JP3247457B2 - Multifunctional protein - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a protein having multifunctional properties.

[0002]

2. Description of the Related Art Various effects that are effective on the human body have been reported for polyphenols contained in tea extracts. For example,
Antibacterial properties against caries, alveolar pyorrhea and food poisoning bacteria,
The effects of improving renal failure, improving the balance of intestinal bacteria, lowering cholesterol, suppressing cancer, and suppressing rotavirus are known. Further, deodorizing effects on various odor components are also known.

[0003] The eggshell membrane present inside the eggshell of birds is a kind of protein, and its powder or protein is used as a therapeutic or cosmetic material for skin diseases such as burns and cuts, and utilizes the heavy metal adsorption capacity of the eggshell membrane. It is known to use as a heavy metal remover. (JP-A-2-182154, JP-A-2-26560, JP-A-2-231426, JP-B-54-22499, JP-A-58-150433)

[0004]

The above-mentioned polyphenols, although having many functions, are water-soluble and therefore have poor sustained effect and are unstable.
In addition, their application range has been limited due to their bitterness and astringency. On the other hand, the application of eggshell membranes is also similarly limited to the fields of medical polymers, cosmetics, resins, and the like, and it is desired to elucidate the functions of the eggshell membranes and to increase the value of the eggshell membranes in order to expand the range of application. Therefore, the appearance of a multifunctional substance that can be applied to various fields has been demanded.

[0005]

Means for Solving the Problems The present inventors have found that polyphenols, which are tea extracts, are adsorbed on eggshell membranes, and that the functions of the polyphenols and eggshell membranes are exhibited as they are. The invention has been completed. That is, the present invention provides a multifunctional protein obtained by adsorbing polyphenols, which are tea extracts, to eggshell membranes. Hereinafter, the present invention will be described in detail.

[0006] The eggshell membrane of the present invention refers to a membrane existing inside the eggshell of an avian egg. When used, the eggshell membrane obtained from the eggshell by a known method is crushed and powdered and chemically and enzymatically solubilized. The protein may be in any form such as an egg shell membrane protein.

The tea extract used in the present invention is extracted from one or more of green tea, oolong tea and black tea, and is usually a powder. Green tea, oolong tea,
One or a mixture of two or more powders obtained by extracting from one kind of black tea may be used. For example, Sanfenone (manufactured by Taiyo Kagaku Co., Ltd.), which is a hot extract of green tea, is known. Furthermore, the polyphenols contained in the tea extract differ depending on the type of tea, but are generally (+) catechin, (−) epicatechin, (+) gallocatechin, (−)
Epigallocatechin, (−) epicatechin gallate,
(-) Epigallocatechin gallate, free theaflavin, theaflavin monogallate A, theaflavin monogallate B, and theaflavin digallate.

[0008] The method for producing the multifunctional protein of the present invention is as follows. That is, after mixing and adsorbing the eggshell membranes of the respective forms to the tea extract solution containing the above-mentioned polyphenols, if the eggshell membrane is insoluble, it is recovered by filtration, and in the case of the solubilized eggshell membrane protein, the solvent is precipitated or acidified. And recovered by precipitation. Thereafter, the non-adsorbed polyphenol can be obtained by washing off with water or a solvent and drying. The solvent used can be selected as appropriate, and can be selected from methanol, ethanol, propanol, isopropanol,
Acetone, ethyl acetate and the like are used. The acid used is not particularly limited, but usually hydrochloric acid, sulfuric acid, acetic acid, citric acid, lactic acid and the like are used.

The content of the polyphenols adsorbed on the eggshell membrane is usually 0.005-15 to the weight of the eggshell membrane from the viewpoint of functionality.
%, Preferably 0.5-10%.

The multifunctional protein thus obtained does not impair the properties of the eggshell membrane, such as hygroscopicity, and the various functions of the polyphenols, such as the antibacterial properties, and is chemically, physically and biologically. It is very stable and has a sustained effect on various functions. It was also found that the bitterness and astringency of polyphenols were reduced. From the above, the use of the multifunctional protein of the present invention has been broadened, and it can be used in the fields of medicine and cosmetics in which eggshell membranes have been conventionally used, foods in which tea extracts have been used, medicines and the like. .

Hereinafter, the present invention will be described with reference to examples and test examples, but the present invention is not limited thereto.

【Example】

Example 1 10 kg of eggshell after breaking eggs was crushed in a mixer together with twice the amount of water, and eggshell membranes were recovered by decantation and dried.
It was further freeze-pulverized to obtain 0.2 kg of eggshell membrane powder.

Example 2 10 kg of the washed egg shell was added to 50 L of a 0.2 M sodium sulfide solution, and heated at 95 ° C. for 1 hour to solubilize the eggshell membrane. 5N sulfuric acid was gradually added to the eggshell membrane protein solution obtained by filtration while stirring to adjust the pH to 4.3. After leaving at room temperature for one hour,
The precipitated protein was recovered by filtration and washed with water. The obtained protein was suspended in 2 L of water, adjusted to pH 10 with sodium hydroxide, and solubilized. Next, the solution was adjusted to pH 4.5 with hydrochloric acid, reprecipitated, and washed with water. These operations of solubilization, precipitation and washing with water were repeated three times. Lyophilize the precipitate after washing with water and powder the eggshell membrane protein
100 g were obtained.

Example 3 100 g of a dried eggshell membrane was dispersed in 1 liter of water. This eggshell membrane dispersion was added to Corolase (Higuchi Shokai Co., Ltd.)
Was added and the mixture was reacted at 50 ° C. for 20 hours with stirring. After the reaction, the mixture was subjected to a heat treatment at 95 ° C. for 30 minutes, and then filtered to remove insolubles, thereby obtaining a transparent decomposition solution of eggshell membranes.
After concentrating under reduced pressure to a volume of 1/3, the same amount of ethanol was added and the solvent was precipitated. The precipitate was separated by filtration, washed with a small amount of ethanol, and dried to obtain 90 g of eggshell membrane protein powder.

Example 4 Sanfenone, a green tea extract (manufactured by Taiyo Kagaku Co., Ltd.) 5
g was dissolved in 200 ml of ethanol. To this solution, 10 g of the eggshell membrane powder obtained in Example 1 was added and stirred at room temperature for 2 hours. The solution was removed by filtration, washed with water, and the non-adsorbed sanphenone was washed off with ethanol and dried to obtain 11.5 g of the multifunctional protein of the present invention. The content of sanphenone per eggshell membrane weight was about 9.8%.

Example 5 10 g of the eggshell membrane protein obtained in Example 2 is dissolved in 100 ml of a sodium hydroxide solution having a pH of 10. Next, 5 g of sanphenone, which is a tea extract, was dissolved in this solution and reacted at 40 ° C. with stirring for 2 hours. After the reaction, the pH was adjusted to 4 with hydrochloric acid, and the obtained precipitate was collected by filtration. After washing with water and drying, 14 g of the multifunctional protein of the present invention was obtained. The content of sanphenone per eggshell membrane protein was about 3.5%.

Test Example 1 (Hygroscopic Test) The hygroscopic property of the product of the present invention obtained in Example 4 was examined. As a control, eggshell membrane powder not adsorbing the tea extract was used. The sample dried and weighed in advance is placed at a specified humidity (65%,
(80%, 90%) in a desiccator for three days and nights, the weight of the sample after moisture absorption was measured, and the moisture absorption was calculated from the difference by the following equation. Moisture absorption rate = (moisture absorption weight−dry weight) / dry weight × 100 The results are shown in Table 1.

[0017]

[Table 1]

Test Example 2 (Antibacterial Test) An antibacterial test of the product of the present invention obtained in Example 4 was conducted by the following method. Bacillus subtilis (B
acillus subtilis IFO3007), Staphylococcus aureus (Staphylococcus aureus IFO12732), Pseudomonas aeruginosa IFO3
080) and Esacherichia coli IFO354
5), Streptococcus mutans (Streputococc
us mutansMT8148), Bacteroides Zingivalius (B
acteroides gingivalis ATCC33277), Propionibacterium acnes GAI5419
Was used to determine the minimum growth inhibitory concentration (%) by the agar dilution method. The medium used was a standard agar medium (Eiken) or GA
M agar medium (Eiken).

An agar medium containing samples of various concentrations is prepared,
One platinum loop of the bacterial solution of each strain, which had been cultured in the enrichment medium beforehand, was inoculated on the agar and cultured under each condition. After the culture, the minimum growth inhibitory concentration was measured from the growth state of the fungus.
Table 2 shows the results.

[0020]

[Table 2]

Test Example 3 (Taste Test) A taste test was carried out by 10 panelists using the product of the present invention obtained in Example 4 and sanfenone. Evaluation method is bitter
The astringency was evaluated on a 4-point scale (very strong, strong, weak and absent). Table 3 shows the results.

[0022]

[Table 3]

Test Example 4 (Deodorizing Test) A deodorizing test was performed on the product of the present invention obtained in Example 5 as follows. Odorous components: methyl mercaptan, trimethylamine, ammonia, test method: 1 g of the product of the present invention and 1 ml of 0.3% malodorous component were added to a 10 ml vial bottle, sealed and kept at 37 ° C. Five minutes later, the gas in the vial bin was extracted, the content of each malodorous component was measured by gas chromatography analysis, and the reduction rate with respect to the blank was calculated by peak area conversion to obtain the deodorization rate.

The conditions for gas chromatography are as follows. Apparatus; Shimadzu GC-14A column; SUPELCOWAX10 (30m, 0.25mmID., 0.25μmdf) Carrier gas; He 1ml / min Sample injection; Split 50: 1, injection volume 1ml Column temperature; 60 ° C Inlet temperature; 200 ° C detector; FID test results are shown in Table 4.

[0025]

[Table 4]

From the above results, the multifunctional protein obtained by adsorbing green tea extract containing polyphenols on eggshell membranes or eggshell membrane proteins retains moisture absorption, antibacterial properties and deodorant properties, and reduces bitterness and astringency. It was shown that.

[0027]

The eggshell membrane used in the present invention is an abundant and very stable protein material, and at the same time, the multifunctional protein of the present invention obtained by adsorbing a tea extract containing polyphenols to the eggshell membrane is a tea extract. It has both the excellent functions of eggshell membranes. Further, since the production method is extremely simple, it can be applied to various fields such as industrial products such as resins, papermaking and fibers, medical polymers, pharmaceuticals, cosmetics and foods.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁷ Identification code FI A61L 9/01 A61P 17/02 A61P 17/02 31/04 31/04 B01D 53/28 B01D 53/28 C07K 14/465 C07K 14 / 465 A61K 37/02 (56) References JP-A-2-202900 (JP, A) JP-A-49-80160 (JP, A) JP-A-62-51971 (JP, A) (58) Fields investigated (Int.Cl. ⁷ , DB name) A61K 35/78 A61K 7/00 A61K 31/35 A61K 38/00 A61L 9/01 A61P 17/02 A61P 31/04 B01D 53/28 C07K 14/465 BIOSIS (DIALOG) CA (STN)

Claims (1)

(57) [Claims] 1. A multifunctional eggshell membrane wherein a tea extract containing polyphenols is adsorbed on an eggshell membrane.