JPH043364B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to a method for effectively and continuously producing medicinal substances contained in plants while the plants are still alive. Plants have been known to contain substances with medicinal properties since ancient times. For example, in Japan, some or all of the plants are dried or powdered under the names of Chinese herbal medicines and herbal medicines. It is widely used in general. The parts of plants that can be used as herbal medicines vary considerably depending on the type of plant. For example, the whole plant is used for genus ginseng, the leaves for foxglove, the roots for ginseng, and the fruit for ginseng. The active ingredients of plants, especially herbal medicines, have been extracted as extracts, tinctures, infusions, and decoctions and used without knowing the active ingredients. However, in recent years, as the active ingredients of many plants have been chemically and pharmacologically identified and elucidated, it has become increasingly common to extract and utilize only the active ingredients. For example, berberine from Oren, digitalis glycoside from digitalis, scopolamine, morphine, etc. are used in their pure state. However, most of the plants used as raw materials are wild or grown outdoors in mountains and fields. Digitalis is the only example that can be found in the literature that is supplied by hydroponic cultivation. However, plants obtained by hydroponics can be used as is or dried and treated with a solvent for extraction in the same manner as the above-mentioned collected plants. Therefore, the plants from which the active ingredients have been extracted are dead and discarded. Therefore, in order to produce new active ingredients, plants must be grown from seeds and cultivated. On the other hand, as a biological recovery method, that is, a method for continuously extracting the active ingredients of plants without killing them, there is a callus cell culture method. However, there are only a few types of plant components secreted by these cells, and only about 10 types have been reported, including berberine from Oriental japonica, protopine from bamboo rush, caffein from coffee, shikonin from Japanese ginseng, ginsenoside from Panax ginseng, glutathione from tobacco, and ubiquinone-10. It is. Additionally, there is knowledge that some components in the plant body may be secreted from the roots. For example, in outdoor cultivation of vegetables, when the soil after harvesting is analyzed, the amounts of acids such as benzoic acid, coumaric acid, and ferulic acid, benzaldehyde, salicylaldehyde, amino acids, and coumarin glycosides are found to be higher than before cultivation. It has been recognized that the amount increases in soil, and this is considered to be the cause of continuous cropping problems and split soil diseases. As a result of various research into medicinal substances contained in plants, active ingredients, properties of plants, and cultivation methods, the present inventors have found that while growing plants without killing them, the medicinal substances and We invented a production method that involves secreting active ingredients from plants into nutrient solutions and recovering them. Hydroponic cultivation includes hydroponic method, aeroponic method (spray cultivation method, etc.), depending on the type of medium, culture solution, and air supply method.
There are fixed culture cultivation methods (sand cultivation method, charcoal cultivation method, etc.), mixed methods, etc., but since the present invention is a method that uses a nutrient solution, that is, a culture solution, it is not possible to use any of the above-mentioned hydroponic cultivation methods. It can also be applied to cultivation methods. The present invention relates to plants containing medicinal substances and other active ingredients in their bodies, such as oren, gentian,
Seedlings of genus ginseng, ginseng, etc. are grown by the above-mentioned hydroponic cultivation, and an appropriate amount of the culture solution is collected at an appropriate time during the growing period, and from the culture solution,
It recovers medicinal substances and other active ingredients. At this time, by adding new culture solution in the same amount as the collected culture solution, the active ingredient is recovered many times at an appropriate time while growing the plant without collecting the plant body. The composition of the culture solution is adjusted as appropriate depending on the type of plant, but cellulase,
Alternatively, hemicellulase-containing enzymes (e.g., Cellulosin (manufactured by Ueda Chemical), Cellulase Onozuka (manufactured by Yakult), etc.), organic solvents (such as methanol,
ethanol, benzene, etc.) organic synthetic polymers (e.g., polyethylene, glycol, polyvinyl alcohol, etc.), surfactants (e.g., sodium lauryl sulfate, sucrose fatty acid ester, monoglyceride, etc.), or two or more thereof as appropriate. When added in sufficient amounts, these additives promote the secretion of medicinal substances, etc. in plants, act as secretion promoters, and significantly improve yields. The amount of the secretion enhancer added varies depending on the type of plant, the composition of the culture solution, and even the type of secretion enhancer, but it seems to be effective in the range of 0.01 to 10% of the culture solution. be. Medicinal substances and other active ingredients secreted from the plants into the culture solution by hydroponics are recovered from the culture solution by collecting an appropriate amount of the culture solution every 10 days, one month, or other appropriate growth period. As a recovery method, any method such as an adsorption method, a molecular sieve method, or a method of concentrating and drying the culture solution can be applied. Next, an experimental example will be shown. (Experiment example) Oren (cultivated outdoors for 2 years, and one plant has 7 to 8 stems with leaves.) 1
Wrap the stem of the plant in urethane foam, tie it loosely with wire, and place it in a wide-mouthed bottle (height: 180 cm) containing the culture solution.
mm, opening diameter 60 mm, volume 800 ml) and cultivated at 25°C and 8500 lux for 30 days. The culture solution had the composition shown in Table 2 by adding various additives to the basic medium shown in Table 1. After culturing for a certain period of time, the culture solution was freeze-dried, methanol was added to the obtained dried product, and after being left at room temperature for 2 hours, it was filtered, and the filtrate was concentrated to dryness under reduced pressure. 0.5 ml of methanol was added thereto again, and the resulting supernatant was used as a sample for analysis.

【table】 This was diluted to 1/2 and used as the basic medium.

[Table] The plant Copis used in this experiment (scientific name: Copis
Japonica) has been used as a herbal medicine for stomach health in Japan since ancient times, and its medicinal substance has berberine as its main component. From the above experimental results, berberine can be secreted and recovered over time (culture days 10, 20, and 30 days) using only the basic medium. Furthermore, when a secretion promoter as described above is added to the basic medium, secretion is promoted and the medicinal substance berberine can be recovered more effectively. Example 1 The cultivation method was M type hydroponic method. In other words, the stems of each of the three plants are wrapped in urethane foam, and then wrapped in a styrofoam plate (thickness 15 cm).
mm, size 380 x 240 mm).
mm) and fix it in the cultivation tank (380 x 240 x depth
140mm). The volume of culture solution was 15, reflux was performed twice a day for 2 hours at a flow rate of about 5/min, the light was 6500 lux, and the temperature was 25°C. 27
After cultivation for one day, culture solution 3 was extracted, and berberine was extracted using the same method as in Experiment 1, and purified by preparative thin layer chromatography. As a result, 80 μg of berberine could be obtained. After replenishing the same amount of new culture solution as the sampled culture solution, culture solution 3 was sampled every month thereafter, and the berberine content was quantified. The monthly berberine content in the culture solution is shown in "Table 3".

[Table] Example 2 The stems of 8 plants (cultivated outdoors for 2 years, with 3 stems with leaves collected as one plant) were wrapped in urethane foam, and then covered with a styrofoam board (thick The Styrofoam plate was fixed in a small hole (20 mm in diameter) drilled in a culture tank (380 mm x 240 mm x 140 mm in depth). A culture solution 20 similar to that in Example 1 was circulated at a flow rate of 1/min at a room temperature of 25°C and an illuminance of 2000 lux, and a weakly acidic ion exchange resin Amberlite IRC-50 (H ⁺ type, Organo Co., Ltd. 50 ml of ion exchange resin was filled with 50 ml of ion exchange resin, and both ends of the tube were connected via a pipe (diameter 4 cm x 15 cm) filled with glass wool, so that the culture solution passed through the ion exchange resin. 1
The ion exchange resin was replaced with a new one every week, and after washing the former with water, the adsorbed matter was eluted with 100 ml of 0.2N hydrochloric acid methanol solution. After the eluate was concentrated to dryness under reduced pressure, 10 ml of methanol was added to the residue to dissolve it, and the berberine content was determined from this solution in the same manner as in Example 1. The amount of berberine adsorbed from the culture solution every week is shown in [Table 4].

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Claims (1)

[Scope of Claims] 1. A method of secreting medicinal substances and other active ingredients from the plant body into the nutrient solution by hydroponically cultivating plants containing active ingredients in the plant body, and recovering the medicinal ingredients. A unique method for producing medicinal substances using hydroponic cultivation. 2. A patent claim characterized in that one type or a suitable combination of two or more of enzymes containing cellulase or hemicellulase, organic solvents, organic synthetic polymers, and surfactants are added to the nutrient solution for hydroponic cultivation. The manufacturing method described in Scope 1.