JPH01146961A - Production of natural red pigment and food - Google Patents

Abstract

PURPOSE:To produce a natural red pigment which can be easily used in food, by removing unnecessary pigments by extracting a red pigment and water-soluble substances from red pigment-producing algae such as red algae or blue-green algae with water or a salt solution. CONSTITUTION:Red pigment-producing algae such as red algae (e.g., Porphyridium culenteum) or blue-green algae (e.g., Calothrix prepicima) is extracted with water or a salt solution. When algae of a strain which cannot be obtained in abundance by raising or from nature, they are used after being cultivated. The extraction of pigments can be performed by using algae as such or after breaking their cells by using frozen wet algae or a dried product. To the extract, a solvent (e.g., methanol) is added to remove unnecessary substances (e.g., chlorophyll and phycocyanin) by precipitation and subjected to centrifugation or filtration to leave natural red pigment in the liquid. The obtained pigment has a bright color tone of pink to red-purple and a high tinting strength, excels in stability against heat, light, etc., and can be used in food.

Description

DETAILED DESCRIPTION OF THE INVENTION <Industrial Application Field> The present invention relates to a method for producing a natural red pigment obtained from red algae or blue-green algae, and to foods to which the pigment is added.

<Prior Art> Conventional natural red pigments for food include carotenoids, red malt pigments, laccaic acid, hibiscus pigments, beet red, and quinone pigments. When using these dyes, since they are food additives, it is desirable to be able to stably color the desired color with a small amount of the dye. However, with conventional dyes, the desired color tone may not be obtained, or the coloring strength and stability may be insufficient. Furthermore, when such natural red pigments are used to color white products such as milk and yogurt, there is a problem in that they do not produce a bright pink color, but only a dull pink color.

The production of such natural red pigments is usually carried out by extracting from each raw material, concentrating it, ammonium sulfate salting out, and purifying it by column chromatography, but these methods are complicated, and reduce workability and production. It was inferior in terms of efficiency.

<Problems to be Solved by the Invention> The present invention has a pink to reddish-purple color tone that cannot be obtained by conventional food coloring alone, and the color tone is bright and has strong coloring power. The present invention aims to present a simple method for producing a water-soluble natural red pigment that is stable against light, solvents, etc. and can be used in foods, and to provide foods containing the pigment.

<Structure of the Invention> As a result of intensive research into red pigments produced by algae, the present inventors have developed a method for producing natural red pigments using red algae and blue-green algae that is simple and has excellent production efficiency, which is different from conventional methods for producing natural red pigments. The present inventors have discovered that when a water-soluble natural red pigment obtained by a manufacturing method is used in foods, it has excellent stability against heat, light, etc., and brightness, and has completed the present invention.

That is, the present invention extracts red pigments and water-soluble substances from red pigment-producing algae such as red algae or blue-green algae with water or a salt solution, and then removes unnecessary substances by treating the extract with a solvent. The present invention provides a method for producing a natural red pigment, which is characterized by obtaining a red pigment, and a food product, which is characterized by containing the obtained natural red pigment.

The properties of the red pigment of the present invention are as follows.

■Thermal stability: When treated at pH 6.0 and 0 to 40°C for 30 minutes, the absorbance at 565 nm remains at 95 cm or more.

(b) Stability range: When treated at 25°C and pH 5.0 to 6.5 for 30 minutes, the absorbance at 565 nm remains at 95 cm or more.

Solubility Soluble in cold water and hot water, but insoluble in organic solvents such as ethanol, acetone, and ether.

The primordium and classification of algae used in the present invention are published by Kyoritsu Shuppan, “Algae Research Method” (1974) co-authored by Toshi Torisawa and Mitsuo Chihara.
The algae used in the production of this red pigment include, for example, red algae;
) R-1, blue-green algae; Calothrix brevissima (Ca1
othr1x br@vimsimm) M-7, Anabaena varia
bills) M-2, Cylindrospermum rmsci (C)rllndrospermum rmsci
cola) M-32 and Ashi, respectively, are preserved as microalgae strain archives at the Institute of Applied Microbiology, University of Tokyo. However, in carrying out the present invention, there is no need to be limited to a specific algae strain, and red algae that are widely cultivated in Japan, such as Asakusanori;
a) A blue-green algae, Aphanothece sp., a species of the genus Aphanothece;
p, ), Hinanori; a species of the genus Porfurobsis (Por
phyropsis sp, ), one species of the blue-green algae Phormidium (Phormidlum3p), one species of the Nostoc genus (No5toc sp, ), Tolypothnix tanuis
) and other algae strains can be used to produce pigments.

For algae strains that cannot be obtained in large quantities through cultivation or from nature, cultivation is carried out in accordance with a culture method suitable for each algae strain, using a medium and culture conditions that allow the algae to grow. As the culture medium, a nutrient medium containing inorganic salts necessary for the growth of algae, seawater, and soil extract may be used as they are, or an appropriate amount of nutrients may be added to these.
There is a culture medium with added vitamins. Carbon sources include bicarbonate, carbonate and carbon dioxide. Further, as nitrogen sources, nitrates, ammonium salts, nitrogen gas, and organic nitrogen such as bezotone, meat extract, yeast extract, and urea can also be used. Phosphorus sources include inorganic phosphates such as sodium phosphate and potassium phosphate, and organic phosphates such as β-glycerophosphate.

Other cations include sodium, potassium, magnesium, calcium, and iron, and anions include chlorine and sulfuric acid. Trace metals include boron, manganese, zinc, copper, molybdenum, vanadium, chromium, nickel, titanium, cobalt, lithium, bromine, strontium, rubidium, and iodine. EDTA if necessary
It is also possible to add trace amounts of vitamin components such as chelating agents such as vitamin B1, vitamin B12, and biotin.

Since red algae and blue-green algae have originally grown under natural environmental conditions, various inorganic and organic compounds can be used for their growth. Therefore, the cultivation of the above-mentioned algae is not limited to only the above-mentioned components. In order to improve the red pigment content, it is preferable to set the nitrogen source in the medium components to be relatively large compared to the carbonate source.

Culture at a temperature of 5-60°C, for most species at 20-35°C, and at a pH of
2 to 11 are usually carried out by shaking culture or deep aeration agitation culture. In order to carry out photosynthesis efficiently, the irradiation light is uniformly applied to the algae so as to maintain a constant intensity. The irradiation light may be either sunlight or artificial light, and stronger irradiation with green light is preferable in order to increase the red pigment content. Furthermore, the content of the red pigment of the present invention can be increased by culturing until the late linear growth phase when the algae approach the maximum concentration. After culturing, harvest by conventional methods such as centrifugation and filtration.

Extraction of the pigment can be carried out by using the harvested algal bodies as they are, or by disrupting the cells using frozen wet algal bodies and dried products. Cell destruction can be done by adding water or salt solution to the algae, using a homogenizer, sonication, freezing/thawing,
There are physical treatment methods that involve osmotic pressure treatment, and biological treatment methods that decompose cell walls with glycolytic enzymes (cellulose hydrolase, mannan hydrolase, etc.) that have the ability to hydrolyze polysaccharides. Depending on the type, these methods may be combined to destroy cells. Extraction is carried out by adding 2 to 20 times the amount (w/'w) of water or a saline solution to the cell disrupted product obtained as described above, and adding a weak alkaline solution or a weak acid solution to a pH of preferably 4.0 to 7.0. , especially p) f5.0-
Adjust the pH to 6.5 and reduce the pH to 1 to 12 with gentle stirring.
Perform time extraction. As the salt solution, an acidic buffer solution preferably has a pH of 4.0 to 7.0, particularly a pH of 5.0 to 6.5.
For example, known buffers such as phosphate buffer and pine kilvain buffer can be used. The concentration of this solution is preferably 1/100 to 1/10 normal.

Further, in order to prevent the dye from being denatured, it is preferable to carry out the reaction in a dark place and at a low temperature of around 4°C. However, care must be taken to prevent denaturation, which is not limited to this temperature. Next, this extracted product is filtered or centrifuged to separate and remove algae residues to obtain an extract. This extract contains red phycoerythrin, back phycocyanin,
and clophyll.

Extraction and separation are particularly advantageous when carried out at pH 5.0 to 6.5, as more than 50% by weight of algae residues such as chlorophyll pigments and cell membrane components can be separated and removed at this stage than when carried out under alkaline conditions.

Furthermore, in order to separate this pigment from the kernel extract, a solvent,
Preferably methanol, ethanol, acetone or a mixture thereof is added to the extract in an amount of 5 to 70% by weight, preferably 25% by weight.
By adding ~50 weight ttIb, unnecessary chlorophyll, phycocyanin, etc. can be precipitated and removed. In this manner, the natural red pigment of the present invention can be obtained in the liquid by centrifugation or filtration again. The obtained dye solution is stored as a dye, either as it is or by drying by a method such as freeze-drying to form a solid powder.

The red pigment of the present invention has the advantage of being a natural product, is highly safe, and exhibits a pink to red color, which is in high demand as a food pigment. When this pigment is used as a red food pigment, it may be in either powder or liquid form.

When used as a powder, a color value adjusting agent such as lactose, a color adjusting agent such as sodium citrate, and a clarifying agent can be added to the chromogen powder.

The amount of lactose added can be adjusted to 40-80% to adjust the color value, and the amount of sodium citrate can be adjusted to 5-1%.
By adding 0%, the sea can be adjusted to 6.0 to 7.0.

When used in liquid form, the present chromogen powder can be used by directly dissolving it in water, but it is preferable to use it by dissolving the above-mentioned color value adjusting agent. Pigment powders and liquids exploded in this manner can be used in a wide variety of applications, but are particularly useful in foods. The food of the present invention can be used as a coloring agent for various sweets such as drinks, frozen desserts, candies, and chewing gum, dairy products such as yogurt, fish paste products, ham, sausages, etc. υ Especially used for white foods. It's good to do that. The amount added is 0.001% to 10%, preferably 0.01% to 0.0% depending on the purpose.
By adding 5%, sufficient pink to red coloring can be obtained.

<Effects of the Invention> As already mentioned, the present invention employs an oil-based method in which necessary pigments are removed by adding a solvent such as ethyl alcohol, rather than complex methods such as ammonium sulfate fractionation and column chromatography. A very red pigment can be obtained. On the other hand, natural red pigments include dulaccaic acid, red malt pigment, and beetroot to ibiscus pigment, but when mixed with colloidal white foods such as milk, they become dull and produce a purplish color that does not turn pink. All I could get was a belt. However, since the red pigment obtained by the present invention exhibits a bright pink color, it is very suitable as a coloring agent for white foods such as milk and ice cream. Furthermore, this dye has excellent solvent resistance and heat resistance compared to other protein-bound dyes, so it can also be used as a general-purpose red dye.

<Example> The present invention will be explained in more detail with reference to Examples below.
The present invention is not limited to these examples.

Example 1 Example of producing a red pigment from blue-green algae (Calothrix genus) Callothrix tulepisima M-7 strain was added to the Fitzgerald medium shown in Table 1 below, and the absorbance at 560gm was 0.1.
Inoculated so that
Aerated culture was carried out while supplying carbon dioxide.

Table 1 Fitzgerald medium composition NaNO s O-496
J'2HPO40,039,9 Mg5O4-7H200,075# Ca C70・2H200-0361 Na2C030, o2oII Na28103'9H20o, ossp Iron citrate
0.006.9 citric acid
0.006.9p) 19.0 Medium trace element solution H3BO43,1,9 MnSO4"4H202-2311 ZrtSO4-7H200,287# (NH4)6Mo, 024"4H200,088#Co
(No,)2-4H200,146,9N12WO4・
2H200,033,9KBr
O,11911KI
O,08:lCd(NO3)2.4H200,154
llNl504(NH4)2So4φ6H200,19
8,! 9VO8O4-2H200,02,9 At2(S04)3 to 2SO4・24H200,474
,91/l0NH280410001! After 1/7 days, the late linear growth phase was reached, and the absorbance of the culture solution was 2.5. This culture g200I! When harvested using a centrifuge, approximately 250 I was obtained in a wet heavy lid.

Three times the amount of 10 mPi11 in this wet algae! Potassium phosphate buffer (pH 6.0) is added, the cells are sufficiently destroyed by ultrasonication, and extraction is performed with gentle stirring for 1 hour. This dye suspension is centrifuged at 5000G for 15 minutes to separate solid and liquid. The obtained supernatant was concentrated using an ultrafilter to obtain 250 g of a concentrated solution. 501 for this
Add 1Ll of ethyl alcohol and stir to make 100OOG
, and centrifuged for 10 minutes to obtain a supernatant.

This supernatant was freeze-dried to obtain red dye 12F'i. This one
When #ill was dissolved in distilled water, the absorption at 565 nm was 1.21. Also, when the color was measured using a colorimeter, L=71.5. a=28.5. It was b-17.1. In addition, in a phosphate buffer solution with a pH of 6.0,
No discoloration was observed when heated at ℃ for 30 minutes.

Example 2 After harvesting the red algae Porphyra tenera f cultivated naturally by a normal method, distilled water gold in an amount three times the wet weight i (10 kg) of the algae was added to the algae, and the pH was adjusted to 6.5 K. adjust. At this time, a portion of the Mi cells is destroyed due to the osmotic pressure difference between the inside and outside of the cell, and the dye is eluted. After the mixture was further fragmented into fine pieces using a homonaive, cellulose-degrading enzyme, mannan-degrading enzyme, and lysozyme were added in amounts of 0.1 to 10 pieces each, and the mixture was reacted for 1 hour. The reaction solution was heated to 5000G.

Centrifuge for 30 minutes to remove cell debris.

This purple color was concentrated to 151 by ultrafiltration and 5! of ethyl alcohol and stirred at 8000G.

Centrifugation was performed for 15 minutes to obtain a red supernatant. This supernatant was freeze-dried to obtain 140 g of a reddish-purple solid. When this IFill was dissolved in distilled water, the absorbance was 1.42. Also, when the color was measured using a colorimeter, L = 63.5, a = 3
3.8, b=18.2. Also, no fading was observed when heated at 40°C for 30 minutes in a phosphate buffer solution with a pH of 6.0.

Application Example 1 When an ice cream was prepared using the red pigment obtained in Example 1 according to the following recipe, a bright pink ice cream was obtained.

Application Example 2 When strawberry-flavored milk was prepared using the red pigment obtained in Example 2 according to the following formulation, a strawberry-flavored milk drink with a bright pink color was obtained.

When heated at a total temperature of 63°C for 30 minutes, almost no fading was observed - Comparative Application Example 1 In the formulation of Application Example 2, the product of the present invention was substituted with beet red (commercial product, hot water extracted concentrate, gold-based However, the color was completely distorted from purple to pink, and a bright pink color similar to strawberry milk could not be obtained.

Agent: Patent Attorney Katsutoshi Takahashi

Claims (1)

[Claims] 1. Red pigment and water-soluble substances are extracted from red pigment-producing algae such as red algae or blue-green algae with water or a salt solution, and then unnecessary substances are removed by treating the extract with a solvent. A method for producing a natural red pigment, which comprises removing the pigment to obtain a red pigment. 2. Red pigments obtained by extracting red pigments and water-soluble substances from red pigment-producing algae such as red algae and blue-green algae with water or a salt solution, and then removing unnecessary substances by treating the extract with a solvent. Food characterized by containing.