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Application of ultrasound for green extraction of proteins from spirulina. Mechanism, optimization, modeling, and industrial prospects
... [ [155][156][157][158][159][160][161][162][163] Irradiation-Assisted Extraction ...
... Another comparative investigation involving three edible macroalgae-Sargassum wightii, Ulva rigida, and Gracilaria edulis-revealed that UAE at 120 min markedly increased phenolic and flavonoid levels in S. wightii, alongside significant rises in antioxidant activity (by ferric-reducing antioxidant power and DPPH radical scavenging) compared to traditional extraction [157]. Research has also indicated that UAE can outstrip other extraction techniques, achieving enhanced protein recovery rates in marine algae [159] and accelerating lipid or carbohydrate extraction in species such as Nannochloropsis oculata and Rhodosporidium toruloides, respectively [158,161,162]. ...
Marine macroalgae (commonly known as seaweeds), a rich yet underexplored resource, have emerged as a promising source of bioactive compounds with potent antioxidant properties. While oxidative stress is a critical factor in the pathogenesis of numerous chronic diseases, including neurodegenerative disorders, cardiovascular conditions, and cancer, macroalgae-derived compounds such as polyphenols, carotenoids, peptides, and sulfated polysaccharides have demonstrated the ability to mitigate oxidative damage through multifaceted mechanisms. These compounds neutralize reactive oxygen species and modulate key cellular pathways involved in inflammation and apoptosis. Despite significant advancements, gaps persist in understanding the pharmacokinetics, bioavailability, and clinical applications of these bioactives. Additionally, the inefficiencies of traditional extraction methods call for adopting innovative, environmentally friendly techniques that preserve bioactivity. This review synthesizes current knowledge on the therapeutic potential of macroalgal bioactives, acknowledges the contributions of other marine algae where relevant, highlights challenges in their extraction, and proposes future directions for research and application.
... Vernès et al. [12] further optimized the UAE process by combining it with an aqueous two-phase extraction system, which led to higher purity levels of phycocyanin. This combined approach also reduces processing times and makes large-scale production more feasible due to its energy efficiency and scalability. ...
... This combined approach also reduces processing times and makes large-scale production more feasible due to its energy efficiency and scalability. The authors showed that this method increased the purity of phycocyanin from 0.42 to 1.31 in a single extraction step [12]. ...
The cyanobacterium Limnospira platensis, vulgarly Spirulina, has gained significant attention due to its high protein content, rich bioactive compounds, and health benefits, making it a valuable resource in biotechnology, nutraceuticals, food supplements, biopharmaceuticals, and cosmetics. Recent advancements in fermentation technology have considerably improved the efficiency, scalability, and cost-effectiveness of L. platensis production while addressing environmental sustainability and enhancing product quality. Based on well-recognized databases (Google Scholar, PubMed, Scopus, Web of Science), this review explores the latest developments in L. platensis fermentation, emphasizing strain improvement, bioprocess engineering, and optimization of fermentation parameters. It also examines key factors such as bioreactor design, downstream processing, and innovative monitoring technologies aimed at maximizing biomass yield and bioactive compound production. Additionally, emerging applications of fermented L. platensis in various industries and future perspectives, including large-scale production, regulatory barriers, and biosafety considerations, are discussed. These insights provide a comprehensive outlook on the future of L. platensis fermentation in biotechnological applications.
... Modern green extraction techniques such as ultrasound-assisted extraction (UAE), high-pressure homogenization (HPH), pressurized water extraction (PWE), and supercritical carbon-dioxide extraction between others eliminate these problems and become popular alternatives due to their environmental friendliness, better extraction efficiency, and faster extraction process [20,21]. UAE, which uses ultrasound for particle disruption, is one of the most widely used green extraction techniques for PBP extraction due to its simplicity and general availability of equipment such as the ultrasonic baths and probes [22][23][24][25][26]. In recent years, there is a number of works that apply pressures up to 500 MPa for PBP isolation [27,28], suggesting the potential of using high pressures for extractions. ...
... From the feasibility point of view, the UAE using probe seems to be a less suitable method than PWE due to the necessary optimization of ultrasonic on/off cycles, the need to externally cool the sample during extraction, and the difficulty of using larger sample amounts and solvent volumes due to the need of larger containers to prevent extract escape. In addition, local overheating of the sample could occur during UAE using probe, which may have caused inadvertent breakage of longer protein chains during intensive sonication [24]. Furthermore, UAE requires additional sample handling such as centrifugation and filtration. ...
Introduction
Cyanobacterium Arthrospira platensis (AP) (Nordstedt) Gomont contains high content of phycobiliproteins (PBP), which are an important source for food industry. Methods effectively extracting proteins contained in AP cells are demanded to provide a supply of the material. Water‐based extraction methods are advisable due to the high solubility of the PBP.
Objectives
Extraction techniques such as ultrasound assisted extraction (UAE) and pressurized water extraction (PWE) are popular due to their environmental friendliness, better extraction efficiency, and faster extraction process. In this paper, efficiency of the two methods is compared.
Materials and Methods
PWE along with UAE is utilized for release of PBP from the AP cells. The extraction parameters including time, temperature, pressure, and ultrasound intensity are tested to obtain the most efficient setup. The methods were evaluated using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS‐PAGE), and the replicates of PWE extracts were further analyzed by capillary isoelectric focusing with laser‐induced fluorescence (cIEF‐LIF).
Results
The developed PWE method using higher pressure treatment at lower temperature was significantly faster than UAE methods, and the SDS‐PAGE results showed a high content of phycobiliproteins in the extracts. cIEF‐LIF analysis showed that the sequential PWE of individual samples was repeatable, and the mild extraction provided a fluorescent profile similar to the commercially available C‐phycocyanin standard.
Conclusion
Pressurized water extraction was shown to be an efficient, rapid, and well‐automated extraction method for AP proteins in general, including bioactive phycobiliproteins. Obtained results encourage the use of PWE in small‐scale analytical applications for primary extraction of proteins.
... In addition, effective protein recovery from Spirulina requires cell disruption to release intracellular proteins (Vernès et al. 2019). Ultrasound-assisted extraction (UAE) has been used in fractionation procedures for microalgae and cyanobacteria, facilitating cell disruption and protein release (Deng et al. 2023). ...
... This result may reflect an increase in the collapse pressure of the bubbles formed during acoustic cavitation, as reported in a previous study . Increasing the pressure also enhances the diffusion of molecules out of cell residues (Vernès et al. 2019). At high power (300 W), an increase in pressure from 0 to 3 bar resulted in a significant decrease in protein extraction yield, from 71.7 to 9.5%, probably due to the combined effects of high pressure and high power, as discussed above. ...
The cyanobacterium Spirulina platensis was subjected to a fractionation process involving ultrasound-assisted extraction and membrane filtration to obtain a pure phycocyanin fraction and a clarified colorless protein fraction free of chlorophyll and carotenoids. The effects of pressure and power on total protein release were assessed. The retention of the extracted proteins was then assessed by ultrafiltration, with and without ammonium sulfate precipitation. Total protein recovery yields reached 97% in aqueous solution, at a low frequency (12 kHz), atmospheric pressure, and with an ultrasonic power of 200 Watts (W). Ammonium sulfate (25% w/v) precipitation was used to remove pigments and impurities from the crude protein extract. Finally, semi-frontal ultrafiltration resulted in high levels of C-phycocyanin recovery in the retentate: 95% and 91% with 10 and 100 kDa-cutoff membranes, respectively. However, the levels of total non-pigmented proteins in the permeate compartment did not exceed 67% with a 100 kDa-cutoff membrane. A fractionation process is proposed here for the valorization of two different protein fractions from Spirulina platensis.
... Specifically, ultrasonication yielded longer roots, but non-ultrasonicated biomass yielded wider leaves, both cases in eggplants. Ultrasonication of microalgae aqueous solutions induces rapidly collapsing, acoustic cavitation bubbles, causing localized, short-term pressure changes and shockwaves that rupture microalgal cells (Vernes et al., 2019). This facilitates the extraction of bioactive substances, including proteins, polyphenols, carotenoids, phycobiliproteins, and more (Bhosle et al., 2015;Poojary et al., 2016). ...
... There are three potential reasons why this hypothesis was not met: (i) Sunday-drying the biomass is sufficient for cell lysis, (ii) the Sunday-drying and ultrasonication methods applied did not sufficiently burst cell walls and membranes, and (iii) more time is needed for treatment differences to become apparent. Regarding the former, while ultrasonication is a preferred method for cell lysis in microalgae, Sunday-drying is also an effective method (Vergel-Suarez et al., 2023;Vernes et al., 2019). In a study comparing effectiveness of drying methods, Madhubalaji et al. (2021) found that Sunday-drying and lyophilization retained more bioactive compounds and antioxidant activities than the alternatives (oven drying, drum drying, and spray drying). ...
Biofertilizers offer a sustainable alternative to chemical fertilizers, notorious for their harmful impact on the environment and human health. Sources of biofertilizers include cyanobacteria, such as those from the genus Arthrospira, which contain bioactive compounds that enhance plant growth directly and indirectly. To promote widespread biofertilizer adoption, experimental trials are essential to compare their efficacy against chemical fertilizers and different biofertilizer varieties. Some biofertilizers rely on ultrasonication for cell rupture, but since A. maxima undergoes sun‐drying to obtain the biomass powder, which also ruptures cells, ultrasonication may be unnecessary. We assessed the biofertilizer capabilities of Arthrospira maxima (Setchell & N.L. Gardner) Geitler biomass on three cash crops, banana (Musa acuminata Colla), cowpea (Vigna unguiculata L. Walp.), and eggplant (Solanum melongena Mill. Dunal). We compared sonicated and non‐sonicated biomass (both 20 g L⁻¹) against a commercially available, brown algae Ascophyllum nodosum (L.) biofertilizer (4 mL L⁻¹ OptiMar), a commercially available chemical fertilizer (1.5 mL L⁻¹ Triple15), and a negative control of tap water. Few differences were observed among A. maxima treatments, suggesting sonication might be unnecessary. OptiMar, Triple15, and the negative control did not yield the highest biometric values of any trait in any crop. If commercially available fertilizers and biofertilizers do not outperform the negative control, it raises crucial questions regarding their effectiveness and the appropriate concentrations at which they should be applied. Non‐sonicated biomass often outperformed OptiMar, Triple15, and the negative control, demonstrating the potential Arthrospira maxima has for stimulating plant growth without ultrasonication.
... Using ultrasound-intensified hydrolysis has been found to enhance extraction yields and decrease the duration of collagen extraction from Japanese sea bass skin and microalgae (Hyun Kyung Kim et al., 2012;Vernès et al., 2019). The results obtained via ultrasonic extraction indicate that the proteins were largely intact, indicating minimal occurrence of protein breakdown. ...
... (Mahali & Sibi, 2019). The research investigation on A. platensis determined that using UAE (20 kHz) with a sodium phosphate buffer significantly enhanced protein extraction by 229% (Vernès et al., 2019). ...
... As extraction power increased, the yield initially increased, then decreased, attaining a maximum at 320 W. When the power was further increased, the yield decreased and fell below the initial level at 400 W. Higher power levels led to the formation of larger vapor bubbles, resulting in more intense implosions. The violent implosion effect releases high-intensity micro-jets and shockwaves, which induce cell wall rupture and facilitate the release of GFG [21]. However, at a certain critical value, ultrasonic power reaches a saturation point, whereby further increases result in the formation of invalid bubbles, which enhance scattering attenuation and reduce cavitation strength [22]. ...
Ultrasound-assisted extraction (UAE) is a technique that can enhance the efficiency of the extraction of bioactive ingredients. In previous work, ginkgo flavonol glycosides (GFG) were extracted using a deep eutectic solvent (DES). To further enhance mass transfer efficiency in this work, ultrasound-assisted deep eutectic solvent (UADES) was employed for GFG extraction. The optimal extraction parameters were determined to be the ultrasonic power 320 W, extraction time 63.6 min, and extraction temperature 32.5℃. The extraction yield of GFG reached 5.60 mg/g, surpassing that achieved through DES extraction. Notably, the extraction time was drastically shortened from 11.8 h to 63.6 min. The analysis of changes in micro-morphology, crystalline structure, and cellulose content of the phytoextraction solid residue revealed that the UADES effectively disrupted the crystalline regions of lignocellulose within the cell wall, and the ultrasound enhanced the solubilization of DES to lignocellulose, thus improving the efficiency of the extraction process.
... However, future studies could explore a direct comparison between MAE and UAE to further assess their relative efficiencies under similar conditions. Table 1 presents a comparative analysis of protein extraction yields from Spirulina and Chlorella biomass using various extraction methodologies, as reported in recent studies [39][40][41][42][43][44]. The results from this study demonstrate that microwave-assisted DES extraction significantly outperforms other methods in terms of protein yield. ...
... The molecular weight of the polypeptides ranges from 35 to 47 kDa, and their isoelectric point falls between pH 4.5 and 5.0 [15]. In addition to traditional extraction methods like alkaline, acidic, and salt extraction [9,[16][17][18][19][20][21][22], alternative techniques such as microwave extraction, ultrasound, and enzyme-assisted extraction have also proven effective. These methods typically require an alkaline environment, which can cause protein denaturation, affecting its nutritional value and functional properties [23]. ...
The growing interest in a plant-based diet leads to the search for new sources of protein in the human diet as an alternative to animal proteins. Plant materials that can supplement protein as additives in food products are being studied. Watermelon seeds (Citrillus lanatus L.) are rich in proteins and waste from the food industry; however, their extraction is not completely cost-free, and the flour production process may involve additional costs related to their extraction and processing. The studies showed that watermelon seed protein concentrate, obtained using the alkaline extraction method, contained 82.52 g/100 g of protein and 1.51 g/100 g of fat. The polyphenol content in the protein preparation from defatted watermelon seeds was 1.9 mg gallic acid/g, and the antioxidant activity of the concentrate was 29.26 µmol Trolox/g (by the ABTS+). The obtained watermelon seed protein concentrate was characterised by solubility of more than 80% (at pH = 10), water absorption at the level of 2.46 (g water/g) and oil absorption equal to 2.1 (ml oil/g), showed poor foaming properties (1.51%), and was characterised by low emulsification.
... Though extraction with ultrasound has multiple benefits, its effectiveness may vary based on the protein sources as well as ultrasound parameters including frequency, chosen extraction solvent and intensity. It is crucial to adjust these parameters correctly for getting the required extraction efficiency and maintaining the protein quality [69]. Henceforth, Fig. 1 depicts the ultrasound technique mechanism for facilitating protein extraction. ...
Food security for the increasing global population is a significant challenge of the current times particularly highlighting the protein deficiencies. Plant-based proteins could be considered as alternate source of the protein. The digestibility and PDCASS value of these proteins are still a concern. Blue proteins, the new approach of utilizing the proteins from aquatic sources could be a possible solution as it contains all the essential amino acids. However, the conjugation of these proteins with fats and glycogen interferes with their techno-functional properties and consequently their applicability. The application of power ultrasound for extraction and modification of these proteins from aquatic sources to break open the cellular structure, increase extractability, alter the protein structure and consequently provide proteins with higher bioavailability and bioactive properties could be a potential approach for their effective utilization into food systems. The current review focuses on the application of power ultrasound when applied as extraction treatment, alters the sulphite and peptide bond and modifies protein to elevated digestibility. The degree of alteration is influenced by intensity, frequency, and exposure time. The extracted proteins will serve as a source of essential amino acids. Furthermore, modification will lead to the development of bioactive peptides with different functional applications. Numerous studies reveal that blue proteins have beneficial impacts on amino acid availability, and subsequently food security with higher PDCAAS values. In many cases, converted peptides give anti-hypertensive, anti-diabetic, and anti-oxidant activity. Therefore, researchers are concentrating on ultrasound-based extraction, modification, and application in food and pharmaceutical systems.
... The estimated amount of CO 2 released in the atmosphere in the case of traditional extraction is higher than that for UAE (Fabiano-Tixier et al., 2016). Conventional methods reject over 4000 9 g of CO 2 whereas ultrasonic extraction yields 200 g in the atmosphere (Vernes et al., 2019). There are some constraints of using ultrasound for extraction. ...
As global protein demand surges, it is urgent to explore novel protein sources and develop advanced extraction technologies that can surpass the technological and scientific limitations of conventional methods. This systematic review summarises the emerging protein sources and sustainable protein extraction techniques, focusing on their role in enhancing food system sustainability. This article consolidates current research on novel extraction techniques, such as ultrasound‐assisted extraction, pulsed electric field‐based extraction, enzyme‐assisted extraction, subcritical water extraction and extraction using eutectic solvents, particularly from agro‐industrial by‐products. These methods assist in optimising protein recovery while aligning with circular economy principles by minimising waste and resource use. By evaluating the sustainability and practicality of these extraction methods, the article aims to guide future innovations and policy decisions, ultimately contributing to a more sustainable and resilient global food system.
... The extraction process is carried out with a ratio of 1: 5 (20 gr of Spirulina platensis powder + 100 mL aquades), which will attract polar compounds contained in Spirulina platensis, one of which is protein. The sonication method for 15 minutes with the help of ultrasonic vibrations can generate significant energy to destroy the cell walls of Spirulina platensis, attracting bioactive compounds (Vernes et al., 2019) and shortening the extraction time. Next, maceration for 24 hours so that it is well extracted. ...
It is essential to treat wastewater before it is discharged into water bodies. The wastewater treatment method that is widely used is coagulation using the synthetic coagulant PAC, whose continuous use can harm human health and reduce the pH value of the water. Therefore, wastewater treatment innovation is needed to overcome this problem. This study will examine the potential of Spirulina platensis as a biocoagulant to reduce turbidity and TSS. This research was conducted in 3 stages, starting from the extraction, protein test, and optimum dose test. The results showed that Spirulina platensis has 0.0114375% protein in 20 gr and can reduce the lowest turbidity at doses of 10-5 ml / L and 10-10 ml / L (in 2 injections) with an effectiveness value of reducing turbidity levels by 63.02% and able to reduce the lowest TSS levels at doses 10-10 ml / L (in 2 injections ) with an effectiveness value of reducing levels by 85%. Based on the Wilcoxon test, it was found that the P-value < 0.05, which means that there is a significant difference in TSS values and turbidity between the results before and after Spirulina platensis biocoagulant treatment.
... Mano-thermosonication (MTS) was applied to extract proteins from the dry mass of Spirulina by Vernès et al. (2019). A 1000 W ultrasonic device with a frequency of 20 kHz, utilizing a biomass/solvent ratio of 1:20 (g/g), was employed for sample treatment. ...
Microalgal, species are recognized for their high protein content, positioning them as a promising source of this macronutrient. Spirulina platensis, in particular, is noteworthy for its rich protein levels (70 g/100 g dw), which are higher than those of meat and legumes. Incorporating this microalgae into food can provide various benefits to human health due to its diverse chemical composition, encompassing high amount of protein and elevated levels of minerals, phenolics, essential fatty acids, and pigments. Conventional techniques employed for protein extraction from S. platensis have several drawbacks, prompting the exploration of innovative extraction techniques (IETs) to overcome these limitations. Recent advancements in extraction methods include ultrasound-assisted extraction, microwave-assisted extraction, high-pressure-assisted extraction, supercritical fluid extraction, pulse-electric field assisted extraction, ionic liquids assisted extraction, and pressurized liquid extraction. These IETs have demonstrated efficiency in enhancing protein yield of high quality while maximizing biomass utilization. This comprehensive review delves into the mechanisms, applications, and drawbacks associated with implementing IETs in protein extraction from S. platensis. Notably, these innovative methods offer advantages such as increased extractability, minimized protein denaturation, reduced solvent consumption, and lower energy consumption. However, safety considerations and the synergistic effects of combined extraction methods warrant further exploration and investigation of their underlying mechanisms.
... Also, the sonication time has a major impact on protein recovery. Sonication is advantageous in microalgal cell disruption which uses cavitation bubbles to release the intracellular compounds (Vernès et al. 2019). Phong et al. (2018) also showed that sonication led to a greater protein recovery from Chlorella sp. ...
Microalgae are photosynthetic microorganisms that have the capacity to synthesize a diverse range of biomolecules depending on their phylogenetic classification. Microalgae hold promise as sustainable and renewable energy resources; however, their full potential remains untapped due to the costliness and time-intensive nature of downstream processing. The objective of this study is to overcome these challenges by utilizing a single processing unit for the extraction of three major biomolecule classes namely, lipids, proteins, and carbohydrates. These biomolecules of marine Chlorella sp. NITT 02 were recovered in a single extraction process using the triphasic system. The parameters defining the triphasic system were optimized for the maximum recovery of biomolecules using Response Surface Methodology (RSM) coupled Genetic Algorithm (GA) optimization. The simultaneous complete recovery of lipids with 86.46 % of total fatty acids, 97 % protein recovery and 97.1 % carbohydrate recovery were obtained with i) 2.5:1 of t-butanol:culture volume, ii) 70 % t-butanol, iii) 32.9 % ammonium sulfate, and iv) 55 min sonication time. All three extracted biomolecule classes were characterized using FTIR, GC-MS, UV-Visible spectrophotometry, and XRD. From the current investigation, the recovery efficiency of the triphasic system is higher than the conventional sequential extraction of biomolecules and the time taken for extraction is 1.25 h with energy consumption of 0.7833 kWh. Hence, the adoption of this biorefinery approach is suitable for making microalgae commercially viable for producing valuable products from the biomolecules.
... This is because proteins become more charged in more alkaline pH environments, resulting in repulsion among the molecules and, consequently, increasing solubility [16]. In addition, the increase of solubilized proteins combined with ultrasound is supported by literature [17][18][19], probably due to the mechanic effects of ultrasound, such as the release of compounds by breaking the cell wall and stirring the solution, which allows higher interactions between the solvent and the proteins [20]. ...
... However, conventional SLE uses a large amount of solvent, generating toxic residues, posing risks of chemical transformation of the extracts and creating challenges in completely removing the solvent from the purified product. Furthermore, traditional SLE methods are characterized by limited efficiency [75]. ...
The heightened interest in healthy dietary practices and the preference for fresh, minimally processed foods with reduced additives have witnessed a significant surge among consumers. Within this context, bioactive compounds have garnered attention as potent agents offering beneficial biological effects when integrated into food formulations. Nevertheless, the efficacy of these bioactive compounds in product development encounters numerous challenges during various processing and storage stages due to their inherent instability. Addressing these limitations necessitates exploring novel technological approaches tailored explicitly to the application of bioactive compounds in food production. These approaches should not only focus on preserving the bioactive compounds within food matrices but also on retaining the sensory attributes (color, taste, and aroma) of the final food products. The impact of microalgae and their bioactive compounds on human health and well-being has been extensively reported in the literature. However, there is still a gap regarding the processing and stability of microalgal bioactive compounds to improve their application in the food industry. The main goal of the present work is to point out how to overcome technological challenges in enhancing the stability of bioactive compounds from microalgae for optimal food applications.
... 9,10 Among these advancements, using cavitation for tissue penetration stands out as a new, efficient, and safe technique developed in recent years. 11 Several notable applications have emerged, harnessing this mechanism for various purposes, including histotripsy, 12 biofilm removal, 13 sonoporation, 14 and others. 15 The principle underlying these applications involves the generation of cavitation bubbles in proximity to biological tissues using methods like ultrasound, electric sparks, and lasers. ...
A high-fidelity multiphase flow computational model is utilized to investigate the interaction mechanism between anti-phase tandem bubbles and tissue materials in a free-field environment. The formation of liquid jets generated by tandem bubble coupling and its effects on tissue deformation are analyzed. Parametric studies are conducted to explore the impacts of bubble–bubble distance ( γ b b), bubble size ratio ( S b b), and bubble–tissue distance ( γ t b). The results indicate that the regime of tissue penetration varies under different γ b b. For small γ b b, the tissue deformation is mainly attributed to the stretching of upper bubbles and liquid jets; whereas for large γ b b, tissue deformation is primarily induced by the jets themselves; and for moderate γ b b values, it is caused by a combined effect involving both jets and the evolution of toroidal bubbles. Comparative analysis shows the significant impact of varying Sbb on bubble coupling dynamics, with larger Sbb values correlating with more potent tissue penetration. Furthermore, the study also reveals that, beyond γtb exceeding 3, penetration ceases to manifest, advocating for the maintenance of γtb below 1.4 for practical applications.
... Mano-thermo-sonication is a type of ultrasonic extraction which works by combining pressure, temperature and ultrasound intensity to facilitate the extraction of water-soluble bioactive compounds from a sample matrix. This is because, the method not only facilitates cell disruption but also enhances mass transfer phenomena or effective diffusivity for better extraction yield [123]. Thus, assuring the optimum interactive effect of these extraction parameters is very important. ...
Extraction techniques of important bioactive molecules from the major concern of Seafood byproducts are getting emphasis for better valorization. Employing green extraction technologies for efficient and quality production of these bioactive molecules is also strictly required. Hence, understanding the extraction process parameters to effectively design an applicable optimization strategy could make this real. In this review, statistical optimization strategies applied for the extraction process parameters of bioactive molecules from Seafood byproducts are focused. The type of experimental designs, and techniques applied to criticize and validate the effects of independent variables on the extraction output was addressed. Dominant parameters studied were enzyme/substrate ratio, pH, time, temperature and power of extraction instruments. The yield of bioactive compounds, chitin and chitosan, proteins and peptides, enzymes and carotenoids (Astaxanthins) were the most studied responses. Efficiency and/or economic and quality considerations and their selected optimization strategies that favour the production of potential bioactive molecules were also reviewed.
... plays a crucial role in understanding environmental impacts due to its high metal binding capacity (Ito and Hori, 1989;Ramelow et al., 1992;Holan and Volesky, 1994). In recent years, the number of studies on algae processing methods and their use as a protein source has rapidly increased (Ghribi et al., 2015;Waghmare et al., 2016;Vernes et al., 2019). These studies aim to isolate high amounts of proteins from algae in waste status and transform them into high-value products. ...
The excessive proliferation of green algae in aquatic ecosystems threatens aquatic life, leading to oxygen depletion and water pollution. This study investigates two common green algae species, Ulva sp. and Cladophora sp., with potential in terms of protein and phenolic compounds. Cladophora sp. and Ulva sp. extracts were analyzed for total phenolic content using the Folin-Ciocalteau method. Despite lower phenolic content compared to specific plant species, both algae species exhibit various phenolic compounds. GC-MS analysis indicates the presence of major compounds such as limonene in Cladophora sp. and Tetradec-1-ene in Ulva sp., suggesting potential applications in the pharmaceutical and cosmetic industries. Despite modest protein amounts, the study emphasizes that algae, aligned with the increasing interest in plant-based nutrition, are a promising source for plant-based protein production. Ulva sp. and Cladophora sp. algae demonstrate potential as alternative protein sources and reservoirs of bioactive phenolic compounds from waste sources. This study pioneers further research in the food, pharmaceutical, and cosmetic industries to contribute to sustainable water resource utilization.
... However, approximately 42% of protein recovery was recorded from Spirulina by using methanol and ethanol solvents [25]. In another study, about 28.4% of protein content was derived from 100 g of Spirulina biomass by using an ultrasound mechanism with a protein recovery rate of 50% [26]. Based on a clinical study, the Spirulina extract rectifies the symptoms of allergic rhinitis patients by suppressing IL-4 and Th2 cell differentiation [27]. ...
Introduction
Among different blue-green algae, Spirulina (Arthrospira platensis) is known for its high protein content and
is hence considered a dietary single-cell protein. In recent decades, Spirulina has been one of the nutritive
additives in food and beverage products to enhance the nutritional value of food products. The aim of this
study was to brew a “Prot-Tea” enriched with antioxidants and protein for nutrition care.
Materials and methods
In combination, both the Spirulina and green tea were brewed together in three different formulations:
formulation A: 0.5 g of Spirulina powder and 1.5 g of green tea; formulation B: 1 g each of Spirulina powder
and green tea; and formulation C: 1.5 g of Spirulina powder and 0.5 g of green tea. All the formulations were
brewed in 100 mL of hot water at 80°C for two minutes. Then, all the formulations were subjected to a 2,2-
diphenylpicrylhydrazyl free radical scavenging assay and a quantitative protein estimation assay (Lowry
method).
Results
Among the three different formulations, formulation B showed greater antioxidant and protein content. The
antioxidant activity of formulation B was directly proportional to the concentration, with an IC50 value of
37.98 µL/mL. Similarly, the protein content was also higher in formulation B, with 287.33 µg/100 µL. The
total antioxidant in formulation B was 47.61 mg of ascorbic acid equivalent. Concurrently, the total protein
content was 229.86 mg in 80 mL of the total volume of Prot-Tea yield.
Conclusion
Based on this study, Prot-Tea is considered a future healthy beverage for nutrition care.
... In the context of Spirulina biomass, the techniques of PEF, SF, HPP, and ultrasonication have found predominant use in the extraction of vital nutrients, such as protein [166][167][168] phycocyanin [169][170][171][172], lipids [173,174], phenolics [175] and other pigments [176][177][178][179] from Arthrospira biomass [159,168]. Interestingly, their primary application appears to be centered around the extraction of valuable compounds rather than the direct control of contamination in Spirulina biomass. ...
... As a non-allergenic and non-animal protein source, it offers a viable alternative to conventional proteins in several industries [5,6]. However, to realize its full potential, the development of efficient protein extraction and purification methods is essential [7,8]. Therefore, this study aimed to evaluate different protein extraction methods -such as agitation, bead milling, and ultrasound -and protein isolation via precipitation using ethanol or ammonium sulfate to optimize the protein yield and purity. ...
... Physical methods are preferred for a large scale, being non-toxic and fast, despite the high-energy cost (Molino et al., 2020). Among the methods frequently reported in the literature for cell rupture, soxhlet, maceration, freeze-thawing, ultrasound, sonication, microwave-assisted, pressurized liquid, supercritical fluid extraction, pulse electric field stand out (Ventura et al., 2017;Mansour et al., 2019;Vernès et al., 2019;Molino et al., 2020). ...
... Conventional microalgae extraction methods, such as liquid-liquid or solid-liquid extraction, usually require long extraction times and massive amounts of solvents and can even cause the loss of some nutrients and bioactive compounds during extraction. Top of Form Extraction using ultrasound (ultrasonic-assisted extraction or UAE) has been extensively studied to extract compounds with high added value from microalgae (Vernes et al., 2019). Ultrasound requires less solvent consumption, short extraction times, and gives good extraction yields. ...
Chlorella sp. microalgae is a potential source of natural antioxidants and other bioactive compounds used in the food and pharmaceutical industries. Ultrasound pretreatments followed by enzymolysis are widely used to improve the functional properties of biological materials. This work aimed to investigate the effects of ultrasound pretreatments followed by bromelain hydrolysis on the proteins, phenolic content, and antioxidant activity of Chlorella sp. crude extract. The experimental design for the study involved two factors: ultrasound pretreatment time (0, 5, and 10 min) and bromelain hydrolysis time (0, 2, 4, and 6 hours). The protein was measured using Lowry assay while the total phenolic content was measured using a Folin–Ciocalteu assay. The highest protein content (7.09 ± 0.44 mg/mL) and phenolic concentrations (0.28 ± 0.01 ìg/mL) were obtained at 5 min ultrasound pretreatment and 6 hours enzymolysis. For antioxidant activity, 5 min ultrasound pretreatment followed by 4 hours of enzymolysis resulted in the highest DPPH (1,1-diphenyl-2-picrylhydrazyl radical) inhibitory activity (81.74%). Meanwhile, ultrasound pretreatment for 10 min followed by 6 hours of enzymolysis resulted in the highest ABTS (2,20 azinobis-(3-ethylbenzthiazoline-6-sulfonic acid) inhibitory activity (94.10%). This study showed that variation in ultrasound pretreatments followed by bromelain hydrolysis could be used to improve the protein content, phenolic content, and antioxidant activity of Chlorella sp. crude extract.
... Extraction was performed for 10, 20, 30, and 40 min using a Fisher Sonic Dismembrator (Model 300, USA) and solid/solvent ratios of 1:20 and 1:30 (w/v). Ultrasound power (P) and ultrasound intensity (UI) were calculated using the following equations by Vernes et al. (2019). ...
This study evaluates the effect of ultrasound-assisted extraction on the extractability of polyphenols from peanut skins (PS) and their antioxidant, and anticancer activities. The extraction was performed with solid/solvent ratios of 1:20 and 1:30 (w/v) at ultrasound intensity ranging from 5.8 to 15.4 W/cm2 for different extraction times (10, 20, 30 and 40 min). The highest polyphenol yield was 167.46 mg GAE/g dried PS. The most abundant polyphenols were catechin, syringic acid, and vanillic acid. The PS ultrasound extract (PSUE) increased the oxidative stability of sunflower oil by four times its initial level. PSUE possessed high inhibitory activity against MCF-7, HepG-2, HCT-116, and PC-3 cancer cell lines, with IC50 ranging from 1.85 ± 0.13 to 6.1 ± 0.43 μg/ml. In addition, the cytotoxicity of PSUE was examined on HFB4 human normal melanocytes using the MTT assay. These results suggest that PSUE can be used as a natural antioxidant and anticancer agent.
... Consequently, UAE increases the extraction yield, rate of extraction and reduces the extraction time without the need of applying high temperatures. [21] Numerous applications of UAE for the extraction of lipids, proteins, carotenoids, and phycocyanin are already reported in the literature [11,15,21,23]. ...
To meet the criteria of modern extraction, it is essential to use solvents that simultaneously reach the standards of green chemistry and allow the extraction of a wide spectrum of bio-compounds. Natural Deep Eutectic Solvents (NaDES), as a green solution, meet this difficult task and represent a satisfactory alternative to organic solvents. Microalgae are recognized as a sustainable resource to produce polar and non-polar metabolites, such as ca-rotenoids, free fatty acids (FFA) and phycobiliproteins. Among them, Spirulina (Arthrospira platensis) is one of the most studied, due to its strong potential for food, health and cosmetic. In this work, two biorefinery scenarios were investigated using polar and non-polar NaDES. The first one is a combination of a polar/non-polar sequential solid/liquid extraction together with a liquid-liquid extraction of the polar extract. The second scenario implements an innovative triphasic solid/liquid/liquid approach, based on the simultaneous use of polar and non-polar NaDES in the solid/liquid extraction step. Several NaDES were screened in this study to identify the best polar/non-polar NaDES pair. According to the results, the use of the triphasic approach allowed an increase of the productivity by a factor 4 for chlorophylls and FFA, and by a factor 2 for carotenoids. It was also interesting to note that the phycocyanin-enriched polar fraction achieves much higher levels of purity than the corresponding single-phase extraction for both applied biorefinery scenarios. These results pave the way for the first time for a microalgae biorefinery entirely based on the use of NaDES and industrially credible.
... Consequently, UAE increases the extraction yield, rate of extraction and reduces the extraction time without the need of applying high temperatures. [21] Numerous applications of UAE for the extraction of lipids, proteins, carotenoids, and phycocyanin are already reported in the literature [11,15,21,23]. ...
... These phenomena allowed larger sizes of vapour bubbles to be produced and implode more violently. The violent implosion of vapour bubbles generated high-intensity of microjet and shockwaves, causing the cell wall to breakdown easily and more target compounds to be released into the solvent [62,63]. However, the extraction yields of cinnamic acid and cinnamaldehyde decreased (p < 0.05) as the ultrasonic power further increased from 160 W to 200 W. ...
Synthetic polymers are considered a threat due to their impact on the environment via disposal and their dependence on fossil sources. The upcoming alternative technology in the packaging industry is green packaging materials obtained from natural sources like plants, animals, and microorganisms. These are considered to be environmentally safe and include carbohydrates like starch, lignin, and cellulose and also proteins like keratin, collagen, and gelatin, as well as microbial-origin biopolymers like polyhydroxy alkanoates and exo-polysaccharides (EPS). Among the various classes of bio-polymers, starch is considered non-toxic, available, and economical and is used in the form of active films in food packaging. Bio-polymeric polyphenolic compounds like gallic acid, which are extracted via supercritical extraction, are added to enhance the anti-oxidant properties of the chitin and gelatin-based films in active food packaging. Nanomaterials of biological origin are also incorporated in active food packaging inclusively bacterial nanocellulose, which is a hydrocolloid nanocellulose substrate that is synthesized from the non-pathogenic strains of Komagataeibacter due to its high crystallinity, thermal stability, and mechanical strength. It is necessary to conduct more basic research on applying biodegradable polymers in food packaging and to make environmental protection efforts.
Fruit based blends are gaining popularity due to their tremendous properties like they are rich in vitamins, minerals, antioxidants, flavonoids, carotenoids and dietary fibres, making them highly able to reduce the risk of certain diseases. Due to high water content fruit juices are highly vulnerable to microbial contamination and spoilage. Therefore it is a challenge for food industry to maintain the integrity and reduce the destruction of these compounds in fruit juices. Thermal treatments can destroy the heat sensitive compounds like vitamins and antioxidants. Therefore ultrasound (US) has gained popularity as a sustainable non-thermal technology as high frequency sound waves from ultrasound cause disruption of the matrix and favour the compounds to seep out of the cell. US facilitate the extraction of bioactive components like ascorbic acid, carotenoids and phenolic compounds as well as nutrients such as carbohydrates, proteins, and lipids and increase their accessibility enhancing the nutritive value of the fruit juices. The current review’s objective is to provide a brief summary of research findings which focus specifically on the current use of US on various fruit based juices. However, there are fewer researches evaluating the impact of ultra-sonication on the macro and micronutrients.
Ultrasonic compaction, also known as ultrasonic agglomeration, is an emerging technology that represents a novel alternative for food agglomeration; it is of great interest to the food industry. This review aims to gather information on the physicochemical, organoleptic, microbiological, and structural changes generated by ultrasound and study the fundamentals of agglomeration and ultrasound in different food matrices. In addition, chemical changes are reported in some nutrients related to conformational changes, such as the disintegration of diacylglycerides into monoacylglycerols, disordering of the crystalline region of starch granules to the amorphous phase, disruption of the membrane in plant cells, and transient or permanent modification of the protein structure (3D folding). The increasing development of patents can provide an insight into the potential of ultrasonic agglomeration applications in the food industry.
In this study, four kinds of Spirulina platensis ( S. platensis) peptides were obtained by enzymolysis of protein after different pretreatment to S. platensis powders. The novel coronavirus main protease (SARS-CoV-2 main protease, Mpro) was successfully prepared by heterologous expression in E. coli and its activity was detected. After identification of S. platensis peptides sequences, highly active peptides were synthesized and their inhibition effects on Mpro was verified. Subsequnently, the mechanism of action between them were analyzed by computer simulations. Results showed that the extraction rate of the protein from S. platensis was 96.11%, the activity of Mpro was 845.90 U mg ⁻¹ , the peptide MQGPNY could inhibit the activity of Mpro with the inhibition rate of 20.21%±3.8% at the concentration of 2 mM. Fuerthermore, results from computer Simulation showed that the peptide MQGPNY forms four pairs of hydrogen bonds with Mpro, which are Gln189, Ser46, Thr26 and Glu166. In addition, there were nine free residues involved in hydrophobic contacts, which were His164, His41, Thr24, Met49, Thr45, Thr25, Gly143, Asn142, and Cys145 and Met165. This study developed natural peptides from S. platensis , which could inhibited the activity of Mpro. Besides, the mechanism of the peptide MQGPN acting on Mpro was revealed by the method of computer Simulation. The foundings could provide theoretical support for the use of protein and its hydrolysates from S. platensis in functional food and supplement formulations in the post-epidemic era.
Significant attention is focused on mushrooms and mushroom protein hydrolysates owing to their sustainability and nutritional profile with different uses in diverse industries. This review examines the composition, functional properties, production methods and applications of mushroom protein hydrolysates. It commenced with a brief overview of protein hydrolysates where their importance and applications in food, pharmaceutical, cosmetic, and agricultural industries were explored. The significance of mushrooms as alternatives to animal proteins applicable to producing protein hydrolysates was discussed. Methods including conventional (chemical and enzymatic) extraction, ultrasound-assisted and microwave-assisted extractions used to extract proteins during protein hydrolysates production were discussed. Using enzyme-assisted-ultrasound-extraction to enhance protein extraction efficiency is exclusively highlighted as a promising technique. To obtain enhanced protein yield and quality, and protein hydrolysates with desired characteristics, crucial factors consisting of pH, temperature, extraction time, solvent type, enzyme concentration and ratio-of-substrate-concentration need to be optimized. The physicochemical and functional properties of mushroom protein hydrolysates are summarized. Variability of mushroom species, extraction efficiency, enzyme selection and cost, standardization, and consumer perception which constitute challenges and limitations for production and application of mushroom protein hydrolysates are
addressed. Research should be conducted to optimize extraction and hydrolysis processes and explore new mushroom species.
Techniques for extracting important bioactive molecules from seafood byproducts, viz., bones, heads, skin, frames, fins, shells, guts, and viscera, are receiving emphasis due to the need for better valorization. Employing green extraction technologies for efficient and quality production of these bioactive molecules is also strictly required. Hence, understanding the extraction process parameters to effectively design an applicable optimization strategy could enable these improvements. In this review, statistical optimization strategies applied for the extraction process parameters of obtaining bioactive molecules from seafood byproducts are focused upon. The type of experimental designs and techniques applied to criticize and validate the effects of independent variables on the extraction output are addressed. Dominant parameters studied were the enzyme/substrate ratio, pH, time, temperature, and power of extraction instruments. The yield of bioactive compounds, including long-chain polyunsaturated fatty acids, amino acids, peptides, enzymes, gelatine, collagen, chitin, vitamins, polyphenolic constituents, carotenoids, etc., were the most studied responses. Efficiency and/or economic and quality considerations and their selected optimization strategies that favor the production of potential bioactive molecules were also reviewed.
Due to factors including population growth, rising incomes, increased urbanization, and aging populations, the need for protein components is continuously expanding on a global scale. Traditionally, foods produced from animals (such as dairy, eggs, and meat) provide most of the dietary protein needs of people. However, the identification of sustainable alternative protein sources is thus required to replace animal proteins. Based on exceptional and widespread ecological adaption, microalgae act as of the edible proteins. Microalgae have a high photosynthetic efficiency and can flourish in disadvantaged environments using non-potable effluent. The purity of the amino acids (AAs) and the technological functional characteristics of microalgae proteins are comparable to those of reference proteins. Recalcitrant cell walls, however, make it difficult to digest and use the microalgae proteins effectively. Furthermore, the palatability and low sensory scores limits the microalgal biomass in the food and feed industries. Meat analogues, emulsifiers, and bioactive peptides are a few novels uses for microalgae proteins. The development of low-cost growing techniques, wet downstream processing based on biomass, with bio-refinery strategies would increase the sustainability ability as human and animal feed applications.
This book aims to inform readers about the latest trends in environment-friendly extraction techniques in food analysis. Fourteen edited chapters cover relevant topics. These topics include a primer green food analysis and extraction, environment-friendly solvents, (such as deep eutectic solvents, ionic liquids, and supramolecular solvents), and different extraction techniques.
Key Features
- Includes topics for basic and advanced readers
- Covers a wide range of green solvents for food analysis
- Emphasizes modern extraction techniques (including supercritical fluid extraction, the gas expanded liquid extraction, pressurized liquid extraction, microwave-assisted extraction, pulse electric field extraction)
- Provides notes on selection of solvents
- Includes references for every chapter
The blend of fundamental and applied information makes this an ideal reference for food chemistry students and research scholars. It also serves as a guide for professional experts working in food analysis and sustainability roles.
Cottonseed meal (CM) is a useful protein resource, but the presence of gossypol (a toxic compound) limits it utilization in industry. To reduce the gossypol content, the use of ultrasonication in extracting protein from CM was explored. Following single-factor experiments, operative limits of independent variables affecting removal
ratio of total-gossypol (RRTG) and extractability of CM protein (CMP) were achieved as temperature (35–55 ◦C), sonication power (250–350 W), time (12–24 min), and NaOH concentration (2–6 g/L). Box-Behnken’s Design centered on a 3–level-4–factor was employed to optimize ultrasonication conditions on the yield and RRTG of
CMP. The predictive models were statistically adequate (p < 0.0001) to reflect the experimental results of the responses as verified by insignificant lack-of-fit (p ≥ 0.1333) and high coefficient of determination (R2 ≥ 0.976) values. The optimized sonication conditions for two responses were 53 ◦C, 320 W, 21 min, and 4.6 g-NaOH/L. Under optimized conditions, the observed removal ratio of free- and total-gossypol and yield values were 93.21 %, 87.56 %, and 58.23 %, which substantially increased by 15.64 %, 12.88 %, and 29.56 %, respectively over control (alkaline extraction). Furthermore, sonicated isolate showed an increase in solubility and emulsion property relative to control, buttressed by particle sizing and zeta potential outcomes. Sodium dodecyl sulfide polyacrylamide gel electrophoresis (SDS-PAGE), Fourier Transform Infrared (FTIR) spectroscopy, atomic force microscopy (AFM), and amino acid data showed notable changes in molecular weight, functional groups, surface topography, and hydrophobic amino acid content of CMP, respectively (i.e., after sonication), suggesting partial unfolding in CMP conformation. Accordingly, CMP isolates (especially the sonicated protein) could be considered edible because of the observed low gossypol content (< 0.045 %) and thus can be utilized as natural additives in food formulations.
Background Spirulina is multicellular and filamentous cyanobacteria that have achieved a considerable popularity in the health sector, food industry and aquacultures. It develops and grows in water, can be harvested and processed easily. It has very high content of macro and micronutrients, essential amino acids, proteins, lipids, vitamins, minerals and anti-oxidants. Spirulina is considered as a complete food supplement to fight against malnutritional deficiencies in developing countries. Spirulina is deemed safe for human consumption as evident by its long history of food use and latest scientific findings. In recent years, Spirulina has gathered enormous attention from research fraternity as well as industries as a flourishing source of nutraceutical and pharmaceuticals. Scope and approach The primary objective of this paper is to review the utilization of Spirulina as a dietary supplement in the food industry. In the present work, the three main area of Spirulina research: growth, harvesting and potential application are presented. Key findings and conclusion The important growth parameters have been studied to enhance Spirulina biomass productivity qualitatively and quantitatively. This review provides useful information on commercially viable technology for Spirulina cultivation. Mass cultivation and Innovative formulations are further needed to fortify conventional foods with Spirulina based protein system.
Population growth combined with increasingly limited resources of arable land and fresh water has resulted in a need for alternative protein sources. Macroalgae (seaweed) and microalgae are examples of under-exploited “crops”. Algae do not compete with traditional food crops for space and resources. This review details the characteristics of commonly consumed algae, as well as their potential for use as a protein source based on their protein quality, amino acid composition, and digestibility. Protein extraction methods applied to algae to date, including enzymatic hydrolysis, physical processes, and chemical extraction and novel methods such as ultrasound-assisted extraction, pulsed electric field, and microwave-assisted extraction are discussed. Moreover, existing protein enrichment methods used in the dairy industry and the potential of these methods to generate high value ingredients from algae, such as bioactive peptides and functional ingredients are discussed. Applications of algae in human nutrition, animal feed, and aquaculture are examined.
Marine microalgae and seaweeds (microalgae) represent a sustainable source of various bioactive natural carotenoids, including β-carotene, lutein, astaxanthin, zeaxanthin, violaxanthin and fucoxanthin. Recently, the large-scale production of carotenoids from algal sources has gained significant interest with respect to commercial and industrial applications for health, nutrition, and cosmetic applications. Although conventional processing technologies, based on solvent extraction, offer a simple approach to isolating carotenoids, they suffer several, inherent limitations, including low efficiency (extraction yield), selectivity (purity), high solvent consumption, and long treatment times, which have led to advancements in the search for innovative extraction technologies. This comprehensive review summarizes the recent trends in the extraction of carotenoids from microalgae and seaweeds through the assistance of different innovative techniques, such as pulsed electric fields, liquid pressurization, supercritical fluids, subcritical fluids, microwaves, ultrasounds, and high-pressure homogenization. In particular, the review critically analyzes technologies, characteristics, advantages, and shortcomings of the different innovative processes, highlighting the differences in terms of yield, selectivity, and economic and environmental sustainability.
The successful commercial exploitation of Arthrospira because of its high nutritional value, chemical composition and safety of the biomass has made it one of the most important industrially cultivated microalgae. Knowledge of its biology and physiology, which is essential for understanding the growth requirements of this alkaliphilic organism, has been used in developing suitable technologies for mass cultivation. The relationships between environmental and cultural factors, which govern productivity in outdoor cultures, are discussed in connection with growth yield and efficiency. The response of Arthrospira and its modification under stress are described, together with the strategy of osmotic adjustment and the mechanism of internal pH regulation to alkalinity. The metabolic plasticity of the response to disparate environmental stimuli is demonstrated in the natural environment, but is also well-expressed in the maintenance of high productive monoculture in intensive outdoor cultivation systems. While the confused taxonomy of Arthrospira and its relationship with Spirulina has been resolved by study of the ultrastructural feature of trichomes and 16S rRNA sequence analysis, the problem of species definition is still ongoing. However, molecular methods such as total DNA restriction profile analyses of a wide range of strains are helping to resolve this. © 2012 Springer Science+Business Media B.V. All rights reserved.
Microalgae are microorganisms that have different morphological, physiological, and genetic traits that confer the ability to produce different biologically active metabolites. Microalgal biotechnology has become a subject of study for various fields, due to the varied bioproducts that can be obtained from these microorganisms. When microalgal cultivation processes are better understood, microalgae can become an environmentally friendly and economically viable source of compounds of interest, because production can be optimized in a controlled culture. The bioactive compounds derived from microalgae have anti-inflammatory, antimicrobial, and antioxidant activities, among others. Furthermore, these microorganisms have the ability to promote health and reduce the risk of the development of degenerative diseases. In this context, the aim of this review is to discuss bioactive metabolites produced by microalgae for possible applications in the life sciences.
This review presents useful and green techniques of solvent-free extraction used in ancient times, such as extraction of olive oil and citrus essential oil, and innovative techniques, such as pulsed electric field, microwave, instantaneous controlled pressure drop, and extrusion. We discuss the devices, their applications, mechanisms, and parameters influencing sample preparation prior to analysis of natural products.
The design of green and sustainable extraction methods of natural products is currently a hot research topic in the multidisciplinary area of applied chemistry, biology and technology. Herein we aimed to introduce the six principles of green-extraction, describing a multifaceted strategy to apply this concept at research and industrial level. The mainstay of this working protocol are new and innovative technologies, process intensification, agro-solvents and energy saving. The concept, principles and examples of green extraction here discussed, offer an updated glimpse of the huge technological effort that is being made and the diverse applications that are being developed.
This paper illustrates the Ultrasound Assisted Extraction (UAE) of β-carotene from Spirulina platensis. Various parameters such as extraction time, solvent type, biomass to solvent ratio, temperature, electrical acoustic intensity, length of the probe tip dipped into the solvent, duty cycle and pre treatment effect were explored for the extraction of β-carotene. From economic point of view, the optimal conditions for the extraction of β-carotene from Spirulina were 1.5g Spirulina (2min pre soaked in methanol) in 50ml n-heptane at 30°C temperature, 167W/cm(2) electrical acoustic intensity and 61.5% duty cycle for 8min with probe tip length of 0.5cm dipped into the extracting solvent from the surface. The maximum extraction achieved under the above mentioned optimum parameters was 47.10%. The pre-treatment time showed a promising effect on the yield as pre-treating the biomass with methanol for 2min before ultrasonication showed 12 times increase in extraction yield of β-carotene.
Fluorescent and red light environments generate greatly different patterns of pigmentation and morphology in Fremyella diplosiphon. Most strikingly, red-illuminated cultures contain no measurable C-phycoerythrin and have a mean filament length about 10 times shorter than fluorescent-illuminated cultures. C-phycoerythrin behaves as a photoinducible constituent of this alga. Spectrophotometric and immunochemical procedures were devised so that C-phycoerythrin metabolism could be studied quantitatively with [(14)C]-phenylalanine pulse-chased cultures. Transfer of red-illuminated cultures to fluorescent light initiates C-phycoerythrin production by essentially de novo synthesis. C-phycoerythrin is not degraded to any significant extent in cultures continuously illuminated with fluorescent light. Transfer of fluorescent-illuminated cultures to red light causes an abrupt cessation of C-phycoerythrin synthesis. The C-phycoerythrin content of cultures adapting to red light decreases and subsequently becomes constant. Loss of C-phycoerythrin is not brought about by metabolic degradation, but rather by a decrease in mean filament length which is effected by transcellular breakage. In this experimental system, light influences intracellular C-phycoerythrin levels by regulating the rate of synthesis of the chromoprotein.
1ère partie : Principes généraux du métabolisme protéique et des besoins en protéines et en acides aminés chez l'homme chap. 4
Toluidine blue (TB) staining either alone or in association with other methodologies has the potential to answer a variety of biological questions regarding the human, animal and plant tissues or cells. In this brief review, we not only report the primary use of TB to detect the anionic substrates and availability of their binding sites, but also unveil the resulting applications of TB staining in biological research. Among these applications, the uses of TB staining to identify the changes in chromatin DNA-protein complexes, nucleolus location, and extracellular matrix proteoglycan complexes associated with different physiological and pathological events are described. The usefulness of TB staining to monitor the effects elicited by environmental insults on chromatin and intercalation of drugs into the DNA is also included.
The potential of microalgae as an alternative energy source has been adequately studied. However, exclusive use of microalgae as an energy feedstocks cannot warrant their scalability and economical sustainability due to the high cost involved in their biomass processing. The co-processing of microalgae biomass with other related bio-refinery applications can offset their cost and improve their sustainability. Thus, it triggers up the need of exploring the potential of microalgae biomass beyond their typical use. Microalgae offer interesting features to qualify them as alternative feedstocks for various bio-refinery applications. Microalgae have unique abilities to utilize them for industrial and environmental applications. Thus, this review discusses to expand the scope of integrating microalgae with other biotechnological applications to enhance their sustainability. The use of microalgae as a feed for animal and aquaculture, fertilizers, medicine, cosmetic, environmental and other biotechnological applications is thoroughly reviewed. It also highlights the barriers, opportunities, developments, and prospects of extending the scope of microalgae. This study concludes that sustained research funding, and a shift of microalgae focus from biofuels production to bio-refinery co-products can qualify them as promising feedstocks. Moreover, technology integration is inevitable to off-set the cost of microalgae biomass processing. It is expected that this study would be helpful to determine the future role of microalgae in bio-refinery applications.
Protein concentrates and isolates were developed from the microalga Spirulina sp. LEB 18 biomass and characterized by functional aspects for food application. Proteins were solubilized at pH 11 and precipitated at pH 4.2 (protein isoelectric point determined by potentiometric titration), with the aid of a high-speed homogenizer. With this procedure, it was possible to obtain a protein concentrate with 83.9 ± 1.7 wt% of protein and a protein isolate with 91.3 ± 1.2 wt%. Protein extraction from this microalga allowed a significant increase in protein solubility and foam stability. Furthermore, proteins from both concentrate and isolate presented higher resistance to thermal denaturation than the original proteins of Spirulina sp. LEB 18 biomass. These results show the application potential of the concentrate and the protein isolate from Spirulina sp. LEB 18 in foods for protein supplementation, since they have >80 and 90% protein, respectively.
Industrial relevance text
Using proteins from microalgal sources can serve as a sustainable alternative to meet world food demand. The isoelectric precipitation extraction method can be applied on a large scale, achieving high yields. Protein extracts present potential application in specific foods for protein supplementation. Protein extraction can be allied to other biocomposites extraction, such as carbohydrates and lipids for biofuels production.
Arthrospira platensis is a species of filamentous cyanobacteria whose high protein content renders it suitable as a nutritive supplement for human and animal diets. Furthermore it is easy to grow as it tolerates a broad range of environmental stresses such as extremes of pH, salinity and heavy metals. In this study, changes in the growth, pigments and proteome of A. platensis were analyzed under two major abiotic stresses limiting productivity, salinity and iron. The lower concentrations of stresses showed varied effects on fresh weight, growth rate, pigment constituents and soluble protein of A. platensis. A proteomic analysis was conducted using SDS-PAGE first to obtain an overview of gross changes, followed by 2-DE and mass spectrometry to identify specific proteins whose abundance was affected by these environmental stresses. Eighteen spots were differentially expressed under the stress conditions. Of these, six were found with increased abundance responding only to Fe²⁺ stress, and five as a result of NaCl stress alone. Using LC–MS/MS, 37 proteins were identified within these spots. These proteins were classified into thirteen categories according to their function annotation. The present analysis could provide important data related to stress-tolerance by microalgae.
There are more than 1300 articles in scientific literature dealing with positive impacts of Ultrasound-Assisted Extraction (UAE) such as reduction of extraction time, diminution of solvent and energy used, enhancement in yield and even selectivity, intensification of diffusion, and eliminating wastes. This present study aims to understand what are the mechanism(s) behind these positive impacts which will help to design a decision tool for UAE of natural products. Different microscopic observations (Scanning Electron Microscopy (SEM), Environmental Scanning Electron Microscopy (e-SEM), Cyto-histochemistry) have been used for spacial and temporal localization of metabolites in rosemary leaves, which is one of the most studied and most important plant for its antioxidant metabolites used in food industry, during conventional and ultrasound extraction. The study permits to highlight that ultrasound impacted rosemary leaves not by a single or different mechanisms in function of ultrasound power, as described by previous studies, but by a chain detexturation mechanism in a special order: local erosion, shear forces, sonoporation, fragmentation, capillary effect, and detexturation. These detexturation impacts followed a special order during ultrasound treatment leading at the end to the total detexturation of rosemary leaves. These mechanisms and detexturation impacts were identified in glandular trichomes, non-glandular-trichomes and the layer adaxial and abaxial cuticle. Modelling metabolites diffusion phenomenon during conventional and ultrasound extraction with the second Fick's law allowed the estimation of diffusivities and solvent penetration into the inner tissues and in meantime to accelerate the release of valuable metabolites.
Biological significance:
Arthrospira platensis, widely known as Spirulina, is a type of cyanobacteria which is cultivated at large scale for it great commercial value. It has become a consensus that continually increasing temperature due to global warming is bringing serious threat to agriculture, including the Spirulina cultivation. High temperature not only limits biomass accumulation by Spirulina, but also changes the composition of nutrition. Therefore there is a greater need than ever before to understand how Spirulina tolerates and cope with high temperature. In this study, we for the first time applied the iTRAQ-based quantitative proteomic technology to investigate the thermotolerance of Spirulina. Our results showed that many biological processes were altered by heat stress. Most significantly, we found that heat stress harmed the photosynthesis ability and caused low energy production, and to deal with this situation, energy demanding processes like protein synthesis were down-shifted and the alternative energy metabolism process glycolysis was up-shifted. Our results also show other important proteins, like the classical heat shock proteins and some antioxidant proteins, are also increased. Thus our study sheds light to our understanding of the mechanism underlying the thermotolerance of Spirulina.
Microalgae are a very interesting and valuable natural source of highly valuable bioactive compounds, such as vitamins, essential amino acids, polyunsaturated fatty acids, minerals, carotenoids, enzymes and fiberfibre. Due to its potential, microalgae have become one of the most promising and innovative sources for new food and functional products., that Moreover, microalgae can be used as functional ingredients in order to enhance the nutritional value of foods and, thus hence, to favourably affecting favourably human health by improving the well-being and quality of life, but also decreasing curtailing diseases and illness risks. This review provides an overview of the current knowledge of the health benefits associated to microalgae consumption, bioactive compounds, functional ingredients, and foods.
The morphology of Spirulina during cultivation is susceptible to external interferences, but the morphogenesis mechanism is still unclear. Here the proteomic changes of linear Spirulina and spiral Spirulina were comparatively investigated via isobaric tag for relative and absolute quantitation (iTRAQ). Totally 165 and 167 differences in proteins expression were screened out from the TJSD2/TJSD3 and TJBC4-1/TJBC4-2 groups, respectively. Gene ontology and metabolic pathway analysis of differences in proteins expression uncovered the metabolic pathways (photosynthesis, carbon fixation, sugar metabolism) that were significantly enriched with the proteins correlated with Spirulina morphogenesis. The results of differences in proteins expression in metabolic pathway were verified by quantitative real-time PCR. We also built a putative model of Spirulina morphogenesis mechanism and thought multiple metabolic pathways interact and take part in Spirulina morphogenesis.
Manothermonication is a recognized and efficient method used for sterilization in food preservation. The synergistic effect of sonication combined with pressure and temperature allows enhancing the cavitation activity. Never employed for extraction, this study is about the transposition of this process as a tool of extraction. In this study, Rhodosporidium toruloides yeast was submitted to extraction by four modes of sonication, with a temperature ranged from 20 to 55 °C and a pressure between 1 to 2 bars. The lipids extraction yields were compared to the conventional maceration. Microbial oils obtained from both processes were analyzed and quantified by HPTLC (High Performance Thin-Layer Chromatography) and GC-FID (Gas Chromatography with flame ionization detector) after transesterification of lipids. Manothermosonication (30 min, 2 bars, 55 °C) permits to enhance of approximately 20% the extraction yield of lipids to compared to conventional maceration. The fatty acid profiles of each pretreatment and extraction by US, MS, TS and MTS do not affect the fatty acid profiles of yeast (majority of oleic acid (C18:1n9), linoleic acid (C18:2n6) and palmitic acid (C16:0)). Manothermosonication technique shows a great potential for lipid extraction from oleaginous microorganisms.
This study was performed to evaluate the responses of Escherichia coli 0157:H7 inoculated in an apple-carrot blended juice to manothermosonication (MTS) treatments. The MTS treatments were conducted in a continuous-flow MTS system. The juice samples were exposed to ultrasound treatment at combinations of three temperatures (60, 50 and 40 °C) and three pressure levels (100, 200, and 300 kPa) for five residence times (15, 30, 45, 60, and 75 s). The results showed that higher treatment temperature (i.e. 60 °C) and hydrostatic pressure in the MTS system significantly enhanced the microbial reduction. A FDA mandated 5-log CFU/ml reduction of E. coli 0157:H7 for juice processing was achieved in 30 s for MTS treatment at 60 °C, in comparison to 60 s at 50 °C. The Weilbull and Log-logistic models provided the best fitting of the inactivation data for the MTS treatments. Extensive damage of E. coli 0157:H7 cells treated with MTS was observed on micro-images of scanning electron microscopy and transmission electron microscopy.
The ethanolic extraction of minor compounds (phospholipids, tocopherols and sugars) present in sunflower collets was studied at 50 and 60 °C in a batch reactor. The extracted material consisted of two phases: a hexane-soluble fraction, comparable to degummed sunflower oils, and a hexane-insoluble fraction high in phospholipids and sugars. Sugars were extracted in large proportion, especially the indigestible raffinose, increasing the nutritional value of the meal. The sugar reduction percentage in the sunflower collets increased over extraction time to up to 60 and 80% at 50 and 60 °C. The effective diffusion coefficient (De) for tocopherols was higher than that for phospholipids (3.950 10⁻⁹ and 2.596 10⁻⁹ m²/s, respectively), both being temperature-independent in the analyzed range. De of sugars was 6.50 10⁻¹⁰ and 1.51 10⁻⁹ m²/s for 50 and 60 °C, respectively. Using ethanol as extraction solvent could improve the oil and meal quality, and help obtain a third phospholipid-rich phase after fractionation.
In the past few years, much interest has focused on the applications of ultrasound in food processing and technology. Many reports suggest effective benefits from combining ultrasound with classical techniques. Applications of ultrasound generally involve processes that can enhance rates, improve quality and/or safety, and reduce processing time. Ultrasound assistance has now become of significant importance when scaling up laboratory processes and can play a key role in implementing sustainable “green” technology.
Here we look at the concepts of hazard analysis and critical control points (HACCP) and hazard and operability (HAZOP) in the design of an ultrasonic food processing operation. These concepts are currently used as total quality management tools for the improvement of industrial operations. The HACCP program ensures the processing and production of safe foods by analyzing health hazards associated with processing, distribution, and consumption of foods, and in which the critical control points of the process are identified. The HAZOP analysis is used to identify hazardous working conditions in specific steps of a processing operation, taking into account the health effects of both contact and airborne ultrasound exposure.
This study investigate the use of ultrasound assisted extraction (UAE), the present work describes the application of ultrasound to improves the extraction efficiency of the classical solvent extraction techniques such as maceration and soxhelt extraction to extract various bioactive compound from natural source like plant and animals. In this review extraction of various compounds i.e.β carotene, flavonoids, anthraquinones, isoflavones, cepharanthine, melanin, ursolic acid, from various plants i.e. morinda citrifolia, mitragyna speciosa, oscimum sanctum, polygonum cuspidatum, auricularia auricula, stephania rotunda lour,iris tectorum maxim, heterophyllaea pustulata hook, hawthorn seed, spirulina platensisand their therapeutic acivities like antiinflammatory, anti-cancer, anti-oxidant, cardioprotective action, immunomodulatory, anti-diabetic, antirheumatic, anti-bacterial, anthelmintic, anti-depressant, hepatoprotective, radioprotective properties was extracted at different temperature 25°, 45°, 90° C, power setting 3, 6, 9 types of solvent used like ethanol methanol, acetone, acetonitrile in different composition and time 15, 30, 45, 60, 90 minute. The effect of various factors such as extraction solvent, solvent concentration, temperature, solvent to solid ratio, ultrasound power, extraction time, and particle size, on the yield of target components were investigated. Under the optimum conditions the extraction process is carried out and the yield obtained with UAE has been compared with batch and soxhelt extraction the result indicated that UAE was an alternative method for extraction. UAE is based upon acoustic cavitation. Cavitation produces physical effects such as liquid circulation currents and turbulence which results in significant increase in the mass transfer rates and thus increased extraction yield. To achieve the same recovery as that achieved by UAE, soxhelt extraction and maceration extraction required much longer time.
In the past few years, much interest has focused on the applications of ultrasound
in food processing and technology. Many reports suggest effective benefits from
combining ultrasound with classical techniques. Applications of ultrasound generally
involve processes that can enhance rates, improve quality and/or safety, and
reduce processing time. Ultrasound assistance has now become of significant
importance when scaling up laboratory processes and can play a key role in
implementing sustainable “green” technology.
Ultrasound-assisted extraction of rapeseed oil was investigated and compared with conventional extraction for energy efficiency, throughput time, extraction yield, cleanness, processing cost and product quality. A multivariate study enabled us to define optimal parameters (7.7 W/cm2 for ultrasonic power intensity, 40 °C for processing temperature, and a solid/liquid ratio of 1/15) for ultrasound-assisted extraction of oil from oilseeds to maximize lipid yield while reducing solvent consumption and extraction time using response surface methodology (RSM) with a three-variable central composite design (CCD). A significant difference in oil quality was noted under the conditions of the initial ultrasound extraction, which was later avoided using ultrasound in the absence of oxygen. Three concepts of multistage cross-current extraction were investigated and compared: conventional multistage maceration, ultrasound-assisted maceration and a combination, to assess the positive impact of using ultrasound on the seed oil extraction process. The study concludes that ultrasound-assisted extraction of oil is likely to reduce both economic and ecological impacts of the process in the fat and oil industry.
In recent decades, microalgae species have focused the attention of several research groups and food industry as they are a great source of nutritionally valuable compounds. The use of environmentally friendly technologies has led to researchers and food industry to develop new alternative processes that can extract nutritionally valuable compounds from different sources, including microalgae. This note describes the potential use of some non-conventional methods including sub- and supercritical fluid extraction, pulsed electric fields, high-voltage electric discharges, high-pressure homogenization, ultrasound- and microwave-assisted extraction, which involve cell disruption to recover nutritionally valuable compounds from microalgae and can help to comply with criteria of green chemistry concepts and sustainability.
Nowadays, there is an increasing trend to develop new extraction methods by using non-conventional technologies as an alternative to classic solid-liquid extraction. Ultrasounds treatment is an alternative cheap, effective and reproducible method for the improved recovery of bioactive compound extracts from plant foods, by-products and algae. The goal of the current article is to revise the impact of ultrasound-assisted extraction on the recovery of polyphenols, carotenoids and chlorophylls from plant and algae materials. At this stage of development, there is a need to optimize ultrasound treatment conditions based on ultrasonic power, the temperature and the sonication duration in order to maximize the antioxidant capacity of the obtained extract, mainly attributed to bioactive compounds. Thereby, the modeling attempts of this technology were also discussed.
The enhancement of release of oak-related compounds from oak chips during wine aging with oak chips may interest the winemaking industry. In this study, the 25-kHz ultrasound waves were used to intensify the mass transfer of phenolics from oak chips into a model wine. The influences of acoustic energy density (6.3-25.8W/L) and temperature (15-25°C) on the release kinetics of total phenolics were investigated systematically. The results exhibited that the total phenolic yield released was not affected by acoustic energy density significantly whereas it increased with the increase of temperature during sonication. Furthermore, to describe the mechanism of mass transfer of phenolics in model wine under ultrasonic field, the release kinetics of total phenolics was simulated by both a second-order kinetic model and a diffusion model. The modeling results revealed that the equilibrium concentration of total phenolics in model wine, the initial release rate and effective diffusivity of total phenolics generally increased with acoustic energy density and temperature. In addition, temperature had a negative effect on the second-order release rate constant whereas acoustic energy density had an opposite effect.
To effectively extract value-added phycocyanin from Spirulina platensis, the effects of processing parameters (pH and temperature) on extraction performance and global kinetics were systematically studied. The extraction kinetics was investigated by varying pH levels (5–8) and temperatures (30–60 °C). An empirical kinetic model incorporating the aforementioned factors was developed. A good agreement between the experimental and fitted data was obtained, which indicated that the extraction process followed second-order kinetics. Furthermore, the model parameters (equilibrium concentration, extraction rate constants, and initial rates of extraction) were calculated and formulated as a function of the operating factors. The activation energy of the extraction was 67.1 kJ mol−1, indicating that the process was endothermic. The predictions obtained from the developed model were compared with the experimental data under the same operating conditions. The predicted and experimental data were consistent, indicating the reliability of the model.
We have adapted the Lowry method to the determination of algal proteins collected on membrane filters. When cells of Euglena gracilis were analyzed the yield of protein was linear with cell amounts to 400 μg. The error of repeatability was 3 μg, and the recovery > 98%.
A number of drying methods studied for the processing of Spirulina (crossflow dried, spray dried and oven dried) resulted in approximately 50% loss of phycocyanin. Therefore fresh biomass was suitable for phycocyanin extraction. Of the extraction methods tested, freezing and thawing of cells, homogenisation using a mortar and pestle in the presence of abrasive material and homogenisation using a blender at 10 000 rpm yielded 19.4±0.4 mg phycocyanin per 100 mg dry weight of Spirulina while water extraction was a slow process. Acid treatment also resulted in phycocyanin leaching. Phycocyanin was stable over a pH range of 5–7.5 at 9±1°C, whereas temperature beyond 40°C lead to instability. The pigment phycocyanobilin was separated from the phycocyanin.
A new method using high pressure processing to extract caffeine from green tea leaves was studied. The effect of different parameters such as high hydrostatic pressure (100–600 MPa), different solvents (acetone, methanol, ethanol and water), ethanol concentration (0–100% mL/mL), pressure holding time (1–10 min) and liquid/solid ratio (10:1 to 25:1 mL/g) were studied for the optimal caffeine extraction from green tea leaves. The highest yields (4.0 ± 0.22%.) were obtained at 50% (mL/mL) ethanol concentration, liquid/solid ratio of 20:1 (ml/g), and 500 MPa pressure applied for 1 min. Experiments using conventional extraction methods (extraction at room temperature, ultrasonic extraction and heat reflux extraction) were also conducted, which showed that extraction using high pressure processing possessed several advantages, such as higher yields, shorter extraction times and lower energy consumption.
Sustainable production of renewable energy is being hotly debated globally since it is increasingly understood that first generation biofuels, primarily produced from food crops and mostly oil seeds are limited in their ability to achieve targets for biofuel production, climate change mitigation and economic growth. These concerns have increased the interest in developing second generation biofuels produced from non-food feedstocks such as microalgae, which potentially offer greatest opportunities in the longer term. This paper reviews the current status of microalgae use for biodiesel production, including their cultivation, harvesting, and processing. The microalgae species most used for biodiesel production are presented and their main advantages described in comparison with other available biodiesel feedstocks. The various aspects associated with the design of microalgae production units are described, giving an overview of the current state of development of algae cultivation systems (photo-bioreactors and open ponds). Other potential applications and products from microalgae are also presented such as for biological sequestration of CO2, wastewater treatment, in human health, as food additive, and for aquaculture.
We have adapted the Lowry method to the determination of algal proteins collected on membrane filters. When cells of Euglena gracilis were analyzed the yield of protein was linear with cell amounts to 400 μg. The error of repeatability was 3 μg, and the recovery > 98%.
Phycocyanin extraction from cyanobacteria Spirulina platensis was optimized using factorial design and response surface techniques. The effects of temperature and biomass-solvent ratio on phycocyanin concentration and extract purity were evaluated to determine the optimum conditions for phycocyanin extraction. The optimum conditions for the extraction of phycocyanin from S. platensis were the highest biomass-solvent ratio, 0.08 gmL(-1), and 25 degrees C. Under these conditions it's possible to obtain an extract of phycocyanin with a concentration of 3.68 mgmL(-1) and purity ratio (A(615)/A(280)) of 0.46.
Spirulina—an overview
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S.K. Ali, A.M. Saleh, Spirulina-an overview, Int. J. Pharm. Pharm. Sci. 4 (2012)
9-15.
Toward a greener process production of spirulina protein extract
- Fig
Fig. 10. Toward a greener process production of spirulina protein extract.
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Ultrasonics -Sonochemistry 54 (2019) 48-60
Besoins en protéines et en acides aminés & qualité des protéines alimentaires
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Ecoextraction du végétal intensifiée par ultrasons
- A.-S Fabiano-Tixier
- N Rombaut
- M Jacottet-Navarro
- A Bily
- F Chemat
A.-S. Fabiano-Tixier, N. Rombaut, M. Jacottet-Navarro, A. Bily, F. Chemat, Ecoextraction du végétal intensifiée par ultrasons, La Sonochimie (2016) 31-35.
Algal proteins: extraction
- Bleakley
Eco-extraction du végétal intensifiée par ultrasons
- Fabiano-Tixier