JPH0699636B2 - Method of producing gardenia blue dye composition - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention is industrially advantageous with a simple operation from a known gardenia blue dye-containing material useful as a natural source dye in foods and drinks, health drugs, cosmetics and the like. The present invention relates to a method for selectively removing a yellowish dye in the material to produce a stable gardenia blueish dye composition having a clear and vivid blue color tone.

More specifically, the present invention relates to an enzyme-treated product of gardenia extract,
A gardenia blue pigment-containing material selected from the group consisting of a microbial culture treatment product of gardenia extract and a reaction product of gardenia extract with a primary amino acid-containing substance is treated with a water-miscible organic solvent, At this time, the water content of the solvent treatment system, expressed as the water content of the water-miscible organic solvent, is about
The present invention relates to a method for producing a gardenia blue dye composition, which comprises treating under the condition of 60% by weight or less, preferably about 5 to about 60% by weight, and collecting a solid content layer from the treated system.

Conventionally, gardenia obtained by allowing an enzyme such as β-glucosidase to act on, for example, a fruit or an extract thereof (generally referred to as gardenia extract in the present invention) of a plant belonging to the gardenia genus Gardenia. Enzymatically treated extract, gardenia extract medium culture containing, for example, gardenia extract obtained by inoculating and culturing a microorganism belonging to the genus Bacillus or Hansenula, further treated with gardenia extract such as amino acid Of the gardenia extract obtained by reacting a primary amino group-containing substance, such as a peptide, a protein and a protein, with a reaction product of the gardenia extract containing the primary amino group-containing substance is contained in the gardenia extract. It is known to be useful as a gardenia blue dye-containing material containing gardenia blue dye converted from the body There.

And, conventionally, the gardenia blue-based dye composition that is industrially produced is a powder of the gardenia blue-based dye-containing material as described above as it is, or is insoluble from the gardenia blue-based dye-containing material. It is usually supplied as a paste-like dye concentrate in which (solid content) is removed and concentrated. Of course, it is also known to apply a refining means for the purpose of increasing the purity of the pigment composition of such gardenia blue pigment-containing material, but a stable gardenia blue pigment composition having a clear blue color tone is produced. Is difficult.

For example, in Japanese Examined Patent Publication No. 52-13971 (JP-A No. 51-6230), a microorganism belonging to the genus Patillus is cultured in an addition medium of an extract of a plant belonging to the gardenia genus Akane family, and the original medium in the medium is used. There has been proposed a method for producing a dye composition, which comprises collecting and producing a dye composition having a color tone different from the color tone and collecting the dye composition.

In this proposal, when collecting the produced green to blue water-soluble dye composition, a known method depending on the purpose of use, that is, in the case of liquid culture, the culture solution as it is, or the solution of the culture solution is used for solid culture. In that case, it is described that the pigment composition can be recovered by appropriately using the ground product of the medium or the solvent extract, and further, for example, in liquid culture, a clear green or blue pigment can be obtained only by separating the cells. When a solution is obtained and it is necessary to increase the purity of the dye composition, a high-purity dye composition using a known purification method such as a solvent extraction method or an adsorption elution method using various adsorbents is further used. It is stated that it is also possible to obtain.

Then, in Example 6 of this proposal, a culture treated product using a gardenia extract-containing solid medium containing a crushed product obtained by crushing dried gardenia fruits with a crusher was sterilized and cooled, and then treated with an 80% ethanol solution. The green pigment composition that was extracted and accumulated in the medium was extracted, and the resulting extract was used as a membrane /
It is described that the aqueous solution of the green dye composition-containing ethanol in the liquid layer is collected by filtration and concentrated under reduced pressure to obtain a water-soluble green dye composition-containing aqueous solution.

Further, in FIG. 1 of this proposal, an absorbance-wavelength curve is shown for the culture solution of the shaking culture using Bacillus subtilis, and the culture solution (pigment solution) 120 hours after the start of the culture has about 600 mμ (590 mμ It is shown that an absorbance peak appears in the vicinity of), and an absorbance peak at about 420 mμ from the start of the culture is present as it is, although the peak height is slightly decreased. Furthermore, the absorbance data are described in all the examples of this proposal, and for any dye composition, 590 mμ and 420 mμ
The presence of an absorbance peak at is shown. The absorbance peaks of all these examples are about
The absorbance at 590 nm is (A) and the absorbance at about 420 nm derived from the yellow dye is (B), and the ratio (A)
The calculation of / (B) can be expressed as follows, for example.

As is clear from this result, the dye compositions obtained by the above proposals each have (A) / (B) of about 1.0, and their color tone is green to greenish blue as described later. It indicates that there is.

According to the studies by the present inventors, the conventional gardenia blue dye-containing material exhibits a clear and vivid blue color tone, including the dye composition described as the blue dye composition-containing aqueous solution or powder in the above proposal. However, there is a problem in that the green to green color is a soot blue, and there is a limitation in the use for a desired application of a clear and vivid blue color tone.

The present inventors have conducted research to provide a production method capable of providing a gardenia blue dye composition exhibiting a vivid blue color tone which overcomes such difficulties.

As a result, the gardenia extract contains, in addition to the iridoid glycoside, a large amount of a yellow pigment such as crocin, and the yellow pigment is treated with an enzyme as described above, a culture treatment with a microorganism, and further. It is considered that even by the conversion treatment of the iridoid glycoside into the blue pigment such as the reaction with the primary amino group-containing substance, the conversion action remains almost without being affected. The gardenia blue pigment-containing material after the treatment has a mixture of a yellow pigment and an unconverted iridoid glycoside (yellow) in addition to the blue pigment, and thus there is a difference depending on the mixture ratio. In any case, it has been found that the green to green color can essentially exhibit only soot blue, and the gardenia blue dye composition having a clear and vivid blue tone cannot be provided.

In Japanese Patent Publication No. 52-13971, the solubility of the composition obtained by collecting a pigment composition having a color tone different from the original color tone produced and accumulated in the medium is very good in water and the presence of water. Below, it is described as being soluble in a hydrophilic organic solvent such as acetone and alcohol, and as described above in Example 6, the green dye-containing extract layer with an 80% ethanol solution is collected.

The inventors of the present invention have conducted research to develop a production method capable of easily obtaining a gardenia blue dye composition having a clear and vivid blue color tone.

As a result, the gardenia extract is treated with an enzyme, a culture with a microorganism, or a reaction with a primary amino group-containing substance to convert the iridoid glycoside contained in the extract into a blue pigment. In the gardenia blue dye-containing material thus formed, a blue dye conversion product contained in the material,
Between the yellow dye such as blackish and the unconverted iridoid glycoside, which has not been changed into a blue dye by the conversion treatment, and their solubility in a water-miscible organic solvent, it is possible to separate them. It was discovered that there was a considerable size difference.

As a result of further research, the gardenia blue dye is well dissolved in water or a water-water-miscible organic solvent mixed system with a relatively large water content, but a water-water miscibility with a small water content. In the organic solvent mixed system, the solubility of the gardenia blue dye is remarkably reduced, while the yellow dye is still dissolved in the mixed system, especially, the water content is about 60% by weight.
In the following, preferably in an amount of about 5 to about 60% by weight of a water-water miscible organic solvent system, most of the blue dye migrates to a solid content layer (insoluble matter layer) and most of the yellow dye. Migrates to the liquid layer, and thus by collecting the solids layer,
It has been discovered that a gardenia blue-based composition having a remarkably improved, clear and vivid blue color tone can be industrially advantageously obtained by an extremely easy means.

According to the study by the present inventors, for example, as shown in FIG. 1 for the gardenia blue dye-containing material (crude blue dye concentrate) in Example 1 described later, the conventional gardenia blue dye composition is about It has an absorbance peak derived from a yellow dye near 440 nm and an absorbance peak derived from a blue dye near about 600 nm, and the ratio of the absorbance (B) near about 440 nm to the absorbance (A) near about 600 nm ( A) / (B) was as small as 1.63, and the obtained dye composition exhibited a green color or a soot-blue color with a rim.
As shown in FIG. 2 in Example 1 which will be described later, in the gardenia blue dye composition obtained by the method of the present invention based on the above-mentioned new findings, the absorbance peak near about 440 nm is substantially disappeared. In addition, the absorbance peak near about 600 nm increases, and the ratio (A) / (B) increases to about 2.10, which is about 2 times, and a gardenia blue dye composition having a clear and vivid blue color tone is obtained. I knew it.

Accordingly, it is an object of the present invention to provide a method for producing a stable gardenia blue dye composition having a markedly improved vivid blue color tone.

These and many other objects and advantages of the present invention will become more apparent from the description below.

According to the method of the present invention, it is known per se selected from the group consisting of an enzyme-treated product of gardenia extract, a culture-treated product of gardenia extract and a reaction product of a primary amino group-containing substance of gardenia extract. The gardenia blue dye-containing material of 1. is treated with a water-miscible organic solvent.

At this time, according to the method of the present invention, the water content of the water-miscible organic solvent treatment system is expressed as the water content of the water-miscible organic solvent, that is, the gardenia blue dye-containing material to be treated contains water. When it is contained, the water content of the water-miscible organic solvent including the water content is not more than about 60% by weight, preferably about 5 to about 60% by weight, more preferably about 5 to about 60% by weight. The yellow pigment and the unconverted iridoid glycoside in the gardenia blue pigment-containing material were treated by treating under the condition of preferably about 10 to about 30% by weight and collecting the solid content layer from the treated system. In the state where the contaminated components mainly consisting of are dissolved in the liquid layer, the objective component mainly consisting of gardenia blue dye is transferred to the solid content layer, and gardenia blue dye having a desired clear and vivid blue color tone. The components can be separated and collected.

The gardenia blue dye-containing material used in the present invention and the method for producing the same are known and can be used in the present invention. For example, gardenia blue dye-containing material, which is an enzyme-treated product of gardenia extract, is known as described in, for example, JP-A-56-92792, JP-B-54-13451, and the like. It can be produced by reacting the fruit of the plant belonging to the above or an extract of the fruit with an enzyme such as β-glucosidase. or,
For example, the gardenia blue dye-containing material, which is a culture-treated product of gardenia extract, is described in, for example,
52-13971, JP-B-59-16751, JP-A-54-96532
As described in JP-A No. 54-152026, JP-A No. 59-20357, and the like, the fruit of the plant or the medium containing the fruit extract contains Bacillus, Hansenula, Satsucaromyces, Monascus. It can be produced by inoculating and culturing a microorganism belonging to the genus, Aspergillus, Rhizopus, and the like. Further, for example, gardenia blue dye-containing material which is a reaction product of gardenia extract with primary amino group-containing material is described in, for example, JP-A-52-53932.
As described in JP-B No. 54-13451, etc., gardenia extracts include proteins such as gelatin, casein, albumin and enzyme proteins; and peptides such as peptone and protein hydrolysates: glycine. , Leucine, alanine, valine, aspartic acid, glutamic acid, and the like, and a primary amino group-containing substance such as amino acids can be used.

These treatments can be used in combination, and in the present invention,
The gardenia blue pigment-containing material selected from the group consisting of enzyme-treated gardenia extract, microbial culture of gardenia extract, and reaction product of gardenia extract with primary amino group-containing substance, It is a name that includes combined treatment products in addition to the above-mentioned treated products.

In the method of the present invention, the gardenia blue dye-containing material known per se as described above is treated with a water-miscible organic solvent, in which case the water content of the water-miscible organic solvent treatment system is as described above. The water content of the water-miscible organic solvent used is about 60% by weight or less. If the water content of the treatment system is too high, exceeding about 60% by weight, expressed as the water content of the water-miscible organic solvent used, the solubility of gardenia blue dye in the treatment liquid layer will increase sharply. The object of the invention cannot be achieved.

Examples of the water-miscible organic solvent used include monohydric or polyhydric alcohols such as methanol, ethanol, n-propyl alcohol, iso-propyl alcohol, ethylene glycol, propylene glycol, and ketones such as acetone. Can be illustrated. These can be used in combination. Among these water-miscible organic solvents, the use of ethanol and acetone is more preferable.

In carrying out the method of the present invention, the water content of the organic solvent treatment system when the gardenia blue dye-containing material is treated with the water-miscible organic solvent as exemplified above is about 60 expressed as the water content of the organic solvent. The organic medium may be used in the form of a water-containing organic solvent or in the form of an organic solvent containing substantially no water as long as the condition of not more than wt% is satisfied. The amount of the water-miscible organic solvent used can be appropriately selected and changed as long as the above water content condition is satisfied, but for example, about 0.5 to about 10 parts by weight, preferably 1 part by weight of the gardenia blue dye-containing material. Amounts of about 2 to about 5 parts by weight can be exemplified.

In carrying out the method of the present invention, the conditions for treating the gardenia blue dye-containing material with the water-miscible organic solvent can be arbitrarily selected. For example, a water-miscible organic solvent is added to a gardenia blue pigment-containing material so as to satisfy the water content condition, and the mixture is stirred, for example, at room temperature to the boiling point of the solvent used for about 10 minutes to about 5 hours, and then allowed to stand, for example. Then, the solvent layer is separated into two layers by decantation or the like, and the sedimented lower solid layer is collected. The above processing may be repeated if desired. The obtained solid content layer does not contain a yellow pigment and exhibits a high purity and vivid blue color. The solid content layer may be diluted by adding water, if desired, and further filtered.

In the gardenia blue pigment composition obtained by the conventionally known method, in addition to the blue pigment, yellow pigments such as crocin and unconverted iridoid glycosides are mixed,
According to the method of the present invention, the blue color conversion product is soluble in water or dilute alcohols, but its solubility is drastically reduced in water-miscible organic solvents having a water content of about 60% or less, and a new finding is found. A gardenia blue dye-containing material is used as a water-miscible organic compound under the condition that the water content of the solvent treatment system is about 60% by weight or less, preferably about 5 to about 60% by weight, as the water content of the solvent. By treating with a solvent, impurities such as a yellow dye are transferred to the solvent phase and removed, and an insoluble portion is collected to obtain a gardenia blue dye composition having a high purity and a clear color tone. it can.

Examples Example 1 8 kg of water was added to 1 kg of coarsely ground dried gardenia fruit, and the mixture was extracted with stirring at 50 ° C. for 8 hours, and then the insoluble matter was separated to obtain an extract.

Next, this extract was concentrated under reduced pressure to obtain 500 g of gardenia fruit extract concentrate having a solid content of 70%.

50 g of this concentrate, 5 g of sodium L-glutamate and 5 parts of water
00 g was put into a 2-volume flask and dissolved, and the pH was adjusted to 6.5 using a 1N-NaOH aqueous solution. Next, add 5 g of the enzyme morphine (Morijin Pharmaceutical Co., Ltd.), stir at about 45 ° C for 20 hours to carry out the blue conversion reaction, and then heat at 90 ° C for 15 minutes to inactivate the enzyme to obtain a crude blue dye. 515 g of solution was obtained. Of this dye solution
The absorbance at 595 nm (A) was 382, or the absorbance at 440 nm (B) was 234.

This dye solution was concentrated under reduced pressure to obtain 170 g of a concentrated liquid. Next, 500 g of 92 wt% ethanol was added to this concentrated liquid (the water content of the treatment system was 25 wt% expressed as the water content of ethanol),
After stirring and mixing uniformly, the mixture was left standing for 24 hours to remove the upper ethanol phase by decantation, and 300 g of water was added to the resulting precipitate to dissolve it, and the desired purified gardenia blue dye solution. Obtained 350 g.

The purified dye solution had an absorbance (A) at 595 nm of 460 or an absorbance (B) at 440 nm of 153. The results are shown in Table 1, FIG. 1 and FIG.

As is clear from the results shown in Table 1, the blue dye obtained by the method of the present invention was a distinctly bright and vivid blue color.

Example 2 Gardenia fruit extract concentrate used in Example 1 (solid content 70
%) 10 g and 590 g of water were placed in a 2-volume flask and dissolved,
The pH was adjusted to 5.0 with N-NaOH. Next, 1 g of the enzyme acylase (Amano Pharmaceutical Co., Ltd.) was added, and the mixture was dissolved with stirring and then left standing for 15 hours. Then, the stirring was restarted, and the stirring was continued at 500 rpm for 40 hours.

The resulting blue-based dye conversion product solution is heat-treated at 90 ° C for 15 minutes to inactivate the enzyme, and then concentrated under reduced pressure.
got g. The absorbance (A) of this concentrated solution at 595 nm is 60.
The absorbance (B) at 0 and 440 nm was 476.

To the obtained concentrated solution was added 130 g of 92% by weight ethanol (expressed as the water content of ethanol, the water content of the treatment system was about 25%).
(% By weight) After stirring for 10 minutes, the mixture was allowed to stand at room temperature for 1 hour. The upper hydrous ethanol layer separated into two layers was colored in yellow green. Remove this ethanol layer by decantation,
Water was added to and dissolved in the lower solid content layer obtained as a candy-like precipitate to obtain 18 g of purified gardenia blue dye solution. The absorbance (A) at 595 nm of this purified gardenia blue dye solution was 1100, and the absorbance (B) at 440 nm was 380, so that the ratio (A) / (B) was 2.89, and a clear and vivid blue color was exhibited. . The results are shown in Table 2.

Example 3 20 g of corn starch, 1.5 g of ammonium nitrate, 1.0 g of corn stay liquor, 0.6 g of monopotassium phosphate, 4.0 g of disodium phosphate and 0.5 g of magnesium sulfate were dissolved in pure water 1 and added to 1 ml of this at 440 nm. Absorbance 1,400
Add 100 ml of gardenia extract.

Adjust the pH of this medium to 6.0 and 50 ml in a 500 ml shoulder shake flask.
They were put in each and sterilized at 120 ° C for 15 minutes.

Hansenula anomala (Ha
nsenla anomala) OUT6316 was inoculated with 2 platinum loops per flask, and shake culture was carried out at 30 ° C for 8 days.

The obtained green culture solution 1 was taken and sterilized at 85 ° C. for 30 minutes, and then diatomaceous earth was used as an auxiliary agent to obtain 900 ml of the solution.
The absorbance per ml of this dye solution is 106, 5 at 440 nm (B).
It was 75 at 95 nm (A).

900 ml of this dye solution was concentrated under reduced pressure using a rotary evaporator to obtain 300 ml of concentrated solution, and then 1,000 ml of 90% by weight ethanol was added while stirring this concentrated solution (about 27% of the treatment system represented as ethanol water content). %) After standing overnight, the ethanol layer was removed by decantation, and 100 g of water was added to the precipitation portion and uniformly dissolved to obtain 135 g of a purified blue dye solution. The absorbance (A) at 595 nm and the absorbance (B) at 428 and 440 nm of this dye solution were 185, and (A) / (B) had a clear bright blue at 2.3. The results are shown in Table 3.

The purified gardenia blue dye composition obtained according to the present invention has a significantly reduced content of the yellow dye, as compared with the conventional gardenia blue dye composition, showing a clear blue color,
Further, since it does not contain an unconverted unstable iridoid glycoside, troubles such as discoloration can be avoided, and it can be suitably used for various foods and drinks, health drugs and the like.

[Brief description of drawings]

FIG. 1 shows the crude blue pigment concentrate obtained by converting the gardenia extract used in Example 1 into blue color by enzymatic treatment, and FIG. 2 shows the crude blue dye concentrate obtained by Method 1 of the present invention. The visible absorption curve of the purified gardenia blue dye was shown (1 ml to 500 ml).

Claims (1)

[Claims] 1. A gardenia blue pigment selected from the group consisting of an enzyme-treated product of gardenia extract, a cultured product of gardenia extract with a microorganism, and a reaction product of gardenia extract with a primary amino group-containing substance. The containing material is treated with a water-miscible organic solvent, wherein the water content of the solvent treatment system is 10 to 30% by weight, expressed as the water content of the water-miscible organic solvent.
A method for producing a gardenia blue dye composition, which comprises treating the gardenia blue dye as a solid layer from a system which has been treated under the conditions of the above.