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1.
Toxins (Basel) ; 16(5)2024 Apr 26.
Article in English | MEDLINE | ID: mdl-38787058

ABSTRACT

Cyanobacterial harmful algal blooms (cyanoHABs) occur in fresh water globally. These can degrade water quality and produce toxins, resulting in ecological and economic damages. Thus, short-term management methods (i.e., algaecides) are necessary to rapidly mitigate the negative impacts of cyanoHABs. In this study, we assess the efficacy of a hydrogen peroxide-based algaecide (PAK® 27) on a Microcystis dominated bloom which occurred within the Pahokee Marina on Lake Okeechobee, Florida, USA. We observed a significant reduction in chlorophyll a (96.81%), phycocyanin (93.17%), and Microcystis cell counts (99.92%), and a substantial reduction in microcystins (86.7%) 48 h after treatment (HAT). Additionally, there was a significant shift in bacterial community structure 48 HAT, which coincided with an increase in the relative abundance of photosynthetic protists. These results indicate that hydrogen peroxide-based algaecides are an effective treatment method for cyanoHAB control and highlight their effects on non-target microorganisms (i.e., bacteria and protists).


Subject(s)
Harmful Algal Bloom , Hydrogen Peroxide , Lakes , Florida , Hydrogen Peroxide/pharmacology , Lakes/microbiology , Microcystis/drug effects , Microcystis/growth & development , Cyanobacteria/drug effects , Microbiota/drug effects , Microcystins , Phycocyanin/pharmacology , Chlorophyll A/metabolism
2.
Int J Mol Sci ; 25(10)2024 May 09.
Article in English | MEDLINE | ID: mdl-38791192

ABSTRACT

The synapses between inner hair cells (IHCs) and spiral ganglion neurons (SGNs) are the most vulnerable structures in the noise-exposed cochlea. Cochlear synaptopathy results from the disruption of these synapses following noise exposure and is considered the main cause of poor speech understanding in noisy environments, even when audiogram results are normal. Cochlear synaptopathy leads to the degeneration of SGNs if damaged IHC-SGN synapses are not promptly recovered. Oxidative stress plays a central role in the pathogenesis of cochlear synaptopathy. C-Phycocyanin (C-PC) has antioxidant and anti-inflammatory activities and is widely utilized in the food and drug industry. However, the effect of the C-PC on noise-induced cochlear damage is unknown. We first investigated the therapeutic effect of C-PC on noise-induced cochlear synaptopathy. In vitro experiments revealed that C-PC reduced the H2O2-induced generation of reactive oxygen species in HEI-OC1 auditory cells. H2O2-induced cytotoxicity in HEI-OC1 cells was reduced with C-PC treatment. After white noise exposure for 3 h at a sound pressure of 118 dB, the guinea pigs intratympanically administered 5 µg/mL C-PC exhibited greater wave I amplitudes in the auditory brainstem response, more IHC synaptic ribbons and more IHC-SGN synapses according to microscopic analysis than the saline-treated guinea pigs. Furthermore, the group treated with C-PC had less intense 4-hydroxynonenal and intercellular adhesion molecule-1 staining in the cochlea compared with the saline group. Our results suggest that C-PC improves cochlear synaptopathy by inhibiting noise-induced oxidative stress and the inflammatory response in the cochlea.


Subject(s)
Cochlea , Intercellular Adhesion Molecule-1 , Noise , Oxidative Stress , Phycocyanin , Synapses , Animals , Oxidative Stress/drug effects , Guinea Pigs , Phycocyanin/pharmacology , Phycocyanin/therapeutic use , Cochlea/metabolism , Cochlea/drug effects , Cochlea/pathology , Synapses/drug effects , Synapses/metabolism , Noise/adverse effects , Intercellular Adhesion Molecule-1/metabolism , Hearing Loss, Noise-Induced/drug therapy , Hearing Loss, Noise-Induced/metabolism , Hearing Loss, Noise-Induced/pathology , Reactive Oxygen Species/metabolism , Male , Spiral Ganglion/drug effects , Spiral Ganglion/metabolism , Spiral Ganglion/pathology , Hydrogen Peroxide/metabolism , Hair Cells, Auditory, Inner/drug effects , Hair Cells, Auditory, Inner/metabolism , Hair Cells, Auditory, Inner/pathology , Antioxidants/pharmacology , Cell Line , Hearing Loss, Hidden
3.
Food Res Int ; 186: 114362, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38729724

ABSTRACT

As food safety continues to gain prominence, phycocyanin (PC) is increasingly favored by consumers as a natural blue pigment, which is extracted from microalgae and serves the dual function of promoting health and providing coloration. Spirulina-derived PC demonstrates exceptional stability within temperature ranges below 45 °C and under pH conditions between 5.5 and 6.0. However, its application is limited in scenarios involving high-temperature processing due to its sensitivity to heat and light. This comprehensive review provides insights into the efficient production of PC from microalgae, covers the metabolic engineering of microalgae to increase PC yields and discusses various strategies for enhancing its stability in food applications. In addition to the most widely used Spirulina, some red algae and Thermosynechococcus can serve as good source of PC. The genetic and metabolic manipulation of microalgae strains has shown promise in increasing PC yield and improving its quality. Delivery systems including nanoparticles, hydrogels, emulsions, and microcapsules offer a promising solution to protect and extend the shelf life of PC in food products, ensuring its vibrant color and health-promoting properties are preserved. This review highlights the importance of metabolic engineering, multi-omics applications, and innovative delivery systems in unlocking the full potential of this natural blue pigment in the realm of food applications, provides a complete overview of the entire process from production to commercialization of PC, including the extraction and purification.


Subject(s)
Microalgae , Phycocyanin , Microalgae/metabolism , Spirulina/chemistry , Spirulina/metabolism , Metabolic Engineering
4.
Nutrients ; 16(9)2024 Apr 26.
Article in English | MEDLINE | ID: mdl-38732545

ABSTRACT

Natural products from mushrooms, plants, microalgae, and cyanobacteria have been intensively explored and studied for their preventive or therapeutic potential. Among age-related pathologies, neurodegenerative diseases (such as Alzheimer's and Parkinson's diseases) represent a worldwide health and social problem. Since several pathological mechanisms are associated with neurodegeneration, promising strategies against neurodegenerative diseases are aimed to target multiple processes. These approaches usually avoid premature cell death and the loss of function of damaged neurons. This review focuses attention on the preventive and therapeutic potential of several compounds derived from natural sources, which could be exploited for their neuroprotective effect. Curcumin, resveratrol, ergothioneine, and phycocyanin are presented as examples of successful approaches, with a special focus on possible strategies to improve their delivery to the brain.


Subject(s)
Curcumin , Neurodegenerative Diseases , Neuroprotective Agents , Resveratrol , Neuroprotective Agents/pharmacology , Humans , Neurodegenerative Diseases/drug therapy , Neurodegenerative Diseases/prevention & control , Curcumin/pharmacology , Resveratrol/pharmacology , Ergothioneine/pharmacology , Biological Products/pharmacology , Biological Products/therapeutic use , Phycocyanin/pharmacology , Animals , Cyanobacteria , Agaricales/chemistry , Microalgae
5.
J Agric Food Chem ; 72(21): 12219-12228, 2024 May 29.
Article in English | MEDLINE | ID: mdl-38747135

ABSTRACT

Phycocyanobilin, an algae-originated light-harvesting pigment known for its antioxidant properties, has gained attention as it plays important roles in the food and medication industries and has surged in demand owing to its low-yield extraction from natural resources. In this study, engineered Corynebacterium glutamicum was developed to achieve high PCB production, and three strategies were proposed: reinforcement of the heme biosynthesis pathway with the introduction of two PCB-related enzymes, strengthening of the pentose phosphate pathway to generate an efficient cycle of NADPH, and fed-batch fermentation to maximize PCB production. Each approach increased PCB synthesis, and the final engineered strain successfully produced 78.19 mg/L in a flask and 259.63 mg/L in a 5 L bioreactor, representing the highest bacterial production of PCB reported to date, to our knowledge. The strategies applied in this study will be useful for the synthesis of PCB derivatives and can be applied in the food and pharmaceutical industries.


Subject(s)
Corynebacterium glutamicum , Metabolic Engineering , Phycobilins , Phycocyanin , Corynebacterium glutamicum/metabolism , Corynebacterium glutamicum/genetics , Phycocyanin/metabolism , Phycocyanin/genetics , Phycobilins/metabolism , Phycobilins/genetics , Fermentation , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Pentose Phosphate Pathway/genetics , Bioreactors/microbiology
6.
Int J Biol Macromol ; 269(Pt 2): 131969, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38697419

ABSTRACT

In this study, different concentrations of sodium alginate were compounded with pectin and phycocyanin to co-prepare composite hydrogel spheres (HP-PC-SA 0.2 %, 0.6 %, 1.0 %, 1.4 %) to evaluate the potential of the composite hydrogel spheres for the application as phycocyanin delivery carriers. The hydrogel spheres' physicochemical properties and bioaccessibility were assessed through scanning electron microscopy, textural analysis, drug-carrying properties evaluation, and in vitro and in vivo controlled release analysis in the gastrointestinal environment. Results indicated that higher sodium alginate concentrations led to smaller pore sizes and denser networks on the surface of hydrogel spheres. The textural properties of hydrogel spheres improved, and their water-holding capacity increased from 93.01 % to 97.97 %. The HP-PC-SA (1.0 %) formulation achieved the highest encapsulation rate and drug loading capacity, at 96.87 % and 6.22 %, respectively. Within the gastrointestinal tract, the composite hydrogel's structure significantly enhanced and protected the phycocyanin's digestibility, achieving a bioaccessibility of up to 88.03 %. In conclusion, our findings offer new insights into improving functionality and the effective use of phycocyanin via pectin-based hydrogel spheres.


Subject(s)
Alginates , Drug Carriers , Hydrogels , Pectins , Phycocyanin , Alginates/chemistry , Pectins/chemistry , Phycocyanin/chemistry , Hydrogels/chemistry , Drug Carriers/chemistry , Drug Delivery Systems , Drug Liberation , Biological Availability , Animals
7.
Arch Microbiol ; 206(6): 258, 2024 May 12.
Article in English | MEDLINE | ID: mdl-38735006

ABSTRACT

Phycocyanin, a blue-coloured pigment, predominantly found and derived from Spirulina sp., has gained researchers' interest due to its vibrant hues and other attractive properties like antioxidant and anti-microbial. However, the lack of reliable and sustainable phycocyanin extraction strategies without compromising the quality has hindered the scaling up of its production processes for commercial purposes. Here in this study, phycocyanin was extracted from wet and dry biomass Spirulina sp., using three different physical cell disruption methods (ultrasonication, homogenization, and freeze-thaw cycles) combined with two different buffers (phosphate buffer and acetate buffer) and water (as control). The result showed that the freeze-thaw method combined with acetate buffer produced the highest yield (25.013 ± 2.572 mg/100 mg) with a purity ratio of 0.806 ± 0.079. Furthermore, when subjected to 30% w/v salt stress, 1.9 times higher phycocyanin yield with a purity ratio of 1.402 ± 0.609 was achieved using the previously optimized extraction method.


Subject(s)
Phycocyanin , Salt Stress , Spirulina , Phycocyanin/metabolism , Phycocyanin/isolation & purification , Spirulina/metabolism , Spirulina/chemistry , Biomass , Freezing
8.
PLoS One ; 19(4): e0300538, 2024.
Article in English | MEDLINE | ID: mdl-38558076

ABSTRACT

PURPOSE: The cryopreservation process damages oocytes and impairs development potential. As a potent antioxidant, C-phycocyanin (PC) regulates reproductive performance. However, its beneficial effects on vitrified human oocytes remain unknown. METHODS: In this study, human GV-stage oocytes obtained from controlled ovarian hyperstimulation (COH) cycles were randomly allocated to three groups: fresh oocyte without freezing (F group), vitrification in medium supplemented with PC (P group), and vitrification in medium without PC as control group (C group). After warming, viable oocytes underwent in vitro maturation. RESULTS: Our results showed that 3 µg/mL PC treatment increased the oocyte maturation rate after cryopreservation. We also found that PC treatment maintains the regular morphological features of oocytes. After PC treatment, confocal fluorescence staining showed a significant increase in the mitochondrial membrane potential of the vitrified oocytes, along with a notable decrease in intracellular reactive oxygen species and the early apoptosis rate. Finally, after in vitro maturation and parthenogenetic activation, vitrified oocytes had a higher potential for cleavage and blastocyst formation after PC treatment. CONCLUSION: Our results suggest that PC improves the developmental potential of cryopreserved human GV-stage oocytes by attenuating oxidative stress and early apoptosis and increasing the mitochondrial membrane potential.


Subject(s)
Cryopreservation , Phycocyanin , Humans , Reactive Oxygen Species/metabolism , Phycocyanin/pharmacology , Cryopreservation/methods , Oocytes , Vitrification
9.
Sci Rep ; 14(1): 9174, 2024 04 22.
Article in English | MEDLINE | ID: mdl-38649495

ABSTRACT

This study aimed to evaluate the efficacy of dielectric barrier discharge treatment (DBD) combined with phycocyanin pigment (PC) in extending the shelf life of Oncorhynchus mykiss rainbow fillets stored at 4 ± 0.1 °C. Microbiological, physicochemical, sensory and antioxidant properties were assessed over an 18-day storage period. The combined DBD and PC treatment significantly inhibited total viable counts and Psychrotrophic bacteria counts compared to the rest of the samples throughout storage. While Total Volatile Nitrogen concentrations remained below international standard until day 18, they exceeded this threshold in control sample by day 9. DBD treatment notably reduced Trimethylamine levels compared to controls (p < 0.05). PC and DBD combined inhibited DPPH and ABTS radical scavenging capacities by 80% and 85%, respectively, while demonstrating heightened iron-reducing antioxidant activity compared to controls. Analysis of 24 fatty acids indicated that PC mitigated DBD's adverse effects, yielding superior outcomes compared to controls. The ratio of n-3 to n-6 fatty acids in all samples met or fell below international standard. Thus, the combined use of DBD and PC shows promise in extending fillet shelf life by over 15 days at 4 °C.


Subject(s)
Food Preservation , Food Storage , Oncorhynchus mykiss , Phycocyanin , Animals , Food Storage/methods , Oncorhynchus mykiss/microbiology , Oncorhynchus mykiss/growth & development , Food Preservation/methods , Phycocyanin/pharmacology , Antioxidants/pharmacology , Plasma Gases/pharmacology , Seafood , Food Packaging/methods
10.
Int J Mol Sci ; 25(8)2024 Apr 11.
Article in English | MEDLINE | ID: mdl-38673801

ABSTRACT

In recent years, marine natural products have become one of the most important resources of novel lead compounds for critical diseases associated with age. Spirulina, a dietary supplement made from blue-green algae (cyanobacteria: scientific name Arthrospira platensis), is particularly rich in phycocyanin, a phycobiliprotein, which accounts for up to 20% of this cyanobacterium's dry weight and is considered responsible for its anti-cancer, anti-inflammatory and antioxidant activities. Although the anti-aging activity of phycocyanin has been investigated, how exactly this compound works against aging remains elusive. The aim of our research is to use the yeast Saccharomyces cerevisiae as a model organism to investigate the anti-aging properties of phycocyanin from A. platensis. Our results show that phycocyanin has a powerful anti-aging effect, greatly extending the chronological life span of yeast cells in a dose-dependent way, as the effect was also pronounced when cells were grown in SD medium under calorie restriction conditions (0.2% glucose). Both ROS and accumulation of dead cells were followed by staining chronologically aged cells with dihydrorhodamine 123 (DHR123) and propidium iodide (PI). Interestingly, we found that most of the aged phycocyanin-treated cells, which were unable to form colonies, were actually ROS+/PI-. Finally, we show that the moment in which phycocyanin is added to the culture does not substantially influence its effectiveness in counteracting chronological aging.


Subject(s)
Phycocyanin , Saccharomyces cerevisiae , Spirulina , Phycocyanin/pharmacology , Spirulina/chemistry , Saccharomyces cerevisiae/drug effects , Reactive Oxygen Species/metabolism , Aging/drug effects , Antioxidants/pharmacology
11.
Theriogenology ; 222: 66-79, 2024 Jul 01.
Article in English | MEDLINE | ID: mdl-38626583

ABSTRACT

In vitro maturation (IVM) and cryopreservation of goat oocytes are important for establishing a valuable genetic bank for domesticated female animals and improving livestock reproductive efficiency. C-Phycocyanin (PC) is a Spirulina extract with antioxidant, antiinflammatory, and radical scavenging properties. However, whether PC has positive effect on goat oocytes IVM or developmental competence after vitrification is still unknown. In this study, we found that first polar body extrusion (n = 293), cumulus expansion index (n = 269), and parthenogenetic blastocyst formation (n = 281) were facilitated by adding 30 µg/mL PC to the oocyte maturation medium when compared with the control groups and that supplemented with 3, 10, 100 or 300 µg/mL PC (P < 0.05). Although PC supplementation did not affect spindle formation or chromosome alignment (n = 115), it facilitated or improved cortical granules migration (n = 46, P < 0.05), mitochondria distribution (n = 39, P < 0.05), and mitochondrial membrane potential (n = 46, P < 10-4). Meanwhile, supplementation with 30 µg/mL PC in the maturation medium could significantly inhibit the reactive oxygen species accumulation (n = 65, P < 10-4), and cell apoptosis (n = 42, P < 0.05). In addition, PC increased the oocyte mRNA levels of GPX4 (P < 0.01), and decreased the mRNA and protein levels of BAX (P < 0.01). Next, we investigated the effect of PC supplementation in the vitrification solution on oocyte cryopreservation. When compared with the those equilibrate in the vitrification solution without PC, recovered oocytes in the 30 µg/mL PC group showed higher ratios of normal morphology (n = 85, P < 0.05), survival (n = 85, P < 0.05), first polar body extrusion (n = 62, P < 0.05), and parthenogenetic blastocyst formation (n = 107, P < 0.05). Meanwhile, PC supplementation of the vitrification solution increased oocyte mitochondrial membrane potential (n = 53, P < 0.05), decreased the reactive oxygen species accumulation (n = 73, P < 0.05), promoted mitochondria distribution (n = 58, P < 0.05), and inhibited apoptosis (n = 46, P < 10-3). Collectively, our findings suggest that PC improves goat oocyte IVM and vitrification by reducing oxidative stress and early apoptosis, which providing a novel strategy for livestock gamete preservation and utilization.


Subject(s)
Cryopreservation , Goats , In Vitro Oocyte Maturation Techniques , Oocytes , Phycocyanin , Vitrification , Animals , Oocytes/drug effects , In Vitro Oocyte Maturation Techniques/veterinary , In Vitro Oocyte Maturation Techniques/methods , Vitrification/drug effects , Cryopreservation/veterinary , Cryopreservation/methods , Phycocyanin/pharmacology , Female , Reactive Oxygen Species/metabolism , Membrane Potential, Mitochondrial/drug effects
12.
Int J Biol Macromol ; 268(Pt 2): 131599, 2024 May.
Article in English | MEDLINE | ID: mdl-38626840

ABSTRACT

Phycocyanin (PC), a protein derived from algae, is non-toxic and biocompatible. Due to its environmental and sustainable properties, it has been studied as an alternative stabilizer for food emulsions. In this sense, the main objective of this work is to evaluate the effectiveness of PC and its use in combination with diutan gum (DG), a biological macromolecule, to prepare emulgels formulated with avocado oil. Z-potential measurements show that the optimum pH for working with PC is 2.5. Furthermore, the system exhibited a structured interface at this pH. The surface tension did not decrease further above 1.5 wt% PC. Interestingly, emulsions formulated with >1.5 wt% PC showed recoalescence immediately after preparation. Although 1.5 wt% had the smallest droplet size, this emulsion underwent creaming due to the low viscosity of the system. DG was used in combination with PC to increase viscosity and reduce creaming. As little as 0.1 wt% DG was sufficient to form an emulgel when incorporated into the previous emulsion, which exhibited pseudoplastic behaviour and viscoelastic properties with very low creaming rates. However, the use of PC in combination with DG resulted in a non-aggregated and stable emulgel with 1.5 wt% PC and 0.1 wt% DG.


Subject(s)
Biocompatible Materials , Emulsions , Phycocyanin , Phycocyanin/chemistry , Emulsions/chemistry , Viscosity , Biocompatible Materials/chemistry , Gels/chemistry , Hydrogen-Ion Concentration , Drug Delivery Systems , Plant Gums/chemistry , Rheology , Surface Tension
13.
Curr Microbiol ; 81(6): 152, 2024 Apr 23.
Article in English | MEDLINE | ID: mdl-38652305

ABSTRACT

Spirulina (Arthrospira and Spirulina spp.) has always been characterized by the helical trichomes, despite the existence of linear forms. A great debate is now open on the morphological flexibility of Spirulina, but it seems that both trichome morphology and C-phycocyanin (C-PC) concentrations are influenced by the culture conditions.This work compared the effect of some key growth factors (medium pH as well as its carbon, potassium, and salt contents) on the growth and C-PC concentration of helical and linear Spirulina strains. Further, two-phase strategies, including light and nitrogen variation, were applied to increase the in vivo C-PC accumulation into the trichomes. Results showed that high pH induced trichomes elongation and improved growth but decreased C-PC content (+ 65 and + 43% vs. -83 and -49%, for helical and linear strains, respectively). Variations in carbon and salt concentrations negatively impacted growth and C-PC content, even if the linear strain was more robust against these fluctuations. It was also interesting to see that potassium increasing improved growth and C-PC content for both strains.The variation of light wavelength during the enrichment phase (in the two-phase strategy) improved by 50% C-PC accumulation in trichomes, especially after blue lighting for 96 h. Similar result was obtained after 48 h of nitrogen reduction, while its removal from the medium caused trichomes disintegration. The current work highlights the robustness of linear Spirulina strain and presents an efficient and scalable way to increase C-PC in vivo without affecting growth.


Subject(s)
Carbon , Culture Media , Phycocyanin , Spirulina , Spirulina/growth & development , Spirulina/metabolism , Spirulina/chemistry , Phycocyanin/metabolism , Culture Media/chemistry , Culture Media/metabolism , Carbon/metabolism , Hydrogen-Ion Concentration , Nitrogen/metabolism , Light , Potassium/metabolism
14.
J Biol Chem ; 300(5): 107262, 2024 May.
Article in English | MEDLINE | ID: mdl-38579990

ABSTRACT

Cyanobacteria harvest light by using architecturally complex, soluble, light-harvesting complexes known as phycobilisomes (PBSs). PBS diversity includes specialized subunit paralogs that are tuned to specific regions of the light spectrum; some cyanobacterial lineages can even absorb far-red light. In a recent issue of the Journal of Biological Chemistry, Gisriel et al. reported the cryo-electron microscopic structure of a far-red PBS core, showing how bilin binding in the α-subunits of allophycocyanin paralogs can modify the bilin-binding site to red shift the absorbance spectrum. This work helps explain how cyanobacteria can grow in environments where most of the visible light has been filtered out.


Subject(s)
Cyanobacteria , Light , Phycobilisomes , Phycobilisomes/metabolism , Phycobilisomes/chemistry , Cyanobacteria/metabolism , Bacterial Proteins/chemistry , Bacterial Proteins/metabolism , Cryoelectron Microscopy/methods , Phycocyanin/chemistry , Phycocyanin/metabolism , Red Light
15.
Bioresour Technol ; 399: 130628, 2024 May.
Article in English | MEDLINE | ID: mdl-38521173

ABSTRACT

The polyextremophilic Galdieria sulphuraria is emerging as a promising microalgal species for food applications. This work explores the potential of heterotrophically cultivated G. sulphuraria as a protein producer for human consumption. To this end, the performances of four G. sulphuraria strains grown under the same conditions were compared. Amino acid profiles varied among strains and growth phases, but all samples met FAO dietary requirements for adults. The specific growth rates were between 1.01 and 1.48 day-1. After glucose depletion, all strains showed an increase of 38-49 % in nitrogen content within 48 h, reaching 7.8-12.0 % w/w. An opposite trend was observed in protein bioaccessibility, which decreased on average from 69 % during the exponential phase to a minimum of 32 % 48 h after stationary phase, with significant differences among the strains. Therefore, selecting the appropriate strain and harvesting time is crucial for successful single-cell protein production.


Subject(s)
Microalgae , Rhodophyta , Humans , Amino Acids/metabolism , Heterotrophic Processes , Phycocyanin/metabolism , Food , Rhodophyta/metabolism , Microalgae/metabolism , Biomass
16.
Biomolecules ; 14(3)2024 Mar 03.
Article in English | MEDLINE | ID: mdl-38540721

ABSTRACT

Phycocyanobilin (PCB) is a natural blue tetrapyrrole chromophore that is found in phycocyanin and plays an essential role in photosynthesis. Due to PCB's antioxidation, anti-inflammatory and anti-cancer properties, it has been utilized in the food, pharmaceutical and cosmetic industries. Currently, the extraction of PCB from Spirulina involves complex processes, which has led to increasing interest in the biosynthesis of PCB in Escherichia coli. However, the PCB titer remains low because of the poor activity of key enzymes and the insufficient precursor supply. Here, the synthesis of PCB was firstly improved by screening the optimal heme oxygenase (HO) from Thermosynechococcus elongatus BP-1(HOT) and PCB: ferredoxin oxidoreductase from Synechocystis sp. PCC6803 (PcyAS). In addition, based on a rational design and the infrared fluorescence method for high-throughput screening, the mutants of HOT(F29W/K166D) and PcyAS(D220G/H74M) with significantly higher activities were obtained. Furthermore, a DNA scaffold was applied in the assembly of HOT and PcyAS mutants to reduce the spatial barriers, and the heme supply was enhanced via the moderate overexpression of hemB and hemH, resulting in the highest PCB titer (184.20 mg/L) obtained in a 5 L fermenter. The strategies applied in this study lay the foundation for the industrial production of PCB and its heme derivatives.


Subject(s)
Escherichia coli , Phycocyanin , Escherichia coli/genetics , Phycobilins/genetics , Heme Oxygenase (Decyclizing) , Heme
17.
J Proteome Res ; 23(4): 1174-1187, 2024 Apr 05.
Article in English | MEDLINE | ID: mdl-38427982

ABSTRACT

Protein homeostasis is essential for cyanobacteria to maintain proper cellular function under adverse and fluctuating conditions. The AAA+ superfamily of proteolytic complexes in cyanobacteria plays a critical role in this process, including ClpXP, which comprises a hexameric ATPase ClpX and a tetradecameric peptidase ClpP. Despite the physiological effects of ClpX on growth and photosynthesis, its potential substrates and underlying mechanisms in cyanobacteria remain unknown. In this study, we employed a streptavidin-biotin affinity pull-down assay coupled with label-free proteome quantitation to analyze the interactome of ClpX in the model cyanobacterium Synechocystis sp. PCC 6803 (hereafter Synechocystis). We identified 503 proteins as potential ClpX-binding targets, many of which had novel interactions. These ClpX-binding targets were found to be involved in various biological processes, with particular enrichment in metabolic processes and photosynthesis. Using protein-protein docking, GST pull-down, and biolayer interferometry assays, we confirmed the direct association of ClpX with the photosynthetic proteins, ferredoxin-NADP+ oxidoreductase (FNR) and phycocyanin subunit (CpcA). Subsequent functional investigations revealed that ClpX participates in the maintenance of FNR homeostasis and functionality in Synechocystis grown under different light conditions. Overall, our study provides a comprehensive understanding of the extensive functions regulated by ClpX in cyanobacteria to maintain protein homeostasis and adapt to environmental challenges.


Subject(s)
Photosynthesis , Synechocystis , Photosynthesis/genetics , Synechocystis/genetics , Synechocystis/metabolism , Adenosine Triphosphatases/genetics , Adenosine Triphosphatases/metabolism , Phycocyanin/metabolism
18.
Toxicol Appl Pharmacol ; 484: 116874, 2024 Mar.
Article in English | MEDLINE | ID: mdl-38428464

ABSTRACT

Melanoma is the most aggressive and deadly skin cancer. The difficulty in its treatment arises from its ability to suppress the immune system, making it crucial to find a substance that increases anti-tumor immunity. C-phycocyanin (C-PC) appears as a promising bioactive, with multifaceted effects against several cancers, but its efficacy against melanoma has only been tested in vitro. Therefore, we investigated C-PC's the anti-tumor and immunomodulatory action in a murine melanoma model. The tumor was subcutaneously induced in C57BL/6 mice by injecting B16F10 cells. The animals were injected subcutaneously with C-PC for three consecutive days. After euthanasia, the tumor was weighed and measured. The inguinal lymph node was removed, and the cells were stained with antibodies and analyzed by flow cytometry. The heart, brain and lung were analyzed by histopathology. C-PC increased the B cell population of the inguinal lymph node in percentage and absolute number. The absolute number of T lymphocytes and myeloid cells were also increased in the groups treated with C-PC. Thus, C-PC showed a positive immunomodulatory effect both animals with and without tumor. However, this effect was more pronounced in the presence of the tumor. Positive immune system modulation may be associated with a reduction in tumor growth in animals treated with C-PC. Administration of C-PC subcutaneously did not cause organ damage. Our findings demonstrate C-PC's immunomodulatory and anti-melanoma action, paving the way for clinical research with this bioactive.


Subject(s)
Melanoma , Skin Neoplasms , Animals , Mice , Phycocyanin/pharmacology , Phycocyanin/therapeutic use , Mice, Inbred C57BL , Skin Neoplasms/drug therapy , Immunomodulation
19.
J Chromatogr A ; 1720: 464801, 2024 Apr 12.
Article in English | MEDLINE | ID: mdl-38479154

ABSTRACT

The high-purity phycocyanin has a high commercial value. Most current purification methods of C-phycocyanin involve multiple steps, which are complicated and time-consuming. To solve the problem, this research was studied, and an efficient affinity chromatography purification for C-phycocyanin from Spirulina platensis was developed. Through molecular docking simulation, virtual screening of ligands was performed, and ursolic acid was identified as the specific affinity ligand, which coupled to Affi-Gel 102 gel via 1-ethyl (3-dimethylaminopropyl)-3-carbodiimide, hydrochloride as coupling agent. With this customized and synthesized resin, a high-efficiency one-step purification procedure for C-phycocyanin was developed and optimized, the purity was determined to be 4.53, and the yield was 69 %. This one-step purification protocol provides a new approach for purifying other phycobilin proteins.


Subject(s)
Phycocyanin , Spirulina , Phycocyanin/chemistry , Molecular Docking Simulation , Spirulina/chemistry , Spirulina/metabolism , Chromatography, Affinity
20.
Poult Sci ; 103(5): 103605, 2024 May.
Article in English | MEDLINE | ID: mdl-38471233

ABSTRACT

We examined the effect of the Persian Gulf algae derivates, phycocyanin (PC) and fucoidan (FUC), on production performance, egg quality, intestinal histomorphology, ileal microflora, and egg yolk biochemistry of laying Japanese quail. A total of 250 six-wk-old Japanese quails with an average body weight of 215 ± 10 g were allocated to 5 treatments, 5 replicates and 10 birds in each replicate in a completely randomized design. The treatment groups received PC (from Spirulina platensis) and FUC (from brown seaweed) in their drinking water while control groups did not. Treatment groups received PC and FUC at 20 or 40 mg/L levels (denoted as PC20, PC40, FUC20, and FUC40, respectively). All birds were fed the same diet. All treatments significantly improved the percentage of hen day egg production (HDEP) (P = 0.002), egg mass (P = 0.002), and feed conversion ratio (FCR) (P = 0.022) but no difference was noted in egg weight (EW) and feed intake (FI). Different levels of PC and FUC significantly increased the thickness of eggshells (P = 0.022); however, the weight of the digestive tract (liver, spleen, proventriculus, gizzard, and pancreas) and oviduct was not affected. Algal derivates improved the villus height (P = 0.007) and crypt depth (P = 0.007) of the duodenum, as well as, the villus height (P = 0.005) and crypt depth (P = 0.026) of the jejunum. Both algal derivates positively affected the intestinal microflora (populations of Lactobacillus (P = 0.017), Coliform (P = 0.005), and Clostridium (P = 0.000)) whereas aerobic bacteria were unaffected. Yolk cholesterol P = 0.012) and yolk malondialdehyde P = 0.050) content were significantly reduced in experimental treatments compared to the control group. In conclusion, our results showed that the treatment of laying Japanese quails with algal derivates positively affects quail performance, intestinal morphology, intestinal microflora, and yolk cholesterol and malondialdehyde. Additional studies exploring optimal dosages and mechanisms of action is warranted to fully understand the scope of the algae derivates in poultry production.


Subject(s)
Animal Feed , Coturnix , Diet , Egg Yolk , Gastrointestinal Microbiome , Polysaccharides , Animals , Coturnix/physiology , Coturnix/anatomy & histology , Diet/veterinary , Animal Feed/analysis , Gastrointestinal Microbiome/drug effects , Female , Polysaccharides/pharmacology , Polysaccharides/administration & dosage , Polysaccharides/chemistry , Egg Yolk/chemistry , Phycocyanin/pharmacology , Phycocyanin/administration & dosage , Dietary Supplements/analysis , Random Allocation , Spirulina/chemistry , Intestines/drug effects , Intestines/anatomy & histology , Intestines/physiology , Ileum/drug effects , Ileum/physiology , Ileum/anatomy & histology
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