JP2897780B2 - Manufacturing method of highly polymerized gelatin - Google Patents

Description

Description: BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention mainly relates to characteristics such as high viscosity and high melting point obtained by cross-linking and polymerizing gelatin with transglutaminase and then stopping the reaction by inactivating the enzyme. A process for producing high molecular weight gelatin having

2. Description of the Related Art Gelatin is a hydrolyzate of collagen, which is a kind of protein contained in skin, bone, etc., and is currently widely used for capsules for photography, food, medicine, cosmetics, etc. ing. However, gelatin has a melting point near room temperature, so it softens at room temperature.Therefore, when transporting other than dried products, or when high melting point and high viscosity are required, the use of natural gelatin is limited. there were.

In order to improve these drawbacks, aldehydes such as glutaraldehyde and gelling agents (or cross-linking agents) such as alum group such as potassium alum have been used, but satisfactory results have not been obtained.

Transglutaminase is an enzyme that forms an ε- (γ-glutamyl) lysine bond between a γ-carboxamide group of a glutamine residue and an ε-amino group of a lysine residue in a protein to cross-link and polymerize the protein. There are literatures in which transglutaminase was allowed to act on solutions of various concentrations of proteins such as casein and soy globulin. In addition, there is an example in which transglutaminase is allowed to act on a high-concentration solution of protein and gelatin to form a gelled product (Japanese Patent Application Laid-Open No. 58-1496).
45, JP-A-1-27471). However, there is no example of producing a polymerized gelatin by reacting transglutaminase on gelatin and then inactivating the enzyme as in the present invention.

Problems to be Solved by the Invention Gelatin, as it is, has a melting point around room temperature,
Even when used for transportation other than dried products, capsules for food medicine, cosmetics, etc., their low melting point and viscosity often pose a problem. Various gelling agents have been used to remedy the above drawbacks, but their main purpose was to obtain a gelled product. Aldehydes among the gelling agents were highly irritating and were not suitable for food and medicine, and the reaction was not sufficient in various alum groups. In addition, since the reaction was rapid, control during production was difficult.

The present inventors have proposed that a transglutaminase of 0.1 units or more, preferably 2 units to 80 units per 1 g of gelatin is allowed to act on a gelatin solution having a concentration of 0.1 to 5% by weight, preferably 1 to 4% by weight to obtain a high melting point, high viscosity, A method for producing gelatin having characteristics such as high molecular weight was established, and the present invention was completed.

Means for Solving the Problems Transglutaminase cross-links and polymerizes a protein by an ε- (γ-glutamyl) lysine bond between a γ-carboxamide group of a glutamine residue and an ε-amino group of a lysine residue in the protein. It is known as an enzyme. The transglutanase may be of any origin as long as it exhibits transglutaminase activity, in addition to the one derived from the actinomycete streptovertirium used in the present invention.

There is no example in which transglutaminase was allowed to act on a gelatin solution to inactivate the enzyme after the reaction, and the reaction was stopped to produce highly polymerized gelatin. With the solution having a gelatin concentration in the range of the present invention, there is almost no change in viscosity over time due to the addition of transglutaminase. Therefore, since the viscosity of the reaction solution does not substantially change, the gelatin solution can be reacted for a sufficient time to obtain a desired polymerization. By changing conditions such as gelatin concentration, transglutaminase addition amount, reaction time, reaction temperature, reaction, and pH, gelatin having various degrees of cross-linking and physical properties can be obtained.

The production method of the present invention is characterized in that a high concentration of transglutaminase is added to a relatively low-concentration gelatin solution and reacted for a long time, and a composition in which an original gelatin component and a crosslinked-polymerized gelatin component are polymerized. The ratio of polymerized gelatin in the product can be increased, and polymerized gelatin can be obtained with a high yield.

The raw material gelatin in the present invention may be either an alkaline method gelatin or an acid method gelatin, but the acid method gelatin having more glutamine residues is easier to form a crosslink.

If the gelatin concentration is 0.1% or less, no sufficient crosslinking is observed, and it is not industrially useful from the viewpoint of cost due to the yield. On the other hand, when the gelatin concentration is 5% or more, the viscosity rises remarkably, and it is somewhat difficult to control the production. The gelatin concentration is 0.1% -5%, preferably 1% -4% (W /
W) is appropriate.

When the amount of transglutaminase added per 1 g of gelatin is 0.1 unit or less, no effective cross-linking is observed, and adding too much is not useful from a cost standpoint.
One or more units, preferably about 2 to about 80 units are suitable.

Regarding the reaction time, a short time does not sufficiently promote the cross-linking reaction, and a long time requires measures against decomposition of gelatin by heat, decomposition by microorganisms, and the like. Therefore, 1
A reaction time of about 30 hours to about 30 hours, preferably 2 hours to 10 hours is appropriate. Regarding the reaction temperature, below room temperature, the reaction speed is slow, and above 60 ° C, the effect of gelatin decomposition by heat becomes large, and from normal temperature, where macromolecules are formed by cross-linking with enzymes and low molecules are also generated by thermal decomposition. Preferably, the reaction is carried out between 60 ° C, 40 ° C-50 ° C
Is optimal. The reaction pH is around neutral, especially pH 6
-7 is optimal, but the reaction proceeds even at pH 4-10. As for the method of inactivating the enzyme, in addition to the thermal inactivation described above, it is also possible to significantly increase or decrease the pH. In addition, an enzyme inhibitor such as EDTA can be used as long as the enzyme activity is lost and the polymerized gelatin is not adversely affected.

 Hereinafter, the present invention will be described with reference to examples.

In all of the following examples, acidic gelatin having a jelly strength of 196 Bloom and a viscosity of 54 mp was used according to the JIS method.
The value measured with a B-type viscometer in this test was 13 cp at 50 ° C,
The melting point (falling ball method) is 31.0 ° C and the freezing point (JIS method) is 25.
At 4 ° C, the ratio of polymers with an α component or higher with a molecular weight of 100,000
8%.

Example 1 Pure water was added to 150 g of gelatin so that the concentration of gelatin became 1.5% by weight.
Adjusted to 0. Then add propylparaben as a preservative in 2.5
g, and 20 units of microbial transglutaminase were added to 1 g of gelatin. The reaction was carried out with stirring at 50 ° C. for 20 hours, and then the temperature was raised to 80 ° C. and maintained at that temperature for 15 minutes to inactivate the enzyme by heat. After concentration, the mixture was concentrated to 10%, cooled, gelled and dried.

The gelatin has a jelly strength of 118 Bloom,
The viscosity was 23 cp, the melting point was 33.0 ° C, and the freezing point was 27.9 ° C.
A gelatin having a higher viscosity and a higher melting point than that of the original gelatin was obtained. Measurement of the molecular weight distribution by liquid chromatography showed that polymers having a molecular weight of 100,000 or more increased to 68% and were highly crosslinked. (Column Asahipak G
S-620: mobile phase 0.1 M phosphate buffer, pH 6.8: temperature 50
° C: detection wavelength 230 nm) (see Fig. 1A).

Example 2 Pure water was added to 50 g of gelatin to a concentration of 1% by weight, and swelling, dissolution and pH adjustment were performed in the same manner as in Example 1.
Five units of transglutaminase were added to 1 g. After reacting at 50 ° C. for 3 hours, the enzyme was inactivated, concentrated to 10%, cooled, gelled and dried.

It was found that those with a high molecular weight of 100,000 or more increased to 60% and that crosslinking was progressing.

The melting point of this gelatin is 31.7 ° C, the freezing point is 26.0 ° C,
Viscosity is 17cp, jelly strength is 197 bloom and high melting point,
A high viscosity gelatin was obtained. (See FIG. 1B) Example 3 Pure water was added to 120 g of gelatin to a concentration of 0.5% by weight, swelling, solvent and pH were adjusted as in Example 1, and 6 g of propylparaben was added as a preservative. Add 80 units of transglutaminase to 1 g of gelatin, and add 5 units at 50 ° C.
After reacting for hours, the reaction was quenched, passed, concentrated to 7.2%, cooled, gelled and dried. From the molecular weight distribution, it was found from the molecular weight distribution that the proportion of high molecular weight gelatin having a molecular weight of 100,000 or more increased to 61%, indicating that crosslinking polymerization had progressed. (See FIG. 1C) Effect of the Invention The production method of the present invention was effective mainly in increasing the viscosity, melting point, and freezing point of gelatin as seen in the following Examples. Thereby, the edible film, the medicinal capsule and the like can be improved by improving the heat resistance and the coagulability of the film, and it is effective for improving the heat resistance of the table jelly and the like. In addition, workability can be improved by increasing the freezing point.

[Brief description of the drawings]

FIGS. 1A to 1D are diagrams showing liquid chromatography charts for analyzing the molecular weight distributions of high molecular weight gelatin and raw material gelatin produced in Examples of the present invention. FIG. 1A is a diagram in which the high molecular gelatin produced according to Example 1 was analyzed. FIG. 1B is a diagram obtained by analyzing a high molecular gelatin produced according to Example 2. FIG. 1C is a diagram in which the high-molecular-weight gelatin produced according to Example 3 was analyzed. FIG. 1D is a chart showing a chart analyzed for raw material gelatin. In FIGS. 1A to 1D, the vertical axis indicates absorbance at a wavelength of 230 nm, and the horizontal axis indicates elution time from the column.

Continuation of the front page (72) Inventor Masao Motoki 1-1, Suzuki-cho, Kawasaki-ku, Kawasaki-shi, Kanagawa Prefecture, Ajinomoto Co., Inc. (56) References JP-A-2-171160 (JP, A) JP-A Heisei 2-86743 (JP, A) JP-A-61-172807 (JP, A) (58) Fields investigated (Int. Cl. ⁶ , DB name) C08J 3/00, 3/24 A23L 1 / 05,1 / 32 A23J 3 / 06,3 / 34 C09H 7/00 A61K 7/00

Claims (1)

(57) [Claims] 1. A method for producing highly polymerized gelatin obtained by adding 0.1 units or more of transglutaminase per 1 g of gelatin to a solution containing 0.1% to less than 2% by weight of gelatin at a concentration of gelatin, and inactivating the enzyme after the reaction.