RESUMEN
Cyanobacteriochromes (CBCRs) are unique cyanobacteria-specific photoreceptors that share a distant relation with phytochromes. Most CBCRs contain conserved cysteine residues known as canonical Cys, while some CBCRs have additional cysteine residues called second Cys within the DXCF motif, leading to their classification as DXCF CBCRs. They typically undergo a process where they incorporate phycocyanobilin (PCB) and subsequently isomerize it to phycoviolobilin (PVB). Conversely, CBCRs with conserved Trp residues and without the second Cys are called extended red/green (XRG) CBCRs. Typical XRG CBCRs bind PCB without undergoing PCB-to-PVB isomerization, displaying red/green reversible photoconversion, and there are also atypical CBCRs that exhibit diverse photoconversions. We discovered novel XRG CBCRs with Cys residue instead of the conserved Trp residue. These novel XRG CBCRs exhibited the ability to isomerize PCB to PVB, displaying green/teal reversible photoconversion. Through sequence- and structure-based comparisons coupled with mutagenesis experiments, we identified three amino acid residues, including the Cys residue, crucial for facilitating PCB-to-PVB isomerization. This research expands our understanding of the diversity of XRG CBCRs, highlighting the remarkable molecular plasticity of CBCRs.
Asunto(s)
Proteínas Bacterianas , Cianobacterias , Ficobilinas , Ficocianina , Ficobilinas/química , Ficobilinas/metabolismo , Ficocianina/química , Ficocianina/metabolismo , Cianobacterias/metabolismo , Cianobacterias/química , Proteínas Bacterianas/química , Proteínas Bacterianas/metabolismo , Proteínas Bacterianas/genética , Isomerismo , Fotorreceptores Microbianos/química , Fotorreceptores Microbianos/metabolismo , Fotorreceptores Microbianos/genéticaRESUMEN
BACKGROUND: Beet filter cake (BFC) is a food-grade solid waste produced by the sugar industry, constituting a permanent source of pollution. Cyanobacteria are considered a sustainable resource for various bioactive compounds such as phycocyanin pigment with valuable applications. This study aimed to use beet filter cake extract (BFCE) as an alternative medium for the economic cultivation of cyanobacterium Leptolyngbya sp. SSI24 PP723083, then biorefined the bioactive component such as phycocyanin pigment that could be used in the production of selenium nanoparticles. RESULTS: The results of the batch experiment displayed that the highest protein content was in BG11medium (47.9%); however, the maximum carbohydrate and lipid content were in 25% BFCE (15.25 and 10.23%, respectively). In addition, 75% BFCE medium stimulated the phycocyanin content (25.29 mg/g) with an insignificant variation compared to BG11 (22.8 mg/g). Moreover, crude phycocyanin extract from Leptolyngbya sp SSI24 cultivated on BG11 and 75% BFCE successfully produced spherical-shaped selenium nanoparticles (Se-NPs) with mean sizes of 95 and 96 nm in both extracts, respectively. Moreover, XRD results demonstrated that the biosynthesized Se-NPs have a crystalline nature. In addition, the Zeta potential of the biosynthesized Se-NPs equals - 17 mV and - 15.03 mV in the control and 75% BFCE treatment, respectively, indicating their stability. The biosynthesized Se-NPs exhibited higher effectiveness against Gram-positive bacteria than Gram-negative bacteria. Moreover, the biosynthesized Se-NPs from BG11 had higher antioxidant activity with IC50 of 60 ± 0.7 compared to 75% BFCE medium. Further, Se-NPs biosynthesized from phycocyanin extracted from Leptolyngbya sp cultivated on 75% BFCE exhibited strong anticancer activity with IC50 of 17.31 ± 0.63 µg/ml against the human breast cancer cell line. CONCLUSIONS: The BFCE-supplemented medium can be used for the cultivation of cyanobacterial strain for the phycocyanin accumulation that is used for the green synthesis of selenium nanoparticles that have biological applications.
Asunto(s)
Ficocianina , Selenio , Ficocianina/biosíntesis , Ficocianina/metabolismo , Selenio/metabolismo , Selenio/química , Cianobacterias/metabolismo , Humanos , Antibacterianos/farmacología , Antibacterianos/biosíntesis , Antibacterianos/química , Nanopartículas del Metal/química , Beta vulgaris/química , Nanopartículas/química , Residuos Industriales/análisisRESUMEN
Cyanobacteria hold promise for simultaneous carbon capture and chemicals production, but the regulation and effect of nitrogen (N) and phosphorus (P) remains unclear. This study investigates major productions of glycogen, protein, and C-phycocyanin (C-PC) in Cyanobacterium aponinum PCC10605 under different N/P levels, alongside changes in light and CO2. Increasing nitrate (NO3-) from 2 to 6 mM resulted in a 9.7-fold increase in C-PC and reduced glycogen to 8.9 %. On the other hand, elevating phosphorus from 0.1 to 2 mM under limited nitrogen enhanced biomass and glycogen through the upregulation of carbonic anhydrase, ADP-glucose pyrophosphorylase, and glycogen phosphorylase. Changes in phosphorus levels and CO2 inlet concentrations affected metabolites accumulation and carbon capture efficiency, leading to the best condition of 76 % uptake capacity in direct air capture (DAC). All findings underscore the trade-off between glycogen and protein, representing the importance of N/P levels in nutrient modulation of PCC10605.
Asunto(s)
Cianobacterias , Glucógeno , Nitrógeno , Fósforo , Glucógeno/metabolismo , Nitrógeno/metabolismo , Cianobacterias/metabolismo , Proteínas Bacterianas/metabolismo , Dióxido de Carbono/metabolismo , Biomasa , Ficocianina/metabolismo , Anhidrasas Carbónicas/metabolismo , Nitratos/metabolismoRESUMEN
Cyanobacteriochromes (CBCRs) are distinctive tetrapyrrole (bilin)-binding photoreceptors exclusively found in cyanobacteria. Unlike canonical phytochromes, CBCRs require only a GAF (cGMP-phosphodiesterase/adenylate cyclase/FhlA) domain for autolyase activity to form a bilin adduct via a Cys residue and cis-trans photoisomerization. Apart from the canonical Cys, which attaches covalently to C31 in the A-ring of the bilin, some GAF domains of CBCRs contain a second-Cys in the Asp-Xaa-Cys-Phe (DXCF) motif, responsible for isomerization of phycocyanobilin (PCB) to phycoviolobilin (PVB) and/or for the formation of a reversible 2nd thioether linkage to the C10. Unlike green/teal-absorbing GAF proteins lacking ligation activity, the second-Cys in another teal-absorbing lineage (DXCF blue/teal group) exhibits both isomerization and ligation activity due to the presence of the Tyr instead of His next to the canonical Cys. Herein, we discovered an atypical CBCR GAF protein, Tpl7205g1, belonging to the DXCF blue/teal group, but having His instead of Tyr next to the first-Cys. Consistent with its subfamily, the second-Cys of Tpl7205g1 did not form a thioether linkage at C10 of PCB, showing only isomerization activity. Instead of forming 2nd thioether linkage, this novel GAF protein exhibits a pH-dependent photocycle between protonated 15Z and deprotonated 15E. Site-directed mutagenesis to the GAF scaffolds revealed its combined characteristics, including properties of teal-DXCF CBCRs and red/green-absorbing CBCRs (XRG CBCRs), suggesting itself as the evolutionary bridge between the two CBCR groups. Our study thus sheds light on the expanded spectral tuning characteristics of teal-light absorbing CBCRs and enhances feasibility of engineering these photoreceptors.
Asunto(s)
Proteínas Bacterianas , Cianobacterias , Optogenética , Fotorreceptores Microbianos , Fitocromo , Fitocromo/química , Fitocromo/metabolismo , Fitocromo/genética , Fotorreceptores Microbianos/química , Fotorreceptores Microbianos/genética , Fotorreceptores Microbianos/metabolismo , Cianobacterias/metabolismo , Cianobacterias/genética , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Optogenética/métodos , Luz , Ficocianina/química , Ficocianina/metabolismo , Ingeniería de Proteínas/métodos , Ficobilinas/química , Ficobilinas/metabolismo , Secuencia de AminoácidosRESUMEN
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.
Asunto(s)
Ficocianina , Estrés Salino , Spirulina , Ficocianina/metabolismo , Ficocianina/aislamiento & purificación , Spirulina/metabolismo , Spirulina/química , Biomasa , CongelaciónRESUMEN
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.
Asunto(s)
Corynebacterium glutamicum , Ingeniería Metabólica , Ficobilinas , Ficocianina , Corynebacterium glutamicum/metabolismo , Corynebacterium glutamicum/genética , Ficocianina/metabolismo , Ficocianina/genética , Ficobilinas/metabolismo , Ficobilinas/genética , Fermentación , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Vía de Pentosa Fosfato/genética , Reactores Biológicos/microbiologíaRESUMEN
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.
Asunto(s)
Carbono , Medios de Cultivo , Ficocianina , Spirulina , Spirulina/crecimiento & desarrollo , Spirulina/metabolismo , Spirulina/química , Ficocianina/metabolismo , Medios de Cultivo/química , Medios de Cultivo/metabolismo , Carbono/metabolismo , Concentración de Iones de Hidrógeno , Nitrógeno/metabolismo , Luz , Potasio/metabolismoRESUMEN
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.
Asunto(s)
Cianobacterias , Luz , Ficobilisomas , Ficobilisomas/metabolismo , Ficobilisomas/química , Cianobacterias/metabolismo , Proteínas Bacterianas/química , Proteínas Bacterianas/metabolismo , Microscopía por Crioelectrón/métodos , Ficocianina/química , Ficocianina/metabolismo , Luz RojaRESUMEN
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.
Asunto(s)
Fotosíntesis , Synechocystis , Fotosíntesis/genética , Synechocystis/genética , Synechocystis/metabolismo , Adenosina Trifosfatasas/genética , Adenosina Trifosfatasas/metabolismo , Ficocianina/metabolismoRESUMEN
Spirulina is a promising feedstock for c-phycocyanin, a blue pigment-protein, commercially incorporated in many food products for its desirable bright blue attributes, exceptional bioavailability, and inherent therapeutic properties. Remarkably, enhancing c-phycocyanin synthesis in Spirulina would facilitate economic viability and sustainability at large-scale production, as the forecasted market value is $ 409.8 million by 2030. Notably, the lighting source plays a key role in enhancing c-phycocyanin in Spirulina, and thus, strategies to filter/concentrate the photons of respective wavelengths, influencing light spectra, are beneficial. Enveloping open raceway ponds and greenhouses by luminescent solar concentrators and light filtering sheets enables solar spectral conversion of the sunlight at desirable wavelengths, emerges as a promising strategy to enhance synthesis of c-phycocyanin in Spirulina. Nevertheless, the conduction of techno-economic assessments and evaluation of scalability at large-scale cultivation of Spirulina are essential for the real-time implementation of lighting strategies.
Asunto(s)
Spirulina , Spirulina/metabolismo , Ficocianina/metabolismo , Luz , Luminiscencia , Luz SolarRESUMEN
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.
Asunto(s)
Microalgas , Rhodophyta , Humanos , Aminoácidos/metabolismo , Procesos Heterotróficos , Ficocianina/metabolismo , Alimentos , Rhodophyta/metabolismo , Microalgas/metabolismo , BiomasaRESUMEN
CpcL-phycobilisomes (CpcL-PBSs) are a reduced type of phycobilisome (PBS) found in several cyanobacteria. They lack the traditional PBS terminal energy emitters, but still show the characteristic red-shifted fluorescence at ~670 nm. We established a method of assembling in vitro a rod-membrane linker protein, CpcL, with phycocyanin, generating complexes with the red-shifted spectral features of CpcL-PBSs. The red-shift arises from the interaction of a conserved key glutamine, Q57 of CpcL in Synechocystis sp. PCC 6803, with a single phycocyanobilin chromophore of trimeric phycocyanin at one of the three ß82-sites. This chromophore is the terminal energy acceptor of CpcL-PBSs and donor to the photosystem(s). This mechanism also operates in PBSs from Acaryochloris marina MBIC11017. We then generated multichromic complexes harvesting light over nearly the complete visible range via the replacement of phycocyanobilin chromophores at sites α84 and ß153 of phycocyanins by phycoerythrobilin and/or phycourobilin. The results demonstrate the rational design of biliprotein-based light-harvesting elements by engineering CpcL and phycocyanins, which broadens the light-harvesting range and accordingly improves the light-harvesting capacity and may be potentially applied in solar energy harvesting.
Asunto(s)
Proteínas Bacterianas , Ficobilinas , Ficobilisomas , Ficocianina , Synechocystis , Ficobilisomas/metabolismo , Ficocianina/metabolismo , Ficocianina/química , Synechocystis/metabolismo , Proteínas Bacterianas/metabolismo , Ficobilinas/metabolismo , Ficobilinas/química , Cianobacterias/metabolismoRESUMEN
Based on the use of CQDs as fluorescent probe and covalent coupling method to detect biological molecules with amino groups, to deeply analysis and detect the metabolism of Microcystis aeruginosa. The metabolic changes of carboxyl biomolecules in Microcystis aeruginosa were analyzed by covalent coupling method, including GSH, phycocyanin and SOD enzyme. The changes of GSH content and its correlation between phycocyanin, SOD were analyzed. The content of phycocyanin and SOD reached the maximum on the 65th day, and GSH was more sensitive to the growth and metabolism of microalgae. GSH plays an important role in reducing the external oxidative damage of microalgae cells. The synthesis of glutathione (GSH), GSH/GSSG mutual transformation, the production of phytochelating peptide (PC), the ASA-GSH cycle, and other physiological processes are interconnected. These interactions are crucial for preserving the antioxidant properties of microalgae and regulating redox-sensitive signal transduction.
Asunto(s)
Microalgas , Puntos Cuánticos , Ficocianina/metabolismo , Microalgas/metabolismo , Puntos Cuánticos/química , Fluorescencia , Carbono/química , Glutatión/análisis , Antioxidantes/metabolismo , Superóxido Dismutasa/metabolismoRESUMEN
INTRODUCTION: Ulcerative colitis (UC) is an incurable and highly complex chronic inflammatory bowel disease (IBD) affecting millions of people worldwide. C-phycocyanin (C-PC) has been reported to possess outstanding anti-inflammatory activities and can effectively inhibit various inflammation-related diseases. Whether C-PC-derived bioactive peptides can inhibit intestinal inflammation is worth research and consideration. METHODS: The inhibition activities of three anti-neuroinflammatory peptides were evaluated using 2-4-6-trinitrobenzen sulfonic acid (TNBS)-induced zebrafish colitis model. Subsequently, the abilities of peptides to promote gastrointestinal motility were also examined. The changes in the intestinal pathological symptoms and ultrastructure of intestinal, reactive oxygen species (ROS) levels, and antioxidant enzymes were then determined after co-treatment with peptides and TNBS. Transcriptome analysis was used to investigate the underlying ameliorating TNBS-induced colitis effects molecular mechanisms of better activity peptide. Furthermore, quantitative reverse-transcription polymerase chain reaction and molecular docking techniques verified the mRNA sequencing results. RESULTS: Three peptides, MHLWAAK, MAQAAEYYR and MDYYFEER, which significantly inhibit macrophage migration, were synthesized. The results showed that these peptides could effectively alleviate the inflammatory responses in the TNBS-induced zebrafish model of colitis. In addition, co-treatment with TNBS and C-PC peptides could decrease ROS production and increase antioxidant enzyme activities in zebrafish larvae. Moreover, MHLWAAK had the most significantly therapeutic effects on colitis in zebrafish. The transcriptome analysis suggests that the effect of MHLWAAK on TNBS-induced colitis may be associated with the modulation of nuclear factor-erythroid 2-related factor 2 (Nrf2) and mitogen-activated protein kinase (MAPK) signaling pathway associated genes. In addition, molecular docking was conducted to study the prospective interaction between peptides and the key proteins that streamline the Nrf2 and MAPK signaling pathways. IL-6, JNK3, TNF-α, KEAP1-NRF2 complex and MAPK may be the core targets of MHLWAAK in treating colitis. CONCLUSION: The results suggested that the three C-PC-derived peptides could ameliorate TNBS-induced colitis in zebrafish, and these peptides might be a promising therapeutic candidate for UC treatment.
Asunto(s)
Colitis , Enfermedades Inflamatorias del Intestino , Animales , Pez Cebra/metabolismo , Factor 2 Relacionado con NF-E2/metabolismo , Ficocianina/metabolismo , Proteína 1 Asociada A ECH Tipo Kelch/metabolismo , Antioxidantes/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Simulación del Acoplamiento Molecular , Estudios Prospectivos , Enfermedades Inflamatorias del Intestino/tratamiento farmacológico , Enfermedades Inflamatorias del Intestino/metabolismo , Colitis/inducido químicamente , Colitis/metabolismo , Colitis/patología , Sistema de Señalización de MAP Quinasas , Inflamación , Modelos Animales de EnfermedadRESUMEN
Hydrogen peroxide (H2O2) is an environmentally-safe algaecide used to control harmful algal blooms and as a disinfectant in various domestic and industrial applications. It is produced naturally in sunny-water or as a by-product during growth, and metabolism of photosynthetic organisms. To assess the impact of H2O2 on Arthrospira platensis, several biochemical components, and antioxidant enzymes were analysed. The growth and biomass of A. platensis were decreased under the effect of H2O2. Whereas, the concentration up to 40 µM H2O2 non-significantly induced (at P < 0.05) the Chl a, C-phycocyanin (C-PC), total phycobiliprotein (PBP), and the radical scavenging activity of A. platensis. The half-maximal effective concentrations (EC50) for H2O2 were 57, 65, and 74 µM H2O2 with regards to the biomass yield, Chl a, and C-PC content, respectively. While, the total soluble protein, and soluble carbohydrates contents were significantly induced. However, the higher concentrations (60 and 80 µM) were lethal to these components, in parallel to the initiation of the lipid peroxidation process. Surprisingly, the carotenoids content was non-significantly increased by H2O2. Despite the relative consistency of catalase (CAT), the activities of superoxide dismutase (SOD) and peroxidase (POD) enzymes were boosted by all of the tested concentrations of H2O2. The relative transcript abundance of selected regulatory genes was also investigated. Except for the highest dose (80 µM), the tested concentrations had almost inhibitory effect on the relative transcripts of heat shock protein (HSP90), glutamate synthase (GOGAT), delta-9 desaturase (desC), iron-superoxide dismutase (FeSOD) and the Rubisco (the large subunit, rbcL) genes. The results demonstrated the importance of the non-enzymatic and enzymatic antioxidants for the cumulative tolerance of A. platensis.
Asunto(s)
Antioxidantes , Spirulina , Antioxidantes/metabolismo , Spirulina/química , Spirulina/metabolismo , Peróxido de Hidrógeno/toxicidad , Peróxido de Hidrógeno/metabolismo , Estrés Oxidativo , Superóxido Dismutasa/metabolismo , Ficocianina/farmacología , Ficocianina/química , Ficocianina/metabolismoRESUMEN
Chlorophyll (Chl) is the most abundant pigment in photosynthetic plants, but it is prone to degradation during processing and storage, limiting its usage in the food industry. This study developed a technique for increasing Chl photostability by light-induced Chl/phycocyanin (PC) triple synergistic aggregation. Under continuous illumination settings, the results revealed that the Chl retention increased to 406% compared to the control. A model of Chl/PC complexes was constructed using multiligand molecular docking, and the aggregation mechanism was investigated by quantum chemistry, which demonstrated that PC could provide an ideal central hydrophobic cavity for Chl aggregates and thus further enhance the aggregation of Chl on the basis of Chl/PC complexes. The core driver of the improved photostability of Chl is photoexcitation-induced Chl aggregates. This study enriches our understanding of the interaction mechanism between PC and Chl, and we hope that this study can provide broader ideas for the development of natural pigment products.
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Clorofila , Ficocianina , Clorofila/metabolismo , Ficocianina/metabolismo , Clorofila A , Simulación del Acoplamiento Molecular , FotosíntesisRESUMEN
Bone regeneration is a multistep and regular physiological process that occurs normally in fracture repair and bone defects. However, some factors such as aging, particular diseases and some drugs prevent or slowdown bone natural healing. Cell therapy using stem cells and differentiation activating factors is an effective treatment method for bone regeneration triggering in unusual conditions. Therefore, in the present study the effect of phycocyanin and phycoerythrin pigments which isolated from Spirulina platensis and Gracilaria gracilis algae was investigate on osteogenic differentiation potency of human Amniotic Mesenchymal Stem Cells (hAMSCs). For this purpose, hAMSCs were exposed to 300, 500, and 700 µg/ml concentrations of phycocyanin and phycoerythrin pigments and then the cells viability was measured with MTT assay in 48 and 72 h after treatment. The osteo-differentiation level of cells was studied by measuring ALP activity using calorimetric method and Alizarin red staining for calcium deposition in 7 and 21 days after treatment. Also, total RNA of cells was extracted in different time periods and then cDNA synthesized with specific primers, and relative expression of Runx2, ß-catenin and Osteocalcin genes were investigated using SYBR Green RT-qPCR technique. Osteogenic differentiation of hAMSCs that treated with pigments was confirmed by mineral deposits staining and increased level of ALP activity. Furthermore, these pigments elevated significantly the expression of osteogenic marker genes compared to control samples and caused hAMSCs to differentiate into osteoblast cells. According to these results, phycocyanin and phycoerythrin may suggest as suitable osteogenic supplements with low toxicity, low cost and high efficiency, although the molecular mechanism of its efficacy is not available yet.
Asunto(s)
Gracilaria , Células Madre Mesenquimatosas , Humanos , Osteogénesis , Ficocianina/farmacología , Ficocianina/metabolismo , Ficoeritrina/metabolismo , Ficoeritrina/farmacología , Diferenciación Celular , Células CultivadasRESUMEN
Due to their unique biochemical and spectroscopic properties, both heme and phycocyanobilin are widely applied in the medical and food industries. Synechocystis sp. PCC 6803 contains both heme and phycocyanin, and is capable of synthesizing phycocyanin using heme as a precursor. The aim of this study was to uncover viable metabolic targets in the porphyrin pathway from Synechocystis sp. PCC 6803 to promote the accumulation of heme and phycocyanin in the recombinant strains of microalgae. A total of 10 genes related to heme synthesis pathway derived from Synechococcus elongatus PCC 7942 and 12 genes related to endogenous heme synthesis were individually overexpressed in strain PCC 6803. The growth rate and pigment content (heme, phycocyanin, chlorophyll a and carotenoids) of 22 recombinant algal strains were characterized. Quantitative real-time PCR technology was used to investigate the molecular mechanisms underlying the changes in physiological indicators in the recombinant algal strains. Among the 22 mutant strains, the mutant overexpressing the haemoglobin gene (glbN) of strain PCC 6803 had the highest heme content, which was 2.5 times higher than the wild type; the mutant overexpressing the gene of strain PCC 7942 (hemF) had the highest phycocyanin content, which was 4.57 times higher than the wild type. Overall, the results suggest that genes in the porphyrin pathway could significantly affect the heme and phycocyanin content in strain PCC 6803. Our study provides novel crucial targets for promoting the accumulation of heme and phycocyanin in cyanobacteria.
Asunto(s)
Porfirinas , Synechocystis , Ficocianina/genética , Ficocianina/metabolismo , Synechocystis/genética , Synechocystis/metabolismo , Hemo/genética , Clorofila A , Ingeniería GenéticaRESUMEN
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 24 h 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.
Asunto(s)
Ficocianina , Synechococcus , Ficocianina/genética , Ficocianina/metabolismo , Ficobiliproteínas/metabolismo , Ficobiliproteínas/farmacología , Synechococcus/genética , Campos MagnéticosRESUMEN
Aquatic photosynthetic organisms evolved to use a variety of light frequencies to perform photosynthesis. Phycobiliprotein phycocyanin 645 (PC645) is a light-harvesting complex in cryptophyte algae able to transfer the absorbed green solar light to other antennas with over 99% efficiency. The infrared signatures of the phycobilin pigments embedded in PC645 are difficult to access and could provide useful information to understand the mechanism behind the high efficiency of energy transfer in PC645. We use visible-pump IR-probe and two-dimensional electronic vibrational spectroscopy to study the dynamical evolution and assign the fingerprint mid-infrared signatures to each pigment in PC645. Here, we report the pigment-specific vibrational markers that enable us to track the spatial flow of excitation energy between the phycobilin pigment pairs. We speculate that two high-frequency modes (1588 and 1596 cm-1) are involved in the vibronic coupling leading to fast (Asunto(s)
Ficobilinas
, Ficocianina
, Ficobilinas/química
, Ficocianina/química
, Ficocianina/metabolismo
, Ficobiliproteínas/química
, Fotosíntesis