RU2315094C9 - Method and device for producing phycoerythrin with high optical density - Google Patents

Abstract

FIELD: biochemistry, in particular, methods and devices for producing coloring substances, possible use in food and cosmetic industry, and also during various biological research.

SUBSTANCE: phycoerythrin protein pigment is produced by extraction from seaweed. It is extracted from seaweed, selected from a group including Galaxaura oblongata, Halymenia ceylanica, Helminthocladia australis and Porphyra dentate.

EFFECT: phycoerythrin has high optical density.

2 cl, 27 dwg, 2 tbl

Description

BACKGROUND OF THE INVENTION

Technical field

The present invention relates to a method and apparatus for producing phycoerythrin with high optical density (OD), and more particularly, to a method and apparatus for producing phycoerythrin with high optical density from a group of plants including Galaxaura oblongata, Halymenia ceylanica, Helminthocladia australis and Porphyra dentata.

Prior art

Natural protein pigments derived from plants are safe for use in food and beverages. They are resistant to moderate heat and are acidic or alkaline solutions. Due to this, they can be used in the food and cosmetic industry as coloring agents. In addition, pure pigments can be used for fluorescein labeling of antibodies used as diagnostic tools in immunological, clinical, cytological and biochemical studies.

Phycocyanin and phycoerythrin are two natural protein pigments that are currently widely used in many fields. Since the main raw material for producing phycocyanin is blue-green algae, such as Spirulina and Microcystis, which are easy to grow, and many methods have been developed to grow algae and produce phycocyanin, the supply of phycocyanin is not a problem. Nevertheless, there is still a shortage of phycoerythrin, in connection with which its price remains high. Such a shortage is explained by the lack of raw materials and the technological difficulties in the production of phycoerythrin on an industrial scale.

Most phycoerythrin is obtained by extraction from red thallic structures of algae, such as Porphyra and Ceramium, and only to a small extent by Porphyridium, which is now obtained by growing in tanks.

Despite the fact that wild red algae and artificially grown Porphyra are used in increasing quantities as raw materials for phycoerythrin, most of these raw materials have a high gel content, which makes the extraction of phycoerythrin from such plants extremely difficult, especially when it comes to dried algae . In addition, the quantity and quality of wild algae depends on the season and ambient temperature. These circumstances make the production of phycoerythrin from wild and artificially grown Porphyra algae even more difficult.

The extraction of phycoerythrin from Porphyridium is also associated with difficulties, since the collection of unicellular plants usually requires a lot of time and labor, and the soluble polysaccharide that is released during the growth of algae slows down the collection and affects the extraction of phycoerythrin.

US Pat. No. 5,358,858 proposes a solution to the aforementioned problems using a method for producing phycoerythrin from Bangia atropurpurea and Porphyra angusta. Since these filamentous plants do not contain gel, they can be stored under conditions such as nutrient media with controlled temperature, degree and duration of illumination. Phycoerythrin can be easily extracted from filamentous plants. During the implementation of the proposed in the named invention method:

1. At sea, the thallus structures of mature Bangia atropurpurea or Porphyra angusta are collected and washed with sterilized sea water. After brief drying in air, they are placed in a nutrient medium (medium SWM-III, see table 1). After a few hours, spores stand out from the thallus structures of Bangia atropurpurea or Porphyra angusta. Then, the released spores are removed from the initial nutrient medium and placed in a germination chamber, in which the temperature corresponds to room temperature, the degree of illumination is 1000-4000 lux, and the duration of illumination is 10-16 hours per day.

Table 1 NaNO ₃ 8.5 g / 100 ml take 2 ml Na ₂ HPO ₄ 0.595 g / 100 ml take 2 ml Na ₂ EDTA 0.5 g / 100 ml take 2 ml 0.01625 g / 100 ml take 2 ml FeCl ₃ (or FeCl ₃ · 7H ₂ O) 0.02885 g / 100 ml 2 ml * metal 1PI 2 ml * Vitamins 2S-3 2 ml Soil extract 50 ml Trisaccharides (10 cc / l) 500 mg Liver extract 10 mg Sea water up to pH 7.5 1 liter

(Get HCl with a higher density and an initial pH of less than 7, then add a NaOH solution to bring the pH to about 7.5. This avoids the formation of precipitate.)

* metal 1PI H ₃ IN ₃ 12.368 g MnCl ₂ 1.385 g ZnCl ₂ 0.109 g add 2 l of distilled water CoCl ₂ · 6H ₂ O 4.479 mg CuCl ₂ · 2H ₂ O 0.034 mg * Vitamins 2S-3 Thiamine NS 0.5 g Capantothenate 0.1 g A nicotinic acid 0.1 g Para-aminobenzoic acid 10 mg Vitamin H (Biotin) 1 mg add 2 l of distilled water Vitamin B8 (Inositol) 5 g Folic acid 2 mg Timin 3 mg AT 12 1 mg

2. After the branched strands sprout from the pores, they are transferred to flasks with SWM-III growth medium and grown under the above conditions to form colonies. The filamentous colonies are then pulverized in a sterilized mill into small pieces and placed in a larger container, such as a reservoir, to facilitate their further growth. Then filamentous colonies are transferred to a larger room, where they continue to grow. Then they are again crushed for further cultivation to the required amount. It is important that when growing filamentous colonies in tanks, it is necessary to supply fresh air (300 ml / min) in them. Then the colonies are collected and filtered through a network with a number of cells from 100 to 400. The nutrient medium is removed and reused.

3. Then the collected and filtered strands of Bangia atropurpurea or Porphyra angusta are quickly dried in vacuum or with heated air and crushed into powder. The powder is added to a phosphate solution or water and mixed thoroughly. Foreign matter is removed by centrifugation and a red, clear pigment solution is obtained. To obtain the crude phycoerythrin, (NH ₄ ) ₂ SO ₄ is added to the solution and a 20% -30% saturated solution is obtained from which unnecessary proteins are removed, after which the solution is settled using a 60% -65% saturated solution (NH ₄ ) ₂ SO ₄ . The obtained phycoerythrin has a ratio of OD ₅₆₅ / OD ₂₈₀ in the range of 1.4-1.6 and can be used for the production of pigments applicable in the food and cosmetic industries.

4. The crude precipitated phycoerythrin is further purified by gel chromatography. For example, after a single purification by Sephadex chromatography G200, the ratio of OD ₅₆₅ / OD _{280 of the} obtained phycoerythrin can reach 5.1-5.2. The purity of phycoerythrin, measured by electrophoresis in the presence of sodium dodecyl sulfate, is about 99%. This is an indication that phycoerythrin obtained by the method of the invention can be used as a reagent for immunoassay.

Given that phycoerythrin with an OD ratio of ₅₆₅ / OD ₂₈₀ in the range of 5.1-5.2 is obtained as a result of complex purification processes carried out twice in stage 4, the cost and duration of production increase. In this regard, it is necessary to create a method using filaments of other plants, in which phycoerythrin with high optical density can be obtained directly from the first red transparent pigment solution, which is obtained in stage 3.

Summary of the invention

In view of the prior art described above, an object of the present invention is to provide a method for producing phycoerythrin with high optical density, which does not have the problems of the above conventional method for producing phycoerythrin from Bangia atropurpurea and Porphyra angusta.

Another objective of the present invention is to provide a method for producing phycoerythrin with high optical density, providing a high weight yield of phycoerythrin in terms of unit weight of algae selected from the group consisting of Galaxaura oblongata, Halymenia ceylanica, Helminthocladia australis and Porphyra dentata, in order to reduce production costs.

An additional objective of the present invention is to provide a method for producing phycoerythrin with high optical density, providing phycoerythrin with high optical density from the first red transparent algae pigment solution selected from the group including Galaxaura oblongata, Halymenia ceylanica, Helminthocladia australis and Porphyra dentate, in order to reduce the number stages of the production process.

In view of the above and other arguments that will become apparent from the description, in accordance with the general principles of the present invention, a method for producing phycoerythrin with high optical density (OD) is proposed, during which a gametophyte with mature spores for tetraspores is grown in a nutrient medium, of which get tetraspores, grow tetraspores at a temperature of 15-30 ° C, illumination of 500-6000 lux and a light / dark ratio of more than 10:14 to germinate the threads, collect the grown threads, add the grown yarn into a liquid solution with a pH level in the range of 5-10, by centrifuging a liquid solution at a speed of 6000 rpm for 10 minutes at 4 ° C, a red transparent protein pigment solution containing phycoerythrin is obtained and salted out from a red transparent protein solution the pigment is a phycoerythrin concentrate in a gel form, while the gametophyte is selected from the group of algae, the life cycle of which includes gamogenesis, asexual reproduction and vegetative reproduction.

In accordance with the above task, algae are selected from the group including Galaxaura oblongata, Halymenia ceylanica, Helminthocladia australis and Porphyra dentata.

In accordance with the above task, FIG. 7B illustrates a spectrogram of a chromatographic analysis of phycoerythrin extracted from grown spore strands of red algae Galaxaura oblongata carried out on a wavelength of 565 nm by high-performance liquid chromatography.

In accordance with the above task, FIG. 8B illustrates a spectrogram of the chromatographic analysis of phycoerythrin extracted from the grown spores of Halymenia ceylanica red algae carried out on a wavelength of 565 nm by high performance liquid chromatography.

In accordance with the above task, FIG. 9B illustrates a spectrogram of a chromatographic analysis of phycoerythrin extracted from grown spore strands of Helminthocladia australis red algae carried out on a wavelength of 565 nm by high performance liquid chromatography.

In accordance with the above task, FIG. 10B illustrates a spectrogram of a chromatographic analysis of phycoerythrin extracted from grown spore strands of red algae Porphyra dentata, carried out on a wavelength of 565 nm by high performance liquid chromatography.

In accordance with the task set above, SWM-III medium is used as a nutrient medium.

In accordance with the above task, the SWM-III medium is an inorganic medium SWM-III

In accordance with the task set above, in the process of growing tetraspores with the aim of germinating the threads, the threads are additionally crushed into small segments and, under the same conditions, their larger containers are grown until they grow to the required size, while the supply air is supplied to the container, so that small pieces of threads are in suspension.

In accordance with the above task, cultivation occurs in improved conditions at a temperature of 20 ° C, illumination of 2000 lux and a light / dark ratio of 12:12.

In accordance with the task set above, at the stage of collecting the grown yarns, they additionally collect the grown yarns using a network with a number of cells from 20 to 400, dry the grown yarns and grind them into powder.

In accordance with the above task, the method of drying the grown yarns is selected from the group including the method of vacuum drying or the method of drying with heated air.

In accordance with the above task, the liquid solution consists of water and potassium phosphate.

In accordance with the task set above, at the stage of salting out phycoerythrin in gel form, a 20% solution of (NH ₄ ) ₂ SO _{4 is} additionally added to the red transparent protein pigment and the red transparent pigment protein solution is centrifuged at a speed of 6000 rpm for 10 minutes at a temperature 4 ° C to remove unwanted proteins and obtain a solution of protein pigment with a higher degree of purity.

In accordance with the task stated above, at the stage of salting out phycoerythrin in gel form, a 60-65% (NH ₄ ) ₂ SO ₄ solution of a higher purity is additionally added to a red transparent protein solution of a pigment, and a red transparent protein solution of a pigment with a higher degree of purity is centrifuged at a speed of 6000 rpm for 10 minutes at a temperature of 4 ° C, to obtain a phycoerythrin concentrate in gel form.

In accordance with the task stated above, at the stage of salting out phycoerythrin in gel form, the phycoerythrin concentrate in gel form is additionally subjected to dialysis and purified by phycoerythrin by gel chromatography.

In accordance with the above task, gel chromatography is a Sephadex gel chromatography G200.

In accordance with the task set above, at the stage of salting out phycoerythrin in gel form, the phycoerythrin concentrate in gel form is further purified by ultrafiltration.

Brief Description of the Drawings

The described features and many of the accompanying advantages of the present invention can be more easily appreciated by reading the following detailed description with reference to the attached drawings, in which:

figure 1 shows a spectrogram of a chromatographic analysis of the optical density and fluorescence of phycoerythrin, measured by high-performance liquid chromatography,

figure 2 shows the life cycle of Bangia atropurpurea,

figure 3 shows the life cycle of Porphyra angusta,

figure 4 shows the life cycle of Nemalion;

on figa-5C shows a spectrogram of a chromatographic analysis of phycoerythrin extracted from Bangia atropurpurea, carried out by liquid chromatography high resolution at 280 nm, 565 nm and 615 nm,

on figa-6C shows a spectrogram of a chromatographic analysis of phycoerythrin extracted from Porphyra angusta, carried out by high performance liquid chromatography at 280 nm, 565 nm and 615 nm,

7A-7C show a spectrogram of a chromatographic analysis of phycoerythrin extracted from Galaxaura oblongata by high-performance liquid chromatography at 280 nm, 565 nm and 615 nm,

on figa-8C shows a spectrogram of a chromatographic analysis of phycoerythrin extracted from Halymenia ceylanica, carried out by high-performance liquid chromatography at 280 nm, 565 nm and 615 nm,

on figa-9C shows a spectrogram of chromatographic analysis of phycoerythrin extracted from Helminthocladia australis, carried out by high-performance liquid chromatography at 280 nm, 565 nm and 615 nm,

10A-10C show a spectrogram of a chromatographic analysis of phycoerythrin extracted from Porphyra dentata by high-performance liquid chromatography at 280 nm, 565 nm and 615 nm,

11-15 show elements of a device for producing phycoerythrin with high optical density proposed in the present invention.

Description of Preferred Embodiment

Some exemplary embodiments of the present invention are described in more detail below. However, it should be recognized that the present invention can be practiced in a number of other embodiments other than those directly described, and the scope of the present invention is not limited by anything other than the attached patent claims.

Phycoerythrins are water soluble fluorescent protein pigments that are obtained from algae. Owing to their special fluorescent properties, they are widely used for fluorescein labeling of antibodies used as diagnostic tools. Since phycoerythrin has the highest fluorescence intensity among phycobiliproteins, it is used in many tests using fluorescin. Figure 1 shows a spectrogram of a chromatographic analysis of the absorption and emission of phycoerythrin, carried out by high performance liquid chromatography. Chromatography was performed under the following conditions:

High Resolution Liquid Chromatography Column:

HYDROCELL DEAE NP10

Column Size: 50 × 4.6 mm

Buffer A: 10 mMK-PBS pH 6.0

Buffer B: 10 mMk-PBS, 0.5 M NaCl pH 6.0

Gradient: 0% buffer B → 12 min. → 50% buffer B

Detection: 565 nm

Fluid flow rate: 1 ml / min.

High resolution liquid chromatography was carried out using a pump, filters, detectors and a recording device.

As shown in FIG. 2 and FIG. 3, throughout the life cycle of Bangia atropurpurea and Porphyra angusta, gamogenesis and asexual reproduction of successive generations alternately occur. In mature male gametophytes, spermatozoa are produced in spermatangia. Such spermatozoa fall into the water and are carried by the course, appearing in the carpogons of female gametophytes. Carpospores are produced in the mature carpogon, from which filamentous thallic structures (spore plants) are formed with spore-receptacles for shell-shaped spores, the spores of which, standing out from mature spore-receptacles, grow into young upright thallus structures. After some time, young upright thallic structures emit monospores, from which new young upright thallic structures or upright thallic structures (gametophytes) many times grow in proper condition. Thus, alternating gamogenesis and asexual reproduction of successive generations continues.

In other algae, such as Nemalion, shown in figure 4, during the life cycle there is an alternation of gamogenesis, asexual reproduction and vegetative propagation. Carpospores are produced in a mature carpogon, from which spore plants with tetraspores grow, which, standing out from mature spore sites, grow into filamentous thallic structures. Filamentous thallic structures grown from tetraspores, and filamentous thallic structures grown from karpospores have various properties. Thus, filamentous thallic structures grown from Karpospores do not contain gel and can thus be preserved under certain controlled conditions, such as a second medium with a given temperature, degree of illumination, and daily duration of illumination. The present invention provides a method for extracting phycoerythrin with high optical density from filamentous thallic structures of tetraspores. For this, algae are selected from the group including Galaxaura oblongata, Halymenia ceylanica, Helminthocladia australis, Porphyra dentate, or any combination thereof. This is shown in more detail in FIGS. 11-15 by the example of Galaxaura oblongata. The method and device proposed in the invention are described below.

1. Gametophytes with mature spore vessels, selected from algae, whose life cycle includes gamogenesis, asexual propagation and vegetative propagation, such as Galaxaura oblongata, Halymenia ceylanica, Helminlhocladia australis and Porphyra denlala, are collected and washed with sterilized water (not the first ) After short-term air drying, they are placed in the second medium 100 (inorganic medium SWM-III). After several hours, the tetraspores 101 are removed from the algae. Then, the tetraspores 101 are removed from the second medium and placed in a growing chamber, such as a sheet glass chamber 103, in which the temperature, degree of illumination, light / dark ratio and daily illumination duration are 15-30, respectively ° С, 500-6000 lux, over 10:14 and 10-16 hours a day. More preferably, the temperature, the degree of illumination and the light / dark ratio are 20 ° C, 2000 lux and 12-12, respectively. See FIG. 11.

2. After branched strands 201 sprouted from tetraspores, strands 201 (not shown) are collected and transferred to a flask with an inorganic medium SWM-III, such as a container 200, and grown under the above conditions to form colonies. Then filamentous colonies are cut into small segments and transferred to a larger capacity tank to facilitate their further growth. Then filamentous colonies are transferred to a larger room, where they continue to grow. After being transferred to a larger space, their further growth occurs. Then the colony is again crushed for further cultivation to the required amount. It is important that when growing filamentous colonies in tanks, it is necessary to supply fresh air (300 ml / min) to them using a pump 202 so that the colonies are suspended in medium 204. Then the colonies are collected and filtered through a network 203 with a number of cells from 100 to 400. The culture medium is recovered and reused. See FIG. 12.

3. Then the filtered filaments 300 are collected by flotation and dried quickly in a dryer, such as a unit 301 for vacuum drying or drying with heated air and pulverized in a mill 302. See Fig. 13. A powder, for example 2.4 g of powder, is added to a solution with a pH in the range of 5-10, such as containing 70 ml and 10 mM potassium phosphate 304 or any liquid, and mixed thoroughly with a stirrer 303. After centrifugation in a centrifuge 305 at a speed of 6000 rpm / min for 10 minutes at a temperature of 4 ° C receive a red transparent pigment solution. See FIG. The crude phycoerythrin is then obtained by adding a 20% -30% saturated solution of (NH ₄ ) ₂ SO ₄ , 307 to remove excess protein precipitate 308 using a precipitant (not shown), and a 60% -65% saturated solution (NH ₄ ) ₂ SO ₄ 309 to obtain crude phycoerythrin with high optical density, suitable for producing pigments 310, which are applicable in the food and cosmetic industries. By a test method, it was found that the ratio of OD ₅₆₅ / OD _{280 of} pigment 310 is 2.66. See FIG. 15. The above-mentioned red transparent pigment solution can also be obtained by directly grinding wet filaments and adding potassium phosphate to them. The pigment can also be obtained by ultrafiltration.

4. Precipitated crude phycoerythrin can be further purified by gel chromatography or ultrafiltration. For example, after a single purification by Sephadex chromatography G200, the ratio of OD ₅₆₅ / OD _{280 of} the resulting phycoerythrin is 4.5. After a multiple purification process, the ratio of OD ₅₆₅ / OD ₂₈₀ of phycoerythrin reaches 5.3. The purity of phycoerythrin is about 99% according to the results of electrophoresis in the presence of sodium dodecyl sulfate. This is an indication that phycoerythrin obtained by the method of the invention can be used as a reagent for immunoassay.

FIGS. 5A-6C show a high resolution liquid chromatography chromatography spectrogram at 280 nm, 565 nm and 615 nm to which phycoerythrin extracted from Bangia atropurpurea and Porphyra angusta was subjected. Figures 7A-10C show a high-resolution liquid chromatography spectrogram at 280 nm, 565 nm and 615 nm that was subjected to phycoerythrin extracted from Galaxaura oblongata, Halymenia ceylanica, Helminthocladia australis and Porphyra dentata. Since the spectrograms of the chromatographic analysis of phycoerythrin differ depending on the type of phycoerythrin extracted from the algae, they make it easy to determine the source of phycoerythrin.

Table 2 shows two values of optical density for six species of algae, which include Bangia atropurpurea, Porphyra angusta, Galaxaura oblongata, Halymenia ceylanica, Helminthocladia australis and Porphyra dentata. The first line of table 2 shows the amount (in milligrams) of phycoerythrin obtained in terms of the same amount (in grams) of algae. The second and third lines show the ratios OD ₅₆₅ / OD ₂₈₀ and OD ₆₁₅ / OD ₅₆₅ for the algae used in the present invention and the algae Bangia atropurpurea and Porphyra angusta. The higher the first ratio, the greater the degree of purity of the obtained phycoerythrin. The phycoerythrin obtained in this way is notable for a sufficiently high degree of purity for the production of food and cosmetic pigments. It is also applicable to immunotherapy for the prevention of diseases. The second ratio is the ratio of the purity of phycocyanin and phycoerythrin. The lower the second ratio, the lower the purity of the obtained phycocyanin and, therefore, the higher the quality of the obtained phycoerythrin. It follows that from algae threads used in the present invention, such as Galaxaura oblongata, Halymenia ceylanica, Helminthocladia australis and Porphyra dentata, phycoerythrin with a higher optical density than is known from the prior art can be obtained.

table 2 Seaweed Wa Pa Go Not On Pd RPE (mg) / algae (g) 53.5 38.89 57.74 46.59 48.1 44.98 OP ₅₆₅ / OP ₂₈₀ 1.40 1,54 2.66 1.44 2,34 1.96 OD ₆₁₅ / OD ₅₆₅ 0.19 0.53 0.14 0.10 0.15 0.21

RPE is a phycoerythrin protein with a maximum ability to absorb ultraviolet rays at a wavelength of 566 nm and fluorescence at a wavelength of 575 nm.

Ba - Bangia atropurpurea

Pa - Porphyra angusta

Go - Galaxaura oblongata

He - Halymenia ceylanica

Ha - Helminthocladia australis

Pd - Porphyra dentata

Although a specific embodiment of the invention has been illustrated and described above, it will be apparent to those skilled in the art that various changes may be made thereto without departing from the scope of the invention, which is limited solely by the attached claims.

Claims (25)

1. A method of producing phycoerythrin with high optical density (OD) from filamentous tetraspores of certain algae, the life cycle of which includes gamogenesis, asexual reproduction and vegetative propagation, in order to obtain phycoerythrin with high optical density (OD), in which the algae is selected from the group including Galaxaura oblongata, Halymenia ceylanica, Helminthocladia australis, Porphyra dentate, and combinations thereof. 2. The method according to claim 1, in which the filamentous tetraspores are filaments of Karpospores. 3. The method according to claim 2, in which phycoerythrin extracted from filamentous carpospores Galaxaura oblongala, Halymenia ceylanica, Helminthocladia australis or Porphyra dentata, is subjected to chromatographic analysis by high performance liquid chromatography at 565 nm and the spectrograms shown in Fig.7B, 8B, 9B and 10B, respectively. 4. The method according to claim 3, during the implementation of which to obtain phycoerythrin with high optical density in a nutrient medium, gametophyte is grown with mature spore vessels, from which tetraspores are obtained, tetraspores are grown at a temperature of 15-30 ° C, illumination of 500-6000 lux and the ratio light / dark above 10:14, in order to germinate the filaments, the grown filaments are collected, the grown filaments are added to the liquid solution with a pH level in the range of 5-10, by centrifuging the liquid solution, a red, transparent protein pigment solution containing phycoerythrin is obtained, and the phycoerythrin concentrate in gel form is salted out from the red transparent protein pigment solution. 5. The method according to claim 1, in which the medium is an inorganic medium SWM-III. 6. The method according to claim 4, in which the grown filaments are collected by flotation method at a temperature, degree of illumination and light / dark ratio, respectively, 20 ° C, 2000 lux and 12:12. 7. The method according to claim 1, in which at the stage of collecting and growing yarns additionally the grown filaments are collected by a net, the grown filaments are dried and the grown filaments are crushed into powder. 8. The method according to claim 7, in which the network has from 20 to 400 cells. 9. The method according to claim 7, in which the grown filaments are dried by a method selected from the group including drying under vacuum or drying with heated air. 10. The method according to claim 4, in which centrifugation is carried out for 10 minutes at a speed of 6000 rpm at a temperature of 4 ° C. 11. The method according to claim 4, in which the liquid solution consists of water and potassium phosphate. 12. The method according to claim 4, in which at the stage of salting out phycoerythrin in gel form, an additional 20% solution of (NH ₄ ) ₂ SO ₄ and red pigment protein solution are additionally added and centrifuged a red transparent protein pigment solution at a speed of 6000 rpm for 10 min at 4 ° C to remove unwanted proteins and obtain a solution of protein pigment with a higher degree of purity. 13. The method according to claim 4, in which at the stage of salting out phycoerythrin in gel form, an additional 60-65% (NH ₄ ) ₂ SO ₄ solution of pigment with a higher degree of purity is further added to the red transparent protein solution and the red transparent protein solution is centrifuged pigment with a higher degree of purity at a speed of 6000 rpm for 10 min at a temperature of 4 ° C to obtain a phycoerythrin concentrate in gel form. 14. The method according to claim 4, in which at the stage of salting out phycoerythrin in gel form, an additional concentrate of phycoerythrin in gel form is purified by ultrafiltration. 15. The method according to claim 4, in which at the stage of salting out phycoerythrin in gel form, the phycoerythrin concentrate in gel form is further subjected to dialysis and purified phycoerythrin by gel chromatography. 16. The method according to clause 15, in which gel chromatography is a Sephadex gel chromatography G200. 17. A device for using filamentous tetraspores of certain algae, the life cycle of which includes gamogenesis, asexual propagation and vegetative propagation, in order to obtain phycoerythrin with high optical density (OD), consisting of the first gametophyte growth medium with mature spore carriers for tetraspores, from which tetraspores located near the first medium of the second medium for growing tetraspores at a temperature of 15-30 ° C, illumination of 500-6000 lux and a light / dark ratio of more than 10:14 to germinate neither yarn located in the second medium of the grown yarn collection means located near the collected means of grinding powder of the grown yarns in the collection means located near the grinding means of the mixing means of the grown milled yarn and grown yarns to obtain a red transparent solution of protein pigment located near the mixing means precipitation agents for the production of phycoerythrin by salting out of a red transparent solution of protein pigment, while hydrogen if selected from the group of algae, including Galaxaura oblongata, Halymenia ceylanica, Helminthocladia australis, Porphyra dentate, and combinations thereof. 18. The device according to 17, in which phycoerythrin extracted from filamentous carpospores Galaxaura oblongata, Halymenia ceylanica, Helminthocladia australis or Porphyra dentata, is subjected to chromatographic analysis by high performance liquid chromatography at 565 nm and the spectrograms shown in Fig.7B, 8B, 9B and 10B, respectively. 19. The device according to 17, in which the medium is an inorganic medium SWM-III. 20. The device according to 17, in which the grown threads are collected by flotation method. 21. The device according to 17, in which the conditions of temperature, degree of illumination and the light / dark ratio are, respectively, 20 ° C, 2000 lux and 12:12. 22. The device according to 17, in which the collection tool is a network with a number of cells from 20 to 400. 23. The device according to 17, in which the drying means is a vacuum device or a device for drying with heated air. 24. The device according to 17, in which the deposition agent operates on the principle of salting out, during which a 60-65% solution of (NH ₄ ) ₂ SO _{4 is} added to the red transparent protein pigment solution with a higher degree of purity. 25. The device according to 17, in which the salting out is carried out by ultrafiltration or gel chromatography, which is a G200 Sephadex gel chromatography.

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