JP2689511B2 - Food - Google Patents

Description

TECHNICAL FIELD The present invention relates to a caries preventive food capable of preventing caries.

Problems to be Solved by Prior Art and Invention Periodontitis, periodontal disease such as alveolar pyorrhea, and caries are said to be two major diseases of the oral cavity. The causes of these oral diseases are:
For example, in the former case, Bacteroides gingivalis, etc.
In the latter case, it is said that bacteria such as Streptococcus mutans colonize in the oral cavity and proliferate in large numbers. Therefore, in order to prevent oral diseases, the colonization of these bacteria in the oral cavity is suppressed. Is effective.

Conventionally, from such a viewpoint, a blood antibody or a milk antibody obtained by immunizing a mammal with whole cells or periodontal disease-causing bacteria such as Bacteroides gingivalis or Streptococcus mutans or a bacterial cell component is used for oral composition. It is known that it is incorporated into foods or foods to prevent colonization of the above bacteria in the oral cavity and prevent oral diseases (JP-A-60-
142915, JP-A-60-38327, JP-A-60-38329, JP-A-61-1
12028, JP 61-112029, JP 61-112030, JP 61-
289024, JP-A-62-417, JP-A-61-277632).

However, development of an active ingredient capable of effectively suppressing colonization of oral bacteria and preventing dental caries is desired from the viewpoint of compounding properties in the case of producing foods.

Means and Actions for Solving the Problems The present inventors, in view of the above circumstances, as a result of further investigation, cell wall fraction of Streptococcus mutans or Streptococcus sanguis, fibrous structure fraction,
When a bacterial cell component selected from the GTF fraction and the protein fraction was used as an antigen and poultry was immunized with this, antibodies were accumulated and present in the yolk of the resulting egg, and carrageenan or xanthan gum was added to this yolk The precipitate was removed, preferably the supernatant was subjected to fractional precipitation with ammonium nitrate, and the water-soluble egg yolk fraction obtained by dialyzing this with water was used as a food material. The inventors of the present invention have found that the oral colonization of the above-mentioned bacteria can be effectively suppressed when food is ingested, and have completed the present invention.

 Hereinafter, the present invention will be described in more detail.

The dental caries preventive food according to the present invention is a cell wall fraction of Streptococcus mutans or Streptococcus sanguis, a fibrous structure fraction, a GTF fraction and a protein fraction as a bacterial component selected as an antigen, and immunized with this. Carrageenan or xanthan gum was added to the yolk obtained from the poultry, the resulting precipitate was removed, and the supernatant was subjected to fractional precipitation with ammonium sulfate, and the water-soluble yolk fraction obtained by dialyzing this with water was used. The material is made from a painting.

Here, Streptococcus mutans bacteria, Streptococcus sanguis bacteria, for example, using the dialyzed external solution of BHI medium as a medium, for example treated with formalin after washing the grown bacteria, those that have been subjected to known culture, pretreatment Can be used. In this case, as the Streptococcus mutans bacteria, those belonging to the serotype classification c which is often present in the human oral cavity are preferable. Such a Streptococcus
As mutans, NCTC10449, Ingbritt, OMZ70, JC
-2 etc. are mentioned. Also, as Streptococcus sanguis, NCTC10556, NCTC10558 and SSH-83, KIH-
T etc. are mentioned.

These Streptococcus mutans and Streptococcus sanguis cell components include the cell wall fraction of the bacterium, the fibrous structure fraction of the bacterium, the glucosyl transferase (GTF) fraction of the bacterium, and the protein of the bacterium. Use fractions.

In this case, the cell wall fractions of Streptococcus mutans and Streptococcus sanguis are determined by the method of Bleiweis et al. (J. Bacteriolo, 88 , 1198-12).
00,1964), using a Brown cell disruptor, perform a disruption process using glass beads having a diameter of 0.17 to 0.18 mm, and treat the resulting cell wall with trypsin to remove the proteins mixed in the cell wall, Those prepared by a method such as washing with distilled water and freeze-drying can be used. The fibrous structure fraction is J.Va.
n Hoate) et al. (Arch.Oral.Bio., 16 , 1131-1141,1
According to 971), 5% sucrose was added to the BHI dialysis medium, cultivated under anaerobic conditions, and 3 times the amount of ethanol was added to the centrifugation supernatant of the culture broth to prepare a precipitate, etc. Can be used. Furthermore, the GTF fraction was calculated by the method of Inoue et al. (Microbial Aspects of dental caries Vol.II).
I, 665-682,1976 (Information Retrieval Inc.))
According to the procedure, inoculate Streptococcus mutans and Streptococcus sanguis in BHI dialysis medium, remove the cells by centrifugation after growth, add ammonium sulfate to the supernatant, and precipitate the 40% ammonium sulfate fraction with 50 mM phosphorus. A product prepared by a method such as dialysis with an acid buffer and using a concentrated solution can be used. In addition, the protein fraction was analyzed by the method of Lehner et al. (J. General M
icrobiology, 122 , 217-225, 1981) and cultured in BHI dialysis medium, and the supernatant obtained by centrifugation was fractionated with 75% ammonium sulfate, and the precipitate was collected in the presence of 6M urea. DE-52 column chromatography is carried out, the protein fraction is further dissolved in physiological saline, and after dialysis, gel filtration is performed on Sepharose CL6B to obtain a fraction.

The present invention uses, as a food material, a water-soluble egg yolk fraction containing an antibody obtained by immunizing poultry with the above-mentioned antigen. In this case, chickens, ducks, quails and the like are used as poultry to be immunized. In addition, as a method for immunizing poultry, it is possible to immunize with an antigen alone or in combination of two or more kinds, and at that time, a method of immunizing subcutaneously or intramuscularly with an adjuvant, a method of orally immunizing with feed or water, etc. The method can be adopted.

As a method of obtaining a water-soluble egg yolk fraction, carrageenan or xanthan gum is added to the egg yolk, the resulting precipitate is removed by a method such as centrifugation, and preferably the supernatant is subjected to fractional precipitation with ammonium sulfate, The method of dialysis with water is adopted.

The food of the present invention is one or two of the above water-soluble egg yolk fractions.
It can be produced by using seeds or more as raw materials, and can be prepared as, for example, chewing gum, ice cream, bavarois, mayonnaise, dressing, chocolate, pudding, fresh cream, etc. In this case, the water-soluble egg yolk fraction is ingested. It is preferable to use it in an amount of 0.0001 to 50 g / kg / day.

The other ingredients of the food are appropriately selected according to the type of the food.

Effects of the Invention According to the food of the present invention, by incorporating an aqueous solution yolk fraction obtained from poultry immunized with Streptococcus mutans or Streptococcus sanguis bacterial component, colonization of the bacteria in the oral cavity It effectively prevents and is effectively used for the prevention of dental caries, and since the compounding properties of these antibody-containing components are good, there are no difficulties in the preparation of foods.

 Hereinafter, the effects of the present invention will be specifically shown by experimental examples.

[Experimental Example 1] Using the following antigen, the antibody fraction was obtained by the following method.

(1) Preparation of antigen Streptococcus mutans NCTC10449, Streptococcus sanguis NCTC10556 BHI medium was used as a dialyzate external solution, and after washing the grown bacteria, formalin-treated one was used as a whole cell antigen.

Cell wall fraction of Streptococcus mutans NCTC10449, Streptococcus sanguis NCTC10556 Method of Bleiweis et al. (J. Bacteriol., 8
8 , 1198-1200, 1964) was used.

Fibrous structure fraction of Streptococcus mutans NCTC10449, Streptococcus sanguis NCTC10556 According to the method of J. Van Hoate et al. (Arch.Oral.Bio., 16 , 1131-1141, 1971) What was prepared was used.

Glucosyl transferase race fraction of Streptococcus mutans NCTC10449, Streptococcus sanguis NCTC10556 Inoue et al. (Miorobial Aspects of dental ca
ries Vol.III, 665-682, 1976 (Information Retrieva
l. Inc.]) was used.

Protein fraction of Streptococcus mutans NCTC10449, Streptococcus sanguis NCTC10556 Method of Lehner et al. (J. General Microbiol
Ogy, 122 , 217-225, 1981) was used.

(2) Method for preparing chicken egg antibody fraction The above antigen was mixed with Freund's complete adjuvant to immunize chickens, and then a mixture of the antigen and Freund's incomplete adjuvant was immunized three times every two weeks to prepare eggs. Was collected. Immunization was performed once a month after the decrease in antibody. After that, the egg was divided into egg yolk and egg white, an equal amount of water was added to the egg yolk, a suspension of 0.5% λ-carrageenan was added, and the mixture was stirred and then centrifuged at 10,000 rpm for 10 minutes to collect the supernatant.
Furthermore, fractional precipitation with 20% ammonium sulfate was repeated 5 times, dialyzed against water, and added with 10 times concentration of physiological saline to obtain 0.15.
An M-NaCl aqueous solution was prepared and then filtered with a 0.45 μm filter to collect the filtrate.

Next, an intraoral fixation test of Streptococcus mutans and Streptococcus sanguis was performed by the following method.

(3) Oral colonization experiment of Streptococcus mutans and Streptococcus sanguis Five hamster 5-week-old males were grouped into five, and 1 × 10 ⁸ Streptococcus mutans NCTC10449 strain or Streptococcus sanguis NCTC10556 strain was inoculated to each. From that day, mix the above chicken egg antibody fraction with drinking water to 1%
After administration as a solution, after 1 week and 4 weeks, the dentition in the oral cavity of the hamster was rubbed with a sterile cotton ball and immersed in a small amount of physiological saline to uniformly disperse the bacteria. BH
Total number of cells and Streptococcus mutans, plated on I plate medium and Mitis salivarium plate medium,
The number of colonies of Streptococcus sanguis was counted. The numbers of Streptococcus mutans and Streptococcus sanguis were expressed as the number of bacteria per 10,000 total bacteria. The concentration of the antibody fraction in the drinking water was 0.025%.

 The results are shown in Tables 1 and 2.

From the results in Tables 1 and 2, it is found that the hen egg antibody fractions of Streptococcus mutans and Streptococcus sanguis suppress the colonization of Streptococcus mutans and Streptococcus sanguis well. Furthermore, it was confirmed that the bacterial cell component was superior to the whole bacterial cell as an immunogen.

Examples will be described below. In addition, all% show weight%.

[Example 1] Chewing gum Gum base 43.9% Calcium carbonate 2.0 Water candy 15.0 Sugar 30.0 Sucrose palmitate 1.0 Fructose 4.0 Maltose 3.0 Fragrance 1.0 Egg yolk anti-cell wall antibody fraction 0.1 (use bacterium: Streptococcus mutans NCTC10449) 100.0% [Example 2 ] Ice cream Cream (fat ratio 50%) 16.84% Milk (fat ratio 3.7%) 42.17 Sugar-free skimmed condensed milk 24.24 Sugar 11.25 Corn syrup 4.65 Stabilizer 0.35 Egg yolk antifibrous structure antibody fraction 0.5 (Used bacteria: Streptococcus mu) Tansu NCTC 10449 or Streptococcus sanguis NCTC 10556) 100.0% [Example 3] Ice cream cream (40% fat percentage) 31.54% Milk (3.7% fat percentage) 32.16 Sugar-free skimmed condensed milk 15.08 Sugar 11.25 Corn syrup 4.67 Stabilizer 0.3 Egg yolk antiprotein Antibody fraction 5.0 (bacteria used: Streptococcus muta Scan NCTC10449) 100.0% Example 4 bavarois dough milk 375ml sugar 60 g 1 this sheath vanilla gelatin 20g cream 75ml yolk anti fibrillar structures antibody fraction 5 (using bacteria: Streptococcus mutans NCTC10449 or Streptococcus Sangisu NCTC10556)

Claims (2)

(57) [Claims] 1. An egg yolk obtained from poultry immunized with a cell component selected from a cell wall fraction, a fibrous structure fraction, a GTF fraction and a protein fraction of Streptococcus mutans or Streptococcus sanguis as an antigen. Carrageenan or xanthan gum is added to, and the water-soluble egg yolk fraction obtained by removing the generated precipitate is used as a raw material. 2. A supernatant obtained after the precipitation of the water-soluble egg yolk fraction produced by adding the carrageenan or xanthan gum is removed, and the supernatant is subjected to fractional precipitation with ammonium sulfate and dialyzed against water. The food according to claim 1, which has been prepared.