JP3141083U - A device that produces phycoerythrin with a high absorption coefficient - Google Patents

Abstract

The present invention relates to an apparatus capable of obtaining phycoerythrin having a high comparative value of extinction coefficient (OD) using specific algae.
[Solution] Among the specific algae, there are Galaxaura oblongata, Ionohana celanica, Helminthocladia australis, and a combination of dwarf and entarara. Using the three generational changes of sexual reproduction, asexual reproduction, and vegetative reproduction in the life history of specific algae, the light, temperature, and nutritional conditions are controlled under the characteristic that various gums are not contained in the follicular filamentous stage. Control and prolong the filament stage, and also extract phycoerythrin having a high extinction coefficient comparison value using it as a material.
[Selection] Figure 1

Description

  The present invention relates to an apparatus using a specific algae, and more particularly to an apparatus capable of obtaining phycoerythrin having a high extinction coefficient (OD) comparison value using the specific algae.

  Regardless of whether it is phycoerythrin, phycocyanin or other natural protein dyes, it usually has practical safety, heat resistance, acid / alkalinity, etc. It can be applied to cosmetics, and at the same time, it can also be applied to clinical diagnosis and cell biochemistry research as a fluorescent dye for antibody markers in immunology. Currently developed and applied in the market are phycocyanin and phycoerythrin, and many of the raw materials for producing phycocyanin are cyanobacteria that are bred and cultured in large quantities, such as Spirulina. And Microcystis, and many of the culture and production methods have been patented. Because phycoerythrin lacks raw materials or the raw materials are disadvantageous for the production of pigments, the production amount is small and expensive. Worldwide, there is a demand for tens of thousands of pounds of edible red pigment every year, and there is a great demand for red pigments that are natural, safe and stable, and also emit special fluorescence, and the price drops after mass production. , Its application range and market supply are expected to expand.

  At present, most of the algae-derived red chromoproteins have been isolated from the large red leaves of red algae, such as Porphyra, Ceramium, etc., and some can greatly control the culture. Extracted from Porphyridium.

  Currently, a large amount of red algae resources exist in the wild, and the cultivated genus Amanori is used as a raw material for phycoerythrin. It is not easy to extract pigments, especially the dried algae raw material, and the wild algae raw material varies seasonally in terms of both quality and quantity. Is not easy.

  Currently, it is extremely difficult to collect cells even when culturing porphyridium, and the collection of unicellular algae is an operation that consumes energy and time consistently, which has a huge impact on production costs. Soluble polysaccharides secreted by cells during culture not only interfere with cell collection, but also affect pigment extraction.

  In order to solve the above-mentioned problem, US Pat. No. 5,358,858 discloses a method for producing phycoerythrin from filamentous bovine kenpur (Bangia atropurpurea) and Kosjinori (Porphyra angusta). Under the characteristic that various gums are not contained in the filamentous spore body stage in the life history of Japan, using the controlled light, temperature and nutrient conditions, the filamentous body stage is extended and one step further Using it as a material, phycoerythrin is extracted. The procedure is as follows.

  1. In the cultivation and reproduction of filamentous bodies, the SWM-III (Table 1) was improved after collecting game mates of bovine kenori or kosujinori that were matured in the field, washing the algal bodies cleanly with sterilized sea water and a brush, and slightly drying them in the shade. Into the culture dish containing A cover glass is laid on the bottom of the culture dish. After the spores are released and attached to the cover glass, they are provided with an illuminance of 1000 lux to 4000 lux and a light irradiation of 10 to 16 hours per day at room temperature. Germinate and grow in a culture box, and finally grow until a filamentous spore that branches into multiple branches is exhibited.

  Table 1 SWM-III

（ｐＨ値を調節する際には、先ず濃ＨＣｌによりｐＨ≦７に調節し、さらに適当な濃度のＮａＯＨによりｐＨ値を７．５に調節する。このようにすれば消毒時に白色の沈殿が発生することを回避することができる）
なお、＊１（ＰＩ−ｍｅｔａｌ）及び＊２（Ｓ−３ Ｖｉｔａｍｉｎｓ）の組成は、以下の通り。

(When adjusting the pH value, first adjust the pH to ≦ 7 with concentrated HCl, and then adjust the pH value to 7.5 with NaOH at an appropriate concentration. This will produce a white precipitate during disinfection. Can be avoided)
In addition, the composition of * 1 (PI-metal) and * 2 (S-3 Vitamins) is as follows.

（Ｓｔｏｃｋ溶液はすべて褐色ビン内に装入して、冷蔵保存する）
２．糸状体を掻き落とした後、培養液の入った三角フラスコに移し、上記の培養条件下で培養し、糸状体の凝集ブロックを育成する。糸状体の凝集ブロックを破砕し、更に大きく成長させるためのメスフラスコに移して引続き更に多くの凝集ブロックとなるように発育させ、併せて順次培養体積を増大させ、より大きな体積の光線照射培養槽内において、空気を流入させて（１分間当り３００ｍｌの清浄な空気）、大量に繁殖させて培養する。それを１００〜４００メッシュの網目状の布により濾過する。濾過された培養液は、回収し再利用が可能であり、後続の培養に対しては何の妨げにもならない。

(All stock solutions are placed in a brown bottle and stored refrigerated)
2. After the filamentous body is scraped off, it is transferred to an Erlenmeyer flask containing a culture solution, and cultured under the above-described culture conditions to grow a filamentous aggregate block. Crush the agglomerate block, transfer it to a volumetric flask for further growth, continue to grow to become more agglomerate blocks, and simultaneously increase the culture volume, light irradiation culture tank of larger volume Inside, air is introduced (300 ml of clean air per minute), and a large amount is propagated and cultured. It is filtered through a 100-400 mesh mesh cloth. The filtered culture medium can be recovered and reused and does not interfere with subsequent culture.

  3. After collecting the algal bodies, the filamentous structure is quickly dried with vacuum or hot air, polished to a powder form, stirred thoroughly with water or a phosphate solution, extracted, centrifuged, and clarified. A deep red algal protein solution is obtained. When concentrating and purifying the algal pigment, some impurity proteins are removed and precipitated with a 20% ammonium sulfate solution, and further, the chromoprotein is precipitated with a 60% to 65% ammonium sulfate solution in the solution, which results in OD565. /OD280=1.4 to 1.6, a crude red pigment that can be used for foods and cosmetics.

  4). The precipitated protein is further purified by gel filtration. At that time, Sephadex G200 is used to separate the resin. In the first stage, a phycoerythrin product having an OD ratio of 3.3 to 3.7 is obtained, and phycoerythrin having an OD ratio of 5.1 to 5.2 is obtained by repeating the same process. Further electrophoresis of the product by electrophoresis (SDS) results in a purity of about 99%, which can be used as an immunoassay reagent.

  In the above method, it is collected because the volume is small in the complicated process of heating and gum separation, which was essential in the conventional pigment extraction method of Amanori and Amica, and the extraction method from single cell plants such as porphyridium. However, the problem that the polysaccharide that it secretes affects the extraction of the pigment is avoided, but the deep red algal protein solution formed directly from bovine or Kosuginori filaments has its OD565 / OD280. Is only 1.4 to 1.6, it is necessary to obtain phycoerythrin with a high OD value by concentration and purification of algal pigment. Therefore, it is necessary to discover other algal species having filaments in which the first formed deep red algal protein solution has a high OD value.

  In the background of the above idea, the crimson algae protein solution directly formed using the known bovine or Kosuginori filaments has a very low extinction coefficient comparison value. The apparatus which can obtain the phycoerythrin provided with the high extinction coefficient comparison value using specific algae of this is disclosed, and generation | occurrence | production of the said situation is avoided. The specific algae are Galaxaura oblongata, Isomenopha celanica, Helminthocladica australis, and Porphyra enthusiast.

  One object of the present invention is to increase the weight comparison value of the phycoerythrin content per unit weight and reduce the unit cost by an apparatus capable of obtaining phycoerythrin using the specific algae. Is to achieve.

  Another object of the present invention is to extract phycoerythrin having a high extinction coefficient comparison value using an apparatus capable of obtaining phycoerythrin using the specific algae, thereby reducing the phycoerythrin purification procedure. To achieve the purpose.

  Based on the above objectives, the present invention has a high extinction coefficient comparison value by using algal species carpospore filaments with three generational alternations in life history, sexual reproduction, asexual reproduction and vegetative reproduction An apparatus for producing phycoerythrin is utilized. It has a culture solution for culturing the specific algae containing mature gametophytes of gamets, a first culture tank for obtaining fruit spores, close to the first culture tank, A second culture tank having an illuminance of 6000 lux, having a light / dark ratio of 10:14 or more and culturing the fruit spores and growing them to obtain a filamentous body; A collector that can be installed in a two-cultivation tank and that is close to the collector and collects the filament that has been removed from the collector and installed in the collector. A polishing machine for the production of a dark chromoprotein solution by introducing an acid / alkaline buffer solution and the filamentous material after polishing, and a proximity of the agitation machine From the dark chromoprotein solution. It is equipped with a precipitator for precipitating the phosphorus.

  According to the present invention, chromatograms at 565 nm obtained by high-performance liquid chromatography of phycoerythrin extracted from the spore filamentous bodies of Ragulus oblongata, Ionohana seiranica, Benimozuku and Oniamanori are shown in FIGS. 7B and 8B, respectively. As shown in FIGS. 9B and 10B.

  According to the present invention, the culture solution is a SWM-III culture solution.

  According to the present invention, the SWM-III culture solution is a SWM-III culture solution from which organic substances have been removed.

  According to the present invention, the method of culturing the spore filamentous body is a swirling suspension culture method.

  According to the present invention, the culture environment is 20 ° C., illuminance of 2000 lux, and the light / dark ratio is 12:12.

  According to the present invention, the collector collects the filaments using a mesh cloth.

  According to the present invention, the mesh cloth is a mesh cloth of 20 to 400 mesh.

  According to the present invention, the polishing machine is also equipped with a dryer.

  According to the present invention, the dryer is a vacuum dryer.

  According to the present invention, the dryer is a hot air dryer.

  According to the present invention, the precipitator precipitates phycoerythrin by salting out.

  According to the present invention, 60 to 65% ammonium sulfate solution is added when phycoerythrin is precipitated by the salting-out method.

  According to the present invention, a purifier is further provided, and the phycoerythrin is purified by gel filtration.

  According to the present invention, the gel filtration method is a gel filtration method using Sephadex G200.

  According to the present invention, a purifier is further provided, and the phycoerythrin is purified by ultrafiltration.

  In the present invention, phycoerythrin having a high extinction coefficient comparison value can be obtained by using another specific algae.

  The preferred embodiment of the present invention is described in detail below. However, since the present invention can be widely applied to other embodiments in addition to the detailed description, the present invention is not limited thereto, and it is based on the scope of the utility model registration application described later.

  Algae protein is a kind of water-soluble fluorescent protein pigment derived from the body of algae. Because it has special fluorescent properties, it is now widely applied mainly on fluorescent reagents in flow cytometers. Among various algal proteins, phycoerythrin is one of them and one of the highest emission intensity among natural products. For this reason, such fluorescent dyes are used in many fluorescence inspection methods, and what is shown in FIG. 1 is a chromatogram of the UV absorption and fluorescence emission layer of phycoerythrin obtained by high performance liquid chromatography (HPLC). . The chromatographic conditions are as follows.

HPLC column: HYDROCELL DEAE NP10
Column size: 50x4.6mm
Buffer A: 10 mM K-PBS pH 6.0
Buffer B: 10 mM K-PBS, 0.5 M NaCl pH 6.0
Gradient: 0% Buffer B → 12min → 50% Buffer B
Detection: 565nm
Flow rate: 1ml / min
A high-speed liquid chromatograph is mainly composed of a high-precision high-pressure pump, a separation tube, a detector and a recorder.

  According to Ushikenori's life history (as shown in Fig. 2) and Koszinori's life history (as shown in Fig. 3), the life history of the two types of algae is generally sexual reproduction and asexual reproduction. In the sexual reproduction, it is a gametophyte forming a fruit organ (female gametophyte) and a sperm (male gametophyte), and after the female gametophyte is fertilized, it develops to form a follicular sac. After the inner fruit spores mature, they are released and become filamentous spores. In asexual reproduction, germination directly from a single spore forms a small alga body, and after a period of time, a single spore is formed again, and a single spore germinates into a small algal body, which circulates under appropriate environmental conditions. Below, single spores formed by small alga bodies germinate into large algal bodies. At the same time, small algae are converted into large algae, and the sexual reproduction generation proceeds. The known technique utilizes the characteristic of not containing various gums at the filamentous spore body stage, and under the condition of controlling the light, temperature, and nutrition, the filamentous body stage is extended and further advanced. As phycoerythrin is extracted.

  In the life history of algae, in addition to two generational changes, sexual reproduction and asexual reproduction, some algae also have many vegetative breeding generations. That is, a spore that forms a tetraspore (ie, a tetraspore), in the process, after the fertilizer is fertilized, the spore (carpospophyte), the spore (carpospore), the tetrasporophyte A tetraspore sac is formed, and the last is a tetraspore, among which the fruit spores and tetraspores germinate to form filaments. However, the properties of the two types of filaments are different, as shown in the life history of Nemion (FIG. 4). Similarly, in the present invention, a specific algae of the vegetative breeding generation is utilized under the condition that light, temperature, and nutrition are controlled by utilizing the characteristic that it does not contain various gums at the stage of the follicular filamentous spore body. While extending the body stage, the phycoerythrin with a high OD value is extracted using it as a material. Among them, the specific algae of the present invention include Galaxaura oblongata, Ionohana seilanika, Helminthocdia australis, and the combination of the enthu It is as follows.

  1. In the cultivation and reproduction of the filamentous body, SWM- containing no organic matter after washing the algae with a sterilized seawater and a brush to clean the gametophyte containing the mature spore of the genus Oblongata, Place in culture dish containing III (as in Table 1). A cover glass is laid on the bottom of the culture dish. After the fruit spores are released and attached to the cover glass, the illuminance of 500 lux to 6000 lux and the light / dark ratio of 10:14 or more per day at 15 to 30 ° C. Germinate and grow in a culture box equipped with light irradiation for 10 hours or longer, and finally grow until a filamentous body branched into multiple branches is exhibited.

  2. After the filamentous body is scraped off, it is transferred to a large culture tank containing a culture solution and cultured under the above-mentioned growth environment conditions to grow a gentle and small aggregate block of the filamentous body. At the same time, the culture volume is sequentially increased, and in a larger volume of light irradiation culture tank, air is introduced by a fan to float small aggregated blocks in the culture solution (referred to as swirling suspension culture) and propagate in large quantities. Incubate. It is filtered through a 20-400 mesh mesh cloth. The filtered culture fluid can be recovered and reused and does not interfere with subsequent culture.

  3. After harvesting the algal bodies, the filamentous structure is quickly dried with a vacuum or warm air machine, and further polished with an automatic polishing machine, and then 2.4 g of powder is taken out and 70 ml, 10 mM potassium phosphate solution Alternatively, add other acid / alkaline buffer, stir well with a stirrer to maintain the pH value of the solution between 5 and 10, and further centrifuge at 6000 rpm, 10 min, 4 ° C. This will give a clear and deep red algal protein solution. When concentrating and purifying the algal pigment, some impurity proteins are removed and precipitated with a 20% ammonium sulfate solution, and further, the chromoprotein is precipitated with a 60% to 65% ammonium sulfate solution in the solution, which results in OD565. /OD280=2.66 is a crude red pigment that can be used for foods and cosmetics. The clear crimson algal protein solution can also be obtained by crushing directly collected wet algal bodies, adding a potassium phosphate solution, and then separating by a centrifuge to concentrate and purify algal pigments. Can also be carried out by ultrafiltration.

  4). The precipitated protein is further purified by gel filtration. In that case, resin separation is performed using Sephadex G200 having a length of 120 cm. In the first stage, a phycoerythrin product having an OD ratio of 4.5 or more is obtained. By repeating the same process, a phycoerythrin having an OD ratio of 5.3 or more is obtained. When electrophoresis is performed by SDS), the result is a purity of about 99%, which can be used as an immunological test reagent.

  Shown in FIGS. 5A-C are chromatograms at 280 nm, 565 nm, and 615 nm obtained by high performance liquid chromatography (HPLC) of pure phycoerythrin extracted from bovine henori. 6A to C are chromatograms at 280 nm, 565 nm, and 615 nm obtained by high performance liquid chromatography of pure phycoerythrin extracted from Kosgenori. In fact, for phycoerythrin extracted from different algal species, there are also differences in the chromatograms obtained by high performance liquid chromatography, similar to human fingerprints. Therefore, based on this, it is possible to identify from which algal species the phycoerythrin is generated. Next, shown in FIGS. 7A to C are chromatograms at 280 nm, 565 nm, and 615 nm obtained by high performance liquid chromatography of pure phycoerythrin extracted from the genus Oblongata. Similarly, FIG. 8A-C, FIG. 9A-C and FIG. 10A-C are obtained by high performance liquid chromatography of pure phycoerythrin extracted from Isonohana seiranica, Benimozuku and Oniamanori respectively in the present invention. The obtained chromatograms at 280 nm, 565 nm, and 615 nm.

  Table 2 is a comparative table of phycoerythrin extracted from bovine kenori, kosuginori and specific algae of the present invention. The first column is a comparative value of phycoerythrin weight (mg) contained per unit alga weight (g), and shows phycoerythrin that can be produced from the same algal weight. According to it, for the specific algae proposed in the present invention, the comparative values of the phycoerythrin content are all the same as when using the well-known bovine kenori, Kosujinori, and among them, The comparison value of is the highest. The second and third columns are comparison values of extinction coefficients of phycoerythrin extracted from algal species (OD565 / OD280) and (OD615 / OD565), and the former shows comparison values of phycoerythrin and the purity of other proteins. The latter indicates the purity ratio between phycocyanin and phycoerythrin. (OD565 / OD280) The higher the comparative value, the higher the purity of phycoerythrin, the higher the added value, the higher the utility value as a food or cosmetic pigment, and also for the subsequent purification process This is advantageous and can be applied as a reagent for immunological tests in medical terms. Phycocyanin absorbs the fluorescence emitted by phycoerythrin, and when used as a food or cosmetic pigment, the fluorescent coloration effect is poor, but the lower the (OD615 / OD565), the lower the phycocyanin content. Therefore, it is more appropriate in terms of the fluorescence coloring effect. Similarly, for the specific algae proposed in the present invention, it is clearly shown that all the phycoerythrin extracted from them have comparatively high extinction coefficient comparison values. Therefore, the phycoerythrin extracted using the filaments of the specific algae in the present invention has a high phycoerythrin content and a high extinction coefficient comparison value when compared with the known bovine kenori and kosginori filaments. It has the advantage of having.

  Table 2 Analysis of pigment components in algae powder of each algae species

ＲＰＥ：フィコエリスリンの一種であり、親水性を備えているとともに水と安定した水溶液を形成する。その最大ＵＶ吸収波長と蛍光発散波長はそれぞれ５６６ｎｍと５７５ｎｍである。

RPE: A type of phycoerythrin that is hydrophilic and forms a stable aqueous solution with water. Its maximum UV absorption wavelength and fluorescence emission wavelength are 566 nm and 575 nm, respectively.

Ba: Bengali apurpurea
Pa: Porphyra angusta
Go: Galaxaura oblongata
Hc: Isonyohana celanica
Ha: Helminthocladia australis
Pd: Porphyra dentata
As described above, the desired effect is achieved in the embodiment of the present invention, and the concrete structure disclosed therein is not recognized in various similar products and is not published before filing. It shall conform to the provisions and requirements of the Utility Model Law and apply for registration of the utility model based on the law. Thank you for your review and approval of utility model rights.

  Although the present invention has been described based on one suitable embodiment, the present invention is not limited to this embodiment. Although specific examples have been described as described above, as is apparent, all equivalent changes or modifications made without departing from the spirit of the present invention are registered as utility models of the present invention. It goes without saying that they are included in the scope of the claims. In addition, all similar and approximate changes or modifications made without departing from the spirit of the present invention are also included in the scope of claims for utility model registration of the present invention. At the same time, the scope of the invention should be construed by the broadest definition, thereby including all modifications and similar structures.

It is a chromatogram of UV absorption and fluorescence emission layer of phycoerythrin obtained by high performance liquid chromatography (HPLC). It is the life history of Ushikenori. It is the life history of Kosujinori. It is the life history of sea urchins. AC are chromatograms at 280 nm, 565 nm and 615 nm, respectively, obtained by high performance liquid chromatography of pure phycoerythrin extracted from bovine henori. AC are chromatograms at 280 nm, 565 nm, and 615 nm, respectively, obtained by high performance liquid chromatography of pure phycoerythrin extracted from Kosgenori. A to C are chromatograms at 280 nm, 565 nm and 615 nm, respectively, obtained by high performance liquid chromatography of pure phycoerythrin extracted from the genus Oblongata. AC are chromatograms at 280 nm, 565 nm, and 615 nm, respectively, obtained by high-performance liquid chromatography of pure phycoerythrin extracted from Isonohana seiranica. AC are chromatograms at 280 nm, 565 nm, and 615 nm, respectively, obtained by high-performance liquid chromatography of pure phycoerythrin extracted from Benimozuku. AC are chromatograms at 280 nm, 565 nm, and 615 nm, respectively, obtained by high-performance liquid chromatography of pure phycoerythrin extracted from Onianori.

Claims (9)

  It has a culture solution for culturing a specific algae containing mature gametophytes, and is close to the first culture tank for obtaining the fruit spores, and has an illuminance of 500 lux to 6000 lux. And a second culture tank for culturing the fruit spores and growing the fruit spores to obtain filaments in an environment having a light / dark ratio of 10:14 or more and 15 to 30 ° C., and the second culture A collector that can be installed in a tank and that collects the filaments, and is close to the collector, and the filaments that are taken out of the collector and installed in the collector are polished. A stirrer that is close to the grinder, introduces the acid / alkaline buffer solution and the filament after the grind, and obtains a dark chromoprotein solution, and the stirrer In close proximity to precipitate phycoerythrin from the dark chromoprotein solution And the specific algae are selected from the group consisting of Rhinoceros oblongata, Ionohana seiranica, Benimozuku, Oniamanori and combinations thereof, characterized by sexual reproduction, asexual reproduction and nutrition in life history An apparatus for producing phycoerythrin having a high extinction coefficient comparison value using algal sporophyte filamentous bodies with three generational alternations for breeding.   Chromatograms at 565 nm obtained by high-performance liquid chromatography of phycoerythrin extracted from the spore filamentous bodies of Ragulus oblongata, Isonohana seiranikika, Benimozu and Oniamanori are shown in FIGS. 7B, 8B, 9B and FIG. The alga sp. Filamentous filamentous body having three generational changes of sexual reproduction, asexual reproduction and vegetative reproduction in the life history according to claim 1, characterized in that it is as shown in 10B. An apparatus used to produce phycoerythrin with high extinction coefficient comparison values.   The life culture according to claim 1, wherein the culture solution is a SWM-III culture solution or a SWM-III culture solution from which organic substances are removed. An apparatus for producing phycoerythrin having a high extinction coefficient comparison value using algal sporophyte filamentous bodies having three generational changes of vegetative propagation.   The method for cultivating the follicular spores is a swirling suspension culture method, wherein the life history according to claim 1 comprises three generational alternations of sexual reproduction, asexual reproduction and vegetative reproduction. An apparatus for producing phycoerythrin having a high comparison value of extinction coefficient using the fruit spores of algae species.   3. The culture environment according to claim 1, wherein the culture environment has an illuminance of 20 [deg.] C., 2000 lux, and a light / dark ratio of 12:12. An apparatus for producing phycoerythrin having a high extinction coefficient comparison value by using algal sporophyte filaments with generational changes.   The life collector according to claim 1, wherein the collector collects the filaments using a mesh cloth, and the mesh cloth is a mesh cloth of 20 to 400 mesh. An apparatus for producing phycoerythrin having a high extinction coefficient comparison value by using algal sporophyte filaments having three generational alternations of sexual reproduction, asexual reproduction and vegetative reproduction.   The life polisher according to claim 1, wherein the polisher is also equipped with a dryer, and the dryer can be a vacuum dryer or a hot air dryer. An apparatus for producing phycoerythrin with high extinction coefficient comparison values using algal sporophyte filaments with three generational alternations of reproduction and vegetative reproduction.   The precipitator precipitates phycoerythrin by a salting-out method, and 60% to 65% ammonium sulfate solution is added when the phycoerythrin is precipitated by the salting-out method. An apparatus for producing phycoerythrin having a high extinction coefficient comparison value using the algal sporophyte filamentous body having three generational alternations of sexual reproduction, asexual reproduction and vegetative reproduction in the described life history.   The phycoerythrin is further filtered by an ultrafiltration method or a gel filtration method, and the gel filtration method is a gel filtration method using Sephadex G200. A device that produces phycoerythrin having a high extinction coefficient comparison value using the algal sporophyte filamentous body having three generational changes of sexual reproduction, asexual reproduction, and vegetative reproduction in the life history of the plant.

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