Heat at 0C for 10 minutes to inactivate the nonnet. After cooling the resulting reaction solution (# elementary decomposition solution) to room temperature, gel filtration and ion exchange chromatography are applied to obtain about 2 polypeptides having the primary structure shown below from the reaction solution.
I got I.

Arg-Pro-Lys-Hl 5-Pro-I 1e
-Lys-Hls-Gln-Gly-Leu-Pro-
Gln-Glu-Val-Leu-Aan-Glu-A
Emulsifying property of gn-Leu-Leu-Arg-Ph@(-Phe) polypeptide in margarine: Using the polypeptide obtained above as an emulsifier, the following f. Margarine having the composition shown in 1 was produced in a conventional manner using a votator. For comparison, margarine was produced in the same manner using monoglyceride and lecithin as emulsifiers instead of the above polypeptide, and the emulsion stability of both margarines was examined.

Table 1 Hydrogenated soybean oil (38C) 24.5 24.5
Palm oil 12.0 12
．． 0 White soybean oil 32.0 32.0 Corn oil 12.0
12.0 Skim milk powder 2.0 2.0 Salt 1.5 1.
5 Polypeptide 0.6- Monoglyceride -0.5 Cytin
0.1 water 15.0 1
5.0 The average diameter of water droplets in the dispersed state of both margarines was measured using a microscope and the results are as follows.

This invention 1.9μ comparison
Example 4.3μ The average diameter of clean water indicates the state of emulsion stability of 1-galin, and the smaller the diameter δ, the better the emulsion stability.

Attorney: Yoshi Kawaguchi & Li procedural amendment, Commissioner of the Patent Office, Kazuo Wakasugi, 1, Indication of case: Patent Application No. 28594, filed in 1982.
, Title of the invention Polypeptide emulsifier 3, Relationship with the case of the person making the amendment Patent applicant name (669) Snow Brand Milk Products Co., Ltd. 4, Agent 5, Date of amendment order 1 Issue 8, Details of the amendment are as follows: Correct accordingly.

(1) No. 13i [Insert a line break between lines 13 and 14 to insert the following description.

[Next, the application of the polypeptide thus obtained to various emulsified foods will be illustrated. (2) IE page 13, line 14: “Emulsifying properties of polypeptides in margarine:
” is corrected by “1) Emulsifying property of polypeptide in margarine:” K.

(3) Add the following statement after line 6 of the 15th Tribute.

A cream having the formulation shown below was prepared by the following procedure using 1 weight of the polypeptide obtained above as an emulsifier. In addition, for comparison, we used one with no emulsifier added, and soda casein (Na-ca@ein) instead of polypeptide.
) 1 wt.
60) Creams were produced in the same manner using 0.4 weight of each.

Cream composition: Salad oil 34 (weight 1 s) Wi-fat milk powder 4 Water 66 Cream manufacturing procedure: Disperse skim milk powder of the above composition in water with each emulsifier,
A mixture prepared by adding salad oil heated to 0° C. and 60° C. was stirred at a rotation speed of 100 rpm using a Polytron to form a cream. Check the size of the oil droplets for each cream obtained! The emulsification state was examined by observing the The results are shown in Section 2.

Table 2 As shown in Table 2, the size of oil droplets in the casein using soda casein was as large as in the casein without emulsifier, and the emulsification state was poor, but in the case using the polypeptide according to the present invention, the oil droplet size was as large as in the casein without emulsifier. The oil droplets are so small that the emulsified state is smooth, comparable to the case of the currently used sucrose m81PPm ester.

The emulsifying properties of polypeptides in the production of cheese-like products by adding butter oil to natural cheese were investigated in accordance with Section F.

Add 29 parts by weight of a 1.591 solution of polypeptide and 23 parts by weight of a 3.8 capsule solution to a formulation having the composition shown below, and heat at 40°C using a food cutter at about 200O.
The mixture was emulsified by stirring at rpm, and the emulsification state of the resulting emulsion was examined.

Composition of raw material mixture: 56% by weight Protein 13 Ash 2.3 Others 0.5 Water 16.2 The results show that when emulsified by adding 1.5% of polypeptide and 29% by weight of Solution III, the fat globules in the emulsion decrease. The size is 2.
4μ, and 3.μ of the polypeptide. When emulsified by adding 23% by weight of aUl solution, 1. It was sμ.

Claims (2)

[Claims] (1) An emulsifier consisting of a polypeptide composed of 5 to 50 amino acids, obtained by acting a proteolytic enzyme on all kain. (2) An emulsifier consisting of a polypeptide composed of 5 to 50 amino acids obtained by allowing a protease to act on α-casein and/or β-casein. 13) The emulsifier according to claim 1 or 2, wherein the polypeptide is composed of 10 to 30 amino acids.