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Effect of pretreatment methods and enzymatic hydrolysis on the protein content of Mastocarpus stellatus. Dry weight = 33.3% of wet algae. a water-soluble proteins (mg g⁻¹ dw); b R-PE yield (mg g⁻¹dw); c R-PE Purity Index (PI) (n = 3, bars = s.d.)
Source publication
This study reports the extraction of R-phycoerythrin (R-PE) from the red seaweed, Mastocarpus stellatus. Phosphate buffer 20 mM with pH 7.1 showed better results for R-PE extraction following classical maceration. The grinding of freeze dried material in liquid nitrogen aided greater extraction of R-PE. The treatment of freeze-dried seaweed with di...
Citations
... However, alternative and innovative methods would be valuable, notably to avoid expensive pre-treatments, and to save time and increase the extraction of other compounds. In this context, enzyme-assisted extraction (EAE) has demonstrated its interest for R-PE extraction from various red seaweed species [28][29][30], but without success on G. turuturu [31]. Over the past two decades, different innovative techniques such as microwave-assisted extraction, supercritical fluid extraction and pressurized liquid extraction, have emerged to retrieve biomolecules from seaweeds [32][33][34]. ...
... In addition, the efficiency of this combined process would be influenced by numerous operating parameters related to enzymes, free or immobilized, and ultrasound devices [38,53,55,56] that need to be systemically investigated and optimized. Previous works have already demonstrated that experimental designs are an efficient tool to improve the R-PE extraction yields from other red seaweeds by enzymatic hydrolysis [28][29][30], to optimize the UAE of compounds from Ascophyllum nodosum [44,57], and the UAEH of compounds of plant and animal origin such as bioactive polysaccharides from a plant leaves and molluscs [58,59]. ...
Enzyme-assisted extraction (EAE) and ultrasound-assisted extraction (UAE) are both recognized as sustainable processes, but little has been done on the combined process known as ultrasound-assisted enzymatic hydrolysis (UAEH), and even less on seaweed. The present study aimed to optimize the UAEH of the red seaweed Grateloupia turuturu for the extraction of R-phycoerythrin (R-PE) directly from the wet biomass by applying a response surface methodology based on a central composite design. Three parameters were studied: the power of ultrasound, the temperature and the flow rate in the experimental system. Data analysis demonstrated that only the temperature had a significant and negative effect on the R-PE extraction yield. Under the optimized conditions, the R-PE kinetic yield reached a plateau between 90 and 210 min, with a yield of 4.28 ± 0.09 mg·g−1 dry weight (dw) at 180 min, corresponding to a yield 2.3 times higher than with the conventional phosphate buffer extraction on freeze-dried G. turuturu. Furthermore, the increased release of R-PE, carbohydrates, carbon and nitrogen can be associated with the degradation of G. turuturu constitutive polysaccharides, as their average molecular weights had been divided by 2.2 in 210 min. Our results thus demonstrated that an optimized UAEH is an efficient method to extract R-PE from wet G. turuturu without the need for expensive pre-treatment steps found in the conventional extraction. UAEH represents a promising and sustainable approach that should be investigated on biomasses where the recovery of added-value compounds needs to be improved.
... Drying the plant biomass prior to extraction has been shown to have a significant impact on the protein yield obtained during subsequent EAE. Dry algal biomass (Mastocarpus stellatus) resulted in a higher protein yield (2.94 mg protein/g dw) compared to when wet biomass was used (1.72 mg protein/g dw) (Nguyen et al. 2017). The higher protein yield from the freeze-dried sample may be linked to the smaller particle sizes obtained following homogenisation in liquid nitrogen, whereas the wet biomass was cut into <1 cm pieces prior to freezing. ...
... In addition, reaction pH was reported to have a major contribution to the extraction yield of specific proteins, e.g., during the extraction of R-phycoerythrin (R-PE) which is a major component of the light-harvesting pigment protein complexes, known as phycobiliproteins, found in red algae. The highest protein (0.32 mg/g dw) and R-PE (0.27 mg/g dw) yields were obtained at pH 7.1 compared to a lower pH extraction, i.e., at pH 6.5 (Nguyen et al. 2017). Furthermore, reaction pH not only affects protein yield but can also influence the functional properties of the recovered proteins (Rommi et al. 2015). ...
Enzyme-assisted extraction (EAE) is an environmentally friendly green processing technique used to aid protein extraction from different plant sources. This is due to its mild operating conditions, reduced waste generation and low energy consumption compared to chemical and physical extraction approaches. A range of food grade carbohydrase and protease preparations have been employed to aid protein extraction/solubilisation from different plant sources by hydrolysis of the plant cell wall and the proteins therein. Different statistical tools can be employed to optimise enzyme treatment parameters including enzyme:substrate, pH, incubation temperature and hydrolysis duration to yield maximal protein recovery. While EAE facilitates protein recovery, it may also enhance the nutritional (digestibility) and techno- and bio-functional properties of the extracted proteins, particularly when using protease-assisted extraction. Combining EAE with physical techniques, e.g., ultrasonic processing, for biomass pre-treatment can enhance plant cell wall disruption with a view to enhancing protein extraction efficiency. This approach can facilitate economic feasibility by reducing the energy required and the quantity of enzyme used and, therefore, the overall cost of the extraction process. An overview of the application of EAE in protein/peptide recovery from different plant sources including oilseeds, nuts, cereals, pulses and algae is provided herein.
... The latter authors also highlighted the notable protein content of this seaweed (21.3%, dry weight), although lower values were identified in other works (15%, dry weight) (Blanco-Pascual et al. 2014a). The red color of this seaweed is attributed to pigments such as phycobiliproteins, considered a light-harvesting protein complex founds in chloroplast of red seaweeds (Nguyen et al. 2017) and their extracts are used as natural dye, food colorant, dietary supplements and are incorporated to functional products (Mysliwa-Kurdziel & Solymosi, 2016;Fleurence et al., 2018). Mastocarpus stellatus is considered a reliable source of hybrid carrageenans. ...
This study focusses on the valorization of the red seaweed Mastocarpus stellatus, collected in the Northern coast of Portugal, as a natural source of high value compounds due to its beneficial properties. An annual monitoring of the seaweed was performed by determining its lipids, ash, carbohydrates, phycobiliproteins, total phenolic compounds, antioxidant capacity and carrageenan from three different rocky shores located in the north of Portugal. The results showed a seasonal and spatial variability of the studied compounds between October 2018 and September 2019 depending on the climatic variables of temperature, precipitation, and solar radiation. The most productive season coincided with the warmest months, except for carbohydrates and phycobiliproteins, which were promoted in the colder season. The spatial variation also could be explained by the proximity to water channel discharges at the sampling sites. Complementary studies on the carrageenan fraction were conducted in one of the shores due to the high biopolymer content, to determine their carrageenan proportion between the summer and winter period and establish their rheological capabilities for the formulation of gelling matrices. The extracted biopolymers exhibited typical structural and viscoelastic characteristics of kappa/iota-hybrid carrageenans, suggesting notably differences depending on the harvest season, which is critically relevant to define future applications.
... Different groups of substances can be extracted depending on the solvent used. Watersoluble proteins and R-phycoerythrin were extracted from macroalgae M. stellatus [219] with solutions such as tap water, pure water, 0.1 mol·L −1 phosphate buffer (pH 6.5), 20 mmol phosphate buffer (pH 7.1), and 50 mmol phosphate buffer (pH 6.45). The best results were observed when extracting with 20 mmol buffer solution. ...
The beneficial properties of algae make them perfect functional ingredients for food products. Algae have a high energy value and are a source of biologically active substances, proteins, fats, carbohydrates, vitamins, and macro- and microelements. They are also rich in polyunsaturated fatty acids, proteins, mycosporine-like amino acids, polysaccharides, polyphenols, carotenoids, sterols, steroids, lectins, halogenated compounds, polyketides, alkaloids, and carrageenans. Different extraction parameters are used depending on the purpose and the substances to be isolated. In this study, the following parameters were used: hydromodule 1:10 and an extraction duration of 1–2 h at the extraction temperature of 25–40 °C. A 30–50% solution of ethanol in water was used as an extractant. Algae extracts can be considered as potential natural sources of biologically active compounds with antimicrobial activity and antiviral properties. The content of crude protein, crude fat, and carbohydrates in U. Prolifera, C. racemosa var. peltata (Chlorophyta), S. oligocystum and S. fusiforme (SF-1) was studied. It was found that C. muelleri (Bacillariophyta), I. galbana (Haptophyta), and T. weissflogii (Bacillariophyta) contain about 1.9 times more omega-3 than omega-6 fatty acids. N. gaditana (Ochrophyta), D. salina (Chlorophyta), P. tricornutum (Bacillaryophyta) and I. galbana (Haptophyta) extracts showed inhibitory activity of varying intensities against E. coli or P. aeruginosa. In addition, algae and algae-derived compounds have been proposed to offer attractive possibilities in the food industry, especially in the meat sector, to evolve functional foods with myriad functionalities. Algae can increase the biological activity of food products, while the further study of the structure of compounds found in algae can broaden their future application possibilities.
... Cellulase, coupled with carrageenase or agarase, was reported to markedly improve the protein yield from Chondrus and Gracilaria compared to the individual application and non-enzymatic process (Fleurence et al., 1995). Similarly, the combined action of cellulase plus xylanase appeared to increase the R-PE yield Nguyen, Morançais, Fleurence, & Dumay, 2017). On the contrary, the cocktail of cellulase and α-amylase did not significantly affect the extraction yield compared to those achieved with individual enzymes on C. chamissoi (Vásquez et al., 2019). ...
Background
With the increase in world population, decreased farmland, and global climate changes, ensuring adequate food supply to maintain food security is of immediate attention. As a key macronutrient for human health, the supply of sufficient dietary protein is undoubted of concern. Animal proteins are good dietary protein sources, but their production incurs a high carbon footprint; this has driven the effort to seek alternative protein sources.
Scope and approach
This review aimed to elaborate on the scientific research progress in red seaweed proteins, including the nutrition, functionalities, methods of extraction, and to explore their prospects as an alternative protein source. Applications of red seaweed protein in food and nutraceutical industries, environmental impact, affordability, and related safety concerns were also discussed.
Key findings and conclusions
Red seaweeds have a comparable essential amino acid profile to ovalbumin, representing a sustainable alternative to terrestrial proteins. Pre-treatment and extraction methods are pivotal in modulating protein digestibility and functionalities; enzymatic extraction approaches appear to improve nutritional value and food functionalities. Red seaweed proteins have a wide range of applications in food based on their physicochemical properties, while their bioactivities can be tailored for nutraceutical purposes. The use of red seaweed proteins as functional food ingredients is emerging, with good potential in bioactive microencapsulation. Efforts are required to improve the seaweed cultivation process to a commercial scale and gain consumer acceptance in the western world. More research is also necessary to enhance seaweed protein extraction and improve their functionalities for food and nutraceutical applications.
... Common solvents applied and tested among researchers, to retrieve PE from red seaweeds, are phosphate buffer [35,46,50,58,69,70,73,78,79,89,90,93] and sodium phosphate buffer [53,103], featured prominently in Table 1. However, solvents such as potassium phosphate buffer [96], distilled water [45,77,83,91], acetic acid-sodium acetate [81,102], citrate buffer [57,74], diethyl ether [74], and EDTA [84], have also been found to be effective. ...
Algae taxa are notably diverse regarding pigment diversity and composition, red seaweeds (Rhodophyta) being a valuable source of phycobiliproteins (phycoerythrins, phycocyanin, and allophycocyanin), carotenes (carotenoids and xanthophylls), and chlorophyll a. These pigments have a considerable biotechnological potential, which has been translated into several registered patents and commercial applications. However, challenges remain regarding the optimization and subsequent scale-up of extraction and purification methodologies, especially when considering the quality and quantity needs, from an industrial and commercial point of view. This review aims to provide the state-of-the-art information on each of the aforementioned groups of pigments that can be found within Rhodophyta. An outline of the chemical biodiversity within pigment groups, current extraction and purification methodologies and challenges, and an overview of commercially available products and registered patents, will be provided. Thus, the current biotechnological applications of red seaweeds pigments will be highlighted, from a sustainable and economical perspective, as well as their integration in the Blue Economy
... The PE concentration obtained from 50-100 µm was similar to the concentration of PE found in Mastocarpus Stellatus (2.0 mg/g dry seaweed) [24]. Repetitive extractions would be needed to determine the total concentration of PE in the Chondrus crispus. ...
To better understand the migration properties of hybrid carrageenan from the seaweed tissue during carrageenan extraction, the effect of increasing the seaweed surface area by the mechanical disintegration of gametophyte Chondrus crispus chips was studied under various temperature and time extraction conditions. Dried Chondrus crispus seaweed chips were milled by a rotor beater mill and classified into eight different size fractions by sieving with varying mesh sizes from 50 to 2000 μm. During extraction at 22 °C, the red color of the filtrate increased significantly with the decreasing particle size of the fraction, correlating with the increasing phycoerythrin concentration (from 0.26 mg PE/g dry seaweed in the >2000 μm size fraction to 2.30 mg PE/g dry seaweed in the <50 μm size fraction). On the other hand, under the same extraction conditions, only a small increase in carrageenan precipitate was obtained with the decreasing size fractions (from no recovery in the >2000 μm size fraction to 2.1 ± 0.1 g/kg filtrate in the <50 μm size fraction). This yield was significantly lower than the ones from extractions at 45 °C (5.4 ± 0.1 g/kg) or at 90 °C (9.9 ± 2.1 g/kg) for the same particle size and time conditions. It could be concluded that hybrid carrageenan extraction is not surface area dependent, while phycoerythrin is. Therefore, it seems that phycoerythrin and carrageenan extraction follow different mechanisms. This creates potential for the selective extraction of each of those two compounds.
... Currently, a limited number of red algae and microalgae species such as Porphyra haitanensis, Porphyridium purpureum, Bangia fusco-purpurea, Synechococcus leopoliensis, and Spirulina platensis are being used to drive commercial applications of PE production (Chaloub et al., 2015). The key factors are cultivation thresholds in indoor systems, productivity, the difficulty of extraction due to high polysaccharide content, and PE stability (Nguyen et al., 2017;Sekar & Chandramohan, 2008). Some research has tried to use other potential species, such as Rhodomonas sp. ...
Phycoerythrin (PE) is a compound with strong potential for both basic research and industrial applications, but short supply and high prices have so far hindered its development. One common problem is a shortage of biomass for extraction. The aim of the present study was to determine a cultivation strategy (optimizing temperature, irradiance, photoperiod, and light quality) to produce greater biomass and higher PE concentrations in the alga Colaconema sp. We found that an optimized culture process could increase algae growth 7–9 fold while allowing extraction of 9–10 mg g⁻¹ total phycobiliproteins, containing 60%–65% PE. Low energy costs make this approach economically feasible and competitive when compared with existing methods. Our results suggest an improved strategy for the large-scale production of PE and offer valuable applications in the algae industry.
... Reports are available on the application of enzymes for the extraction of various biomolecules, which is also applicable for microalgae [48][49][50][51][52]. A few pieces of literature on primary extraction of PBP (R-phycoerythrin, R-PE) from macroalgae are available [12,53,54]. The consortia of enzymes could able to digest the matrix of polysaccharides resulting in a higher degree of extraction. ...
Algae (both micro and macro) have gained huge attention in the recent past for their high commercial value products. They are the source of various biomolecules of commercial applications ranging from nutraceuticals to fuels. Phycobiliproteins are one such high-value low volume compounds that are mainly obtained from micro and macroalgae. In order to tap the bioresource, a significant amount of work has been carried out for large scale production of algal biomass. However, work on downstream processing aspects of phycobiliproteins (PBPs) from algae is scarce, especially in the case of macroalgae. There are many difficulties in cell wall disruption of both micro and macro algae because of their cell wall structure and compositions. At the same time, there are several challenges in the purification of phycobiliproteins.
The current review article focuses on the recent developments in downstream processing of phycobiliproteins (mainly phycocyanins and phycoerythrins) from micro and macroalgae. The current status, recent advancements and potential technologies (that are under development) are summarised in this review article besides providing future directions for the present research area.
... Response surface methodology (RSM), utilizing a complete quadratic polynomial to demonstrate the relationships, is a regularly used and efficient biotechnology optimization method. A central composite design (CCD) in RSM, a statistical approach that fully considers the interaction and influence between variables, has been utilized, which is widely applied in media conditions [21][22][23] and enzyme production of media components [24][25][26]. ...
The strains capable of degrading cellulose have attracted much interest because of their applications in straw resource utilization in solid-state fermentation (SSF). However, achieving high spore production in SSF is rarely reported. The production of spores from Streptomyces griseorubens JSD-1 was investigated in shaker-flask cultivation in this study. The optimal carbon, organic nitrogen and inorganic nitrogen sources were sucrose, yeast extract and urea, respectively. Plackett–Burman design (PBD) was adopted to determine the key medium components, and the concentration levels of three components (urea, NaCl, MgSO4·7H2O) were optimized with the steepest ascent path and central composite design (CCD), achieving 1.72 × 109 CFU/g of spore production. Under the optimal conditions (urea 2.718% w/v, NaCl 0.0697% w/v, MgSO4·7H2O 0.06956% w/v), the practical value of spore production was 1.69 × 109 CFU/g. The determination coefficient (R2) was 0.9498, which ensures an adequate credibility of the model.
