TWI224135B - Method of Helminthocladia australis for producing high OD phycoerythrin - Google Patents

Abstract

The invention relates to a method of Helminthocladia australis for producing high OD phycoerythrin. It has been found that Helminthocladia australis has three generations in the life cycle sexual reproduction, asexual reproduction and vegetative propagation. Then, carpospore filamentous plants thereof do not contain gel and can be maintained under some controlled conditions. The invention takes advantage of these findings to provide a method for producing high OD phycoerythrin from carpospore filamentous phase of Helminthocladia australis.

Description

1224135 Case No. 91133930 V. Description of the invention (1) [Technical field to which the invention belongs] The present invention relates to a method for discovering Helminthocladia aus ▲ a tra 1 is. Special brothers can obtain Method with high absorbance (0 D) ratio of phycoerythrin. [Previous technology] As far as phycoerythrin and phycocyanin have practical safety, they can be used in food and makeup for fluorescent pigments, but should be developed in the market and phycoerythrin should be mass-produced. Microcystis (Microcystis). Phycoerythrin has low yield and expensive demand due to raw materials. A natural and safe substance is necessary. It is expected to have a wide range of applications and market supply. It is not usually qualitative, and antibodies are standard. Most of the phycoerythrin can be patented with patina. The red color of the pigment is large. It should be a stable product in terms of § or other natural protein pigments. It can also be used for immunology. For the diagnosis of g bed and cytobiochemical research, "phycocyanin" is a green algae, such as spirulina (Spirunna), which is the raw material for phycocyanin production. Lack of or unfavorable pigment due to raw materials. After tens of thousands of pounds of edible red, stable, and special fluorescent products are produced worldwide each year, the price will be reduced & more: such as Porphyra, most of the phycoerythrin protein is isolated from the large leafy, celery algae of red algae (Ceramium), etc., a small part: take

Page 6 1224135 _ Case No. 91133930 _ Year Month Day Xiu Tun _ V. Description of the invention (2) Porphyridium which can be controlled and cultivated by the giant salamander. Although there are a lot of wild red algae resources and cultivated laver as raw material for phycoerythrin ', most of the red algae are rich in glial matter (agar gum, carrageenan, red bath gum), which is difficult to cause the pigment to die out, especially dry. The latter algae materials are even more so. In addition, the wild algae materials have seasonal differences in quality and quantity, which is difficult for humans to master. The cultivation of purple ball baths also has significant difficulties in collecting cells. Because the collection of single-cell baths has always been an energy-consuming and time-consuming operation, the production cost is very large and the soluble polysaccharides secreted by algal cells during culture In addition to blocking the collection of cells, it also affects the extraction of pigments. In order to grow and control the angus t dish for daily use, 1. The dried leaves of the purple leaf are put into the incubator at room temperature and the incubator at room temperature solves the above problems. US Pat. No. 5,358,858 proposes a dish (Bangia atropurpurea) and a narrow dish. The method of producing phycoerythrin from the filamentous body of porphyra a, using the characteristics of the filamentous sporophyte stage in the hair history and purple leaf history of purple leaf does not contain various gums, which benefits the light, temperature and nutritional conditions. Next, continue its filamentous phase, and use it as a material to extract phycoerythrin. The steps are as follows: Culture and reproduction of the body, collect mature hair dishes or gametophytes from the wild, and clean the algae with sterilized seawater and brush. The body is clean and slightly overcast into a petri dish containing modified swm-nr (as shown in Table 1). The cover is covered with coverslips. After the spores are released and attached to the coverslip, f is at 1 0 0 0 lux. Illumination of ~ 40 0 0 1 UX and germination and growth under 10 ~ 16 hours of daily light 'finally developed a branched filamentous sporophyte.

II 1 face 1224135 _Case No. 91133930_ Year Month Day Amendment V. Description of Invention (3) Table 1 SWM- Π

NaNO3 8.5 g / 100 ml 2m 1 Na2HP04 0.595 g / 100 ml 2m 1 Na2EDTA 0.5 g / 100 ml 2m 1 0.01625 g / 100 ml 2m 1 FeCl3 (or FeCl3 • 7H20) 0. 0 2885 g / 100 ml 2ml wood PI -meta 1 2m 1 3 Vitamins 2m 1 Soil extract 50ml Tr is (1 0 cc / 1) 5 0 Omg Liver extract 1 Omg Sea Water up to pH = 7.5 11 (When adjusting the pH: first adjust to concentrated HC1 to pH < = 7, and then with appropriate concentration of NaOH to pH7.5, so as to avoid white precipitation during disinfection) * PI -me ta 1 H3 BO3 1 2.3 68 g MnCl2 1.385 g ZnCl2 0.109 g re-distilled water 21 CoCl2 · 6H20 4.479 mg CuCl2 · 2H20 0.0 34 mg

Page 8

1224135 η of case number 91133 V. Description of the invention (4) ** S-3 Vitamins Thiamin HC1 Capantothenate

0 · 0. g g

Nicotinic acid P-am i nobenzo ic acid Biotin Inositol Folic acid Thymine B12 (All St ◦ ck solutions are stored in 0.1 g 10 mg 1 mg 5 g 2 mg 3 mg 1 mg brown water 2 1 Refrigerated storage) 2. After the filaments are scraped off, they are moved to the environment containing the cultured 'bred filamentous group' and moved to larger growth flasks. Therefore, the volume of the culture should be gradually expanded and should be pumped (30 0 per minute).胄 # | 乞 ~ 40 0 mesh filtering of the mesh, without any hindrance to the continued cultivation. The conical flask of the culture solution, the gathering group of the above-mentioned growth bodies. The gathering of broken filaments continues to develop into more gatherings Group, and in a larger volume of light culture tank, clean air) 1 large breeding culture. The culture liquid can be recycled and reused. After the 3. After the algal body is harvested, the filamentous structure can be quickly dried by vacuum or warm air, and then ground into powder, and then fully stirred and extracted with water or phosphate solution. After centrifugation, a clarified deep red algae protein solution can be obtained. Concentration and purification of algal pigment can be removed by 20% ammonium sulfate solution to remove some impurities in the protein sink, and then precipitated in the solution by 60% ~ 65% ammonium sulfate solution. The output of pigment protein is 0D565 / 0D28G = 1 · 4 ~ 1 · 6 is a thick red pigment, which is food-grade and cosmetic. ~ ---- ------ _

Page 9 1224135 __ Case No. 91133930 _ Year Month __ Day _ Amendment V. Description of Invention (5) ^ Pigments used. 4. The precipitated protein is further purified by gel filtration and separated by Sephadex G20 0 resin. The first procedure can obtain a phycoerythrin product with an OD ratio of 3 · 3 ~ 3 · 7, and repeat again. A phycoerythrin having a ratio of 5.1 to 5.2 can be obtained, and the fruit obtained by gel electrophoresis (SDS) electrophoresis shows a purity of about 99%, and can be used as a reagent for immunoassay. Although the above methods can avoid the traditional hand-made laver and celery algae extract = complex hand-to-hand separation of gel value, continuation, or cyanococcus, etc .: It is difficult to affect the extraction set of chroman and its secretion, directly Envy into the μ--ask the question, but the edible laver and laver laver filaments still need to be reed solution, its OD565 / 〇D28. Only h4 ~ 16, so we-; hairpin i; chen shrink and purification to obtain high OD value of phycoerythrin, because of the protein solution has a high OD value; :: body, its first formation of deep red algae [Content of the invention ] A < praise brother back φ

The filamentous body directly opens the 4T portal temporal crest of the deep red algae egg. The method of the present invention is described by Yau 1, a method and a thin photometric ratio. It is known that the use of Hydrilla verticillata and Porphyra sclerotiorum liquid, whose absorptivity ratio is relatively low, can be obtained with high morphology. Wood ,,, and engineering, avoid on

1224135

The weight ratio Bit ::::: The weight of the cyanocyanine-containing compound is another method, which is to use Spirulina algae to obtain eosin. With its extractable, it has the following functions: to swim, and to achieve the purpose of reducing the phycoerythrin purification step. The phylloxanthin is the same as the above. In the present invention, a method for extracting algae seeds is used. The life history has sexual reproduction and asexual reproduction. The nutrition and reproduction are alternated. Obstetric suction = three Shisu. According to the above conception, the algal red of Absalpium chinense has an absorbance ratio. The spore filaments are carpospore filaments. According to the above concept, the algae species is Helmfl: nthocla ^ dia. The spectrogram is shown in Figure VII of Figure B of the patent specification. The chromatographic light is based on the above conception, in which the latitude The phycoerythrin with photometric ratio comprises the following steps: cultivating the garments containing mature fruit sporangia and obtaining the fruit spores σ _ ^ under the control of worms, 5gg1ux ~ 6_1ux , Zhaodu fruit spores were cultivated under the environment, so that they grow into exhalations and the " darkness is more than 10:14

Page 11 1224135 _ Case No. 91133930 _ year and month amended _ V. Description of the invention (7) Acid test buffer, stirred: stir, centrifuge to get a dark pigment protein solution; and remove the dark color by salting out method Impurities in the pigment protein solution and precipitation of pigment proteins. According to the above concept, the culture medium is SWM-III culture medium. According to the above concept, the SWM-II culture medium is a swmm culture medium without organic matter. According to the above concept, a method in which the fruit spore filaments are cultured is a spin-floating culture method. According to the above idea, the culture environment is 20 ° C, and the illumination and light-dark ratio of 20000 lux is 1 2: 12. According to the above concept, the step of collecting the filaments further comprises: collecting the filaments by a mesh collection method; drying the filaments; and grinding the filaments into a powder. According to the above idea, the net collection method uses 20 to 400 mesh nets. According to the above concept, a method in which the filament is dried is a vacuum method. According to the above concept, a condition in which the filament is dried is a warm air method.

Page 12 1224135

The rotation speed is 6000 rpm, according to the above concept, wherein the centrifugation time is 10 m i η and the temperature is 4 ° C. According to the above concept, the pH of the solution is maintained between 5 and 10. Youchong solution is a potassium phosphate solution, which is soluble in money: According to the above conception, in which the salting-out method removes impurities by adding 20% sulfuric acid. According to the above conception, where: The Y dish analysis method / Erdian pigment protein is added with a 60% b 5 / ammonium sulfate solution. According to the above-mentioned concept, which further includes purification and separation of phycoerythrin by ultrafiltration (ul traf i 1 tration). According to the above-mentioned concept, which It further includes purification and isolation of phycoerythrin by gel filtration chromatography. According to the above concept, the gel filtration chromatography is a gel filtration chromatography using Sephadex G2 0. According to the above-mentioned purpose, the present invention finds a method for using Life history has three generations of sexual reproduction, asexual reproduction and vegetative reproduction. Carpospore filaments of algae species alternate to produce phycoerythrin with high absorbance ratio.

Page 13 1224135 _Case No. 91133930_Year_Month_Revision V. The device of the invention description (9) 'contains: a first culture tank with a culture medium for the cultivation of clade algae (Helmint ho cladia australis) containing mature fruit sporangia Gametophyte and get fruit spores; a second culture tank with an illuminance and light-dark ratio of 500iux ~ 600iux is 10: 14 or more and 15 ~ 3 0 ° C in an environment for Cultivate the fruit spores to grow into filaments; a collector to collect the filaments; a grinder to grind the filaments; a stirrer, add an acid-base buffer solution and grind the The filaments are used to obtain a dark pigmented protein solution; a precipitator is used to precipitate phycoerythrin from the dark pigmented protein solution. Chromatographic spectrum of 5 6 5nm extracted from the fruit spore filaments—according to the above idea, in which the bath red pigment from Cladosporium sp As shown in FIG. 7B of the book, according to the above concept, the culture medium is swm_m culture medium. Swm-m broth according to the above concept. Among them, the SWM ~ m culture solution was deorganized. According to the above concept, the floating culture method was used. Among them, 1 the method of feeding the fruit spore filaments is to spin. According to the above concept, the illumination and light-dark ratio is 12:12. . The breeding environment is 20 ° C, 2 0 0 0 lux

^ 4135 5 Description of the invention Case No. 9113 刈 30 (10)

According to the above concept, wherein the mesh cloth is 20 to 40 0 mesh mesh. According to the above concept, wherein the grinder further includes a drying cry dryer according to the above concept, wherein the dryer is vacuum-drying according to the above concept, wherein the dryer is warm Air dryer. According to the above-mentioned concept, wherein the precipitator is Shen Xingping using a salting-out method. According to the above-mentioned concept, wherein the salting-out method precipitates phycoerythrin by adding 60% 6 5% ammonium sulfate solution. According to the above conception, it further includes a purifier, and the phycoerythrin is purified by gel filtration chromatography. According to the above concept, the gel filtration chromatography method is a gel filtration chromatography method using Sephadex G2 0 0. According to the above-mentioned concept, a purifier is further included, and the phycoerythrin is purified by ultrafiltration (u 11 r a f i 11 r a t i ο η).

Page 15 1224135 ___Case No. 91133930_Year Month Chromium π:, V. Description of the Invention (11) [Embodiment] The preferred embodiment of the present invention will be described in detail as follows. However, in addition to the detailed description, the present invention can be widely implemented in other embodiments, and the scope of the present invention is not limited, which is subject to the scope of subsequent patents. Phycobiliprotein is a water-soluble fluorescent protein pigment from algae. Due to its special fluorescent properties, it is currently widely used in fluorescent reagents for flow cytometers. Among the various phycobiliproteins, phycoerythrin is one of them, and it is also the one with the highest fluorescence intensity in natural materials. Therefore, there are many fluorescent detection methods using such pigments. UV absorption and fluorescence emission chromatograms obtained by a chromatograph (HPLC). The chromatographic conditions are as follows: HPLC column: HYDROCELL DEAE NP10

Column size: 50 * 4.6mm

Buffer A: lOmMK-PBS pH 6.0

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

Gradient :. % Buffer B—12min—50% Buffer B

Detection: 5 65nm

Flow rate: lml / min and high performance liquid chromatography is mainly composed of high precision high pressure pump, separation tube, detector and recorder. Observing the life history of the hair algae (as shown in the second picture) and the life history of the laver (as shown in the third picture), we can know the life history of the two algae.

Page 16 1224135

Generally, when the female spouse with a fruiting organ matures, the spores are direct), isotype algae, or monomorphic or transformed into sperm produced by material reproduction through the use of filamentous threads and temperature. Sporozoite is the straight algae shown in the single and germinated into small and small algae. It can also be known that the technique is the light of control and further alternates with two generations of sexual reproduction and asexual reproduction. There are ( Female spermatophores and sperm sacs (male partners) After fertilization, sporangia will develop into sporangia, which will diffuse out and become filamentous sporophytes. Probiotics germinate into small algae (such as the second or third picture to a certain time can produce monospores, the monospores circulate until the appropriate environmental conditions, the seeds can germinate into large algae (such as the first The fronds shown in the second and third pictures). At the same time, the small bathing body 'reentered the sexual reproduction generation. The spore body stage does not contain the characteristics of various gums, and it continues its filamentous shape under nutritional conditions. Extraction of phycoerythrin at the body stage. In addition to the algae's life history alternated between two generations of sexual reproduction and asexual reproduction, the other part of the algae also has one more vegetative reproduction generation, that is, the sporophyte of the owner's tetraspore (ie four Sporophyte), the process of which is to produce carposporephyte, carppore, tetrasporophyte, tetrasporangium, and finally It is tetraspore, in which both fruit spores and tetraspores will germinate into filaments', but the characteristics of these two filaments are different, please refer to the life history of the sea (Nemali〇n) (as shown in the fourth picture ).with The present invention is to use undifferentiated cladospira with vegetative breeding generation, which contains tetraspores and fruit spores. The characteristics of its filamentous stage do not contain various gums, in the controlled light, temperature, nutritional conditions Next, continue its filamentous stage, and further 1224135 I No. 9139393 (1 V. Description of the invention (13) The steps are as follows: Using this as a material, a phycoerythrin with a high 0D value is extracted, and its spore 1 cyst branch curtain contains The mature quarters are slightly overcast and dried, and then put in SWM_m culture medium containing no organic matter. = The inner bottom is covered with glass flakes' to be released by the quarter brussels and fruit spores = attached = after the glass sheet, at 15 ~ 3 (rc , I of 50 0 1 ux ~ 600 〇iux i = 1 dark, the growth of hair teeth in the incubator with more than 10:14 and more than 10 light per day 'finally develop branched filamentous bodies. Body scraping Move down to a large culture tank containing the culture medium, and culture under the conditions of the above long-term brothers, the looseness of the filaments can be reduced: gradually, expand the volume of the culture, and culture in a larger volume of light: Zhongya Xinyu Milking makes the small poly group suspended in the culture medium . Borrowed by 2.0, the net is filtered and removed. Culture: Recycling and reuse, it does not hinder subsequent cultivation. Oral nourishment is available. 3. After the algal body is harvested, due to the filamentous structure It can be dried quickly by vacuum or warm air, and then pulverized by an automatic mill, then take 2.4 g of powder, dry add ^ 70 ml, 10 mM potassium phosphate solution or other acid-base buffer solution Disperse it to maintain the pH of the solution between 5 ~ 10. Stir it thoroughly, and then centrifuge it at 6,000 = m, 10 min, 4 to obtain a clear dark red ^ < protein. Solution. Concentration and purification of algae pigments can be obtained by 20% ammonium sulfate solution = to remove part of the impurity protein precipitation, and then dissolve the pigment protein in the solution by 60% ~ 65% ammonium sulfate, which is OD ^ / OD ^ = 2 · 66 thick red pigment, pigment of food grade and cosmetics. The clarified deep red algae protein can be dissolved in the wet algal body directly harvested and then added with potassium phosphate solution, and then centrifuged overnight....

Page 18 1224135 Case No. 91133930 V. Description of the invention (14) The concentration and purification of the algal pigment can be carried out by ultrafiltration. 4. Purify Shen Dian's protein by gel filtration chromatography (gel flltr = tion), and use Sephadex G200 resin with 120 cm length to separate L. The first procedure can obtain phycoerythrin with an OD ratio of 4.5 or higher. The product can be weighed again to obtain phycoerythrin with an 0D ratio of 5.3 or more. The result obtained by gel electrophoresis (SDS) electrophoresis shows a purity of about 99%, which can be used as a reagent for immunoassay. After analyzing the spectrum with high efficiency, it can be seen by humans that the algae have been extracted from the snail branch 2 8 0 nm &# < ίΓρ / ήΓ. The pure phycoerythrin extracted from Shufa vegetable is 280nm, 565nm, The 615nm chromatogram, younger brothers, A ~ C are the diagrams of the huoshan—oon * < ,,, and galvanic erythrocyanin from the purple leaf liquid chromatography. Actually, the layer effects of 28nm, 565nm, and 615 ° are "== the phycoerythrin extracted from the algae species, and the fingerprints of the finger-im spectrum are also different. Take out the algae: production system. Seventh The a to c diagrams are the chromatographic spectra of the present invention, 565 ·, 6, and the "Table 1 paid by Jinyi is the known head color # Comparison table of taking phycoerythrin * Two leaf purple mining and the worm of the present invention The ratio of the weight of the extracted algae (mg) is the unit of algae weight (g), which contains phycoerythrin and phycoerythrin, and we can clear the ratio of phycoerythrin that can be produced under the same bath weight. Second side: 2 Spirulina of the invention 1 contains a high row of phycoerythrin extracted from vine species 1224135

Photometric ratio (OD56V0D280) and (〇D 阳 / 〇D56M, 1 ratio of protein purity, the latter means algae) blue; two primes and its ratio, (0D56V0D), the higher the ratio is the red / = The higher the purity value, 'the use of pigments in food or cosmetics;' its additional value is made purified, because it can be used in medical immunity: 2: the more favorable the back-end phycocyanin will absorb the fluorescent light emitted by phycoerythrin, the greater the 2 1 ^ 丄 二 Because # σ 纟 | g vs Μ ^ is used as food or makeup. Fluorescent color rendering is not good when using 迚 mouth pigment. Therefore, the less the phycocyanin content, the less visible the color rendering effect m. The Spirulina sp. 1 extract of the present invention has a value. Therefore, using the snail = body =: U of the present invention, the advantages of the ratio of red pigment content and high absorbance in the filamentous body of Porphyra spp. Table 2 Analysis of algae pink pigment of each algae species Algae RPE mg / g Algal flour Absorptivity A5 6 5 / A280 Absorptivity A615 / A565

Ba Pa Ha 53 · 5 38.89 48. 1 1. 40 1.. 54 2. 34 0. 19 0. 53 0. 15 RPE: — a phycoerythrin that is hydrophilic and forms a stable aqueous solution in water '# t AUV absorption wavelength and fluorescence emission wavelength are 566nm and 1224135 銮 No. 91133930

V. Description of the invention (16) 5 75nm 〇

Ba ·· Hair dish (Bangia atropurpurea)

Pa: Porphyra angusta

Ha: Helminthocladia australis In summary, 'the embodiments of the present invention can achieve the expected effect, and the specific structure disclosed is not only not seen in similar products, nor disclosed before the application, Since it has fully complied with the requirements of the Patent Law = requirements, I love to submit an application for an invention patent in accordance with the law. Although the present invention has been described by citing a preferred embodiment, the invention is not limited to the illustrated embodiment. Although specific examples have been cited and described previously, it is obvious that other equivalent changes or modifications made without departing from the spirit disclosed by the present invention should be included in the scope of patent application of the present invention. In addition, all other similar and approximate changes or modifications' without departing from the spirit disclosed by the present invention are also included in the scope of patent application of the present invention. At the same time, the scope of the present invention should be construed in the broadest definition so as to encompass all modifications and similar structures.

Case No. 91133930 Simple illustration of the drawing [Simplified illustration of the drawing] In order to make the technical means used in the present invention easy to understand, here are the features, β ^ characteristics, achieved goals and effects. The explanation is as follows: ^ ^ ^ ^ 疋 Chromatographic spectrum of algae red absorption and fluorescence divergence obtained by ^ performance liquid chromatography (HPLC); the second picture shows the life history of hair dish; the third picture The life history of Porphyra angustifolia is shown; the life history of the sea surface is shown; younger brothers A ~ C picture drawing + 3 w — the liquid layer fights broadly, the steamed bun 1 ^ the pure phycoerythrin meridian taken out High performance map. The chromatographic spectra of 280nm, 565nm, and 615nm obtained by Yiyi show that the pure phycoerythrin extracted from P. annua is highly efficient.

Brother Liu A ~ G can be applied to the liquid phase layer ~~ Sakai City 不 Shipi Mountain Heart Shellfish Ice Bucket $ ,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,. The 280nm, 565nm, 615nm Chromatographic Spectrum Figure 7: A ~ C final liquid phase layer; I: shows the chromatographic spectrum of 280nm, 565nm, 615nm of pure phycoerythrin extracted by Spirulina algae by high-performance 9 instrument

Page 22 1224135 Case No. 91133930 Month, Day Amendment

Page 23

Claims (1)

1224135-^^ 3393Q _Year Month Day Six, application for patent scope: 1 · — A method for producing high-absorbance bath red pigment from 4 ^ stick bath (Helminthocladia australis), its life The history has three generations of sexual reproduction, asexual reproduction and vegetative reproduction, and the filamentous body cultured from its carpospore and tetraspor is used to produce the phycoerythrin with an absorbance ratio back, The method includes the following steps: culturing the gametophyte containing mature fruit sporangia and tetrasporangium gametophyte with a culture medium to obtain fruit spores and tetraspores; and placing the composition of fruit spores and tetraspores at 15 ~ At 30 ° C, culture under an environment where the illuminance and light-dark ratio of 5 0 0 1 ux ~ 6 0 0 0 1 ux is above 〇: 丨 4 or more to grow into filaments; collect the filaments; add acid The buffer solution is tested, stirred, and centrifuged to obtain a dark pigmented protein solution; and the impurities in the dark pigmented protein solution are removed by salting out and the pigmented protein is precipitated. 2. The method for producing high-absorbency phycoerythrin from Cladophyllum as described in item 1 of the patent scope of Shenying ', wherein the culture solution is m culture solution. 3 · As described in item 2 of the patent application, a method for producing high-absorbency phycoerythrin from spirulina is used, wherein the melon nutrient solution is SWM-III culture without organic matter. 1224135 _Case No. 91133930_ Year Month Amendment _ Sixth, the scope of patent application Liquid. 4. The method for producing high-absorbency phycoerythrin from cladosporium as described in item 1 of the scope of the patent application, wherein the method of cultivating the fruit spores and the filaments of the tetraspores is a rotary float culture method. 5. The method for producing high-absorbency phycoerythrin from Cladosporium as described in item 1 of the scope of the patent application, wherein the culture environment is 20 ° C, and the illumination and light-dark ratio of 20000 1 ux is 1 2: 1 2. 6. The method for producing high-absorbency phycoerythrin from Cladosporium as described in item 1 of the scope of the patent application, wherein the step of collecting the filaments further comprises: collecting the filaments by a net collection method; and drying the filaments A body; and grinding the filament into a powder. 7. The method for producing high-absorbency phycoerythrin from cladospira as described in item 6 of the scope of the patent application, wherein the net collection method uses 20 to 400 net mesh. 8. The method for producing high-absorbency phycoerythrin from cladosporium as described in item 6 of the scope of patent application, wherein the method for drying the filaments is a vacuum method. 9. The method for producing high-absorbency phycoerythrin from cladospira as described in item 6 of the scope of the patent application, wherein the condition for drying the filaments is the warm air method. Page 25 1224135 Case No. 91133930 ± _ Ά Amendment VI. Patent application scope 10. The method for producing high-absorbency phycoerythrin from cladosporium as described in item 1 of the patent application park, wherein the condition of centrifugation is the speed of 6000 rPm , Time 10 minutes and temperature 4 ° C. 11. The method for producing high-absorbency phycoerythrin from Cladosporium as described in Item 1 of the patented patent garden, wherein the acid buffer solution is a potassium phosphate solution 'to maintain the pH of the solution between 5 and 10. 12. The method for producing high-absorbency phycoerythrin from cladospira as described in item 1 of the scope of the patent application, wherein the salting-out method removes impurities by adding a 20% ammonium sulfate solution. 13. The method for producing high-absorbency phycoerythrin from cladosporium as described in item 1 of the scope of the patent application, wherein the salting-out method is to add 60% to 65% ammonium sulfate solution. 14. The method for producing high-absorbance laccase from cladosporium as described in item 1 of the scope of the patent application, which further comprises purifying and separating phycoerythrin by ultrafiltratin. 15 · If the scope of patent application is the first! The method for producing high-absorbency phycoerythrin from Cladospira, item I further includes purification and separation of phycoerythrin by gel filtration chromatography. Page 26 1224135 Page 27