JPH0356714B2 - - Google Patents

Description

[Detailed description of the invention]

[Industrial Application Field] The present invention relates to water chlorella in which an iodine component is enriched and immobilized as a compound in freshwater chlorella cells, and an anticancer agent containing the same as a main component. [Prior Art] Freshwater unicellular green algae (hereinafter simply referred to as Chlorella), such as Chlorella, Scenedesmus, and Chromydomonas, have extremely high growth rates and light utilization efficiency compared to ordinary cultured plants, and they also contain proteins, lipids, carbohydrates,
Contains many nutrients such as vitamins and minerals, making it an excellent food and feed. In addition, the physiologically active substances it contains are known to have the effect of promoting the growth of microorganisms, animals and plants, and the medical effects of chlorella are also attracting attention. There are two types of chlorella: natural marine-produced chlorella and industrially cultivated freshwater-produced chlorella, but the freshwater version is currently produced and sold in large quantities as a health food, while the marine-produced version has problems with productivity and has not been industrialized. I haven't reached it yet. Chlorella is expected to be used as a gastrointestinal ulcer treatment agent, hypertension treatment agent, cholesterol regulator, anticancer agent, etc., and research on its ingredients is being carried out. There are subtle differences in the medicinal ingredients, and although marine chlorella is said to be significant, the cause has not yet been fully researched. Normally, chlorella contains about 7% mineral components, the main ones being phosphorus, potassium, magnesium, calcium, sulfur, iron,
The fact that it contains many of these ingredients is one of the factors that increases the value of chlorella as a health food and in terms of its medical effects. [Problems to be Solved by the Invention] Although chlorella has attracted attention as a health food in recent years, its main effects are unknown and it has not yet been commercially available as a drug. This is thought to be because marine chlorella has problems with industrial productivity, and freshwater chlorella has extremely small amounts of medicinal components, making it impossible to extract and separate the components. [Other Means for Solving the Problems] The present inventors conducted various studies focusing on the differences between marine chlorella and freshwater chlorella, and found that natural marine chlorella contains about 80 ppm in terms of iodine. They found that commercially available freshwater chlorella does not contain appreciable amounts of iodine. By culturing under these conditions, chlorella with a higher iodine content than that of marine chlorella can be obtained, and the inventors have found that its pharmacological effects are also greater than that of marine chlorella, leading to the completion of the present invention. It was expected that iodine ions would inhibit the growth of freshwater chlorella, as they are used as disinfectants, but test results showed that contrary to that expectation, even in a medium containing a relatively high concentration of iodine ions, freshwater chlorella It was found that the growth of Chlorella was hardly inhibited. Furthermore, freshwater chlorella cultured in a medium containing iodine ions contains a higher proportion of iodine than marine chlorella, and even if this chlorella is thoroughly washed, there is almost no decrease in iodine, so iodine atoms are absorbed into chlorella cells. It was confirmed that the iodine ions were fixed in a form that bound to certain components within the chlorella, and that the iodine ions were not simply absorbed or adsorbed by the chlorella. The present invention relates to chlorella enriched with the above-mentioned iodine component, that is, iodine ions and/or
Or freshwater chlorella enriched with iodine components, characterized by culturing short-cell green algae in a medium containing 0.1 ppm or more of iodate ions in the presence of light, and uses thereof. Seawater normally contains about 0.06 ppm of iodine ions. As mentioned above, marine chlorella grown in seawater contains about 80 ppm of iodine. On the other hand, according to the present invention, freshwater chlorella having an iodine component content (per dry weight) of 100 ppm or more in terms of iodine can be easily obtained. A more preferable range of the iodine content per dry weight of the iodine-enriched freshwater chlorella obtained in the present invention is approximately
It is 100 to 2000 ppm, but it does not necessarily have to be within this range. The iodine component content in chlorella is usually easily influenced by the concentration of iodine ions and/or iodate ions (hereinafter both referred to as iodine ions, etc.) in the culture medium. That is, the higher the concentration of iodine ions, etc. in the medium, the higher the iodine component content in chlorella. however,
In a medium containing too high a concentration of iodine ions, the growth of chlorella is likely to be inhibited. Therefore,
Although it depends on the type of chlorella, the appropriate concentration of iodine ions, etc. in the medium is about 0.5 to 800 ppm, particularly preferably in the range of about 1 to 800 ppm. In the present invention, Chlorella such as Chlorella pyrenoidosa, Chlorella vulgaris, and Chlorella elliplyidea are preferably suitable; however, Chlorella such as Scenedesmus, Clomydomonas, Selenastrum,
Other chlorella may also be used. The above chlorella is a freshwater chlorella that usually grows in fresh water.
The salt-tolerant variant is called marine chlorella. A medium containing iodine ions and the like can be easily obtained by adding water-soluble iodide, iodate, etc. to a chlorella culture medium. For example, potassium iodide, sodium iodide, calcium iodide and other inorganic iodides, iodine ion sources such as hydrogen iodide, potassium iodate, sodium iodate, calcium iodate, and other iodates and iodates. An iodate ion source may be used. Further, a culture medium can also be produced by adding a nutrient source such as the carbon source described below to underground brine containing iodine ions or the like. The concentration of iodine ions, etc. in the medium is preferably maintained within the above concentration range over the entire range of iodine culture, but is not necessarily limited to this. In particular, it is preferable to adopt a method in which iodine is cultured to some extent in a medium that does not contain iodine ions, etc., and then iodide, which is an ion source such as iodine ions, is added to the medium and the culture is continued.
Also. Due to reasons such as the concentration of iodine ions, etc. increasing incorporation into iodine during culture.
There may be cases where the concentration is 0.5 ppm or less.
However, in the medium containing iodine as a seed, the concentration of iodine ions, etc. should always be 0.1 ppm or more.
The most preferred method is to carry out culturing while maintaining the concentration preferably from 0.5 to 800 ppm. For this reason, it is preferable to adopt a method of adding an ion source to the culture medium continuously or intermittently during the culture. Further, the ion source may be added only once, and it is of course possible to maintain the above concentration without adding an ion source. In any case, the iodine culture in a medium containing iodine ions, etc. is carried out for at least 1 hour.
Preferably, the duration is 1 day, especially at least several days. As the iodine culture medium, a normal iodine culture medium can be used except for the presence of the above-mentioned iodine ions. The usual iodine culture medium is fresh water containing the following carbon sources, nitrogen sources, inorganic sources, and others, but depending on the above-mentioned circumstances, salt water such as seawater or brine may be used.
As a carbon source, carbon dioxide gas; acetic acid, citric acid, succinic acid, and other organic acids and their salts; glycol, sucrose, galactose, and other sugars; ethanol, amino acids, peptides, and other organic substances can be used. As the nitrogen source, ammonium salts, nitrates, ammonia, urea, amino acids, etc. can be used. Inorganic sources include compounds such as phosphorus, sulfur, potassium, magnesium, iron, manganese, cobalt, molybdenum, zinc, and copper. Additionally, other nutritional sources and growth promoters may be used, such as yeast extract, molasses, hormones, etc. These components may be included in the compound used as an ion source such as iodine ions. For example, if potassium is required, potassium iodide can be used as a potassium source. Currently, in commercial iodine cultivation, organic compounds such as acetic acid are used as carbon sources along with carbon dioxide gas. However, culture using only carbon dioxide gas as a carbon dioxide source is also known. In the present invention, it is preferable to use an organic acid such as acetic acid, a salt thereof, or a saccharide together with carbon dioxide gas as the carbon source, but carbon dioxide gas alone may be used as the carbon source.
The concentration of carbon sources in the culture medium, excluding carbon gas, is
Approximately 0.1 to 10% by weight is appropriate. The nitrogen source, along with the inorganic source, is not required in particularly large amounts, and 1% by weight or less of each is usually sufficient. It is not limited to this. It is preferable that the concentration of the carbon source in the culture medium is maintained at a substantially constant level. For example, taking acetic acid as an example, the acetic acid concentration in the medium is preferably maintained at 5±3% by weight during the culture period. This is because acetic acid in this case can be used not only as a carbon source but also as a PH regulator of the medium. Adjusting the pH of the culture medium is important in suppressing the growth of various bacteria. Normally, chlorella can be cultured at a pH of 4 to 7, but if the pH is alkaline, bacteria can easily grow. The preferred pH is approximately 4.5 to 6. When acetic acid is used as a carbon source, this PH range can be maintained at the above acetic acid concentration. In addition, if acetic acid is not used, the pH can be adjusted with other acids. After the cultivation is completed, chlorella and water are separated by centrifugation or the like, and at the same time, washing is performed by washing with water to obtain chlorella with a low water content. This chlorella is then dried into products using methods such as spray drying and freeze drying. Drying is important in preventing deterioration of chlorella, and currently spray drying is considered the most preferable method.
It is said that the freeze-drying method tends to cause product deterioration because the enzyme activity is maintained. However, when extracting and using not only Chlorella itself but also components such as biologically active substances in Chlorella, the freezing method is more effective because it allows extraction of components without deterioration and without drying. considered preferable. EXAMPLES The present invention will be specifically explained below with reference to Examples, but the present invention is not limited to these Examples. Example 1 Chlorella pyrenoidosa and Chlorella ellipridea were cultured using a culture solution having the composition shown in Table 1 below. Table 1 Tap water 20 Acetic acid 1Kg KNO ₃ 5g KH ₂ PO ₄ 5g MgSO ₄・7H ₂ O 10g Iron citrate 0.04g MnCl ₂ 0.02g Add the above culture solution 40 to a 60-liter culture tank, and add about 5g of Chlorella seed stock (in terms of dry weight) and cultured under a light irradiation intensity of 500 to 20,000 lux while performing aeration at room temperature. The PH of the culture solution was measured, and when the PH increased during the culture period, acetic acid was repeatedly added to keep the PH of the culture solution in the range of 4.5 to 6.5. 0.8g of potassium iodide was added to the culture solution 10 days after the start of culture, and then 13 days after the start of culture.
0.8g was added to the culture solution. The algae were harvested 20 days after the start of culture, and the algal bodies were separated by centrifugation, thoroughly washed with water, and freeze-dried. The yield was about 15g. When the iodine content in the dried chlorella obtained was analyzed using a fluorescent X-ray analyzer, it was found to be approximately 100 ppm.
It contained iodine. On the other hand, when the iodine content of Chlorella cultured in the same manner as above without adding potassium iodide to the culture solution was analyzed, it was confirmed that it did not contain iodine. Example 2 One 500 ml portion of the basic medium for chlorella culture shown in Table 2 was poured into Erlenmeyer flask 1 equipped with a PH electrode, a medium supply tube, and a culture solution sampling tube, and the mixture was heated at 120°C for 15 minutes. The potassium iodide solution was sterilized under aseptic conditions through a Millipore filter (HA 0.45 μm), and the iodide ion concentration was 0.
It was added to the medium at concentrations of 100, 300, 500, and 800 ppm. Next, the culture medium shown in Table 2 was cultured at 10Klux and 30°C with light irradiation while aerating air containing 5% carbon dioxide gas.
pyrenoidosa C-28) was planted in the Ellenmeyer flask (1) adjusted above, and irradiated with light at 10Klux at 30°C. Shaking culture was performed while automatically adding supplementary medium (PH3.5). Figures 1 to 5 show the growth of Chlorella over time and the amount of iodine taken into the algae.
It is a diagram. Chlorella growth does not contain iodine
The best control was -=0 ppm, but when the iodine in the medium was in the range of 100 to 800 ppm, both the growth rate of Chlorella and the yield of algae were almost the same.
However, the amount of iodine uptake increased as Chlorella multiplied, and the iodine concentration in the medium was high, resulting in a large amount of iodine uptake. For iodine measurement, chlorella collected over time is thoroughly washed by centrifugation, dried by freeze-drying, oxidatively decomposed with a mixture of chromic acid, chloric acid, and perchloric acid, and the decomposed liquid is treated with thiocyanic acid. The amount of iodine taken up was measured by the iron()-iodine contact reaction method.

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[Table] Pharmacology test anti-cancer effect (effect on Sarcoma-180 solid tumor) (Method) 1 x 10 ⁶ Sarcoma-180 were subcutaneously transplanted into ICR mice (5-week-old male), and from the next day, they were treated according to the present invention. 5 g/Kg of the obtained freshwater chlorella freeze-dried product was mixed into the feed, and after 4 consecutive stages, 5 consecutive stages were performed again from the 4th day. Tumor weight was measured on the 21st day after transplantation to determine the effect.

[Table] (Results) As shown in Table 4 below, the chlorella obtained according to the present invention showed an antitumor effect of about 37%. On the other hand, common freshwater chlorella promoted tumor growth, and marine chlorella slightly inhibited tumor growth.

[Table] Although it is desirable to administer this freshwater chlorella to humans in a form suitable for gastrointestinal medicine and absorption, it can also be used as an iodine enhancer and as animal feed. The dosage for adult treatment is 0.1
It ranges from g to 100 g. The toxicity of the chlorella of the present invention is >1000 mg/Kg in ICR mice, and it is considered to have almost no toxicity to humans. Production Example Oral tablets (1) Chlorella of the present invention 350mg (2) Mannitrate 250mg (3) Starch 40mg (4) Magnesium stearate 10mg Tablets were produced according to the above formulation.

[Brief explanation of drawings]

Figures 1 to 5 are graphs showing the amount of chlorella proliferation (PCV) and the amount of iodine uptake in the grown chlorella against the culture time, and Figure 1 shows the amount of iodine ions in the culture solution [I ^- ] is 0, Figure 2 is 100ppm, Figure 3 is 300ppm,
Figure 4 shows the case of 500ppm, and Figure 5 shows the case of 800ppm.
The case is shown below.

Claims (1)

[Scope of Claims] 1. Freshwater chlorella characterized by having a high content of iodine of 100 ppm or more. 2 Iodine ion and/or iodate ion
The freshwater chlorella according to claim 1, which contains 0.1 ppm to 800 ppm and is cultured in the presence of light at a pH of 4 to 7. 3. An anticancer agent whose main ingredient is freshwater chlorella, which is characterized by a high content of iodine of 100 ppm or more.