JP2001149022A - Proteolysis product of connective tissue - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a new method for obtaining a proteolysis product of connective tissue, and to provide a health food or health drink for applying the obtained proteolysis product for the prevention and convalescence of various diseases. SOLUTION: This method for obtaining the proteolysis product of connective tissue by combining a connective tissue protein such as collagen or elastin with Bacillus natto or a Bacillus natto fermentation product and then subjecting the mixture to a heat-insulting treatment, and a health food or health drink using the proteolysis product.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a connective tissue protein decomposed product characterized by keeping warm by combining natto or fermented natto with a connective tissue protein such as collagen or elastin. This degraded connective tissue protein has good solubility in water or a dilute alcohol solution, is easily absorbed due to its small molecular weight, can be prepared relatively inexpensively, and is considered to have high utility value as a health food or a health drink.

[0002]

2. Description of the Related Art Proteins constituting connective tissue fibers in meat include collagen, which accounts for 90% of them, and reticulin and elastin (edited by Fujimaki et al., Food Industry,
p. 649, Koseisha Koseikaku, Tokyo, 1985). There have been many reports on collagen and elastin as health foods. However, these proteins had a drawback that they had a rather high molecular weight to be used and were hardly soluble in water. As for the enzymatic decomposition method, there was a method using a vegetable enzyme such as papain (Kajio, Food and Science, 1:95, 1997). There have been no reports so far. .

[0003]

The present inventors have discovered novel proteases such as nattokinase or pro-urokinase activator in natto and reported their properties (H. Sumi et al.,
Experientia, 43: 1110, 1987;
Acta Haematol. , 84: 139,199.
0; Food Chemistry, 43: 1124, 1996; Handbook of Bioactive Substances from Plant Resources, p. 579, 1998; Fo
od Style 21, 37, 1999). The present invention intends to degrade connective tissue proteins using natto bacteria and natto fermented products containing these enzymes.

[0004]

Means for Solving the Problems Foods contain various physiologically active substances, and the discoverers have so far found that nattokinase or pro-urokinase, a thrombolytic enzyme, has been found in natto, a traditional fermented food in Japan. Activators have been discovered and their properties and ingestion effects have been clarified (Sumi et al., Edited by the Industrial Technology Society, research and development on functional food materials, food-derived physiologically active substances, pp. 88, 198).
9; Monthly Food Chemical, 12:72, 1990; Invitation to Food Functional Science, p. 36, Sankyo Publishing, 1996; Asahi Shimbun 8/3, 1996). Since they are derived from fermented foods, they have the greatest advantage in that they have few safety issues even if ingested for many years. The present invention provides a method for decomposing collagen and elastin, which are extremely easy connective tissue proteins discovered during the course of such research, and uses the decomposed product as a health food or a health drink.

[0005]

Next, the present invention will be described in detail with reference to examples.

First Example Each test tube was obtained by thoroughly stirring 150 mg of fish collagen (with Uchidaya, Yonago) washed with physiological saline, and 50 g of natto with Takano Foods (Ibaraki) and 150 ml of water. The supernatant was added to 0.2 ml, and 0.17 M borate buffer-saline, pH 7.8 was added to adjust to 2.0 ml.
Incubation was performed at 37 ° C. Under these conditions, the fish collagen sank down, but it was found that the amount decreased with the incubation time. In addition, as a result of measuring the absorbance of the supernatant at 280 nm over time, it was found that fish collagen was degraded from the increase as shown in FIG.

Second Example As a fermented product of Bacillus natto, Japanese cultivated Bacillus natto, whose cultivated extract of Bacillus natto has already been mass-produced by the present inventors' technology. Biozyme-NSK (2,508FU /
g), and a commercially available dried shellfish, sea cucumber, and prawn shrimp (Pro-Foods, Okayama) as a protein derived from fish and shellfish, and the powder was used. Fish collagen isolated from fish scales was provided by Yozo Ishizuka, Ryu Institute (Yonago). The Bacillus natto culture extract showed a decomposing activity for all fish and shellfish proteins. The photograph (FIG. 1) shows a 0.17 M borate buffer solution of a Bacillus natto culture extract in 100 mg of fish collagen, 66 mg of scallop, 73 mg of sea cucumber, and 55 mg of prawn shrimp-saline, pH
1.0 ml of a 20-fold diluted solution of 7.8 was added, and the mixture was incubated at 37 ° C. It can be seen that the amount of precipitated protein decreases with the incubation time. Also, the reaction was concentration and temperature dependent. 1 mg of Bacillus natto culture extract in a borate buffer pH 7.8, 37 ° C, 1 hour incubation up to 25 m
g or more was found to decompose.

Example 3 Bovine collagen (Sigma) well ground in a test tube
50 mg or elastin (Sigma) 16
0 mg, 0.17 M borate buffer-saline,
1.0 ml of pH 7.8, and Bacillus natto culture extract (Bi
ozyme-NSK, 2,508FU / g, strain, Japan.
1.0 ml of a 10-fold diluted solution of the same buffer (Bioscience Research Institute, Kyoto) was added and incubated at 37 ° C.
As a result, both substrates were dissolved, and an increase in the absorbance (280 nm) of the reaction supernatant was confirmed (FIG. 2), but it was found that elastin was particularly well degraded.

Fourth Example In a 300 ml Erlenmeyer flask, 10 g of elastin (Si
gma) into a mixer.
05M ammonium bicarbonate and 0.5 ml of Bacillus natto culture extract (Biozyme-NSK, 2,508FU /
g, stocks, Japan. Institute for Biological Sciences, Kyoto) and 37
The mixture was shaken and kept at 100 rpm for 24 hours at 100 ° C. As a result, it was found that elastin was completely dissolved. The reaction product was centrifuged at 3,000 rpm for 10 minutes to remove white cells, and then freeze-dried to obtain 8.44 g of a yellow-white powder. When this was subjected to SDS-polyacrylamide electrophoresis, it was found that the major protein band stained with Coomassie brilliant blue was smaller than the molecular weight of cytochrome C (about 12,500).

Fifth Example In a 300 ml Erlenmeyer flask, 1.5 g of bovine collagen (Sigma) and 150 ml of 5% polypeptone were added.
S (Wako Pure Chemical Industries)
Sterilized for 5 minutes. By this operation, the collagen was precipitated in the lower layer without dissolving, but this solution was further inoculated with a platinum loop natto fungus (Miyagino Natto Mill, Sendai) obtained from a slant that had been separately cultured, and 100 rpm at 37 ° C. As a result of shaking culture, it was found that collagen was completely decomposed and turned into a solution after 43 hours. This solution is
The supernatant obtained by removing the Bacillus natto by centrifugation at 00 rpm for 10 minutes was lyophilized to obtain a yellow-white powder. This is based on the fibrin plate method and HPL
Method C (Sumi et al., J. Brew. So
c. Japan, 88: 482, 1993, and Japanese Home Economics, 50: 309, 1999).
And 10 μg / g or more of vitamin K2 (menaquinone-
7) It was confirmed that it showed a content, had little odor, was easy to eat, and was well soluble in water and dilute ethanol solution.

Sixth Example 50 g of beef collagen (Nippi, Tokyo, Japan) and 5% Sorpey NY (Nisshin Cosmo Foods, Tokyo, Japan) were placed in a 1-liter jar fermenter, and sterilized at 120 ° C. for 30 minutes. And 0.1 g of Bacillus natto (Meguro fungus: 5 × 10 ¹¹ cells / g, strain / Meguro Research Laboratories, Osaka), and cultured at 40 ° C. for 60 hours. Centrifuge at 3,000 rpm for 10 minutes,
Galacto-oligosaccharide, calcium lactate, ascorbic acid, fructose, glucose, fragrance (plum essence), vitamins B ₁ , B ₂ , B ₆ , B ₁₂ were added to a 10-fold water dilution of the obtained supernatant.
And then bottled with citric acid to adjust to pH 4.0, and the fired one has a high nutritional value and a refreshing taste,
Suitable for health drinks.

[0012]

According to the present invention, it is possible to easily produce masses of collagen and elastin hydrolyzate, which are connective tissue proteins, and to provide a health food which is inexpensive and safe even when eaten and drunk.

[Brief description of the drawings]

FIG. 1 is a graph showing the degradation of fish collagen by a fermented product of Bacillus natto measured by the absorbance (280 nm) of the supernatant.

FIG. 2 is a graph showing the degradation of beef collagen and elastin by a fermented product of Bacillus natto measured by absorbance (280 nm).

FIG. 3 is a decomposed photograph of a fermented product of natto of fish collagen, scallop, sea cucumber, and prawn with time.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) A23L 2/66 A23L 2/00 J

Claims (2)

[Claims] 1. A collagen which is a connective tissue protein,
A method for preparing an enzymatically decomposed product of connective tissue protein, comprising combining elastin or the like with natto or a fermented product of natto and keeping the mixture warm. 2. A health food or a health drink obtained by using the obtained connective tissue protein hydrolyzate as it is or in combination with an appropriate food material.