Extraction Of Phycobiliproteins Research Articles

Optimization of phycobiliprotein extraction from triple algal co-culture

Optimization of phycobiliprotein extraction from triple algal co-culture

Optimized extraction of phycobiliproteins from Arthrospira platensis: quantitative and qualitative assessment of C-Phycocyanin, Allophycocyanin, and Phycoerythrin

Optimized extraction of phycobiliproteins from Arthrospira platensis: quantitative and qualitative assessment of C-Phycocyanin, Allophycocyanin, and Phycoerythrin

Extraction yield and biological activity of phycobiliproteins from Porphyridium purpureum using atmospheric cold plasma discharge and jet systems

Phycobiliproteins (PBPs) extracted from Porphyridium purpureum (P.p) have bioactive properties and are widely used as ingredients in nutraceutical and food applications. This study investigated the use of two cold plasma systems, namely cold plasma discharge system (CPDS) and cold plasma jet system (CPJS), for the aqueous extraction of PBPs from P. p. Three types of PBPs, namely phycoerythrin (PE), phycocyanin (PC) and allophycocyanin (APC) were identified in the crude extracts obtained. The highest PBPs extraction yield (11.31 ± 1.02 mg/g DW P. p) was obtained from CPDS treated samples at 25 kV using N2 for 9 min. CPDS treatments were also shown to be more effective than CPJS treatments in increasing antioxidant activities of the PBPs crude extracts obtained. PBPs crude extracts obtained using CPDS (25 kV; 6 min; N2) had the highest DPPH (69.44 ± 0.10%) and FRAP (207.34 ± 12.96 μmol/L) antioxidant activities observed. PBPs obtained from samples treated with CPDS (25 kV; 9 min; N2) exhibited the highest cytotoxicity potential in Caco-2 human colorectal adenocarcinoma cell lines. This study demonstrates that cold plasma treatments increase the extraction yield of PBPs obtained from P. p and also enhance antioxidant and cytotoxic properties of the PBP crude extracts. However, an increase in treatment time beyond 6 min for both plasma systems was shown to reduce the level of antioxidant activity in PBPs.

Extraction and Native PAGE Separation of Phycobiliproteins from Some Cyanobacteria Collected from Their Natural Habitats

Phycobiliproteins are a group of coloured proteins present in cyanobacteria and algae. They can be divided into three types based on their absorption spectra. These are phycocyanin, phycoerythrin and allophycocyanin. These pigment proteins are used as potential natural colorant in chewing gums, candies, soft drinks, dairy products and cosmetics like lipstick and eyeliners. They are also extensively commercialized for fluorescent applications in clinical immunological analysis. The phycobiliproteins from cyanobacteria have also been reported to have ant-cancerous, anti-inflammatory and antioxidant properties. In view of the increasing demand of these fluorescent pigments, it becomes important to find new species of cyanobacteria and exploit them for their phycobiliprotein content. In present work some commonly available cyanobacteria were collected from their natural habitats and analysed for their phycobiliprotein content. The extraction of phycobiliprotein was done in phosphate buffer and quantitative analysis of the pigment components was done. The study showed that all the cyanobacteria are the potential source of phycocyanin whereas phycoerythrin is significantly present in species of Lyngbya and Oscillatoria. The phycobiliprotein components were separated on Native PAGE which can be partially purified by electroelution. The percentage loss of phycobiliprotein content in stored cyanobacterial biomass for two months showed that phycoerythrin was more stable in Lyngbya and Oscillatoria as compared to the phycocyanin. The species of Scytonema showed good amount of phycocyanin content whereas allophycocyanin was significantly present and was stable in Aulosira sp.

Colorful Diversity─Modified Methods for Extraction and Quantification of Photopigments and Phycobiliproteins Isolated from Phototrophic Micro- and Macroalgae

In the course of combining the Sustainable Development Goals (SDGs) with the science education curriculum, the relevance of the micro- and macroalgae in education is based on the biotechnological future-oriented significance and the ever-growing trend toward plant-based nutrition. So, the micro- and macroalgae are finding their way onto the food shelves and creating biotechnological solutions with regard to climate change (SDG4; SDG13). Their colored photopigments and phycobiliproteins are already established as important natural dyes in the food and textile industries. In addition to being essential in photosynthesis, photopigments have a variety of functions and effects that influence almost all aspects of our lives. The article presents experimental protocols developed based on the established methods for the extraction of photopigments from plants and optimized for the use of phototrophic micro- and macroalgae (Chlorella vulgaris, Arthrospira platensis, and Palmaria palmata). Besides the green chlorophylls and yellow-orange carotenoids in plants, cyanobacteria and red algae developed additional light antenna complexes, so-called phycobilisomes, consisting of different phycobiliproteins. For this purpose, experiments that are simple to execute have been developed to make the colorful world of photopigments visible to the students from the upper secondary level and can be used in both university and school educational settings. Furthermore, a column chromatography was developed, which allows the preparation of the pigment and phycobiliprotein extracts from A. platensis. This procedure is based on the established “supermarket column” and was optimized according to the use of powdered amounts of A. platensis. Additionally, results from a first implementation in a classroom setting will be discussed.

Extraction and purification of phycobiliproteins from algae and their applications.

Microalgae, macroalgae and cyanobacteria are photosynthetic microorganisms, prokaryotic or eukaryotic, living in saline or freshwater environments. These have been recognized as valuable carbon sources, able to be used for food, feed, chemicals, and biopharmaceuticals. From the range of valuable compounds produced by these cells, some of the most interesting are the pigments, including chlorophylls, carotenoids, and phycobiliproteins. Phycobiliproteins are photosynthetic light-harvesting and water-soluble proteins. In this work, the downstream processes being applied to recover fluorescent proteins from marine and freshwater biomass are reviewed. The various types of biomasses, namely macroalgae, microalgae, and cyanobacteria, are highlighted and the solvents and techniques applied in the extraction and purification of the fluorescent proteins, as well as their main applications while being fluorescent/luminescent are discussed. In the end, a critical perspective on how the phycobiliproteins business may benefit from the development of cost-effective downstream processes and their integration with the final application demands, namely regarding their stability, will be provided.

Efficient extraction of phycobiliproteins from dry biomass of Spirulina platensis using sodium chloride as extraction enhancer

An efficient strategy for phycobiliprotein extraction from Spirulina platensis dry biomass has been developed by using NaCl as an enhancer. Different sodium ion and chloride ion salts were screened, and NaCl was selected as the most appropriate solvent for phycobiliprotein extraction. The extraction parameters with NaCl were optimized using response surface methodology. Under optimal operating conditions, a phycobiliprotein extraction rate of 74.8 % and a phycocyanin extraction yield of 102.4 mg/g with a purity of 74.0 % were achieved. Adding NaCl resulted in smaller fragments and destroyed the cell integrity of S. platensis, facilitating phycobiliprotein exudation. The secondary structure and antioxidant activity of phycobiliproteins were not affected by NaCl extraction. The stability of the phycobiliproteins was improved by adding NaCl. This study provides a potential method for phycobiliprotein extraction with high efficiency and good quality using an inexpensive extraction enhancer.

Improvement in the Sequential Extraction of Phycobiliproteins from Arthrospira platensis Using Green Technologies.

Arthrospira platensis (commercially known as Spirulina) is an excellent source of phycobiliproteins, especially C-phycocyanin. Phycobiliproteins are significant bioactive compounds with useful biological applications. The extraction process plays a significant role in downstream microalga production and utilisation. The important pigments found in A. platensis include chlorophyll and carotenoids as nonpolar pigments and phycobiliproteins as polar pigments. Supercritical fluid extraction (SFE) as a green extraction technology for the high-value metabolites of microalgae has potential for trends in food and human health. The nonpolar bioactive compounds, chlorophyll and carotenoids of A. platensis, were primarily separated using supercritical carbon dioxide (SC-CO2) solvent-free fluid extraction pressure; the temperature and ethanol as cosolvent conditions were compared. The residue from the A. platensis cells was subjected to phycobiliprotein extraction. The phosphate and water extraction of A. platensis SFE residue were compared to evaluate phycobiliprotein extraction. The SFE results exhibited higher pressure (350 bar) and temperature extraction (50 °C) with ethanol-free extraction and increased nonpolar pigment. Phycobiliprotein yield was obtained from A. platensis SFE residue by ethanol-free buffer extraction as a suitable process with antioxidant properties. The C-phycocyanin was isolated and enhanced to 0.7 purity as food grade. This developed method can be used as a guideline and applied as a sustainable process for important pigment extraction from Arthrospira microalgae.

Combined extraction methodology for simultaneous recovery of phycobiliproteins and agar from the red alga Gracilaria chilensis C. J. Bird, McLachlan & E. C. Oliveira

The red alga Gracilaria chilensis C. J. Bird, McLachlan & E. C. Oliveira ( Agarophyton chilense Gurgel, J.N. Norris & Fredericq) is one of the few algae commercially farmed in Chile, where this alga is commonly named “Pelillo”. G. chilensis main by-product is agar, a gelling agent used in the food and pharmaceutical industries. This alga is also a valuable feedstock for the biorefinery of phycobiliproteins (PBPs), colored and fluorescent water-soluble proteins with industrial applications. This work aimed to valorize G. chilensis by sequentially extracting PBPs and agar. After freeze-thaw treatment, we successfully extracted two PBPs from alga: R-phycoerythrin (R-PE) and R-phycocyanin (R-PC). After two purification steps, we recovered 45 % of both PBPs (R-PE = 0.20 mg g −1 DW and R-PC = 0.23 mg·g −1 DW). After PBPs extraction, we successfully recovered agar from the residual algal matter. There were no significant differences between the physical parameters of agars obtained after PBP extraction and those from agars directly extracted from G. chilensis (melting and gelling temperature, and agar deformation). The agar yield after PBPs extraction was 24.9 ± 1.0 % and showed a gel strength of 726 ± 182.9 g·cm −2 , whereas agar directly extracted from alga yielded 23.1 ± 2.0 % with a gel strength of 730.5 ± 96.9 g·cm −2 . The agar obtained after PBPs extraction showed no inhibitory effects on microbial growth and allowed the effective separation of nucleic acids without agarose purification. Thereby, the method proposed in this work allowed the simultaneous extraction of PBPs and agar from G. chilensis . • The red seaweed Gracilaria chilensis ( Agarophyton chilense ) is a good source of phycobiliproteins and agar. • Phycobiliproteins are purified by salting-out and anion-exchange chromatography. • Purified phycobiliproteins (R-PE and R-PC) may be used in the food industry. • High-quality agar was successfully recovered from the residual algal matter after phycobiliproteins' extraction. • Agar extracted after phycobiliproteins' purification was useful for microbial growth and nucleic acid separation.

Recent advances in non-conventional techniques for extraction of phycobiliproteins and carotenoids from microalgae

Recent advances in non-conventional techniques for extraction of phycobiliproteins and carotenoids from microalgae

Extraction and Purification of Phycocyanin from Arthrophira platensis Microalgae Using a Green Solid‐Liquid Extraction Technology (RSLDE)

AbstractSpirulina is a blue alga belonging to the class of Cyanobacteria. This alga is characterized by a high content of phycocyanin (PC), a blue photosynthetic pigment belonging to the phycobiliprotein family. As natural dye, PC has a lot of industrial applications, in several fields such as the cosmetic, food, and textile sectors. It is also used in the biomedical field as a fluorescent marker. Anti‐inflammatory, nephroprotective, and hepatoprotective properties are also reported, probably related to its antioxidant activity. However, the extraction of phycobiliproteins from cyanobacteria is a very complicated procedure due to the extreme resistance of the wall of the cellular aggregates that make up spirulina. In literature, several procedures have been used to cause rupture of cell envelopes, through cycles of freezing/thawing, sonication, and rupture of the cell wall with lysozyme. However, all these methods do not guarantee a high quality of the extract, due to the complexity of the mixture obtained. In order to improve the yield of PC and to reduce the extraction process time, in this work, the conventional extraction by sonication is compared with a green innovative extraction technique, the rapid solid liquid dynamic extraction (RSLDE). This technique can generate a pressure and a consequent depression on the matrix to be extracted. The final extract was immediately frozen at −20°C and then lyophilized. UV–Vis spectrophotometric analysis and acrylamide gel electrophoresis are performed to evaluate the extraction efficiency and purity of PC.

Phycobiliproteins-A Family of Algae-Derived Biliproteins: Productions, Characterization and Pharmaceutical Potentials.

Phycobiliproteins (PBPs) are colored and water-soluble biliproteins found in cyanobacteria, rhodophytes, cryptomonads and cyanelles. They are divided into three main types: allophycocyanin, phycocyanin and phycoerythrin, according to their spectral properties. There are two methods for PBPs preparation. One is the extraction and purification of native PBPs from Cyanobacteria, Cryptophyta and Rhodophyta, and the other way is the production of recombinant PBPs by heterologous hosts. Apart from their function as light-harvesting antenna in photosynthesis, PBPs can be used as food colorants, nutraceuticals and fluorescent probes in immunofluorescence analysis. An increasing number of reports have revealed their pharmaceutical potentials such as antioxidant, anti-tumor, anti-inflammatory and antidiabetic effects. The advances in PBP biogenesis make it feasible to construct novel PBPs with various activities and produce recombinant PBPs by heterologous hosts at low cost. In this review, we present a critical overview on the productions, characterization and pharmaceutical potentials of PBPs, and discuss the key issues and future perspectives on the exploration of these valuable proteins.

Assessment of the potential of algae phycobiliprotein nanoliposome for extending the shelf life of common carp burgers during refrigerated storage

This study is focused on the effect of phycobiliprotein extraction of Gracilaria on the quality of common carp burgers, and the application of nanoliposomes containing pigment in the improvement of its antimicrobial and antioxidant activity of burgers during refrigerated storage in 18 days. Burgers were incorporated with phycobiliprotein and liposomal phycobiliprotein (2.5% and 5% w/w), and their chemical and microbial changes in terms of pH, peroxide value (PV), thiobarbituric acid (TBA), total volatile basic nitrogen (TVB-N), total viable counts (TVC), psychrotrophic bacterial counts (PTC), and sensory characteristics were evaluated. Results presented a nanoliposome size of about 515.5 nm with capable encapsulation efficiency (83.98%). Our results showed non-encapsulated phycobiliprotein could delay the deterioration of common carp burgers, as a reduction in PV, TBA, and TVB-N, TVC, and PTC values in burgers treated with free and nano encapsulated phycobiliprotein. Moreover, the potential of phycobiliprotein was improved when it was encapsulated into chitosan coated liposomes. Burgers treated with 5% nanoliposomes displayed the lowest amount of lipid oxidation and microbial deterioration in comparison to others during storage. According to chemical, microbial and sensory evaluation, the shelf life of common carp burgers was increased in samples treated with encapsulated phycobiliprotein at 2.5% and 5%, as compared to the control (p ≤ 0.05).

Uncovering Research Trends of Phycobiliproteins Using Bibliometric Approach.

Phycobiliproteins are gaining popularity as long-term, high-value natural products which can be alternatives to synthetic products. This study analyzed research trends of phycobiliproteins from 1909 to 2020 using a bibliometric approach based on the Scopus database. The current findings showed that phycobiliprotein is a burgeoning field in terms of publications outputs with “biochemistry, genetics, and molecular biology” as the most related and focused subject. The Journal of Applied Phycology was the most productive journal in publishing articles on phycobiliproteins. Although the United States of America (U.S.A.) contributed the most publications on phycobiliproteins, the Chinese Academy of Sciences (China) is the institution with the largest number of publications. The most productive author on phycobiliproteins was Glazer, Alexander N. (U.S.A.). The U.S.A. and Germany were at the forefront of international collaboration in this field. According to the keyword analysis, the most explored theme was the optimization of microalgae culture parameters and phycobiliproteins extraction methods. The bioactivity properties and extraction of phycobiliproteins were identified as future research priorities. Synechococcus and Arthrospira were the most cited genera. This study serves as an initial step in fortifying the phycobiliproteins market, which is expected to exponentially expand in the future. Moreover, further research and global collaboration are necessary to commercialize phycobiliproteins and increase the consumer acceptability of the pigments and their products.

A Review of Phycobiliproteins of Cyanobacteria: Structure, Function and Industrial Applications in Food and Pharmaceutical Industries

Phycobiliproteins are accessory photosynthetic pigments with a tetrapyrrole structure derived from bacterial strains that are organized on a thylakoid membrane inside a structure called phycobilisomes. Phycobiliproteins have been extensively commercialized in the manufacture of fluorescent probes for clinical and immunological analysis, in addition to their ability to dye with proven antioxidant and medicinal properties. Phycocyanin, Phycoerythrin and Allophycocyanin are the main types of phycobiliproteins that are widely used as useful food supplements today; however, in Iran, the true value of this natural pigment with bioactive properties has not been realized. Todays, the use of artificial colors and antioxidants in food product has led to an increase in cancer in many people. Therefore, awareness of the presence of natural food pigments of natural origin is of particular important. On the other hand, since so far no review article on extraction, separation and purification as well as evaluation of the biological activity of the pigment Phycoerythrin and Phycocyanin, has been published in Iran, so such review articles can pave the way for the introduction of natural edible pigments from cyanobacteria are considered to be usable in the food industry. Therefore, the purpose of this review article is to introduce the structure, function, biosynthesis and different methods of extraction of Phycobiliproteins in industrial dimensions along with different applications of Phycocyanin in food and pharmaceutical industries.

A successful method for phycocyanin extraction from Arthrospira platensis using [Emim] [EtSO4] ionic liquid

AbstractResearch into the extraction of phycobiliprotein from Arthrospira platensis is currently ongoing. Most traditional methods of phycobiliprotein extraction include the use of organic solvents, which negatively affect the sustainability of the process. Ionic liquids (ILs) are a promising alternative for phycobiliprotein extraction due to their properties as green solvents. In the present work an imidazolium‐based IL and sonication were studied for phycobiliprotein and carbohydrate extraction. A factorial experimental design was used to optimize the amount of extracted phycocyanin. The maximum extraction yield was achieved by using low biomass/solvent ratios combined with high IL/water ratios and sonication powers, and long operation times. The recovery of IL was studied under these conditions using a dialysis‐based process to separate the IL from the extracted phycobiliproteins. The results reveal the possibility of using the recovered IL for seven consecutive extraction cycles with an acceptable phycocyanin extraction amount, from 75 mg g−1 (fresh IL) to 60 mg g−1 (7 cycles of reused IL). The use of [Emim] [EtSO4] + ultrasound is a promising alternative for phycocyanin extraction, enhancing the results obtained with other IL in terms of extracted phycocyanin and the number of times the IL can be reused. © 2021 Society of Chemical Industry and John Wiley & Sons, Ltd

Bioprospecting of cyanobacterium in Chilean coastal desert, Geitlerinema sp. molecular identification and pressurized liquid extraction of bioactive compounds

This study presents a new cyanobacterium of the genus Geitlerinema sp. isolated from the Chilean coastal desert as a rich source of bioactive compounds. Phylogenetic tree is shown along with a broad biochemical characterization using conventional and pressurized liquid extraction. Variables as extraction yield, phycobiliproteins, lipids and methyl palmitate (C16:0) were studied by means of two experimental designs based on a response surface methodology (RSM) employing two factorial design 32. Factors as temperature (20–200 °C) and percentage of solvents were selected resulting in Factorial Design A with 0–100% (v/v) of ethanol–water and Factorial Design B with 0–100 (v/v) of ethanol–limonene solvents. The main bioactive compounds were total proteins (∼52% wt), phycobiliproteins (especially C-phycocyanin) and methyl palmitate (C16:0). Data obtained showed that water and ethanol:water (1:1, v/v) improved the extraction yield and phycobiliproteins recovery. As for ethanol:limonene extracts, they were better for lipids and methyl palmitate extractions. In general, high temperatures only improved the extraction yield whereas low temperatures favor the extraction of phycobiliproteins. Therefore, results indicated that Geitlerinema sp. could be one of the several cyanobacteria genera that display functional properties for nutritional, pharmaceutical or industrial use as sustainable and cost-effective bioactive compounds in algal biorefinery model.

Production of Phenyllactic Acid from Porphyra Residues by Lactic Acid Bacterial Fermentation

The concept of algae biorefinery is attracting attention because of the abundant valuable compounds within algal biomass. Phenyllactic acid (PhLA), a broad-spectrum antimicrobial organic acid that can be produced by lactic acid bacteria (LAB), is considered a potential biopreservative. In this study, a cascading biorefinery approach was developed to harvest PhLA from Porphyra residues by LAB fermentation. LAB strains were cultivated in de Man, Rogosa and Sharpe (MRS) broth to screen their ability to produce PhLA. As the strains of Lactobacillus plantarum KP3 and L. plantarum KP4 produced higher concentrations of PhLA at 0.09 mg/mL, these two strains were employed for fermentation. After phycobiliprotein extraction, the Porphyra residues, ultrafiltration eluate, phenylalanine (Phe) and yeast extract with a volume of 20 mL were inoculated with LAB strain KP3 and fermented at 37 °C for 120 h. The PhLA content of the fermented broth was 1.86 mg. To optimize the biorefinery process, the ultrafiltration eluate was replaced by commercial cellulase. Up to 4.58 mg of PhLA, which was 2.5 times greater than that produced from KP3 cultured in MRS broth, could be harvested. Taken together, the findings provide an optimized process for LAB fermentation, which is shown to be a feasible algae biorefinery approach to obtaining PhLA from Porphyra residues.

Enzyme-Assisted Method for Phycobiliproteins Extraction from Porphyra and Evaluation of Their Bioactivity

Due to the poor protein availability of algae in their unprocessed form, development of extraction methods for phycobiliproteins is of great significance. This study aimed to extract phycoerythrin (PE) and phycocyanin (PC) from Porphyra via bacterial enzymatic hydrolysis and to evaluate their bioactivity. To induce enzyme production, Porphyra powder was added into the culture medium of two marine bacterial strains. The pH and enzyme activity of the cultured supernatant, namely crude enzyme solution, were significantly raised. For PE and PC extraction, Porphyra were incubated within crude enzyme solution with homogenization and ultrasonication followed by ultrafiltration process. After distinguishing by fast performance liquid chromatography (FPLC), three major fractions were observed and identified as R-PE, R-PC and small molecular PE by high-performance liquid chromatography (HPLC) and polyacrylamide gel electrophoresis (PAGE) analysis. With respect to bioactivity, these three fractions exhibited free radical scavenging and antioxidant activities in a various degree. In addition, the angiotensin-converting-enzyme (ACE) inhibitory activity of both R-PE and R-PC fractions was observed in a concentration-dependent manner. Taken together, the employed process of bacterial enzymatic hydrolysis is suggested to be a feasible method to obtain PE and PC from Porphyra without limiting their bioactivity.

Evaluation of novel 3D-printed monolithic adsorbers against conventional chromatography columns for the purification of c-phycocyanin from Spirulina

Extraction and purification of high-grade phycobiliproteins is a multistep process that typically involves costly chromatographic separation techniques. 3D printing approaches and materials can now be tailored to fabricate low-cost anion exchange monolithic adsorbers for chromatography. In this work, a new printing formulation was used to produce monolithic discs with quaternary amine functionality. The 3D-printed discs were tested for the purification of the phycobiliproteins c-phycocyanin and allophycocyanin from extracts of the cyanobacterium Arthrospira platensis (Spirulina). Analytical grade c-phycocyanin with 41 ± 7% recovery was obtained with the 3D-printed discs in a single step. The 3D-printed discs also demonstrated similar or higher separation properties when compared with commercial Q-Sepharose and commercial monolithic discs. Reproducibility tests showed that the fabrication of the 3D-printed discs was robust and potentially scalable for industry-scale purification. Overall, the operative flexibility, robustness and separation performance of the 3D-printed discs demonstrated the promising potential of monolithic adsorbers for the separation of a wide range of biomolecules.