Production Of Phycobiliproteins Research Articles

Enhanced recovery of R-phycoerythrin using recombination cell-assisted pressurized fluid extraction: yield, characterization, and antioxidant evaluation

The recovery of phycobiliproteins, with an emphasis on R-phycoerythrin (R-PE), from Solieria filiformis macroalgae was investigated in the current study. This is accomplished using an environmentally friendly method known as pressurized fluid extraction. Phycobiliprotein production was analyzed over time, considering four different parameters: i) solvents (ultrapure water and phosphate buffer), ii) macroalgae particle sizes, iii) stirring, and iv) pressure of 200 to 800 bar. Notably, the highest R-PE yield reached an impressive 22.53 mg/g dry biomass. The best extraction conditions were identified as a macerated structure, employing a combination of 500 bar pressure and agitation facilitated by a recombination cell. R-phycocyanin (R-PC) was also extracted with a 3.70 mg/g yield. Two distinct extracts, crude and precipitate, underwent comprehensive characterization. Extracted R-PE exhibited an α-helix structure (80.51–90.39 %), suggesting the existence of other proteins in both extracts. Three different subunits, α (15 kDa), β (21 kDa), and γ (29 kDa), were segregated under SDS-PAGE. Moreover, both extracts demonstrated antioxidant capacity, achieving a 99.55 % reduction in 2,2-diphenyl-1-picryl-hydrazil (DPPH) radicals and a 13.94 % chelation of ferrous ions. This study reports the extraction of biomolecules using the recombination cell for the first time in the literature, proving an appealing alternative for non-aggressive extraction with inherent biological activity. These findings underscore the potential applications of these extracted phycobiliproteins in various fields, including biotechnology and health.

Single-step extraction/pre-formulation process for B-phycoerythrin using glycerol-based eutectic solvents: A step toward more sustainable production of phycobiliproteins

B-Phycoerythrin (B-PE) is a protein pigment of significant interest to the food and cosmetic industries. B-PE is obtained from algal or microalgal biomass, such as Porphyridium cruentum, after complex pre-treatment, extraction, and pre-formulation schemes. This work investigated the use of a single step process for the extraction and pre-formulation of B-PE from P. cruentum without complex biomass pretreatment. Five hydrophilic NaDES based on glycerol with varying water contents (20–40%) were implemented by ultrasound-assisted extraction using a one-step extraction procedure. The yields obtained were satisfaying, ranging from 5 to 15 mg of B-PE per gram of dry matter. Additionally, a purity index suitable for use in cosmetics and food products was achieved, with an average of >1.5. Furthermore, the extracts obtained in a eutectic medium demonstrated enhanced stability for B-PE, with an average loss of 10% after accelerated aging, allowing storage at room temperature without loss of quality. In particular, NaDES based on glycerol and glucose and/or betaine appeared to be the most effective in preserving the integrity and antioxidant activity of B-PE, with a range of 15–30% quenched DPPH. The ComplexGAPI tool demonstrated that our single-step NaDES process is more environmentally friendly than conventional extraction processes, thanks to lower energy consumption, reduced unitary operations, and less waste production. These results provide a solid foundation for sustainable management of the valorization of P-BE for cosmetic or food industries.

Magnetic Fields as Inducers of Phycobiliprotein Production by Synechococcus elongatus PCC 7942.

This study aimed to analyze the effect of magnetic field (MF) application on the metabolism of Synechococcus elongatus PCC 7942. Concentrations of biomass, carbohydrate, protein, lipid, and photosynthetic pigments (chlorophyll-a, C-phycocyanin, allophycocyanin and phycoerythrin) were determined. In cultures with MF application (30 mT for 24h d-1), there were increases of 47.5% in total protein content, 87.4% in C-phycocyanin, and 332.8% in allophycocyanin contents, by comparison with the control. Allophycocyanin is the most affected pigment by MF application. Therefore, its biosynthetic route was investigated, and four genes related to its synthesis were found. However, the analysis of the gene expression showed no statistical differences from the control culture, which suggests that induction of such genes may occur soon after MF application with consequent stabilization over time. MF application may be a cost-effective alternative to increase production of compounds of commercial interest by cyanobacteria.

Phycobiliprotein recovery coupled to the tertiary treatment of wastewater in semi-continuous photobioreactors. Tracking contaminants of emerging concern.

This study evaluated a tertiary wastewater treatment technology using cyanobacteria to recover value-added phycobiliproteins. The presence of contaminants of emerging concern (CECs) in wastewater, cyanobacterial biomass and pigments recovered were also analyzed. For this, a wastewater-borne cyanobacterium (Synechocystis sp. R2020) was used to treat secondary effluent from a municipal wastewater treatment plant, with and without nutrients supplementation. Then, the stability of phycobiliprotein production was assessed by operating the photobioreactor in semi-continuous mode. Results showed similar biomass productivity with and without nutrients supplementation (153.5 and 146.7 mg L−1 d−1, respectively). Upon semi-continuous operation, the phycobiliprotein content was stable and reached up to 74.7 mg gDW−1. The phycocyanin purity ratio ranged from 0.5 to 0.8, corresponding to food grade (>0.7). Out of 22 CECs detected in secondary effluent, only 3 were present in the phycobiliprotein extracts. In order to identify applications, prospective research should focus on CECs removal during pigment purification.

Microalgae-Derived Pigments for the Food Industry.

In the food industry, manufacturers and customers have paid more attention to natural pigments instead of the synthetic counterparts for their excellent coloring ability and healthy properties. Microalgae are proven as one of the major photosynthesizers of naturally derived commercial pigments, gaining higher value in the global food pigment market. Microalgae-derived pigments, especially chlorophylls, carotenoids and phycobiliproteins, have unique colors and molecular structures, respectively, and show different physiological activities and health effects in the human body. This review provides recent updates on characteristics, application fields, stability in production and extraction processes of chlorophylls, carotenoids and phycobiliproteins to standardize and analyze their commercial production from microalgae. Potential food commodities for the pigment as eco-friendly colorants, nutraceuticals, and antioxidants are summarized for the target products. Then, recent cultivation strategies, metabolic and genomic designs are presented for high pigment productivity. Technical bottlenecks of downstream processing are discussed for improved stability and bioaccessibility during production. The production strategies of microalgal pigments have been exploited to varying degrees, with some already being applied at scale while others remain at the laboratory level. Finally, some factors affecting their global market value and future prospects are proposed. The microalgae-derived pigments have great potential in the food industry due to their high nutritional value and competitive production cost.

Production of phycobiliproteins, bioplastics and lipids by the cyanobacteria Synechocystis sp. treating secondary effluent in a biorefinery approach.

Cyanobacteria have been identified as promising organisms to reuse nutrients from waste effluents and produce valuable compounds such as lipids, polyhydroxyalkanoates (PHAs), and pigments. However, almost all studies on cyanobacterial biorefineries have been performed under lab scale and short cultivation periods. The present study evaluates the cultivation of the cyanobacterium Synechocystis sp. in a pilot scale 30 L semi-continuous photobioreactor fed with secondary effluent for a period of 120 days to produce phycobiliproteins, polyhydroxybutyrate (PHB) and lipids. To this end, the harvested biomass from the semi-continuous photobioreactor was transferred into 5 L vertical column batch photobioreactors to perform PHB and lipid accumulation under nutrient starvation. Three hydraulic retention times (HRT) (6, 8 and 10 days) were tested in the semi-continuous photobioreactor to evaluate its influence on biomass growth and microbial community. A maximum biomass concentration of 1.413 g L-1 and maximum productivity of 173 mg L-1 d-1 was reached under HRT of 8 days. Microscopy analysis revealed a shift from Synechocystis sp. to Leptolyngbya sp. and green algae when HRT of 6 days was used. Continuous, stable production of phycobiliproteins in the semi-continuous photobioreactor was obtained, reaching a maximum content of 7.4%dcw in the biomass. In the batch photobioreactors a PHB content of 4.8%dcw was reached under 7 days of nitrogen and phosphorus starvation, while a lipids content of 44.7%dcw was achieved under 30 days of nitrogen starvation. PHB and lipids production was strongly dependent on the amount of nutrients withdrawn from the grow phase. In the case of lipids, their production was stimulated when there was only phosphorus depletion. While Nitrogen and phosphorus limitation was needed to enhance the PHB production. In conclusion, this study demonstrates the feasibility of cultivating cyanobacteria in treated wastewater to produce bio-based valuable compounds within a circular bioeconomy approach.

Effects of herbicide on various life forms with special reference to the paddy fields in the Eastern Belts of India

Effect of external pH on cyanobacterial phycobiliproteins production and ammonium excretionOjit Singh Keithellakpam, Tiwari Onkar Nath, Avijeet Singh Oinam, Indrama Thingujam, Gunapati Oinam, Sharma Gauri Dutt

The Effect of LEDs on Biomass and Phycobiliproteins Production in Thermotolerant Oscillatoria sp.

This study evaluates the role of different LED lights (white, blue/red), intensity (µmol m−2 s−1), and photoperiod in the production of biomass and phycocyanin-C, allophycocyanin and phycoerythrin (C-PC, APC, and PE respectively) from a novel thermotolerant strain of Oscillatoria sp. Results show that a mixture of white with blue/red LEDs can effectively double the biomass concentration up to 1.3 g/L, while the concentration of the selected phycobiliproteins increased proportionally to biomass. Results also indicate that high light intensities (>120 µmol m−2 s−1) can diminish the final concentration of C-PC, APC, and PE, significantly reducing the overall biomass produced. Finally, the photoperiod analysis showed that longer light exposure times (18:6 h) improved both biomass and phycobiliproteins concentration. These results demonstrate that the application of LEDs to produce a novel strain of Oscillatoria sp can double the biomass concentration, and the photoperiod regulation can eventually enhance the final concentration of specific phycobiliproteins such as APC and PE.

Impact of light wavelength on the transcriptome of Porphyridium cruentum and culture yield

Changes in light wavelength can alter the phycobiliprotein content in photosynthetic organisms . These molecules are involved in harvesting light energy for photosynthesis and are interesting microalgal products. Furthermore, increasing or modifying phycobiliprotein content in eukaryotic microalgae is possible. Specifically, exposure to different wavelengths triggers their ability to switch their metabolism using light. Moreover, molecular findings support light modulation of phycobiliproteins in the microalgae Porphyridium cruentum . Therefore, this research aimed to describe how two light spectra affects the transcriptome in P. cruentum cultures. Notably, blue and white wavelengths did not significantly modify the phycobiliprotein and macromolecules content by biomass unit. Nevertheless, a differential expression profile was found in the transcriptome; a PHO80-like cyclin PC3933011 was up-regulated, likely connected to the higher cell number obtained under the white wavelength. Finally, we established the viability of manipulating light for phycobiliprotein production in this P. cruentum and confirmed that light acts as a cell cycle regulator . • P. cruentum phycobiliprotein gene pathway expression was not affected by light. • Narrow white-light induced the overexpression of the PHO80-like cyclin PC3933011. • Cultures of P. cruentum grown under white light showed increased cell concentration. • Modulation of light can be effectively used for P. cruentum growth manipulation. • Macromolecular cell content is not affected by light changes.

Freeze-thaw-assisted aqueous two-phase system as a green and low-cost option for analytical grade B-phycoerythrin production from unicellular microalgae Porphyridium purpureum

Microalgae are promising alternative sources for natural phycobiliprotein production since they grow rapidly and can be cultured on a large scale. However, the overall economic viability of analytical grade phycoerythrin production from microalgae is limited by the high cost of the pretreatment required to break down the cell wall, as well as the expensive reagents and equipment used during purification. This study developed a novel extraction and purification process to recovery B-phycoerythrin (B-PE) in a freeze-thaw-assisted aqueous two-phase system (FT-ATPS) from Porphyridium purpureum. The effects of culture strategy, extraction method and purification factors on B-PE production were studied systematically. The results revealed that low light induced B-PE synthesis, whereas hindered the growth. The stepwise light supplement strategy alleviated the growth limitation under low-light conditions and increased the final B-PE production, up to 182.1 ± 4.1 mg/L (increasing by 73.1 %). Compared with the traditional electricity highly-consumed ultrasonication method, the green freeze-thaw method could extract B-PE without damaging the cell structure and save much more power (>98 %), and the extraction rate reached the highest when frozen at −80 °C. The purity and concentration of B-PE in crude extract were proved to have a significant effect on ATPS purification, and the higher the former, the better the latter. A total production cost of 0.045 $/mg (decreasing by 96.2 %) and purity of 5.6 ± 0.2 (A545/A280) for analytical grade B-PE production were achieved by FT-ATPS based on 4 % polyethylene glycol (PEG2,000) and 20 % ammonium sulfate. Relative to the conventional method of phycoerythrin production, the method developed herein has greater safety, lower cost, and more environmental benefits, which can be function appropriately in the expensive purification of analytical grade phycobiliproteins.

Colaconema formosanum, Sarcodia suae, and Nostoc commune as Fermentation Substrates for Bioactive Substance Production

Bioactive compounds extracted from natural renewable sources have attracted an increased interest from both industry and academia. Recently, algae have been highlighted as promising sources of bioactive compounds, such as polyphenols, polysaccharides, fatty acids, proteins, and pigments, which can be used as functional ingredients in many industrial applications. Therefore, a simple green extraction and purification methodology capable of recovering biocompounds from algal biomass is of extreme importance in commercial production. In this study, we evaluated the application of three valuable algae (Colaconema formosanum, Sarcodia suae, and Nostoc commune) in combination with Pseudoalteromonas haloplanktis (type strain ATCC 14393) for the production of versatile compounds. The results illustrate that after 6 h of first-stage fermentation, the production of phycobiliproteins in C. formosanum was significantly increased by 156.2%, 188.9%, and 254.17% for PE, PC, and APC, respectively. This indicates that the production of phycobiliproteins from algae can be enhanced by P. haloplanktis. Furthermore, we discovered that after S. suae and N. commune were fermented with P. haloplanktis, mannose was produced. In this study, we describe a feasible biorefinery process for the production of phycobiliproteins and mannose by fermenting marine macroalgae with cyanobacteria. We believe it is worth establishing a scale-up technique by applying this fermentation method to the production of phycobiliproteins and mannose in the future.

Commercial Potential of the Cyanobacterium Arthrospira maxima: Physiological and Biochemical Traits and the Purification of Phycocyanin.

Simple SummaryArthrospira maxima is an unbranched, filamentous cyanobacterium rich in important cellular products such as vitamins, minerals, iron, essential amino acids, essential fatty acids, and protein, which has made it one of the most important commercial photoautotrophs. To optimize the growth conditions for the production of target compounds and to ensure profitability in commercial applications, the effects of pH and temperature were investigated. A. maxima has been shown to be tolerant to a range of pH conditions and to exhibit hyper-accumulation of phycoerythrin and allophycocyanin at low temperatures. These traits may offer significant advantages for future exploitation, especially in outdoor cultivation with fluctuating pH and temperature. Our study also demonstrated a new method for the purification of phycocyanin from A. maxima by using by ultrafiltration, ion-exchange chromatography, and gel filtration, producing PC at 1.0 mg·mL−1 with 97.6% purity.Arthrospira maxima is a natural source of fine chemicals for multiple biotechnological applications. We determined the optimal environmental conditions for A. maxima by measuring its relative growth rate (RGR), pigment yield, and photosynthetic performance under different pH and temperature conditions. RGR was highest at pH 7–9 and 30 °C. Chlorophyll a, phycocyanin, maximal quantum yield (Fv/Fm), relative maximal electron transport rate (rETRmax), and effective quantum yield (ΦPSII) were highest at pH 7–8 and 25 °C. Interestingly, phycoerythrin and allophycocyanin content was highest at 15 °C, which may be the lowest optimum temperature reported for phycobiliprotein production in the Arthrospira species. A threestep purification of phycocyanin (PC) by ultrafiltration, ion-exchange chromatography, and gel filtration resulted in a 97.6% purity of PC.

Assessment of novel halo- and thermotolerant desert cyanobacteria for phycobiliprotein production

Four indigenous cyanobacteria isolates identified as QUCCCM 34: Chroococcidiopsis sp., QUCCCM 54: Pleurocapsa sp., QUCCCM 77: Euhalothece sp., and QUCCCM 129: Cyanobacterium sp. were investigated during this study. Temperatures and salinities observed in outdoor were reproduced indoor, using small-scale photobioreactors, and culture conditions were optimized for maximum biomass and phycobiliprotein productions. The strains showed their halo and thermotolerance capacity. The highest biomass productivity was 125 ± 1.1 mg x L−1 d−1 for Pleurocapsa sp.at 30 °C–40 ppt. The major phycobiliproteins were phycocyanin, and the content was strain and age dependent. Pleurocapsa sp., Euhalothece sp., and Cyanobacterium sp reached their highest phycocyanin content (up to 160 ± 2.6 mgPC gx-1) after 4, 8, and 10 days, respectively, while it was only up to 100 ± 3.5 mgPC gx-1 for Chroococcidiopsis sp. at day 4, 40 °C–60 ppt. Increasing temperature and salinity stimulated the phycocyanin synthesis in Chroococcidiopsis sp, Pleurocapsa sp. and Euhalothece sp., whereas only salinity increment enhanced the pigments production(both phycoerythrin and phycocyanin) for Cyanobacterium sp. Finally, all the pigment extracts exhibited an antioxidant and radical scavenging activity which were maximal for the extracts from Pleurocapsa sp., with ≈ 60 mM Trolox equivalent gx−1 and 50%, respectively.

Effects of Climate Change on the Production of Polysaccharides and Phycobiliproteins by Nostoc commune Vaucher ex Bornet et Flahault

Nostoc commune synthesizes polysaccharides and phycobiliproteins under natural conditions, but little is known about how environmental changes could affect their production. In this study, colonies of N. commune were subjected to increases in ultraviolet radiation, ammonium concentration, electrical conductivity, and temperature, to assess the potential changes in the concentrations of polysaccharides and phycobiliproteins. The results indicate that UVB radiation significantly increased the synthesis of polysaccharides (F = 62.691; p < 0.01), while UVA radiation caused a significant increase in the production of total phycobiliproteins (F = 22.472, p < 0.01) phycocyanin (F = 8.546, p < 0.01), phycoerythrin (F = 12.876, p < 0.01), and allophycocyanin (F = 58.143, p < 0.001). Also, 50 µM NH4Cl significantly increased the synthesis of polysaccharides (F = 45.706; p < 0.01) while increased near significant total phycobiliproteins (F = 5.043, p < 0.1), phycoerythrins (F = 4.57, p < 0.1), allophycocyanin (F = 4.892, p < 0.1), and phycocyanin (F = 4.921, p < 0.1). Furthermore, a conductivity value of 4 mScm−1 enhanced near significant the production of polysaccharides (F = 4.816; p < 0.1) and phycocyanin (F = 9.728, p < 0.1). Nevertheless, a significant effect of total phycobiliproteins was observed (F = 23.686, p < 0.01), as well as allophycocyanin (F = 57.092, p < 0.001), and phycoerythrin (F = 13.928, p < 0.01). Finally, the optimal temperature for the synthesis of polysaccharides was 30 °C. Also, 30 ºC significantly increased the synthesis of total phycobiliproteins (F = 292.211, p < 0.001), as well as on phycocyanin (F = 126.433, p < 0.001) and allophycocyanin (F = 7.991, p < 0.05). These data indicate the ability of N. commune to modify its synthesis of polysaccharides and phycobiliproteins in response to extreme environmental conditions related to climate change, underscoring the interest in N. commune for future applied research on the biotechnological and pharmaceutical production of both types of compounds.

Cyanobacterial biodiversity from Thai karstic caves as a potential source for phycobiliprotein production

Cyanobacteria are highly adaptive microorganisms that can survive in harsh conditions, with have outstanding biotechnology potential as the ability to produce value-added compounds. Karst caves are extreme environments with low light and nutrient availability as interesting areas to investigate cyanobacterial diversity with phycobiliprotein production potential. The native cyanobacteria were isolated from 23 karst caves in Thailand. All isolates were morphotype and identified as 12 genera and 86 species, with the Leptolyngbya species commonly abundant. Six isolated cyanobacteria, Leptolyngbya sp. SCOM01, Phormidesmis sp. RK02, Leptolyngbya sp. SWL01, Scytolyngbya sp. LKK05, Leptolyngbya sp. LKK14 and Nostoc sp. SW02 produced phycobiliprotein content at over 400 mg g protein−1. The highest yield of phycobiliprotein (31.9%) was obtained from Nostoc sp. SW02, with the ratio of phycoerythrin:phycocyanin:allophycocyanin 3.4:1.9:1.0. All extracted phycobiliproteins were purified and revealed different spectral characteristics. Knowledge of cyanobacterial isolate distribution in Thai karstic cave habitats has potential as a new source of phycobiliprotein production for further applications.

Production of Phycobiliproteins, Bioplastics and Lipids by the Cyanobacterium Synechocystis Sp Using Secondary Effluents in a Biorefinery Approach

Production of Phycobiliproteins, Bioplastics and Lipids by the Cyanobacterium Synechocystis Sp Using Secondary Effluents in a Biorefinery Approach

Tunisian inland water microflora as a source of phycobiliproteins and biological activity with beneficial effects on human health

Abstract Ten monoclonal microalgal cultures were obtained from several Tunisian inland water bodies, and their dichloromethane and methanolic extracts were screened for antibacterial, antileishmanial, and antioxidant properties, as well as phycobiliprotein production capacity. Cylindrospermopsis raciborskii has been shown to synthesize high levels of phycocyanin and may be an effective alternative source to other sources used for commercial production of phycocyanin. Chroococcus sp. and Leptolyngbya sp1. exhibited the strongest radical scavenging activity against DPPH (IC50 = 212.15 and 263.91 μg ml−1, respectively), indicating their promising potential for use as new effective and non-toxic antioxidants. Furthermore, Dunaliella sp. showed an interesting antileishmanial activity against the pathogens Leishmania infantum and Leishmania major (IC50 = 151 and 284 μg ml−1, respectively), thus representing a good candidate for use against cutaneous and visceral leishmaniasis in Tunisia, a country endemic to these diseases where thousands of new cases are registered every year. These results suggest that the strains of microalgae featured in this work have the potential to serve as natural alternative, safe and sustainable sources of high value-added products that could be used to improve the final biomass value.

Effect of glucose and sodium nitrate on the cultivation of Nostoc sp. PCC 7423 and production of phycobiliproteins

Effect of glucose and sodium nitrate on the cultivation of Nostoc sp. PCC 7423 and production of phycobiliproteins

Biodiversity of cyanobacteria in fresh water ponds of Pudukkottai district, Tamil Nadu, India

Effect of external pH on cyanobacterial phycobiliproteins production and ammonium excretionOjit Singh Keithellakpam, Tiwari Onkar Nath, Avijeet Singh Oinam, Indrama Thingujam, Gunapati Oinam, Sharma Gauri Dutt

Diazotrophs modulate phycobiliproteins and nitrogen stoichiometry differently than other cyanobacteria in response to light and nitrogen availability

AbstractHarmful algal blooms (HABs) frequently occur in eutrophic water bodies, which are often dominated by either non‐diazotrophic or diazotrophic strains of cyanobacteria. The occurrence of HABs is associated with multiple environmental factors that affect cyanobacterial growth, though adaptations of cyanobacteria to respond to diverse growing conditions are largely unknown. One factor affecting HAB persistence is the ability of cyanobacteria to synthesize and metabolize large biomolecules such as the light harvesting pigments phycobiliproteins that can also act as nitrogen (N) storage. Here, we examined how cyanobacteria varied phycobiliprotein content in response to different light intensity and N availability in two HAB forming species, Microcystis aeruginosa (non‐diazotrophic) and Dolichospermum flos‐aquae (diazotrophic). Our results revealed both HAB forming species responded similarly to light intensity, with decreasing light causing an increased production of phycobiliproteins, although at different rates. However, we observed dissimilar phycobiliprotein responses to varied N availability. In low N conditions, M. aeruginosa degraded phycobiliproteins primarily due to the decreased need for photosynthesis, whereas D. flos‐aquae, increased phycobiliproteins production under N‐limited conditions likely caused by the increased energetic requirements for N2‐fixation. Our results indicate that M. aeruginosa stored as much as 30% of cellular N in phycobiliproteins under high N conditions, while D. flos‐aquae only allocated 5% of the cellular N in phycobiliproteins. This study reveals a fundamental physiological difference between non‐diazotrophic and diazotrophic cyanobacteria, highlighting the critical role of phycobiliproteins on the persistence of cyanobacteria blooms under scenarios with varied light intensity and N availability.