CN114702561A - Method for comprehensively extracting phycobiliprotein and carrageenan from delicate solieria - Google Patents
Method for comprehensively extracting phycobiliprotein and carrageenan from delicate solieria Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/405—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from algae
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- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
- C08B37/0036—Galactans; Derivatives thereof
- C08B37/0042—Carragenan or carragen, i.e. D-galactose and 3,6-anhydro-D-galactose, both partially sulfated, e.g. from red algae Chondrus crispus or Gigantia stellata; kappa-Carragenan; iota-Carragenan; lambda-Carragenan; Derivatives thereof
Abstract
The invention relates to a method for comprehensively extracting phycobiliprotein and carrageenan from weak solieria, which comprises the following steps of (1) crushing the weak solieria and repeatedly freezing and thawing and breaking cells to obtain crude phycobiliprotein extract; (2) removing impurities from the crude extract by salting out, dialysis and other modes; (3) purifying by phenyl-agarose expansion column and dextran G-150 gel chromatography column to obtain analytically pure phycoerythrin; (4) adding potassium hydroxide with corresponding concentration into the algae residue separated in the step (1) according to a certain material-liquid ratio for soaking; (5) pickling the algae residues, bleaching, and washing with deionized water until the pH value is 7-8; (6) adding deionized water in a certain proportion into the algae residue cleaned in the step (5), and heating and extracting in a water bath; (7) filtering while hot to obtain filtrate, freezing and thawing after cooling, removing water, and drying to constant weight to obtain carrageenan. The invention takes the weak plumeria as the raw material, improves the economic value of the weak plumeria and realizes the high-value utilization of the weak plumeria.
Description
Technical Field
The invention relates to the technical field of seaweed biochemical preparation, in particular to a method for comprehensively extracting phycobiliprotein and carrageenan from delicate Solieria.
Background
Solieria tenuis (Solieria tenuis) belongs to the Rhodophyta (Rhodophyta), Rhodophyceae (Rhodophyceae), Rhodophyceae (Florideae), Gigartinales (Gigartinales), Solieriaceae (Solieriaceae), Solieria (Solieria). The delicate plumeria is mainly distributed in the rocks or marshes of the medium tide zone in China and the coastal areas of Japan and is often used as food for cold dishes or breeding feed. At present, the functional active substances in the delicate feather are lack of systematic research.
Phycobiliprotein is a natural water-soluble protein with fluorescence, and is divided into phycoerythrin, phycocyanin, allophycocyanin and phycoerythrocyanin. Phycobiliprotein is classified into food grade, pharmaceutical grade and reagent grade according to purity, and the value is gradually improved along with the improvement of the purity. Phycobiliprotein can be used as natural colorant in food and chemical industry, and can also be used as antioxidant, fluorescence labeling reagent, tumor inhibitor, and photosensitizer in medicine and health promotion, biological detection, and photodynamic therapy. The phycobiliprotein has wide application range and high economic value, and the domestic high-purity phycoerythrin still depends on import and is expensive at present, so that the extraction source of the phycoerythrin is expanded, the large-scale extraction of the high-purity phycoerythrin is optimized, and the high-valued utilization of seaweed is facilitated.
Solieria tenuifolia as one of Solieriaceae contains carrageenan. The substance is a hydrophilic colloid, and is prepared from sulfateThe product is prepared by connecting galactose and 3, 6-anhydrogalactose via alpha-1, 3 glycosidic bond and beta-1, 4 bond alternately, and can be classified into kappa-type, iota-type, lambda-type, gamma-type, v-type, xi-type and mu-type carrageenans according to the bonding position of sulfate ester bond, and the kappa-type, iota-type and lambda-type carrageenans are commonly used. Compared with agar, the carrageenan has stable performance and wide application. In food industry and daily chemical industry, it can be used asThickening agent、Gelling agentCan be used for jelly, canned food, sausage product, etc.; as clarifying flocculants for beer, wine, etc.; asSuspending agent、EmulsifierAndstabilizing agentCan be used in toothpaste, paint, pesticide spray, etc.; in the aspects of bioengineering and biomedicine, the polysaccharide can replace agar to be used for preparing a culture medium, and can also be used as soluble dietary fiber to regulate intestinal balance. The price of the carrageenan is gradually increased in recent years, besides the shortage of supply of the carrageenan, the carrageenan is also related to the rising of the price of the seaweed for extracting the carrageenan, the carrageenan is mainly extracted from eucheuma, gelidium amansii and the like at present, and the large production of the carrageenan leads partial algae resources to be tense, so the delicate Solieria is used as a new extraction source of the carrageenan, the extraction source of the carrageenan is expanded, the tense of partial extracted algae resources is relieved, and the economic value of the delicate Solieria is also improved.
At present, a certain bioactive substance is mostly extracted singly, and few documents exist for simultaneously extracting multiple bioactive substances. The prior art does not have the method for comprehensively extracting phycobiliprotein and carrageenan from weak solieria, so that the method comprehensively extracts phycobiliprotein and carrageenan from the weak solieria, enriches the raw material sources of phycoerythrin and carrageenan, is favorable for realizing high-value utilization of the weak solieria and promotes the development and research of the weak solieria. Before the subject group, phycoerythrin with the purity of 2.95 is only obtained by extracting phycoerythrin and agar from asparagus serving as raw materials. Compared with the authorized method for extracting polysaccharide from the method for extracting the bioactive substances from the algae, which is carried out by alkali extraction, enzymolysis and alcohol precipitation, the method for extracting the polysaccharide has the advantages of simple steps, short extraction time, low extraction cost and contribution to large-scale extraction.
Disclosure of Invention
The invention aims to solve the problems and provides a method for comprehensively extracting phycobiliprotein and carrageenan from weak solieria floribunda.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows:
a method for comprehensively extracting phycobiliprotein and carrageenan from delicate Solieria veronica, which comprises the following steps:
A. extracting and purifying phycobiliprotein
(1) The weak solieria is treated according to the material-liquid ratio of 1: 1-1: 1.5 (mass-volume ratio g: mL) adding deionized water to break the wall and grind, putting into a refrigerator at-20-30 ℃, repeatedly freezing and thawing for 3-5 times, filtering with 50-200 mesh (preferably 100 mesh) fine-mesh silk to remove the algae residue, and obtaining the filtrate as the crude phycobiliprotein extract;
(2) adding ammonium sulfate which is 60 to 100 percent of the volume of the crude phycobiliprotein extract into the crude phycobiliprotein extract for salting out;
(3) centrifuging the salting-out solution obtained in the step (2) for 5-10 min at the rotation speed of 4000-6000 rpm and the temperature of 2-8 ℃, removing supernatant, collecting precipitates, adding 100-300 mL of deionized water for dissolving, putting the obtained solution into a dialysis bag for dialysis, and changing water every 2-4 h until the dialysate does not contain sulfate radicals;
(4) loading the dialysate of the step (3) into a phenyl-agarose expansion column at the speed of 1-3 mL/min, eluting with 0.5M ammonium sulfate until colorless, cleaning the impurity proteins adsorbed on the expansion column and the phycoerythrin not adsorbed, eluting the phycoerythrin with 0.2-0.025M ammonium sulfate in a gradient manner, and collecting the eluent;
(5) dialyzing the eluent to remove salt, concentrating until the volume of a dextran G-150 gel chromatographic column is 1-5%, loading, eluting by using 0.05M phosphate buffer solution, and collecting phycoerythrin eluent; thus obtaining purified phycoerythrin;
(6) measuring ultraviolet absorption spectrum, fluorescence spectrum, SDS-PAGE and Native-PAGE of the purified phycoerythrin;
B. extracting carrageenan from phycobiliprotein-extracted phycoresidue
(1) Putting the algae residue with the phycobiliprotein extracted into a beaker, and mixing the algae residue (wet weight) and the potassium hydroxide feed liquid according to the ratio of 1: 1-1: 5 (mass volume ratio g: mL) adding alkali liquor with the mass concentration of 4-8%, and carrying out water bath for 1-3 h at 50-80 ℃;
(2) filtering with a bolting silk of 50-200 meshes (preferably 100 meshes), washing the algae residue with 0.5% acetic acid till the pH is 7-8, bleaching the algae residue with 0.5% sodium hypochlorite, and washing the algae residue with deionized water till the algae residue is neutral;
(3) and then mixing the algae residue with deionized water according to the material-liquid ratio of 1: 1-1: 5 (mass-to-volume ratio g: mL), performing water bath for 2-4 h at 70-100 ℃;
(4) filtering the extracting solution obtained in the step (3) while the extracting solution is hot, freezing the extracting solution for 24-72 hours after the extracting solution is solidified at normal temperature, melting the extracting solution to remove water, and drying the extracting solution at 50-70 ℃ to obtain carrageenan;
(5) the obtained carrageenan is subjected to yield, sulfate group and infrared spectrum determination.
The following describes some aspects of the invention:
the pH of the phosphate buffer was 7.
The glucan G-150 gel is soaked in deionized water for more than 24 hours and fully swells.
Compared with the prior art, the invention has the beneficial technical effects that:
1. the extraction raw material sources of phycoerythrin and carrageenan are expanded, and the research on active substances in the delicate Solieria is enriched.
2. The existing phycoerythrin separation and purification cost is high, and large-scale production does not exist in China, the phycoerythrin with the reagent grade purity of 4.19 is obtained by two times of chromatographic column separation and purification, the yield is 0.06mg/g, the recovery rate is high, the method is suitable for large-scale extraction and purification of phycoerythrin, and the operation is simple and convenient.
3. The extraction rate of the extracted phycobiliprotein residue is 19.60 percent (dry weight), the operation is simple and convenient, the comprehensive utilization is realized, two active substances are extracted simultaneously, and the utilization rate and the economic value of the delicate peacock feather are improved.
Drawings
FIG. 1 shows the absorption spectrum of the crude extract of thin plumes of example 1 at 250-700 nm.
FIG. 2 shows the absorption spectrum and fluorescence spectrum of 250-700 nm of reagent-grade phycoerythrin obtained by purifying Solieria deliciosa in example 1.
FIG. 3 shows SDS-PAGE and Native-PAGE of purified margarita solium in example 1 to obtain phycoerythrin.
FIG. 4 is the FTIR spectrum of carrageenan extracted from the fine and weak Solieria speciosa residue of example 1.
Detailed Description
In order to better understand the essence of the present invention, the method for extracting phycobiliprotein and carrageenan from weak solieria will be described in the following examples, but the present invention is not limited by these examples.
Example 1: method for comprehensively extracting phycobiliprotein and carrageenan from weak solieria
The preparation method comprises the following steps:
A. extracting and purifying phycobiliprotein
(1) Weighing 500g of fresh thin and weak Solieria veronica, cleaning, pulverizing in a wall breaking machine, adding 600mL of deionized water, placing in a refrigerator at-20 deg.C, repeatedly freezing and thawing for 3 times, filtering with 100 mesh fine screen silk to remove algae residue to obtain crude extract of phycobiliprotein, and keeping the algae residue in the refrigerator at-20 deg.C.
(2) Adding ammonium sulfate 70% of the crude extractive solution into the crude extractive solution of phycobiliprotein for salting out.
(3) Centrifuging the salting-out solution obtained in the step (2) at the rotation speed of 5000rpm and the temperature of 4 ℃ for 6min, removing supernatant, collecting phycobiliprotein precipitate, adding a certain amount of water for dissolving, putting the solution into a dialysis bag for dialysis, changing water every 2h until no precipitate is added by barium chloride, indicating no sulfate ions, obtaining 350mL of crude extract, and calculating 112mg phycoerythrin-containing solution according to a formula, wherein the purity is 0.35.
(4) Loading the dialysate of (3) to a phenyl-agarose expansion column at a speed of 2.5mL/min, eluting with 0.5M ammonium sulfate until colorless, washing off impure proteins adsorbed on the expansion column and non-adsorbed phycoerythrin, eluting phycoerythrin with 0.2M, 0.15M, 0.1M, 0.05M and 0.025M ammonium sulfate in a gradient manner, collecting the eluate, calculating to obtain 25.09mg of phycoerythrin in the eluate, wherein the yield is 0.07mg/g, the purity is 2.77 and the recovery rate is 30.43%.
(5) Dialyzing the eluent to remove salt, performing ultrafiltration concentration on 13mL, loading the eluent to a glucan G-150 gel chromatographic column, eluting with 0.05M phosphate buffer solution, collecting phycoerythrin eluent, and calculating to obtain 20.7mg of phycoerythrin in the eluent, wherein the yield is 0.06mg/G, the purity is 4.19, and the recovery rate is 26.09%.
(6) The purified phycoerythrin was analyzed by ultraviolet absorption spectroscopy, fluorescence spectroscopy, SDS-PAGE, and Native-PAGE.
B. Extracting carrageenan from phycobiliprotein-extracted phycoresidue
(1) Extracting the fine and weak solieria residue of phycoerythrin according to the material-liquid ratio of 1: 2 adding 6.5 percent potassium hydroxide solution, and carrying out alkali treatment in a water bath at 62 ℃ for 2 h.
(2) Filtering (1), washing the fine and weak feather algae residues with 0.5% acetic acid until the pH value is 7-8, bleaching the algae residues with 0.5% sodium hypochlorite for 5min, washing the algae residues with deionized water until the algae residues are neutral, filtering the algae residues to be dry, and mixing the algae residues according to a material-liquid ratio of 1: 2 adding deionized water, and carrying out water bath at 90 ℃ for 3 h.
(3) Filtering the solution in the step (2) by using a silk screen, and collecting filtrate.
(4) And (3) solidifying the filtrate at normal temperature, freezing overnight, thawing to remove water, and drying at 60 ℃ to obtain the carrageenan. The yield was weighed to 19.60% (dry weight).
(5) And (4) carrying out sulfate radical content and FTIR characterization determination on the dried carrageenan. The sulfate content was 7.89%. According to FTIR spectrogram, characteristic peaks of the carrageenan are extracted: the absorption peak of CH is 2920cm-1The absorption peak of-OH is 3400cm for stretching vibration on six-membered ring-1Nearby, the absorption peak of the 3, 6-galactoside group was 930cm-1The characteristic peak of COC group is 1100cm-1Total SO of4Has an absorption peak of 1214cm-1At 847cm-1Has characteristic absorption peak, tableSO at C4 of (1,3) -beta-D-galactose of gelatin4It shows that the main component is k-carrageenan which is 803cm-1Has obvious characteristic peak, which indicates that SO is arranged on C2 of 3,6 inner ether-D-galactose4Iota-carrageenan is present. Comparative example 1:
the preparation method comprises the following steps:
A. extracting and purifying phycobiliprotein
(1) Weighing 500g of fresh thin and weak Solieria veronica, cleaning, pulverizing in a wall breaking machine, adding 650mL of deionized water, repeatedly freezing and thawing in a refrigerator at-20 deg.C for 3 times, filtering with fine bolting silk to remove algae residue to obtain crude phycobiliprotein extract, and keeping the algae residue in the refrigerator at-20 deg.C.
(2) Adding ammonium sulfate 70% of the crude extractive solution into the crude extractive solution of phycobiliprotein for salting out.
(3) And (3) centrifuging the salting-out solution in the step (2) at the rotation speed of 5000rpm and the temperature of 4 ℃ for 6min, removing supernatant, collecting phycobiliprotein precipitate, adding a certain amount of water for dissolving, putting the solution into a dialysis bag for dialysis, and changing water every 2h until no precipitate is formed when barium chloride is added, which indicates that no sulfate ions exist. 300mL of the crude extract is obtained, and the phycoerythrin content of 115mg and the purity of 0.35 are calculated according to a formula.
(4) And (3) loading the dialysate of the step (3) to a phenyl-agarose expansion column at a speed of 2.5mL/min, wherein the speed is too high, part of phycoerythrin is washed out without being combined with the column, eluting with 0.5M ammonium sulfate at a speed of 2.5mL/min until the phycoerythrin is colorless, eluting the phycoerythrin by using 0.1-0.0025M ammonium sulfate in a gradient manner, mixing the impurity protein and the phycoerythrin to be eluted, collecting eluent according to the volume of the column, measuring an absorption spectrum after dialysis, calculating to obtain the total phycoerythrin content in the eluent, namely 23.74mg, the yield is 0.05mg/g, the purity is 2.43, and the recovery rate is 21.74%. The sample loading and elution are carried out at a higher speed, the purification effect is greatly reduced, the purity is reduced, and the phycoerythrin loss rate is high.
(5) Dialyzing the eluent to remove salt, performing ultrafiltration concentration to the column volume of 5%, loading to a dextran G-150 gel chromatographic column, eluting with 0.05M phosphate buffer solution, collecting phycoerythrin eluent, dialyzing, determining absorption spectrum, and calculating to obtain phycoerythrin 17.50mg, yield of 0.035mg/G, purity of 3.62, and recovery rate of 14.00%.
B. Extracting carrageenan from phycobiliprotein-extracted phycoresidue
((1) adding 6.5% potassium hydroxide solution into the fine and weak solieria residue after the phycoerythrin is extracted according to the material-liquid ratio of 1: 2, and carrying out alkali treatment for 2 hours in a water bath at 90 ℃.
(2) Filtering (1), washing the fine and weak feather algae residues with 0.5% acetic acid until the pH value is 7-8, bleaching the algae residues with 0.5% sodium hypochlorite for 5min, washing the algae residues with deionized water until the algae residues are neutral, filtering the algae residues to be dry, and mixing the algae residues according to a material-liquid ratio of 1: 2 adding deionized water, and carrying out water bath at 90 ℃ for 3 h.
(3) Filtering the solution in the step (2) by using a bolting silk, and collecting filtrate. (4) And (3) solidifying the filtrate at normal temperature, freezing overnight, thawing to remove water, and drying at 60 ℃ to obtain the carrageenan. The yield was weighed as 10.04% (dry weight), too high temperature alkali extraction resulted in degradation of carrageenan during the alkali treatment, resulting in a decrease in yield.
Comparative example 2:
the preparation method comprises the following steps:
A. extracting and purifying phycobiliprotein
(1) Weighing 500g of fresh thin and weak Solieria veronica, cleaning, pulverizing in a wall breaking machine, adding 600mL of deionized water, placing in a refrigerator at-20 deg.C, repeatedly freezing and thawing for 3 times, filtering with 100 mesh fine screen silk to remove algae residue to obtain crude extract of phycobiliprotein, and keeping the algae residue in the refrigerator at-20 deg.C.
(2) Adding ammonium sulfate 70% of the crude extractive solution into the crude extractive solution of phycobiliprotein for salting out.
(3) Centrifuging the salting-out solution obtained in the step (2) at the rotation speed of 5000rpm and the temperature of 4 ℃ for 6min, removing supernatant, collecting phycobiliprotein precipitate, adding a certain amount of water for dissolving, putting the solution into a dialysis bag for dialysis, changing water every 2h until no precipitate is formed by adding barium chloride, indicating no sulfate ions, obtaining 350mL of crude extract, and calculating 112mg of phycoerythrin with the purity of 0.35 according to a formula.
(4) Loading the dialysate of (3) to a phenyl-agarose expansion column at a speed of 2.5mL/min, eluting with 0.5M ammonium sulfate until colorless, washing off impure proteins adsorbed on the expansion column and non-adsorbed phycoerythrin, eluting phycoerythrin with 0.2M, 0.15M, 0.1M, 0.05M and 0.025M ammonium sulfate in a gradient manner, collecting the eluate, calculating to obtain 25.09mg of phycoerythrin in the eluate, wherein the yield is 0.07mg/g, the purity is 2.77 and the recovery rate is 30.43%.
(5) Dialyzing the eluate for desalting, loading to Q-anion chromatographic column, gradient eluting with 0.05M PBS buffer solution (containing 0.05M, 0.1M, 0.15M, and 0.2M sodium chloride) with pH of 7, collecting phycoerythrin eluate, measuring absorption spectrum, and calculating to obtain phycoerythrin 21.13mg in eluate with yield of 0.04mg/g, purity of 3.85, and recovery rate of 17.39%.
B. Extracting carrageenan from phycobiliprotein-extracted phycoresidue
(1) The material liquid ratio of the fine and weak solieria residue after the phycoerythrin extraction is 1: 2 adding 6.5 percent potassium hydroxide solution, and carrying out alkali treatment in water bath at 60 ℃ for 6 h.
(2) Bleaching the algae residues with 0.5% sodium hypochlorite for 5min, washing the algae residues with deionized water to neutrality, filtering the algae residues to dry, and mixing the algae residues according to a material-liquid ratio of 1: 2 adding deionized water, and carrying out water bath at 90 ℃ for 3 h.
(3) Filtering the solution in the step (2) by using a bolting silk, and collecting filtrate.
(4) And (3) solidifying the filtrate at normal temperature, freezing overnight, thawing to remove water, and drying at 60 ℃ to obtain the carrageenan. The yield was 8.00% (dry weight) and the gum obtained contained some pigment material. Prolonged alkali extraction can degrade carrageenan during the alkali treatment process, resulting in reduced yields.
Claims (4)
1. A method for comprehensively extracting phycobiliprotein and carrageenan from delicate Solieria solifolia is characterized by comprising the following steps: the method comprises the following specific steps:
A. extracting and purifying phycobiliprotein
(1) The weak solieria is treated according to the material-liquid ratio of 1: 1-1: 1.5 (mass-volume ratio g: mL) adding deionized water to break the wall and grind, putting into a refrigerator at-20-30 ℃, repeatedly freezing and thawing for 3-5 times, filtering with 50-200 meshes to remove the algae residue, and obtaining the filtrate as the crude extract of phycobiliprotein;
(2) adding ammonium sulfate which is 60 to 100 percent of the volume of the crude phycobiliprotein extract into the crude phycobiliprotein extract for salting out;
(3) centrifuging the salting-out solution obtained in the step (2) for 5-10 min at the rotation speed of 4000-6000 rpm and the temperature of 2-8 ℃, removing supernatant, collecting precipitates, adding 100-300 mL of deionized water for dissolving, putting the obtained solution into a dialysis bag for dialysis, and changing water every 2-4 h until the dialysate does not contain sulfate radicals;
(4) loading the dialysate obtained in the step (3) into a phenyl-agarose expansion column at the speed of 1-3 mL/min, eluting with 0.5M ammonium sulfate until colorless, cleaning the impure protein adsorbed on the expansion column and the phycoerythrin not adsorbed, then eluting the phycoerythrin with 0.2-0.025M ammonium sulfate in a gradient manner, and collecting the eluent;
(5) dialyzing the eluent to remove salt, concentrating until the volume of a dextran G-150 gel chromatographic column is 1-5%, loading, eluting by using 0.05M phosphate buffer solution, and collecting phycoerythrin eluent; thus obtaining purified phycoerythrin;
(6) measuring the purified phycoerythrin by ultraviolet absorption spectrum, fluorescence spectrum, SDS-PAGE and Native-PAGE;
B. extracting carrageenan from phycobiliprotein-extracted phycoresidue
(1) Placing the algae residue with the phycobiliprotein extracted into a container, and mixing the algae residue (wet weight) with the liquid-liquid ratio of 1: 1-1: 5 (mass volume ratio g: mL) adding 4-8% by mass of potassium hydroxide alkali liquor, and carrying out water bath for 1-3 h at 50-80 ℃;
(2) filtering with a bolting silk of 50-200 meshes (preferably 100 meshes), washing the algae residue with acetic acid with the concentration of 0.5% to the pH value of 7-8, bleaching the algae residue with sodium hypochlorite of 0.5%, and washing the algae residue with deionized water to be neutral;
(3) and then mixing the algae residue with deionized water according to the material-liquid ratio of 1: 1-1: 5 (mass-to-volume ratio g: mL), performing water bath for 2-4 h at 70-100 ℃;
(4) filtering the extracting solution obtained in the step (3) while the extracting solution is hot, freezing the extracting solution for 24-72 hours after the extracting solution is solidified at normal temperature, melting the extracting solution to remove water, and drying the extracting solution at 50-70 ℃ to obtain carrageenan;
(5) the obtained carrageenan is subjected to yield, sulfate group and infrared spectrum determination.
2. The method for comprehensively extracting phycobiliprotein and carrageenan from solieria tenuissima as claimed in claim 1, wherein the method comprises the following steps: the glucan G-150 gel is soaked in deionized water for more than 24 hours and fully swells.
3. The method for comprehensively extracting phycobiliprotein and carrageenan from solieria tenuissima as claimed in claim 1, wherein the method comprises the following steps: the pH of the phosphate buffer was 7.
4. The method for comprehensively extracting phycobiliprotein and carrageenan from solieria tenuissima as claimed in claim 1, wherein the method comprises the following steps: the filtration in the step (1) is preferably performed by filtering with a 100-mesh fine-mesh silk screen to remove algae residues.
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| CN117343215B (en) * | 2023-12-06 | 2024-02-09 | 中国海洋大学 | Method for extracting agar from agar |
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