CN111978383A - Method for synergistically extracting phycoerythrin and grease from wet algae - Google Patents

Method for synergistically extracting phycoerythrin and grease from wet algae Download PDF

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CN111978383A
CN111978383A CN202010914299.7A CN202010914299A CN111978383A CN 111978383 A CN111978383 A CN 111978383A CN 202010914299 A CN202010914299 A CN 202010914299A CN 111978383 A CN111978383 A CN 111978383A
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何勇锦
陈必链
王明兹
黄键
黄子诚
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Fujian Normal University
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Abstract

The invention discloses a method for synergistically extracting phycoerythrin and grease from wet algae, which comprises the following steps: s1, culturing microalgae; s2, collecting wet algae; s3, three-phase separation and synergistic extraction of phycoerythrin and grease; s4, collecting and purifying oil; s5, collecting and purifying phycoerythrin; s6, drying phycoerythrin. The invention takes wet algae as raw material, does not need to break the wall of algae cells, adopts three-phase separation technology, and can synergistically extract phycoerythrin and algae oil containing ARA and EPA by one stepThe extraction rate of the algae oil obtained by the invention can be 98 percent at most, and the purity A of the phycoerythrin565/A280The maximum value can be 6, and the method has the advantages of short time, high efficiency, green safety and energy conservation.

Description

Method for synergistically extracting phycoerythrin and grease from wet algae
Technical Field
The invention belongs to the technical field of marine microalgae, and particularly relates to a method for synergistically extracting phycoerythrin and grease from wet algae.
Background
The red algae is an important biological group in the algae, and has great value in the aspects of marine ecology, biological evolution, food, drug utilization and the like. Red algae can be classified into macroalgae (such as laver) and microalgae (such as porphyridium). Compared with the algae, the microalgae has the advantages of high growth speed, high efficiency of fixing carbon dioxide and high content of active products (such as phycoerythrin, grease and the like), and is concerned by scientific and technological workers. Therefore, the exploitation of active products from marine microalgae biomass is a hot area for the research of algae.
A large number of researches show that phycoerythrin has various biological functions of resisting tumors, improving human immunity, promoting hematopoiesis and the like, and red algae such as porphyridium, rosa multiflora and nostoc can synthesize phycoerythrin. Chenvian et al found that phycoerythrin contents of Porphyridium cruentum and Rhodophyta were 2.98% and 3.60%, respectively (Chenvian, Chenbigan, Zhang Yan-Yan. comparison of three methods for microalgae cell disruption. Biotechnology, 2008, 1: 55-58.). Xu et al found that under optimal conditions, 3.05% phycoerythrin can be synthesized by Porphyridium (Porphyridium purpureum) in the optimum conditions (Xu Yuanchao, Jian Kailin, Zhong Huichang, Wu Shengshan, Ho Shih-Hsin, Zeng Xianhai, Li Jinglong, Tang Xing, Sun Yong, Lin Lu. induced multiplication pattern enhanced the phyceryl protein production, red alga Porphyridium purpureum. bioprocess and Biosystems engineering.2020,43:347 one 355.).
In addition to phycoerythrin, red algae microalgae also synthesize polyunsaturated fatty acids such as eicosatetraenoic acid (C20:4 omega-6, ARA) and eicosapentaenoic acid (C20:5 omega-3, EPA). The ARA and/or EPA can be taken to improve the ARA and/or EPA content of organism tissues, enhance the immunity of the organism, inhibit the growth of pathogenic bacteria and viruses, and prevent and/or treat cardiovascular diseases, tumors, apoplexy, gout and other diseases. It was found that the most suitable time periods for the synthesis of ARA (1.07%, biomass) and EPA (1.94%, biomass) by Porphyridium cruentum were pre-log and end-log, respectively. Phosphorus deficiency favors the synthesis of lipids (666.38mg/L) and ARA (159.74mg/L) (Su Gaomin, Jiao Kailin, Li Zheng, Guo Xiaoyi, Chang Junyu, Ndikubumana Theonest, Sun Yong, Zeng Xianhai, Lu Yinghua, Lin Lu. phosphor yield promoters unsalted fatty acids and aracopical acids Biosystems by microbial Porphyridium purpureum, bioprocess and Biosystems Engineering,2016,39(7): 1129) 1136) by Porphyridium purpureum. In the research on the temperature for synthesizing ARA and EPA by Porphyridium purpureum (Porphyridium purpureum), the higher temperature (35 ℃) is favorable for microalgae to synthesize ARA; however, microalgae tend to synthesize EPA at a temperature of 25 deg.C (Chang Jingyu, Le Kai, Son Xiaoqiang, Jiano Kailin. Scale-up cultivation enhanced aromatic acid accumulation by red microalgal porous microorganism, bioprocess and Biosystems Engineering,2017,40(12): 1-11.).
Phycoerythrin, ARA and EPA are well known as intracellular products. In order to obtain these active products, various extraction methods have been developed by technologists, such as supercritical fluid methods, solvent extraction methods, aqueous two-phase extraction methods, microwave-assisted extraction methods, ultrasonic extraction methods, and the like. In the aspect of phycoerythrin extraction, the porphyra haitanensis is crushed by Zhaoli and the like by a repeated freeze thawing method, and then the porphyra haitanensis phycoerythrin is extracted by using a two-aqueous-phase system, wherein the secondary extraction purity can reach 1.55 (Zhaoli, Pengyelian, Paokuang, Chua Weimin, R-phycoerythrin purified by aqueous two-phase separation, the academic journal of Harbin industry university, 2015,6: 54-58.). Tang et al disrupted the cells of Porphyridium cells by osmotic shock method, concentrated phycobiliproteins by 50kDa ultrafiltration membrane from crude phycobiliproteins, and finally separated by SOURCE 15Q anion exchange chromatography column to obtain phycoerythrin with purity of 5.1 and phycoerythrin recovery of 68.5% (Tang, Zhong, Zhoo Jilu, Ju Bao, Li Wenjun, Wen Shaohong, Pu Yang, Qi n Song. Martiniz et al adopt electric pulse penetration to assist in cell disruption and then carryThe phycoerythrin from porphyridium was taken, the phycoerythrin extraction rate was 3.2% (Mart i yenez Juan m., Delso Carlota,
Figure BDA0002664458920000021
ignacio, Raso Javier. pulsed electric field polymerization and extraction of phyrythrin from Porphyridium cruentum. Algal Research,2019,37: 51-56.). In the extraction of ARA and EPA, Kwangdina et al used ultrasonic wave assisted technology to extract the total oil and fat of Porphyridium (Kwangdina Raymond, ray Inda Innah Zakir Muhammad.Production of biodisel from lipid of Porphyridium cruentum third scientific method, 2014,2014: 1-6.). Asghanur et al, chloroform methanol extraction of Porphyridium purpureum (Porphyridium purpureum) ARA and EPA (Asghanur Maryam, Rodgers Brigitte, Hestekin Jamie A., Eicosaphenaceae organic acid from Porphyridium cruentum: inducing growth and development of microbial for pharmaceutical products, Energies,2015,8(9): 10487-. It is worth noting that the existing methods for extracting phycoerythrin and ARA and EPA only consider extracting 1 intracellular active substances, and do not comprehensively utilize biomass containing active products, which results in waste of resources. The modern fine and further processing requirements are on the premise of ensuring the product quality of a target product, the equipment investment is saved, the production requirements of 'green, high efficiency and safety' are met, the comprehensive utilization rate of byproducts is improved, and zero emission is realized in the processing process, so that the aims of saving resources and improving the added value of the product are fulfilled.
Chinese patent CN 10142946B discloses a method for simultaneously extracting oil and protein from microalgae, which takes wet algae mud as a raw material, adjusts the pH value to be alkaline or alkalescent, and carries out wall breaking of microalgae cells and dissolution of the oil and the protein through steam. Filtering the obtained microalgae slurry to remove cell residues to obtain a mixture of oil and protein; and (3) performing oil-water separation by using a hydrocyclone to obtain microalgae grease, and performing nanofiltration concentration and isoelectric precipitation on a protein aqueous solution, and then performing freeze drying or spray drying to obtain protein powder. The invention takes the algae cells containing water as raw materials, thus saving the energy consumption for drying the algae cells; the weak alkaline or alkaline algae cell sap is adopted for steam wall breaking to enable protein and grease to be dissolved out simultaneously, the wall breaking effect is good, and the melting rate is high, but a steam jet atomizer and a reaction tank are required to be arranged in the steam wall breaking process in the method, the equipment requirement is high, and a conventional laboratory cannot meet the equipment requirement; meanwhile, the extraction process in the invention needs three processes of pH adjustment, and the operation is complicated.
Chinese patent CN 109022523A discloses a process for extracting DHA algae oil and algae protein from Isochrysis galbana, wherein the algae oil and algae protein extraction adopts dry algae powder as raw material, the microwave-ultrasonic successively-assisted chloroform-methanol extraction process is carried out on algae cells, the algae oil extraction and purification are carried out by combining the processes of ion exchange, decoloration and the like, the oil recovery rate reaches more than 95.1%, and the extraction effect is good. However, the processes of algae powder drying, microwave-ultrasonic synergistic extraction and the like in the invention are high-energy consumption processes, and the process extraction cost is high.
In view of the above, it is important to develop a method that is efficient, green and energy-saving in a short time and can simultaneously extract phycoerythrin and grease.
Disclosure of Invention
In order to solve the problems, the invention aims to provide a method for synergistically extracting phycoerythrin and grease from wet algae, which takes the wet algae as a raw material and synergistically extracts microalgal phycoerythrin and grease containing ARA and EPA by a three-phase separation technology, and is short-time, efficient, green and energy-saving.
The technical scheme of the invention is as follows:
a method for synergistically extracting phycoerythrin and grease from wet algae comprises the following steps:
s1, microalgae culture: adding the prepared culture medium into a photobioreactor, inoculating microalgae seeds according to the proportion of 0.05-0.2 g/L, and culturing the microalgae under the illumination condition;
s2, wet alga collection: centrifuging cultured algae cells, removing supernatant, and washing with distilled water for 2 times to obtain wet algae;
s3, three-phase separation and synergistic extraction of phycoerythrin and grease: adding 10-50 times (v/w) volume of distilled water into wet algae obtained by centrifugation, and then adding salts and alcohol reagents, wherein the salt concentration is controlled to be 5-40% (w/v), and the alcohol reagent concentration is controlled to be 10-60% (w/v); stirring and extracting, standing for 1-2 h, and layering the solution; the extraction system is divided into three layers, wherein the upper layer is an alcohol reagent, the middle layer is an alga body, and the lower layer is phycoerythrin;
s4, collecting and purifying oil: collecting the upper layer solution in the step S3, putting the upper layer solution into a rotary evaporator for decompression and concentration, and evaporating the solvent to obtain the algae oil containing ARA and EPA;
s5, collecting and purifying phycoerythrin: collecting the lower layer solution in the step S3, and filtering the lower layer solution by using an ultrafiltration membrane under the conditions that the temperature is 20-45 ℃ and the pressure is 0.02-0.15 MPa, wherein the treatment time is 1-5 h;
s6, drying phycoerythrin: and (3) putting the phycoerythrin solution after the membrane treatment into a spray dryer, and carrying out spray drying treatment under the conditions that the air inlet temperature is 120-180 ℃ and the air outlet temperature is 60-100 ℃ to obtain the phycoerythrin powder.
Further, the culture medium in the step S1 is an f/2 culture medium, an artificial seawater culture medium or a KOCK culture medium, and the adding proportion accounts for 50% -90% of the working volume of the photobioreactor; the illumination culture conditions are as follows: the temperature is 23-40 ℃, the illumination intensity is 1000-10000 Lux, and the culture time is 6-10 days.
Further, in the step S2, the algal cell centrifugation conditions are set as: rotating speed is 3000-8000 rpm, and centrifuging time is 3-10 min.
Further, the salt in step S3 is ammonium sulfate, magnesium sulfate, sodium sulfate, potassium sulfate, dipotassium hydrogen phosphate, or disodium hydrogen phosphate; the alcohol is ethanol, propanol, isopropanol, n-butanol, isobutanol, tert-butanol, n-pentanol, 2-pentanol, 3-pentanol or tert-pentanol.
Further, in step S3, the stirring and extracting conditions are: the stirring speed is 100-1000 rpm, and the processing time is 5-60 min.
Further, in the step S5, the cut-off molecular weight of the ultrafiltration membrane is 30-50 kDa.
Further, in step S1, the photobioreactor is a column photobioreactor, a pipeline photobioreactor, or a raceway pond photobioreactor.
Further, the microalgae in the step S1 is porphyridium, rosa multiflora or nostoc.
Further, the artificial seawater comprises the following components: 5-50 g NaCl, 1-10 g MgSO4·7H2O，1～8g MgCl·6H2O，0.1～5g CaCl2·2H2O，0.1～3g KNO3，0.01～0.2g KH2PO4，0.01～0.2g NaHCO320mL of 0.1-2 mol/L Tris-HCl (pH 7.6), 1mL of trace element mother liquor a, 1mL of FeEDTA solution b, adding distilled water to 1L, and sterilizing at 121 ℃ for 20 min; and (3) a microelement mother liquor a: 4mg ZnCl2，60mg H3BO3，1.5mg CoCl2·6H2O，4mg CuCl2·2H2O，40mg MnCl2·4H2O，37mg(NH4)6Mo7O24Adding distilled water to 100mL, and sterilizing at 121 deg.C for 20 min; FeEDTA solution b: 100mL of 0.05mol/L Na2EDTA(pH 7.6)，240mg FeCl3·4H2O。
The f/2 culture medium comprises the following components: 0.01 to 1g of NaNO3，0.001～0.2g NaH2PO4·H2O，0.001～0.1g Na2EDTA，0.001～0.1g FeCl3·6H2O，1.0～9.9×10-5g CuSO4·5H2O，1.0～9.9×10-5g ZnSO4·7H2O，1.0～9.9×10-5g CoCl2·6H2O，1.0～9.9×10-4g MnCl2·4H2O，1.0～9.9×10-5g Na2MoO4·2H2O，1.0～9.9×10-3g Vitamin B1，1.0～9.9×10-6g Vitamin B12，1.0～9.9×10-6g Vitamin H。
The KOCK medium comprises the following components: 0.05-3 g KNO3，1～50mg KH2PO4，1～10mg MgSO4·7H2O (0.2g/L), 1mL ferric citrate solution (0.025g/L), 10mL soil extractionLiquid, 484mL distilled water, 484mL sea water. Soil extract: the soil extract is prepared by placing 200g of un-fertilized garden soil into a beaker or a triangular flask, adding 1000mL of distilled water, sealing the opening with a vent plug, heating in water bath for 3 hours with boiling water, cooling, precipitating for 24 hours, continuously performing the process for 3 times, filtering, collecting the supernatant, sterilizing in an autoclave, and storing in a refrigerator at 4 ℃ for later use.
The invention has the following beneficial effects:
1. the traditional method for extracting phycoerythrin and algae grease usually adopts dry algae powder, and the process of changing wet algae cells into algae powder is a high-energy-consumption process.
2. In the prior art, the supercritical fluid method and the solvent extraction method can only extract microalgae oil containing ARA and EPA, but cannot extract phycoerythrin; the method adopts wet algal cells, does not need independent wall breaking treatment, only adds water, salts and alcohol reagents to perform oil and protein extraction while breaking the walls of the algal cells, dissolves the oil in an upper alcohol phase, deposits the phycoerythrin in a lower water phase, enables an extraction system to perform three-phase separation, and cooperatively extracts the phycoerythrin and the oil by a one-step method, and has the advantages of simple and convenient operation, low instrument performance requirement and short extraction time. Phycoerythrin purity A extracted by the method of the invention565/A280The extraction rate of the algae oil containing ARA and EPA is 0.8-6%, the extraction rate of the algae oil containing ARA and EPA is 80-98%, the extraction rate of the oil extracted by the method is obviously higher than that of a chloroform methanol method and an ultrasonic-assisted chloroform methanol method, and the extraction effect is good.
3. In the present invention, the salts to be added include ammonium sulfate, magnesium sulfate, sodium sulfate, potassium sulfate, dipotassium hydrogen phosphate and disodium hydrogen phosphate, and the alcohol reagents include ethanol, propanol, isopropanol, n-butanol, isobutanol, t-butanol, n-pentanol, 2-pentanol, 3-pentanol and t-pentanol. In the extraction process of the salt and the alcohol, the salt can effectively precipitate the protein with large molecular weight, improve the solubility of the phycoerythrin in the water phase (namely the lower layer), and avoid the condition that the phycoerythrin can be obtained only by further separation and purification. The oil is dissolved in the alcohol reagent and floats in the upper alcohol phase along with the alcohol reagent, the algal cell residues are positioned in the middle water layer, the three phases are separated, and the extraction efficiency is high.
Drawings
FIG. 1 is a process flow diagram of the present invention.
Detailed Description
The present invention will be described in detail with reference to specific examples.
The method for phycoerythrin detection and grease detection in the examples is as follows:
(1) the phycoerythrin detection comprises the following steps:
a. respectively measuring the absorbances of the extracted phycoerythrin solution at 280nm, 565nm, 620nm and 650nm by using a spectrophotometer;
b. and (3) calculating:
Figure BDA0002664458920000071
Figure BDA0002664458920000072
wherein M isPEIs the mass concentration (mg/mL) of phycoerythrin in the concentrated solution,
MPE＝0.123×A565-0.068A620+0.015A650；
VPEvolume (mL) of phycoerythrin concentrate;
MMis the mass (g) of the microalgae cells.
(2) The grease detection method comprises the following steps:
a. and (3) measuring the quality of the extracted algae oil: mixing a certain amount of wet algae (dry algae powder weight is m)1) Extracting the obtained algae oil, removing organic solvent by rotary evaporator, performing methyl esterification with sodium hydroxide-methanol solution, and purifying with methanolMeasuring fatty acid methyl ester with phase chromatograph, and obtaining the extracted algae oil mass (m) by peak area integration with methyl heptadecanoate as standard2)。
b. And (3) measuring the total lipid content of microalgae: mixing a certain amount of dry microalgae powder (m)3) Adding sodium hydroxide-methanol solution for methyl esterification, analyzing fatty acid methyl ester with gas chromatograph, and obtaining total oil mass (m) by peak area integration with methyl heptadecanoate as standard4)。
c. And (3) calculating:
Figure BDA0002664458920000073
example 1
A method for synergistically extracting phycoerythrin and grease from wet algae comprises the following steps:
s1, microalgae culture: adding a prepared artificial seawater culture medium into a 1L column type photobioreactor, wherein the adding proportion accounts for 50% of the working volume of the reactor, Porphyridium cruentum (Porphyridium cruentum) is taken as a culture strain, microalgae species are inoculated according to the proportion of 0.05g/L, and the illumination culture conditions are as follows: the temperature is 23 ℃, the illumination intensity is 1000Lux, and the culture time is 6 days;
the artificial seawater culture medium comprises the following components: 5g NaCl, 1g MgSO4·7H2O，1g MgCl·6H2O，0.1g CaCl2·2H2O，0.1g KNO3，0.01g KH2PO4，0.01g NaHCO320mL of 0.1mol/L Tris & HCl (pH 7.6), 1mL of trace element mother liquor a, 1mL of FeEDTA solution b, adding distilled water to 1L, and sterilizing at 121 ℃ for 20 min; and (3) a microelement mother liquor a: 4mg ZnCl2，60mg H3BO3，1.5mg CoCl2·6H2O，4mg CuCl2·2H2O，40mg MnCl2·4H2O，37mg(NH4)6Mo7O24Adding distilled water to 100mL, and sterilizing at 121 deg.C for 20 min; FeEDTA solution b: 100mL of 0.05mol/L Na2EDTA(pH 7.6)，240mg FeCl3·4H2O。
S2, wet alga collection: centrifuging the cultured algae cells at 3000rpm for 3min, removing supernatant, and washing with distilled water for 2 times to obtain wet algae;
s3, three-phase separation and synergistic extraction of phycoerythrin and grease: taking centrifuged wet algae (dry weight, 1g), adding 10 times volume (v/w) of distilled water, and then adding salts and alcohol reagents, wherein the salt concentration is controlled to be 5% (w/v), and the alcohol reagent concentration is controlled to be 10% (w/v); stirring and extracting for 5min under the condition that the stirring speed is 100rpm, standing for 1h until the solution is layered; the extraction system is divided into three layers, wherein the upper layer is an alcohol reagent, the middle layer is an alga body, and the lower layer is phycoerythrin; the salt is ammonium sulfate, and the alcohol is ethanol;
s4, collecting and purifying oil: collecting the upper layer solution in the step S3, putting the upper layer solution into a rotary evaporator for decompression and concentration, and evaporating the solvent to obtain the algae oil containing ARA and EPA; the extraction rate of algae oil is 80%.
S5, collecting and purifying phycoerythrin: collecting the lower layer solution in the step S3, and filtering the lower layer solution by adopting an ultrafiltration membrane with the molecular weight cutoff of 30kDa at the temperature of 20 ℃ and the pressure of 0.02MPa for 1 h; purity of phycoerythrin (A)565/A280) The extraction rate of phycoerythrin was 0.8, and the extraction rate of phycoerythrin was 6.5%.
S6, drying phycoerythrin: and (3) putting the phycoerythrin solution after the membrane treatment into a spray dryer, and carrying out spray drying treatment under the conditions that the air inlet temperature is 120 ℃ and the air outlet temperature is 60 ℃ to obtain the phycoerythrin powder.
Example 2
A method for synergistically extracting phycoerythrin and grease from wet algae comprises the following steps:
s1, microalgae culture: adding a prepared f/2 culture medium into a 50T raceway pond photobioreactor, wherein the adding proportion accounts for 90% of the working volume of the reactor, Porphyridium purpureum (Porphyridium purpureum) is taken as a culture strain, microalgae seeds are inoculated according to the proportion of 0.2g/L, and the illumination culture conditions are as follows: the temperature is 40 ℃, the illumination intensity is 10000Lux, and the culture time is 10 days;
the f/2 culture medium comprises the following components: 0.01g NaNO3，0.001g NaH2PO4·H2O，0.001g Na2EDTA，0.001g FeCl3·6H2O，1.0×10-5g CuSO4·5H2O，1.0×10-5g ZnSO4·7H2O，1.0×10-5g CoCl2·6H2O，1.0×10-4g MnCl2·4H2O，1.0×10-5g Na2MoO4·2H2O，1.0×10-3g Vitamin B1，1.0×10-6g Vitamin B12，1.0×10-6g Vitamin H。
S2, wet alga collection: centrifuging the cultured algae cells at 8000rpm for 10min, removing supernatant, and washing with distilled water for 2 times to obtain wet algae;
s3, three-phase separation and synergistic extraction of phycoerythrin and grease: taking centrifuged wet algae (dry weight, 10kg), adding 50 times volume (v/w) of distilled water, and then adding salts and alcohol reagents, wherein the salt concentration is controlled to be 40% (w/v), and the alcohol reagent concentration is controlled to be 60% (w/v); stirring and extracting for 60min under the condition that the stirring speed is 1000rpm, and standing for 12h until the solution is layered; the extraction system is divided into three layers, wherein the upper layer is an alcohol reagent, the middle layer is an alga body, and the lower layer is phycoerythrin; the salt is magnesium sulfate, and the alcohol is tert-butyl alcohol;
s4, collecting and purifying oil: collecting the upper layer solution in the step S3, putting the upper layer solution into a rotary evaporator for decompression and concentration, and evaporating the solvent to obtain the algae oil containing ARA and EPA; the extraction rate of algae oil is 98%.
S5, collecting and purifying phycoerythrin: collecting the lower layer solution in the step S3, and filtering the lower layer solution by adopting an ultrafiltration membrane with the molecular weight cutoff of 50kDa at the temperature of 45 ℃ and the pressure of 15MPa for 5 hours; purity of phycoerythrin (A)565/A280) The extraction rate of phycoerythrin was 5.8%.
S6, drying phycoerythrin: and (3) putting the phycoerythrin solution after the membrane treatment into a spray dryer, and carrying out spray drying treatment under the conditions that the air inlet temperature is 180 ℃ and the air outlet temperature is 100 ℃ to obtain the phycoerythrin powder.
Example 3
A method for synergistically extracting phycoerythrin and grease from wet algae comprises the following steps:
s1, microalgae culture: adding a prepared KOCH culture medium into a 500L pipeline type photobioreactor, wherein the adding proportion accounts for 70% of the working volume of the reactor, rose algae (Rhodella reticulata) is taken as a culture strain, microalgae seeds are inoculated according to the proportion of 0.1g/L, and the illumination culture conditions are as follows: the temperature is 30 ℃, the illumination intensity is 5000Lux, and the culture time is 8 days;
the KOCH medium comprises the following components: 0.05g KNO3，1mg KH2PO4，1mg MgSO4·7H2O (0.2g/L), 1mL of ferric citrate solution (0.025g/L), 10mL of soil extract, 484mL of distilled water, 484mL of seawater. Soil extract: the soil extract is prepared by placing 200g of un-fertilized garden soil into a beaker or a triangular flask, adding 1000mL of distilled water, sealing the opening with a vent plug, heating in water bath for 3 hours with boiling water, cooling, precipitating for 24 hours, continuously performing the process for 3 times, filtering, collecting the supernatant, sterilizing in an autoclave, and storing in a refrigerator at 4 ℃ for later use.
S2, wet alga collection: centrifuging the cultured algae cells at 5000rpm for 7min, removing supernatant, and washing with distilled water for 2 times to obtain wet algae;
s3, three-phase separation and synergistic extraction of phycoerythrin and grease: taking centrifuged wet algae (dry weight, 5kg), adding 30 times volume (v/w) of distilled water, and then adding salts and alcohol reagents, wherein the salt concentration is controlled to be 20% (w/v), and the alcohol reagent concentration is controlled to be 30% (w/v); stirring and extracting for 30min under the condition that the stirring speed is 500rpm, standing for 6h until the solution is layered; the extraction system is divided into three layers, wherein the upper layer is an alcohol reagent, the middle layer is an alga body, and the lower layer is phycoerythrin; the salt is sodium sulfate, and the alcohol is tert-amyl alcohol;
s4, collecting and purifying oil: collecting the upper layer solution in the step S3, putting the upper layer solution into a rotary evaporator for decompression and concentration, and evaporating the solvent to obtain the algae oil containing ARA and EPA; the extraction rate of algae oil is 90%.
S5, collecting and purifying phycoerythrin: collecting the lower layer solution in the step S3, and filtering the lower layer solution by adopting an ultrafiltration membrane with the molecular weight cutoff of 40kDa at the temperature of 30 ℃ and the pressure of 0.1MPa for 3 h; purity of phycoerythrin (A)565/A280) The content of the organic acid is 3.2,the extraction rate of phycoerythrin is 2.5%.
S6, drying phycoerythrin: and (3) putting the phycoerythrin solution after the membrane treatment into a spray dryer, and carrying out spray drying treatment under the conditions that the air inlet temperature is 150 ℃ and the air outlet temperature is 80 ℃ to obtain the phycoerythrin powder.
Example 4
A method for synergistically extracting phycoerythrin and grease from wet algae comprises the following steps:
s1, microalgae culture: adding a prepared artificial seawater culture medium into a 500L column type photobioreactor, wherein the adding proportion accounts for 60% of the working volume of the reactor, Nostoc paludosum is taken as a culture strain, microalgae seeds are inoculated according to the proportion of 0.075g/L, and the illumination culture conditions are as follows: the temperature is 28 ℃, the illumination intensity is 3000Lux, and the culture time is 7 days;
the artificial seawater culture medium comprises the following components: 50g NaCl, 10g MgSO4·7H2O，8g MgCl·6H2O，5g CaCl2·2H2O，3g KNO3，0.2g KH2PO4，0.2g NaHCO320mL of 2mol/L Tris-HCl (pH 7.6), 1mL of trace element mother liquor a, 1mL of FeEDTA solution b, adding distilled water to 1L, and sterilizing at 121 ℃ for 20 min. And (3) a microelement mother liquor a: 4mg ZnCl2，60mg H3BO3,1.5mg CoCl2·6H2O，4mg CuCl2·2H2O，40mg MnCl2·4H2O,37mg(NH4)6Mo7O24Adding distilled water to 100mL, and sterilizing at 121 deg.C for 20 min. FeEDTA solution b: 100mL of 0.05mol/L Na2EDTA(pH 7.6)，240mg FeCl3·4H2O。
S2, wet alga collection: centrifuging the cultured algae cells at 4000rpm for 5min, removing supernatant, and washing with distilled water for 2 times to obtain wet algae;
s3, three-phase separation and synergistic extraction of phycoerythrin and grease: taking centrifuged wet algae (dry weight, 3kg), adding 20 times volume (v/w) of distilled water, and then adding salts and alcohol reagents, wherein the salt concentration is controlled to be 15% (w/v), and the alcohol reagent concentration is controlled to be 20% (w/v); stirring and extracting for 20min under the condition that the stirring speed is 300rpm, standing for 3h until the solution is layered; the extraction system is divided into three layers, wherein the upper layer is an alcohol reagent, the middle layer is an alga body, and the lower layer is phycoerythrin; the salt is potassium sulfate, and the alcohol is propanol;
s4, collecting and purifying oil: collecting the upper layer solution in the step S3, putting the upper layer solution into a rotary evaporator for decompression and concentration, and evaporating the solvent to obtain the algae oil containing ARA and EPA; the algae oil extraction rate is 95%.
S5, collecting and purifying phycoerythrin: collecting the lower layer solution in the step S3, and filtering the lower layer solution by adopting an ultrafiltration membrane with the molecular weight cutoff of 35kDa at the temperature of 25 ℃ and the pressure of 0.07MPa for 2 h; purity of phycoerythrin (A)565/A280) The extraction rate of phycoerythrin is 1.8, and the extraction rate of phycoerythrin is 4.2%.
S6, drying phycoerythrin: and (3) putting the phycoerythrin solution after the membrane treatment into a spray dryer, and carrying out spray drying treatment under the conditions that the air inlet temperature is 130 ℃ and the air outlet temperature is 70 ℃ to obtain the phycoerythrin powder.
Example 5
A method for synergistically extracting phycoerythrin and grease from wet algae comprises the following steps:
s1, microalgae culture: adding a prepared KOCH culture medium into a 50L raceway pond photobioreactor, wherein the adding proportion accounts for 80% of the working volume of the reactor, Porphyridium (Porphyridium purpureum) is taken as a culture strain, microalgae seeds are inoculated according to the proportion of 015g/L, and the illumination culture conditions are as follows: the temperature is 35 ℃, the illumination intensity is 8000Lux, and the culture time is 9 days;
the KOCH medium comprises the following components: 3g KNO3，50mg KH2PO4，10mg MgSO4·7H2O (0.2g/L), 1mL of ferric citrate solution (0.025g/L), 10mL of soil extract, 484mL of distilled water, 484mL of seawater. Soil extract: the soil extract is prepared by placing 200g of un-fertilized garden soil in a beaker or a triangular flask, adding 1000mL of distilled water, sealing the opening with a gas-permeable plug, heating in water bath with boiling water for 3 hr, cooling, precipitating for 24 hr, continuously performing the process for 3 times, filtering, collecting the supernatant, sterilizing in an autoclave, and freezing at 4 deg.CAnd storing in the box for later use.
S2, wet alga collection: centrifuging cultured algae cells at 7000rpm for 8min, removing supernatant, and washing with distilled water for 2 times to obtain wet algae;
s3, three-phase separation and synergistic extraction of phycoerythrin and grease: taking centrifuged wet algae (dry weight, 10kg), adding 40 times volume (v/w) of distilled water, and then adding salt and alcohol reagents, wherein the salt concentration is controlled to be 35% (w/v), and the alcohol reagent concentration is controlled to be 45 (w/v); stirring and extracting for 45min under the condition that the stirring speed is 800rpm, and standing for 8h until the solution is layered; the extraction system is divided into three layers, wherein the upper layer is an alcohol reagent, the middle layer is an alga body, and the lower layer is phycoerythrin; the salt is dipotassium hydrogen phosphate, and the alcohol is isobutanol;
s4, collecting and purifying oil: collecting the upper layer solution in the step S3, putting the upper layer solution into a rotary evaporator for decompression and concentration, and evaporating the solvent to obtain the algae oil containing ARA and EPA; the extraction rate of algae oil is 85%.
S5, collecting and purifying phycoerythrin: collecting the lower layer solution in the step S3, and filtering the lower layer solution by adopting an ultrafiltration membrane with the molecular weight cutoff of 40kDa at the temperature of 28 ℃ and the pressure of 0.12MPa for 4 hours; phycoerythrin (A)565/A280) The purity of (2) was 4.7. The extraction rate of phycoerythrin was 5.4%.
S6, drying phycoerythrin: and (3) putting the phycoerythrin solution after the membrane treatment into a spray dryer, and carrying out spray drying treatment under the conditions that the air inlet temperature is 160 ℃ and the air outlet temperature is 90 ℃ to obtain the phycoerythrin powder.
Example 6
A method for synergistically extracting phycoerythrin and grease from wet algae comprises the following steps:
s1, microalgae culture: adding a prepared artificial seawater culture medium into a 5T raceway pond photobioreactor, wherein the adding proportion accounts for 75% of the working volume of the reactor, rose algae (Rhodella reticulata) is taken as a culture strain, microalgae seeds are inoculated according to the proportion of 0.12g/L, and the illumination culture conditions are as follows: the temperature is 33 ℃, the illumination intensity is 7000Lux, and the culture time is 9 days;
the artificial seawater culture medium comprisesThe components are as follows: 35g NaCl, 8g MgSO4·7H2O，6g MgCl·6H2O，3g CaCl2·2H2O，2g KNO3，0.15g KH2PO4，0.15g NaHCO320mL of 1.5mol/L Tris & HCl (pH 7.6), 1mL of trace element mother liquor a, 1mL of FeEDTA solution b, adding distilled water to 1L, and sterilizing at 121 ℃ for 20 min. And (3) a microelement mother liquor a: 4mg ZnCl2，60mg H3BO3,1.5mg CoCl2·6H2O，4mg CuCl2·2H2O，40mg MnCl2·4H2O,37mg(NH4)6Mo7O24Adding distilled water to 100mL, and sterilizing at 121 deg.C for 20 min. FeEDTA solution b: 100mL of 0.05mol/L Na2EDTA(pH 7.6)，240mg FeCl3·4H2O。
S2, wet alga collection: centrifuging cultured algae cells at 4500rpm for 9min, removing supernatant, and washing with distilled water for 2 times to obtain wet algae;
s3, three-phase separation and synergistic extraction of phycoerythrin and grease: taking centrifuged wet algae (dry weight, 500g), adding 35 times volume (v/w) of distilled water, and then adding salt and alcohol reagents, wherein the salt concentration is controlled to be 35% (w/v), and the alcohol reagent concentration is controlled to be 35% (w/v); stirring and extracting for 26min under the condition that the stirring speed is 900rpm, standing for 6.5h until the solution is layered; the extraction system is divided into three layers, wherein the upper layer is an alcohol reagent, the middle layer is an alga body, and the lower layer is phycoerythrin; the salt is disodium hydrogen phosphate, and the alcohol is 2-pentanol;
s4, collecting and purifying oil: collecting the upper layer solution in the step S3, putting the upper layer solution into a rotary evaporator for decompression and concentration, and evaporating the solvent to obtain the algae oil containing ARA and EPA; the extraction rate of algae oil is 92%.
S5, collecting and purifying phycoerythrin: collecting the lower layer solution in the step S3, adopting an ultrafiltration membrane with the molecular weight cutoff of 45kDa, and filtering the solution at the temperature of 32 ℃ and the pressure of 0.09MPa for 3.5 h; phycoerythrin (A)565/A280) The purity of (A) was 2.7, and the extraction rate of phycoerythrin was 3.7%.
S6, drying phycoerythrin: and (3) putting the phycoerythrin solution after the membrane treatment into a spray dryer, and carrying out spray drying treatment under the conditions that the air inlet temperature is 140 ℃ and the air outlet temperature is 75 ℃ to obtain the phycoerythrin powder.
The following comparative examples 1 and 2 are a method for extracting microalgae algal oil by adopting a chloroform methanol method and an ultrasonic-assisted organic solvent extraction method, and the comparative example 3 is a method for extracting microalgae phycoerythrin by adopting a freeze-thaw method, and the specific examples are as follows:
comparative example 1
1. Obtaining wet porphyridium algae: porphyridium (Porphyridium cruentum) culture and centrifugation collection were performed as in example 1;
2. extracting algae oil: wet algal bodies (dry weight, 1g) were taken, and treated with 20 times volume (v/w) of a chloroform methanol solution (v/v, 2:1) at room temperature for 1 hour with magnetic stirring (400 rpm). Then, 3mL of physiological saline is added, after fully shaking up, the mixture is centrifuged at low speed (1000rpm) for 3min, and a chloroform layer is collected; repeating the treatment for 2 times to obtain middle layer algae; uniformly mixing the chloroform layer samples collected for 3 times;
3. obtaining the crude algae oil: removing chloroform by using a rotary evaporator at 50 deg.C and rotation speed of 100rpm to obtain porphyridium oil.
4. And (3) detecting the purity of the algae oil: the extraction rate of the Porphyridium cruentum oil extracted by the chloroform-methanol method is 62 percent through detection and calculation by a gas chromatograph.
Comparative example 2
1. Obtaining nostoc wet algae: nostoc paludosum (Nostoc paludosum) was cultured and collected by centrifugation as in example 4;
2. ultrasonic wall breaking of nostoc cells: the algae mud is re-suspended by 15mL of normal saline, the ultrasonic output power is 300W, and the processing time is 20 min. Subsequently, the algal bodies were collected by centrifugation.
3. Extracting algae oil: the wet algal bodies (dry weight, 1g) after wall breaking were taken and treated with 25 volumes (v/w) of chloroform methanol solution (v/v, 2:1) under magnetic stirring (300rpm) at room temperature for 1 hour. Subsequently, 4mL of physiological saline was added thereto, the mixture was thoroughly shaken, and then centrifuged at a low speed (800rpm) for 4min to collect a chloroform layer. The middle layer of algal cells was treated repeatedly 2 times. The chloroform layer samples collected 3 times are mixed evenly.
4. Obtaining the crude algae oil: and removing chloroform by adopting a rotary evaporator at the temperature of 50 ℃ and the rotating speed of 100rpm to obtain the nostoc oil.
5. And (3) detecting the purity of the algae oil: the extraction rate of the Nostoc paludosum oil extracted by the ultrasonic-assisted organic solvent method is 68 percent through detection and calculation of a gas chromatograph.
Comparative example 3
1. Obtaining wet rose algae: culture and collection by centrifugation of rose algae (Rhodella reticulata) were carried out as in example 3.
2. Freeze thawing and wall breaking: wet algal bodies (dry weight, 1g) were taken, 150-fold volume (v/w) of physiological saline was added, and treated at-20 ℃ for 5 hours, followed by leaving the algal cells at room temperature for 5 hours. Repeatedly freezing and thawing for 3 times to break cell wall, centrifuging at 8000rpm for 10min, and collecting supernatant;
3. ammonium sulfate precipitation: adding ammonium sulfate solid into the collected supernatant to a final concentration of 20%, salting out for 12h, centrifuging at 8000rpm for 10min, removing foreign proteins, adding ammonium sulfate solid into the supernatant to a final concentration of 45%, salting out for 12h, centrifuging at 8000rpm for 10min, and removing the supernatant. Dissolving the precipitate, dialyzing with dialysis bag with relative molecular weight of 3000 for 24 hr, dialyzing with distilled water for 24 hr, and changing distilled water every 4 hr;
4. and (3) detecting the purity of phycoerythrin: the purity and extraction rate of phycoerythrin extracted from Rosa multiflora (Rhodella reticulata) are respectively 0.9% and 1.7% after calculation by spectrophotometer detection.
From the above examples and comparative examples, it can be seen that in examples 1 to 6 of the present invention, the purity coefficient of the phycoerythrin extracted is 0.8 to 6, and the extraction rate is 2.5% to 6.5%, compared with comparative examples 1 and 2, the purity coefficient and the extraction rate of the phycoerythrin extracted by the method of the present invention are significantly higher than those of the two comparative examples except example 1; the extraction rate of the algae oil in the embodiments 1 to 6 is 80 to 98 percent, compared with the comparative example 3, the extraction efficiency of the algae oil extracted by the method is obviously higher than that in the comparative example, and the method can simultaneously extract phycoerythrin and grease, has short time, high efficiency, simple and convenient operation and low instrument performance requirement.
The above description is only an embodiment of the present invention, and not intended to limit the scope of the present invention, and all modifications of equivalent structures and equivalent processes, which are made by the present specification, or directly or indirectly applied to other related technical fields, are included in the scope of the present invention.

Claims (8)

1. A method for synergistically extracting phycoerythrin and grease from wet algae is characterized by comprising the following steps: the method comprises the following steps:
s1, microalgae culture: adding the prepared culture medium into a photobioreactor, inoculating microalgae seeds according to the proportion of 0.05-0.2 g/L, and culturing the microalgae under the illumination condition;
s2, wet alga collection: centrifuging cultured algae cells, removing supernatant, and washing with distilled water for 2 times to obtain wet algae;
s3, three-phase separation and synergistic extraction of phycoerythrin and grease: adding 10-50 times (v/w) volume of distilled water into wet algae obtained by centrifugation, and then adding salts and alcohol reagents, wherein the salt concentration is controlled to be 5-40% (w/v), and the alcohol reagent concentration is controlled to be 10-60% (w/v); stirring and extracting, standing for 1-2 h, and layering the solution; the extraction system is divided into three layers, wherein the upper layer is an alcohol reagent, the middle layer is an alga body, and the lower layer is phycoerythrin;
s4, collecting and purifying oil: collecting the upper layer solution in the step S3, putting the upper layer solution into a rotary evaporator for decompression and concentration, and evaporating the solvent to obtain the algae oil containing ARA and EPA;
s5, collecting and purifying phycoerythrin: collecting the lower layer solution in the step S3, and filtering the lower layer solution by using an ultrafiltration membrane under the conditions that the temperature is 20-45 ℃ and the pressure is 0.02-0.15 MPa, wherein the treatment time is 1-5 h;
s6, drying phycoerythrin: and (3) putting the phycoerythrin solution after the membrane treatment into a spray dryer, and carrying out spray drying treatment under the conditions that the air inlet temperature is 120-180 ℃ and the air outlet temperature is 60-100 ℃ to obtain the phycoerythrin powder.
2. The method of claim 1, wherein the method comprises the steps of: the culture medium in the step S1 is f/2 culture medium, artificial seawater culture medium or KOCK culture medium, and the adding proportion accounts for 50-90% of the working volume of the photobioreactor; the illumination culture conditions are as follows: the temperature is 23-40 ℃, the illumination intensity is 1000-10000 Lux, and the culture time is 6-10 days.
3. The method of claim 1, wherein the method comprises the steps of: in step S2, the algal cell centrifugation conditions are set as: rotating speed is 3000-8000 rpm, and centrifuging time is 3-10 min.
4. The method of claim 1, wherein the method comprises the steps of: the salts in the step S3 are ammonium sulfate, magnesium sulfate, sodium sulfate, potassium sulfate, dipotassium hydrogen phosphate or disodium hydrogen phosphate; the alcohol solvent is ethanol, propanol, isopropanol, n-butanol, isobutanol, tert-butanol, n-pentanol, 2-pentanol, 3-pentanol or tert-pentanol.
5. The method of claim 1, wherein the method comprises the steps of: in step S3, the stirring and extracting conditions are as follows: the stirring speed is 100-1000 rpm, and the processing time is 5-60 min.
6. The method of claim 1, wherein the method comprises the steps of: in the step S5, the cut-off molecular weight of the ultrafiltration membrane is 30-50 kDa.
7. The method of claim 1, wherein the method comprises the steps of: in the step S1, the photobioreactor is a column type photobioreactor, a pipeline type photobioreactor or a raceway pond photobioreactor.
8. The method of claim 1, wherein the method comprises the steps of: in the step S1, the microalgae is porphyridium, rose algae or nostoc.
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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113476891A (en) * 2021-08-06 2021-10-08 中国科学院南海海洋研究所 Method for simultaneously extracting multiple active substances from porphyridium
CN114015493A (en) * 2021-09-26 2022-02-08 上海师范大学 Method for extracting grease from algae

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1587275A (en) * 2004-07-13 2005-03-02 福建师范大学 Separating and purifying technology for micro algae phycoerythrin
WO2008030076A1 (en) * 2006-09-06 2008-03-13 Instituto Tecnológico y de Estudios Superiores de Monterrey Recovery and purification of b-phycoerythrin produced by porphyridium cruentum using two-aqueous-phase systems and isoelectric precipitation
CN101942014A (en) * 2010-08-23 2011-01-12 中国科学院烟台海岸带研究所 Food grade phycocyanin and preparation method thereof
WO2019234614A2 (en) * 2018-06-06 2019-12-12 Consiglio Nazionale Delle Ricerche A process for the separation and purification of phycobiliproteins
CN111280446A (en) * 2018-12-06 2020-06-16 国投生物科技投资有限公司 EPA extract and preparation method and application thereof

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1587275A (en) * 2004-07-13 2005-03-02 福建师范大学 Separating and purifying technology for micro algae phycoerythrin
WO2008030076A1 (en) * 2006-09-06 2008-03-13 Instituto Tecnológico y de Estudios Superiores de Monterrey Recovery and purification of b-phycoerythrin produced by porphyridium cruentum using two-aqueous-phase systems and isoelectric precipitation
CN101942014A (en) * 2010-08-23 2011-01-12 中国科学院烟台海岸带研究所 Food grade phycocyanin and preparation method thereof
WO2019234614A2 (en) * 2018-06-06 2019-12-12 Consiglio Nazionale Delle Ricerche A process for the separation and purification of phycobiliproteins
CN111280446A (en) * 2018-12-06 2020-06-16 国投生物科技投资有限公司 EPA extract and preparation method and application thereof

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
YONGJIN HE等: "Sustainable biodiesel production from the green microalgae Nannochloropsis:Novel integrated processes from cultivation to enzyme-assisted extraction and ethanolysis of lipids", 《ENERGY CONVERSION AND MANAGEMENT》 *
ZHUBO LI等: "Lipid extraction from non-broken and high water content microalgae Chlorella spp. By three-phase partitioning", 《ALGAL RESEARCH》 *
陈伟平: "紫球藻的廉价培养及藻红蛋白纯化的研究", 《中国优秀硕士学位论文全文数据库 基础科学辑》 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113476891A (en) * 2021-08-06 2021-10-08 中国科学院南海海洋研究所 Method for simultaneously extracting multiple active substances from porphyridium
CN114015493A (en) * 2021-09-26 2022-02-08 上海师范大学 Method for extracting grease from algae
CN114015493B (en) * 2021-09-26 2024-02-20 上海师范大学 Method for extracting grease from algae

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