JP2602966B2 - Method for producing silicon complex protein and silicon complex polysaccharide protein - Google Patents

Description

Description: TECHNICAL FIELD The present invention relates to a method for producing a novel antimutagenic substance, a silicon complex protein and a silicon complex polysaccharide protein, and more particularly to a genus Aspergillus or Lentinus (Shiitake). The present invention relates to a method for assimilating silicon by utilizing the metabolism of a filamentous fungus selected from the group, and producing a silicon complex protein and a silicon complex polysaccharide protein having antimutagenic properties.

[Background of the Invention]

Advances in medical technology have reduced mortality from infectious diseases, and the average life expectancy of Japanese has grown to be ranked at the top of the world.

However, the number of deaths from cancer continued to increase, surpassing the brain defect disorder in 1980, becoming the number one cause of death, and if this trend continues, by the turn of the 21st century, one in two people will die from cancer. It is believed to die. The causes of cancer are still uncertain, but 80-90% are chemicals,
It is thought that the remaining 10 to 20% is due to biological factors such as viruses and physical factors such as ultraviolet rays.

Many chemical carcinogens are present in living environments such as food and air. Protecting from these chemical carcinogens is an effective way to prevent cancer.

These chemical carcinogens are present in foods and luxury goods and we continue to consume them in our daily diet. On the other hand, it has been confirmed that substances that detoxify these exist in foods.

The present inventors have a substance that does not adversely affect the human body and widely detoxifies chemical carcinogens, and if the substance is sufficiently ingested in a daily diet, the carcinogenesis rate can be reduced to a considerably high rate. I researched on the idea that I can do it.

[Object and structure of the invention]

Therefore, the present inventors have conducted intensive studies with the aim of providing a substance that detoxifies a chemical carcinogen existing in the living environment or an anti-mutagenic food made of such a substance that is safe and can be ingested daily. Was.

As a result, it is a mineral that is relatively high in foods, is highly safe, and is a typical semiconductor,
Oxidation in the living body, assimilation utilizing the metabolism of filamentous fungi selected from the genus Aspergillus or Lentinus (Shiitake) generally used for food or food production, which is considered to have a catalytic action of the reduction reaction, It has been found that a substance having anti-mutagenic activity can be obtained for a wide range of chemical carcinogens, and it has been confirmed that a highly stable and highly active substance can be obtained by using complex organosilicon, especially as silicon. Thus, the present invention has been completed.

Thus, the present invention provides a method for producing a silicon-complex protein and a silicon-complex polysaccharide protein, wherein silicon is assimilated in a culture solution of a filamentous fungus selected from the genus Aspergillus or Lentinus (Shitatake). It is.

[Specific description of the invention]

The method for the silicon complex protein and the silicon complex polysaccharide protein of the present invention will be specifically described below.

In the present invention, as described above, silicon is assimilated in a culture solution of a filamentous fungus selected from the genus Aspergillus or the genus Lentina (Shitake).

As the silicon, various compounds can be used. In the present invention, it is particularly preferable to use the organic complex salt thereof. For example, β-carboxyethyl silicon sesquioxide is preferably used. This organosilicon complex (SiCH
₂ CH ₂ COOH) ₂ O ₃ , which is disclosed in JP-A-56-18992.
Japanese Patent Application Laid-Open Publication No. HEI 7-209139 describes the manufacturing method in detail. This compound has excellent affinity for living cells, strong antimutagenic activity, and low production cost, and can usually be produced in the form of a beautiful crystal.

As the filamentous fungus selected from the genus Aspergillus or Lentinus (Shiitake) used in the present invention,
In particular, Asp.Oryzae (Aspergillus), Asp.niger (Black Aspergillus), L.Edodes (Shiitake) and the like are preferably used.

Such silicon, particularly an organosilicon complex, is assimilated by culturing in a culture solution of a filamentous fungus selected from the genus Alpergillus or Lentina (Shitatake) at an appropriate pH and temperature for about 15 days.

For example, the following culture solutions (1) and (2) are used for Asp.
Oryzae and L. Edodes are each cultured to assimilate silicon.

The silicon complex salt used here is the above-mentioned β-carboxyethyl silicon sesquioxide.

(1) Assimilation of silicon by Asp.Oryzae Medium composition glucose 100 g peptone 6 g KH ₂ PO ₄ 0.1 g MgSO ₄ · 7H ₂ O 0.1 g NaCl small amount CaCl ₂ small amount FeCl ₃ small amount Silicon complex 5 g water 1 liter Medium pH: 3.0 culture Conditions: 30 ° C, 15 days (2) Use of silicon by L.Edodes Medium composition Malt extract 100g Yeast extract 2.5g Ammon tartrate 2.0g Silicon complex 5g Water 1L Medium pH: 4.5 Culture condition: 20 ° C, 15 days culture After completion, the cells are separated from the culture filtrate, and the cells are freeze-pulverized, added with water, extracted with stirring, and centrifuged to remove insolubles. Add ammonium sulfate to the centrifugal supernatant to 50% saturation to produce a precipitate. The precipitate is centrifuged and dried to obtain a powder of the silicon complex protein. On the other hand, 2
A double volume of ethanol is added to form a precipitate, which is separated by filtration and dried to obtain a powder of the silicon complex polysaccharide protein.

Hereinafter, the silicon complex according to Asp. Oryzae is described as silicon complex protein-AO and silicon complex polysaccharide protein-AO, and the silicon complex according to L. Edodes is described as silicon complex protein-LE and silicon complex polysaccharide protein-LE. The respective physicochemical properties are shown in Table 1 below.

When used as a food material, the cells can be frozen and pulverized, mixed with a filtrate, and used as it is, or, depending on the application, concentrated and dried.

The antimutagenic substance comprising the silicon complex protein or the silicon complex polysaccharide protein obtained as described above can be used as a nutritional supplement or as a nutritional enhancer in other foods. These can be ingested in various forms, but it is generally preferable to solidify them into powders rather than liquid ones. For example, it is effective to continue sampling every meal every day. The effect is relatively fast acting and is excreted quickly.

As described above, most of the causes of cancer occurrence are thought to be due to secondary carcinogens in the living environment, but the above-mentioned anti-mutagenic substances composed of the silicon complex protein or the silicon complex polysaccharide protein have the following test results. As is clear from the above, the chemical carcinogen can be detoxified to prevent cancer at a high rate.

In general, these secondary carcinogens do not themselves have carcinogenicity, but are metabolized in the living body and exhibit carcinogenicity, and are introduced by hydrolysis, reduction, oxidation, conjugation and the like.

The activity of the anti-mutagenic substance obtained as described above is
Previously known secondary carcinogens in the living environment will be examined.

In the present invention, the above-mentioned silicon complex pale white is subjected to a mutagenicity test using bacteria for three kinds of carcinogens such as azo dye dimethylaminoazobenzene (DAB), benzopyrene contained in scorch, and dialkylnitrosamine contained in tobacco tar. And the antimutagenic activity of the silicon-conjugated polysaccharide protein are measured as follows.

Experiment using Ames test Among Salmonella strains isolated by Ames et al., TA100
The strain was used.

The TA100 strain stored at -80 ° C was thawed, centrifugally washed, and then suspended in the same volume of a 1 / 15M phosphate buffer (pH 7.0). 0.5 mM in soft agar solution (0.6% agar powder, 0.6% NaCl)
Biotin-0.5 mM, histidine solution was added at 1/10 volume.
SD rat male (6 weeks old, average)
178 g) of the liver trituration solution was prepared according to a conventional method and added to the reaction solution.

The composition in 1 ml of the reaction solution (S-9Mix) used in the experiment is shown below.

0.3 ml Liver attrition 8 mM MgCl ₂ 33 mM KCl 5 mM Glucose 6-phosphate 4 mM NADP ⁺ 100 mM Na-phosphate buffer (pH 7.4) Bacterial solution 0.1 ml, Mutagen solution 0.2 ml, Silicon complex 0.2 ml, S-9
After adding 0.5 ml of Mix and sensitizing at 37 ° C. for 20 minutes, 2 ml of soft agar was added and spread on a minimal agar medium.

After culturing at 37 ° C for 2 days, the number of revertant colonies was measured. As a control, 100 mM Na-
A phosphate buffer was used.

 Table 2 shows the results.

From the above results, the number of revertant colonies is significantly smaller in any of the silicon composites and in any of the carcinogens than in the control, and the tendency increases as the concentration increases. Therefore, it is clear that each of the silicon complexes shows strong antimutagenic activity against each mutagen.

 These effects modify and inactivate mutagens.

 Adsorbs mutagenic substances and suppresses mutagenic effects.

And so on.

〔The invention's effect〕

As is apparent from the above detailed description, the silicon complex protein and silicon complex polysaccharide obtained by assimilating the silicon of the present invention in the culture medium of Aspergillus layer or a filamentous fungus selected from Lentinus (Shiitake). Proteins have excellent antimutagenicity and can be taken at all times to detoxify carcinogens in the living environment, so they are very useful.

Continuation of the front page (51) Int.Cl. ⁶ Identification number Agency reference number FI Technical display location C12R 1:69) (C12P 21/00 C12R 1: 645)

Claims (2)

(57) [Claims] 1. A method for producing a silicon complex protein and a silicon complex polysaccharide protein, wherein silicon is assimilated in a culture solution of a filamentous fungus selected from the genus Aspergillus or Lentinus (Shitatake). 2. The method according to claim 1, wherein the silicon assimilated by the filamentous fungus is a complex salt.