JP2571060B2 - Increasing sales of red pigments in Beni flowers - Google Patents

Description

Description: TECHNICAL FIELD The present invention relates to a method for increasing the yield of red pigment of a red flower.

More specifically, the present invention increases the amount of dissolved oxygen by, for example, blowing oxygen or an oxygen-containing gas into the liquid medium when cultivating calli of the flower of the red flower, and efficiently increasing the red pigment of the red flower. How to do it.

Since the present invention enables a large amount of red pigment of Beni flower, which is a natural pigment, to be produced in large quantities, there are great benefits to industries utilizing natural pigments, such as the cosmetics industry and the food industry.

(Prior art) In general, Beni flowers are plants of the family Chrysanthemum which are often cultivated in the Akita region, and the flowers harvested contain beautiful red pigments (cartamine), yellow pigments, etc. Dried flowers are prized as tea for inclusion. The red pigment extracted from the flower is sold as a red dye and a natural lipstick.

Since the red pigment contained in the Beni flower is extremely useful as a natural pigment, the present inventors have previously conducted extensive research on calli culture of the Beni flower in order to mass-produce the red pigment of the Beni flower. In two-stage cultivation of flower cells in solid culture and liquid culture, the concentration of at least one of the components was reduced, thereby producing red-colored callus.

However, even with this method, the amount of red pigment produced is small, and mass production has not yet been achieved.

(Problems to be Solved) The amount of red pigment obtained by culturing the calli of Beni flower with a culture solution is extremely small, so that it has not reached the stage of industrial mass production. The present invention makes it possible to industrially mass-produce red flower pigment by increasing the yield of red pigment by improving the liquid culture method.

(Means for Solving the Problems) The present inventors have conducted research on a method of mass-producing the red pigment of the red flower, and found that when the cell group of the red flower was cultured in a liquid medium, the red pigment of the red pigment was removed. In the coloring step, the amount of dissolved oxygen is reduced by blowing oxygen or an oxygen-containing gas into the medium.
By increasing the concentration to 4.0 ppm or more, the yield of the red pigment was significantly increased.

Further, in the present invention, except for ammonium ion and / or calcium ion and / or magnesium ion, D
It is preferable that the cell group of the flower of Beni is cultured in a liquid medium to which an aromatic amino acid of body and / or DL body or a content thereof is added.

In addition, the yield of the reddish pigment can be further increased by adding a color development promoting substance selected from cellulose powder, microcrystalline cellulose, filter paper, cellulose, chitin, cotton, and silk during culturing.

The present invention is characterized by increasing the amount of dissolved oxygen in a culture medium of a cell group of red flower.

In order to increase the amount of dissolved oxygen in the culture medium, it is preferable to blow sterile air into the culture medium.

Simply insert a single nozzle that blows air into the swirling culture vessel, and then gently blow sterile air from outside into the culture medium. The dissolved oxygen in the culture medium can be reduced from about 1-2 ppm to 7 ppm.
88 ppm, so that the yield of the reddish pigment in the amount of 2-5 liters is increased.

In the present invention, only oxygen or an oxygen-containing gas may be blown.

Further, in the present invention, in order to increase the amount of dissolved oxygen in the culture medium, if a cover-type silicon stopper or the like having excellent air permeability is used for the stopper of the culture vessel, fresh air is blown into the culture medium during the swirling culture, and air is released. As if blown, the amount of dissolved oxygen increases from about 2.2 to 2.5 ppm to about 4.5 ppm when a conventional silico stopper is used, and the red pigment can increase its yield two to three times. .

The cell group of the Beni flower used in the present invention is a leaf of the young plant,
It is collected from stem tissue and the like, but can also be collected from flower buds. The flower bud as referred to herein is a young tissue that has the ability to become a flower in the axil of a flowering plant of Veneae in the future. It has the ability to become a flower by itself when the bud on the top is gone.

In the present invention, the cell group of the flower of the safflower is first cultured in a solid medium, and the obtained callus is deprived of ammonium ions and / or calcium ions and / or magnesium ions to obtain D-form and / or DL-form aromatic amino acids or The cells are cultured in a liquid medium containing the contents. In this culture, culturing only when the color is developed is sufficient. During this culture, a color-promoting substance can be added from the beginning or from the middle.

Generally, Murashige-Skoog medium, white medium, Gamborg medium, and the like used for plant tissue culture have ammonium ions, calcium ions, and magnesium ions necessarily added thereto, but the present invention does not include any or all of them. Nogai. The presence of ammonium and / or calcium and / or magnesium ions significantly reduces the formation of red pigments.

The D-form aromatic amino acids include D-phenylalanine, D-tyrosine, D-tryptophan, and the like, and the addition thereof increases the production of red pigment. D-form aromatic amino acids are generally a mixture of DL-forms when synthesized, but if they contain D-forms,
A mixture of DL-forms or aromatic amino acids of D-form alone may be used, and the same effect can be obtained.

The D-form and / or DL-form aromatic amino acid is preferably added to a liquid medium at about 0.1 mM to 10 mM. Outside this range, the production of reddish pigments tends to decrease.

The color-promoting substance may be at least one selected from cellulose powder, microcrystalline cellulose, filter paper, cellulose, chitin, cotton, and silk. Any substance that promotes the production of a red pigment may be used. 0.1-10 for medium
%, Preferably about 1 to 5%. After the cultivation, the red pigment can be easily separated from the color-promoting substances such as cellulose powder, microcrystalline cellulose, and cellulose, which have become red, with an organic solvent such as pyridine, and a beautiful red pigment can be isolated. is there.

In the present invention, the method for increasing the amount of dissolved oxygen in the medium may be employed in all cultures, but it is effective and sufficient to adopt it only in the final step of coloring the red pigment.

Next, Examples of the present invention will be described. As an improved medium of Murashige-Skoog used here, KBC-G of a column culture medium was used.
1 and KBC-P5, KBC-P1, KBC-
The compositions of P2 and KBC-P3 are shown in Table 1 below.

Example 75 KBC-G1 medium was placed in a 300 ml Erlenmeyer flask at 75 ° C.
Sterilized for 5 minutes. After cooling, the above-mentioned wet calli of Beni flower 3.5
g of the mixture and cultivated by swirling at 25 ° C. for 3 days. The cells grown in about 2 cultures are obtained by suction filtration, and 3.5 g of the cells are added to the KBC-G1 medium in the same manner as above, followed by further culturing for 3 days. This operation is repeated to obtain a vigorous callus.

On the other hand, 75 ml of each KBC-P5 medium was placed in five Erlenmeyer flasks for coloring each set in the manner shown in Table 2, 3.5 g of the callus obtained above was placed, and the cells were cultivated at 25 ° C. for 3 days.

In this example, 1 mM of D-phenylalanine and 4% of cellulose powder were added.

After the culture, the callus and the colored cellulose powder were separated, and the adsorbed dye was eluted with pyridine, and the absorbance at 520 nm was used as the degree of color development. The results are shown in Table 2 below.

Claims (1)

(57) [Claims] 1. A method for increasing the amount of red pigment of a flower of Beni, wherein the amount of oxygen dissolved in the culture medium is increased to 4.0 ppm or more in the step of coloring the red pigment when culturing the cell group of the flower of red safflower in a liquid medium. Law.