JPH0741687A - Production of concentrated natural red pigment oil for food and concentrated natural red pigment oil for food - Google Patents

Abstract

PURPOSE:To obtain a natural red pigment oil having excellent color tone, light- resistance, coloring property, oxidation resistance, anti-inflammatory action and immunostimulating action by extracting yeast of the genus Phaffia with a mixture of hexane and ethanol and concentrating the extract. CONSTITUTION:This pigment oil is produced by extracting Phaffia yeast capable of producing red pigment using a mixture of hexane and ethanol as an extraction solvent, concentrating the obtained crude pigment oil and preferably deodorizing the product e.g. by steam distillation.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for producing a concentrated natural red pigment oil from a Phaffia yeast producing a red pigment, and the obtained concentrated natural red pigment oil. Specifically, astaxanthin-producing yeast Phaffia rhodozyma (Ph
The present invention relates to a method for producing a concentrated natural red pigment oil from a crude extracted pigment oil obtained by extracting a pigment contained in affia rhodozyma) and the obtained concentrated natural red pigment oil. The concentrated natural red pigment oil obtained is not only excellent in color tone, light resistance, colorability and heat resistance, but also has antioxidant activity,
It is used as a new functional edible natural pigment having physiological activities such as anti-inflammatory action and immunostimulatory action.

[0002]

2. Description of the Related Art Various coloring agents have been used for food dyes. Synthetic colorants include red, yellow, green, and blue tar dyes and their aluminum chelates, iron oxides, titanium dioxide, and copper chlorophyllin sodium.
Iron chlorophyllin sodium, water-soluble anato, and β-carotene are approved according to their intended use. Although these colorants are widely used due to diversification of consumer demand and convenience of coloring, in recent years, these synthetic colorants have been replaced with natural pigments as consumer interest in health and safety has increased. is there.

As such a natural pigment, β-
Carotenes, paprika pigments, annatto pigments, red yeast rice pigments, cochineal, safflower pigments, etc. are on the market and are often used, but most of the raw materials are imported and many are unstable in supply and price. In addition, there is nothing satisfactory in light resistance, heat resistance, dye concentration, odor, etc., and there is a demand for the development of new food dyes in the food industry.

[0004] To extract the pigment from the asphyxanthine-producing Phaffia yeast, a solvent is used from the crushed yeast, and a highly polar solvent such as acetone or ethanol is suitable for the solvent used for this extraction. However, these solvents have the drawbacks that they cannot be used for extraction of food additives, are very expensive, and that the use of highly polar solvents increases the amount of contaminants in the extracted pigment oil.

The natural pigments are mainly those obtained by simple operations such as extraction, squeezing, concentration, drying and crushing from natural products such as plants and animals. Since it is obtained from natural products, it is common for extracted oil to contain many components. Commercially available natural pigments often have a pure pigment concentration of several percent, and their use may be limited in terms of the concentration. Since the red pigment produced by Phaffia yeast has a specific color tone that is redr than paprika, demand for marine products, jelly, chocolate, etc. is expected, but in order to expand its versatility in beverages, etc. Higher concentration of natural pigments has become a major issue in pigment development because it is necessary to further increase the concentration.

[0006]

SUMMARY OF THE INVENTION It is an object of the present invention to provide a method for producing concentrated natural red pigment oil for food and concentrated natural red pigment oil for food. More specifically, the object of the present invention is to provide a new food dye which is excellent in coloring properties, stability and dye concentration, which have been the drawbacks of natural dyes, without impairing the characteristics of natural dyes.

[0007]

In the present invention, a red dye-producing Phaffia yeast is subjected to an extraction treatment using a specific extraction solvent, and then the obtained crude extracted pigment oil is concentrated and, if necessary, further deodorized. As a result, a concentrated natural red dye oil for food is produced. The pigment contained in Phaffia yeast producing red pigment is extracted using a mixed solution of hexane and ethanol as an extraction solvent. That is, in order to solve the problems of the above-mentioned prior art, a mixed solvent of hexane and ethanol is adopted as an extraction solvent, and astaxanthin is extracted from the Phaffia yeast using it, and various concentration treatments are performed, and further deodorization treatment is carried out if necessary. By doing so, the above goal was achieved.

As the concentration treatment means, preferably, a low temperature fractionation treatment, a saponification treatment, or a combination of a low temperature fractionation treatment and a saponification treatment is adopted. When the low temperature fractionation treatment and the saponification treatment are used together, a method of performing the low temperature fractionation treatment and then the saponification treatment, and a method of simultaneously performing the low temperature fractionation treatment and the saponification treatment are adopted. As the saponification treatment, it is preferable to apply a method of forming a fatty acid alkali soap or a method of applying a fatty acid metal soap. As the above deodorizing treatment, steam distillation is preferably adopted.

The Phaffia yeast producing a red pigment used as a raw material yeast in the present invention is a yeast which belongs to an incomplete fungus in terms of microbiological classification and produces a red pigment, for example, Phaffia rhodozyma. In the present invention, the Phaffia yeast used for extraction is cultured, then collected by centrifugation or filtration and washed well with water, and then, as it is or after drying, is subjected to physical crushing treatment such as ball milling, freeze crushing, compression crushing, ultrasonic treatment or cellulase. Pretreatment by the enzymatic treatment used is preferred. When pigment oil is extracted from wet bacterial cells using an organic solvent, contaminants are significantly mixed, and the extraction rate is uniformly reduced, so it is generally industrially preferable to perform solvent extraction from dry bacterial cells. Is.

Acetone, ethyl acetate, hexane, ethanol and the like can be considered as the extraction solvent used when extracting the pigment from Phaffia yeast. However, acetone and ethyl acetate, which have a good extraction rate, cannot be used to extract food additives. Hexane is low in price, but has a problem in terms of extraction rate, and ethanol has a good extraction rate, but has a problem of increase in extraction cost and concentration cost due to increase in highly polar components.

As a means for solving these problems, the present invention improves the disadvantages of hexane by mixing with ethanol and provides an inexpensive and excellent extractant for concentrated natural red dye oil for foods. That is, the present invention is to extract astaxanthin more efficiently by appropriately selecting the mixing ratio of hexane and ethanol from crushed Phaffia yeast when extracting the astaxanthin pigment from Phaffia yeast.

With respect to the mixing ratio of hexane and ethanol, the extraction ratio improves as the ratio of ethanol increases. However, if the proportion of ethanol increases, the extraction cost increases and the amount of highly polar substances increases, and the subsequent concentration operation becomes complicated. Therefore, the mixing ratio was judged from the extraction ratio of the dye, the extraction cost, the azeotropic composition of hexane and ethanol, and the like. Hexane: ethanol is 90-50: 10-5
It is used in a proportion of 0, preferably 85-80: 15-20. As for the extraction method, the treatment can be carried out more smoothly by feeding the treatment liquid obtained by stirring and mixing the extraction solvent and the dry cells to the compression grinder while stirring with a pump. It is desirable to perform the crushing process multiple times depending on the extraction.

Triglycerides and free fatty acids play a major role in the lipid composition in Phaffia yeast, and efficient removal of these major components is necessary in order to increase the pigment concentration of the extracted pigment oil. Triglyceride, which is one of the main components, can be removed efficiently by low temperature fractionation. Low temperature fractionation is performed at -60 ° C to 10 ° C, preferably at -50 ° C with hexane, acetone,
It can be performed using an organic solvent such as ethanol, and astaxanthins can be concentrated and purified. For example,
When a material yeast containing a large amount of triglyceride is used, it can be said that low temperature fractionation is an excellent concentration treatment.

Free fatty acid, which is one of the main components, can be efficiently removed by saponification treatment. In the saponification treatment, astaxanthins can be concentrated and purified by removing an alkaline soap of a fatty acid produced by adding an alkali such as sodium hydroxide, potassium hydroxide or calcium hydroxide. For example, when a raw material yeast containing a large amount of free fatty acids is used, the saponification treatment can be said to be an excellent concentration treatment. The saponification treatment can be carried out in an organic solvent such as acetone or hexane or a fat and oil, but the most efficient one is in acetone.

Concentration by low temperature fractionation has a free fatty acid removal rate of about 40%, and in the concentration method utilizing the soap formation method, only the fatty acid is selectively removed. , It is possible to concentrate pigments to a higher degree from crude extract pigment oils that have multiple major lipid components.

Further, the concentrated natural red pigment oil is used for foods, and it is preferable to perform deodorizing treatment for removing yeast odor and solvent removal. Steam distillation is used as a deodorizing treatment means, steam distillation Is preferably usually 100 to 200
℃, preferably 120 ~ 150 ℃, the degree of vacuum is 30tor
It is performed below r. Dye oil extracted from Phaffia yeast generally has an irritating odor peculiar to fermentation products. This pungent odor is particularly severe when the dye is used at a high concentration, and it significantly limits the range of use of the dye oil.

Since this fermentation odor is derived from the medium used for culturing Phaffia yeast, it is considerably reduced by thoroughly washing the collected bacterial cells with water.
This alone is insufficient to deodorize the pigment oil. Further, hexane used as an extraction solvent and acetone used for concentration should not remain in the dye oil as the final product. Therefore, the pigment oil after extraction is usually added to 1
Fermentation odor is remarkably reduced by steam remaining at 0 to 200 ° C., preferably 120 to 150 ° C. and a vacuum degree of 30 torr or less, and hexane and acetone, which are extracted from the pigment oil and purified solvents, are used together with water. It could be completely removed by boiling.

[0018]

The details of the present invention will be described with reference to examples. The present invention is not limited to the embodiments. Extraction example Phaffia rhodozyma
The strains belonging to Zyma) were cultured, the obtained cultures were collected by centrifugation and filtration, and dried bacterial cells were obtained by a spray dryer. Each 300g of this dried bacterial suspension has ethanol,
Hexane, acetone, ethyl acetate, and 3 L of a mixed solvent of each ratio of hexane and ethanol were added, and the mixture was stirred and mixed, and compression pulverization was performed using a compression pulverizer (15MR manufactured by APV GAULIN). The results are shown in Table 1 (dye extraction rate by solvent).

Acetone and ethyl acetate are unsuitable for extraction of food dyes, but both have good extraction ratios of 90% or more, and thus can be sufficiently used in other fields of application. The pigment extraction rate of only hexane was as low as 65.8%, but as the content of ethanol increased, the extraction rate improved almost linearly to 87% when the ethanol content was 15%. The ethanol content must be 75% or more for the extraction rate to be 90% or more, but the proportion of highly polar components other than astaxanthins increases, and subsequent processing becomes complicated, and coexistence of hexane and ethanol From the boiling composition ratio, it was revealed that hexane: ethanol = 85: 15 was the most desirable value.

[0020]

[Table 1]

Example 1 A low temperature fractionation treatment was carried out to remove free fatty acids and triglycerides from crude pigment oil. Crude extract dye oil 10,5
Three times the amount of acetone or n-hexane / ethanol (85:15) was added, and the mixture was stirred at -50 ° C for 1 hour, filtered with 4A filter paper (ADBANTEC), and the filtrate was concentrated under reduced pressure to prepare lipids. The amount and carotenoid amount were determined. When the amount of solvent added was 3 times that of the extracted oil, the concentration of precipitates in the solution was so high that it could not be fractionated because it solidified during low temperature fractionation, but triglyceride was obtained by performing low temperature fractionation with 5 volumes of acetone. , Sterols and phospholipids were removed, and the dye concentration was concentrated about 1.5 times. The results are shown in Table 2 (concentration of pigment oil by low temperature fractionation),

[0022]

[Table 2]

Example 2 The amount of potassium hydroxide equivalent to the free fatty acid in the crude pigment oil was adjusted to 0.
Stir to dissolve in 20 ml of 8% hydrous acetone at 40 ° C., and
g oil (75, 39% free fatty acid) was added and potassium soap was formed with stirring for 10 minutes at room temperature. In order to form a metal soap, CaCl ₂ in an amount equivalent to four times the free fatty acid was added as a powder, and the mixture was sufficiently stirred at room temperature until metal soap was formed and filtered. The results of obtaining the recovery rates of fatty acids and pigments remaining in the oily substance obtained by concentrating the filtrate under reduced pressure are shown in Table 3 (removal of fatty acids from pigment oil). 90% or more of the free fatty acids were removed, and the pigment concentration was concentrated 2.9 times.

[0024]

[Table 3]

Example 3 1 g of pigment oil (free fatty acid: 75.39%) was dissolved in 20 ml of acetone, and the free fatty acid in the pigment oil and an equivalent amount of potassium hydroxide were added until the fatty acid was sufficiently precipitated. The mixture was stirred, the generated potassium soap was filtered, and concentrated.
Acetone was distilled off from the filtrate under reduced pressure, and the recovery rate of fatty acid and dye remaining in the oily substance obtained was obtained. As shown in Table 4 (removal of fatty acids in pigment oil), the free fatty acids were 92.1%.
Removal of dye improved to 90.2%, and dye concentration was 2.
The effect on the lipid composition, fatty acid composition, and pigment composition of the extracted pigment oil by the alkali treatment, which is increased eight times, is shown in Table 5 (composition change due to concentration treatment). Fatty acids were selectively removed from lipids by alkali treatment, palmitic acid removal rate was highest among free fatty acids, followed by oleic acid and linoleic acid in that order. Had no effect.

[0026]

[Table 4]

[0027]

[Table 5]

The same treatment was carried out with hexane, hexane: ethanol (85:15), and ethanol, and the fat recovery rate and dye recovery rate were determined. The lipid removal rate and dye recovery rate of these solvents were Significantly lower than acetone,
The concentration effect was not seen.

[0029]

[Table 6] Removal of fatty acids in pigment oil with hexane and ethanol

Example 4 A case where the alkali treatment and the low temperature fractionation were carried out continuously or simultaneously was shown. For continuous treatment, 10 g of crude extracted pigment oil was dissolved in 5 times the amount of acetone, potassium hydroxide equivalent to the free fatty acid in the pigment oil was added, and the mixture was stirred until potassium soap was sufficiently formed, and then potassium soap was filtered once. Aside,
Furthermore, low temperature fractionation was performed at -50 ° C for 1 hour. Simultaneous treatment was carried out by adding 5 times the amount of acetone to 10 g of crude extracted pigment oil, adding free fatty acid in the pigment oil and an equivalent amount of potassium hydroxide,
Low temperature fractionation was performed at -50 ° C for 1 hour. When the low temperature fractionation was performed after the alkali treatment, a small amount of free fatty acid, triglyceride, sterol, etc. remaining after the alkali treatment were removed, and the dye was concentrated about 4.4 times. The dye was 4.2 times more concentrated when the two treatments were performed simultaneously. Table 6 shows the results of continuous treatment and simultaneous treatment of alkali treatment and low temperature separation.
Shown in.

[0031]

[Table 7]

Example 5 About 20 kg of pigment oil was left under steam under the following conditions. Vacuum degree: 30 Torr Temperature: Tower top 96 ° C., Tower bottom 140 ° C. Vapor amount: 5 kg / hr Feed amount: 20 kg / hr The residual solvent was completely removed from 0.3% before the steam distillation. . In addition, the deodorizing effect was lowered to a level at which there was no problem in use by sensory evaluation. Since steam distillation is carried out at a relatively high temperature, there is a concern that the dye may be lost, but due to the countercurrent contact with steam, there was little decrease in the amount of dye before and after the treatment due to the short residence time at high temperature.

[0033]

EFFECT OF THE INVENTION The red pigment produced by Phaffia yeast can be provided as a concentrated pigment oil for foods which is free from contaminants and is odorless and contains no solvent. Provided concentrated natural red pigment oil for foods, which is used as a new functional natural edible pigment having not only excellent color tone, light resistance and colorability but also physiological activity such as antioxidant activity, anti-inflammatory effect and immunostimulatory effect. can do.

Claims (9)

[Claims] 1. A concentrated natural red pigment for foods, characterized by subjecting Phaffia yeast producing a red pigment to an extraction treatment using a mixture of hexane and ethanol as an extraction solvent, and then concentrating the crude pigment oil obtained. Oil production method. 2. The red dye-producing Phaffia yeast is subjected to an extraction treatment using a mixed solution of hexane and ethanol as an extraction solvent, and then the obtained crude pigment oil is concentrated and then deodorized. A method for producing a concentrated natural red dye oil for food. 3. The method for producing a concentrated natural red dye oil for food according to claim 2, wherein the deodorizing treatment is steam distillation. 4. The method for producing a concentrated natural red dye oil for food according to claim 1, 2 or 3, wherein the concentration treatment is a low temperature fractionation treatment and / or a saponification treatment. 5. The method for producing a concentrated natural red dye oil for foods according to claim 4, wherein when the low temperature fractionation treatment and the saponification treatment are used in combination, the low temperature fractionation treatment is first performed and then the saponification treatment is performed. 6. The method for producing a concentrated natural red dye oil for food according to claim 4, wherein when the low temperature fractionation treatment and the saponification treatment are used in combination, the low temperature fractionation treatment and the saponification treatment are carried out at the same time. 7. The method for producing a concentrated natural red dye oil for food according to claim 4, 5 or 6, wherein the saponification treatment is an application of an alkali soap forming method of a fatty acid. 8. The method for producing a concentrated natural red dye oil for food according to claim 4, 5 or 6, wherein the saponification treatment is an application of a metal soap forming method of a fatty acid. 9. A concentrated natural red dye for food obtained by the manufacturing method according to claim 1, claim 2, claim 3, claim 4, claim 5, claim 6, claim 7 or claim 8. oil.