Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 379
Filtrar
1.
Nutrients ; 16(11)2024 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-38892686

RESUMO

Arthrospira platensis, commonly known as Spirulina, is a photosynthetic filamentous cyanobacterium (blue-green microalga) that has been utilized as a food source since ancient times. More recently, it has gained significant popularity as a dietary supplement due to its rich content of micro- and macro-nutrients. Of particular interest is a water soluble phycobiliprotein derived from Spirulina known as phycocyanin C (C-PC), which stands out as the most abundant protein in this cyanobacterium. C-PC is a fluorescent protein, with its chromophore represented by the tetrapyrrole molecule phycocyanobilin B (PCB-B). While C-PC is commonly employed in food for its coloring properties, it also serves as the molecular basis for numerous nutraceutical features associated with Spirulina. Indeed, the comprehensive C-PC, and to some extent, the isolated PCB-B, has been linked to various health-promoting effects. These benefits encompass conditions triggered by oxidative stress, inflammation, and other pathological conditions. The present review focuses on the bio-pharmacological properties of these molecules, positioning them as promising agents for potential new applications in the expanding nutraceutical market.


Assuntos
Suplementos Nutricionais , Ficocianina , Spirulina , Spirulina/química , Ficocianina/farmacologia , Humanos , Ficobilinas/farmacologia , Ficobiliproteínas , Estresse Oxidativo/efeitos dos fármacos
2.
J Agric Food Chem ; 72(21): 12219-12228, 2024 May 29.
Artigo em Inglês | MEDLINE | ID: mdl-38747135

RESUMO

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.


Assuntos
Corynebacterium glutamicum , Engenharia Metabólica , Ficobilinas , Ficocianina , Corynebacterium glutamicum/metabolismo , Corynebacterium glutamicum/genética , Ficocianina/metabolismo , Ficocianina/genética , Ficobilinas/metabolismo , Ficobilinas/genética , Fermentação , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Via de Pentose Fosfato/genética , Reatores Biológicos/microbiologia
3.
J Phycol ; 60(3): 695-709, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38558363

RESUMO

Crustose coralline algae (CCA) are a highly diverse group of habitat-forming, calcifying red macroalgae (Rhodophyta) with unique adaptations to diverse irradiance regimes. A distinctive CCA phenotype adaptation, which allows them to maximize photosynthetic performance in low light, is their content of a specific group of light-harvesting pigments called phycobilins. In this study, we assessed the potential of noninvasive hyperspectral imaging (HSI) in the visible spectrum (400-800 nm) to describe the phenotypic variability in phycobilin content of an Antarctic coralline, Tethysphytum antarcticum (Hapalidiales), from two distinct locations. We validated our measurements with pigment extractions and spectrophotometry analysis, in addition to DNA barcoding using the psbA marker. Targeted spectral indices were developed and correlated with phycobilin content using linear mixed models (R2 = 0.64-0.7). Once applied to the HSI, the models revealed the distinct phycoerythrin spatial distribution in the two site-specific CCA phenotypes, with thin and thick crusts, respectively. This study advances the capabilities of hyperspectral imaging as a tool to quantitatively study CCA pigmentation in relation to their phenotypic plasticity, which can be applied in laboratory studies and potentially in situ surveys using underwater hyperspectral imaging systems.


Assuntos
Ficobilinas , Rodófitas , Regiões Antárticas , Ficobilinas/análise , Ficobilinas/metabolismo , Imageamento Hiperespectral/métodos , Pigmentos Biológicos/análise , Pigmentos Biológicos/metabolismo , Código de Barras de DNA Taxonômico
4.
Biomolecules ; 14(3)2024 Mar 03.
Artigo em Inglês | MEDLINE | ID: mdl-38540721

RESUMO

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.


Assuntos
Escherichia coli , Ficocianina , Escherichia coli/genética , Ficobilinas/genética , Heme Oxigenase (Desciclizante) , Heme
5.
Plant J ; 118(4): 1207-1217, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38319793

RESUMO

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.


Assuntos
Proteínas de Bactérias , Ficobilinas , Ficobilissomas , Ficocianina , Synechocystis , Ficobilissomas/metabolismo , Ficocianina/metabolismo , Ficocianina/química , Synechocystis/metabolismo , Proteínas de Bactérias/metabolismo , Ficobilinas/metabolismo , Ficobilinas/química , Cianobactérias/metabolismo
6.
Crit Rev Biotechnol ; 44(3): 373-387, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-36775664

RESUMO

Porphyrins, phycobilins, and their proteins have abundant π-electrons and strongly absorb visible light, some of which bind a metal ion in the center. Because of the structural and optical properties, they not only play critical roles as an essential component in natural systems but also have attracted much attention as a high value specialty chemical in various fields, including renewable energy, cosmetics, medicines, and foods. However, their commercial application seems to be still limited because the market price of porphyrins and phycobilins is generally expensive to apply them easily. Furthermore, their petroleum-based chemical synthesis is energy-intensive and emits a pollutant. Recently, to replace petroleum-based production, many studies on the bioproduction of metalloporphyrins, including Zn-porphyrin, Co-porphyrin, and heme, porphyrin derivatives including chlorophyll, biliverdin, and phycobilins, and their proteins including hemoproteins, phycobiliproteins, and phytochromes from renewable carbon sources using microbial cell factories have been reported. This review outlines recent advances in the bioproduction of porphyrins, phycobilins, and their proteins using microbial cell factories developed by various microbial biotechnology techniques, provides well-organized information on metabolic regulations of the porphyrin metabolism, and then critically discusses challenges and future perspectives. Through these, it is expected to be able to achieve possible solutions and insights and to develop an outstanding platform to be applied to the industry in future research.


Assuntos
Metaloporfirinas , Petróleo , Porfirinas , Ficobilinas , Engenharia Metabólica
7.
Int J Mol Sci ; 24(11)2023 Jun 04.
Artigo em Inglês | MEDLINE | ID: mdl-37298688

RESUMO

The phycobilisome (PBS) is the major light-harvesting apparatus in cyanobacteria and red algae. It is a large multi-subunit protein complex of several megadaltons that is found on the stromal side of thylakoid membranes in orderly arrays. Chromophore lyases catalyse the thioether bond between apoproteins and phycobilins of PBSs. Depending on the species, composition, spatial assembly, and, especially, the functional tuning of different phycobiliproteins mediated by linker proteins, PBSs can absorb light between 450 and 650 nm, making them efficient and versatile light-harvesting systems. However, basic research and technological innovations are needed, not only to understand their role in photosynthesis but also to realise the potential applications of PBSs. Crucial components including phycobiliproteins, phycobilins, and lyases together make the PBS an efficient light-harvesting system, and these provide a scheme to explore the heterologous synthesis of PBS. Focusing on these topics, this review describes the essential components needed for PBS assembly, the functional basis of PBS photosynthesis, and the applications of phycobiliproteins. Moreover, key technical challenges for heterologous biosynthesis of phycobiliproteins in chassis cells are discussed.


Assuntos
Ficobilissomas , Rodófitas , Ficobilissomas/química , Ficobilissomas/metabolismo , Ficobilinas , Ficobiliproteínas/química , Ficobiliproteínas/metabolismo , Fotossíntese , Rodófitas/química
8.
J Phys Chem B ; 127(20): 4460-4469, 2023 05 25.
Artigo em Inglês | MEDLINE | ID: mdl-37192324

RESUMO

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 (

Assuntos
Ficobilinas , Ficocianina , Ficobilinas/química , Ficocianina/química , Ficocianina/metabolismo , Ficobiliproteínas/química , Fotossíntese
9.
Int J Mol Sci ; 24(6)2023 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-36982707

RESUMO

Thermophilic cyanobacteria are cosmopolitan and abundant in the thermal environment. Their light-harvesting complexes, phycobilisomes (PBS), are highly important in photosynthesis. To date, there is limited information on the PBS composition of thermophilic cyanobacteria whose habitats are challenging for survival. Herein, genome-based methods were used to investigate the molecular components of PBS in 19 well-described thermophilic cyanobacteria. These cyanobacteria are from the genera Leptolyngbya, Leptothermofonsia, Ocullathermofonsia, Thermoleptolyngbya, Trichothermofonsia, Synechococcus, Thermostichus, and Thermosynechococcus. According to the phycobiliprotein (PBP) composition of the rods, two pigment types are observed in these thermophiles. The amino acid sequence analysis of different PBP subunits suggests several highly conserved cysteine residues in these thermophiles. Certain amino acid contents in the PBP of thermophiles are significantly higher than their mesophilic counterparts, highlighting the potential roles of specific substitutions of amino acid in the adaptive thermostability of light-harvesting complexes in thermophilic cyanobacteria. Genes encoding PBS linker polypeptides vary among the thermophiles. Intriguingly, motifs in linker apcE indicate a photoacclimation of a far-red light by Leptolyngbya JSC-1, Leptothermofonsia E412, and Ocullathermofonsia A174. The composition pattern of phycobilin lyases is consistent among the thermophiles, except for Thermostichus strains that have extra homologs of cpcE, cpcF, and cpcT. In addition, phylogenetic analyses of genes coding for PBPs, linkers, and lyases suggest extensive genetic diversity among these thermophiles, which is further discussed with the domain analyses. Moreover, comparative genomic analysis suggests different genomic distributions of PBS-related genes among the thermophiles, indicating probably various regulations of expression. In summary, the comparative analysis elucidates distinct molecular components and organization of PBS in thermophilic cyanobacteria. These results provide insights into the PBS components of thermophilic cyanobacteria and fundamental knowledge for future research regarding structures, functions, and photosynthetic improvement.


Assuntos
Cianobactérias , Ficobilissomas , Ficobilissomas/genética , Ficobilissomas/metabolismo , Filogenia , Cianobactérias/genética , Cianobactérias/metabolismo , Ficobilinas , Complexos de Proteínas Captadores de Luz/genética , Proteínas de Bactérias/metabolismo
10.
Chembiochem ; 24(5): e202200455, 2023 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-36538283

RESUMO

The blue biliprotein phycocyanin, produced by photo-autotrophic cyanobacteria including spirulina (Arthrospira) and marketed as a natural food supplement or "nutraceutical," is reported to have anti-inflammatory, antioxidant, immunomodulatory, and anticancer activity. These diverse biological activities have been specifically attributed to the phycocyanin chromophore, phycocyanobilin (PCB). However, the mechanism of action of PCB and the molecular targets responsible for the beneficial properties of PCB are not well understood. We have developed a procedure to rapidly cleave the PCB pigment from phycocyanin by ethanolysis and then characterized it as an electrophilic natural product that interacts covalently with thiol nucleophiles but lacks any appreciable cytotoxicity or antibacterial activity against common pathogens and gut microbes. We then designed alkyne-bearing PCB probes for use in chemical proteomics target deconvolution studies. Target identification and validation revealed the cysteine protease legumain (also known as asparaginyl endopeptidase, AEP) to be a target of PCB. Inhibition of this target may account for PCB's diverse reported biological activities.


Assuntos
Cisteína Proteases , Spirulina , Ficocianina/farmacologia , Ficocianina/química , Ficobilinas/farmacologia , Ficobilinas/química , Spirulina/química , Suplementos Nutricionais
11.
J Pharm Sci ; 112(1): 213-224, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36087776

RESUMO

Phycocyanin (C-PC) is a constitutive chromoprotein of Arthrospira platensis, which exhibits promising efficacy against different types of cancer. In this study, we cleaved C-PC's chromophore phycocyanobilin (PCB) and demonstrated its ability as an anti-cancer drug for Colorectal cancer (CRC). PCB displayed an anti-cancer effect for CRC (HT-29) cells with IC50 of 108 µg/ml. Assessing the transcripts levels of some biomarkers revealed that the PCB caused an upregulation in the anti-metastatic gene NME1 level and downregulation of the COX-2 level. The flow cytometric results showed the effect of PCB on the arrest of the cell cycle's G1 phase. In addition, we successfully synthesized the UiO-66 (Zr-MOF). We incorporated the PCB into UiO-66 nanoparticles with a loading percentage of 46 %. Assessment of the cytotoxic effects of UiO-66@PCB showed a 2-fold improvement in the IC50 compared to the free PCB. In conclusion, we have shown that PCB displayed a promising potential as an anti-cancer agent. Yet, it is considered a safe and natural substance that can help to mitigate cancer spread and symptoms. In the meantime, UiO-66 can be used as a safe nano-delivery tool for PCB.


Assuntos
Antineoplásicos , Estruturas Metalorgânicas , Neoplasias , Humanos , Ficocianina/farmacologia , Ficobilinas/farmacologia , Antineoplásicos/farmacologia
12.
Anticancer Res ; 42(10): 5035-5041, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-36191985

RESUMO

Photosynthesis is the basis of almost all life on Earth. In addition to providing energy, plants and algae provide a plethora of secondary substances useful in the treatment of a number of illnesses including a wide array of cancer maladies. The first organisms on Earth used chemosynthesis for their energy needs. Photosynthetic bacteria utilize one of two different photosystems whereas cyanobacteria, eukaryotic algae and plants combine two photosystems in a linear electron transport chain. Accessory pigments such as various chlorophylls, carotenoids and phycobilins absorb the energy of impinging photons and funnel it to the reaction centers (P680 in photosystem II and P700 in photosystem I). Water is split photochemically, electrons are transported to reduce NADPH, oxygen is discarded as waste product, and protons accumulate inside the thylakoid vesicles in the chloroplasts. The resulting electrochemical gradient across the membrane is used to drive an ATPase. The produced ATP and NADPH+H+ are utilized in the Calvin cycle to fix CO2 and to produce fructose.


Assuntos
Complexo de Proteína do Fotossistema I , Complexo de Proteína do Fotossistema II , Adenosina Trifosfatases , Trifosfato de Adenosina , Dióxido de Carbono , Carotenoides , Transporte de Elétrons , Frutose , NADP , Oxigênio/metabolismo , Fotossíntese , Complexo de Proteína do Fotossistema I/metabolismo , Complexo de Proteína do Fotossistema II/metabolismo , Ficobilinas , Prótons , Água
13.
Environ Sci Pollut Res Int ; 29(52): 78942-78959, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-35705761

RESUMO

Owing to the increase of pollutant sources in oceans, seas, and lakes, there is an expected effect on growth and metabolism of planktonic algae which are considered primary producers in the ecosystem. Therefore, it becomes urgent to carry out laboratory studies to test to what extent these pollutants can affect the growth of algae which is necessary as a food for marine fishes. Spirulina is considered the most important algal species due to its high nutritional value for humans and animals. Therefore, this work investigated the effect of different concentrations of Ni2+, Zn2+, and Cu2+ metal ion pollutants on growth of the blue-green alga Spirulina platensis. EC50 was identified to be around 2 mg/l for the three heavy metals. The suitability of Idku Lake for Spirulina platensis growth was investigated using multi-criteria spatial modeling integrated with remotely sensed data processing. Spatial distribution maps of turbidity, water nutrients, and phytoplankton were the input criteria used to assess Idku Lake's suitability. The results obtained proved that low concentrations of the tested heavy metals stimulated growth and pigment fractions (chlorophyll a, carotenoids, and total phycobilins content) but to different degrees. The inhibitory effect was more prominent in the case of copper ions than zinc and nickel ions with all concentrations used. The overall suitability map of Spirulina platensis in Idku Lake showed that the whole lake is suitable for growth and proliferation except for the northwestern corner due to the high salinity levels. The present paper helps to understand the behavior of algae responding to environmental pollution, which supports environmental planners with the necessary baseline for investigating the fate of pollutants and the potential risk.


Assuntos
Poluentes Ambientais , Metais Pesados , Spirulina , Humanos , Carotenoides/metabolismo , Clorofila A/metabolismo , Cobre/metabolismo , Ecossistema , Poluentes Ambientais/metabolismo , Íons/metabolismo , Lagos , Metais Pesados/análise , Níquel/metabolismo , Ficobilinas/metabolismo , Ficobilinas/farmacologia , Spirulina/metabolismo , Zinco/metabolismo
14.
J Immunol Res ; 2022: 4008991, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35726224

RESUMO

Phycocyanobilin (PCB) is a linear open-chain tetrapyrrole chromophore that captures and senses light and a variety of biological activities, such as anti-oxidation, anti-cancer, and anti-inflammatory. In this paper, the biological activities of PCB are reviewed, and the related mechanism of PCB and its latest application in disease treatment are introduced. PCB can resist oxidation by scavenging free radicals, inhibiting the activity of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, and delaying the activity of antioxidant enzymes. In addition, PCB can also be used as an excellent anti-inflammatory agent to reduce the proinflammatory factors IL-6 and IFN-γ and to up-regulate the production of anti-inflammatory cytokine IL-10 by inhibiting the inflammatory signal pathways NF-κB and mitogen-activated protein kinase (MAPK). Due to the above biological activities of phycocyanobilin PCB, it is expected to become a new effective drug for treating various diseases, such as COVID-19 complications, atherosclerosis, multiple sclerosis (MS), and ischaemic stroke (IS).


Assuntos
Ficobilinas , Ficocianina , Spirulina , Anti-Inflamatórios/farmacologia , Isquemia Encefálica , Humanos , Sistema de Sinalização das MAP Quinases , NADPH Oxidases , NF-kappa B , Ficobilinas/farmacologia , Ficocianina/farmacologia , Spirulina/química
15.
ACS Synth Biol ; 11(6): 2089-2097, 2022 06 17.
Artigo em Inglês | MEDLINE | ID: mdl-35580338

RESUMO

Phycocyanobilin (PCB) is a kind of light-harvesting pigment which naturally exists in algae and plays important roles in absorbing and transferring energy. Based on its antioxidant and optical properties, PCB has been applied in food, medicine, and cosmetics. Currently, PCB is mainly extracted from Spirulina through complicated steps; thus, the biosynthesis of PCB in Escherichia coli has attracted more attention. However, due to the lower catalytic efficiency of synthetic enzymes and the deficiency of precursors and cofactors, the titer of PCB remains at a low level. Here, we report the efficient synthesis of PCB by the expression of heme oxygenase-1 from Thermosynechococcus elongatus and PCB: ferredoxin oxidoreductase (PcyA) from Synechocystis sp. using a high-copy number plasmid with an inducible T7lac promoter and the assembly of these two enzymes at a suitable ratio of 2:1 with DNA scaffolds. Additionally, the synthesis of PCB was further enhanced by direct supplementation of 5-aminolevulinic acid (ALA), moderate overexpression of key enzymes in the heme biosynthetic pathway (hemB and hemH), and accelerated cycle of cofactors (NADPH) through the expression of NAD+ kinase and the addition of a reducing agent. Finally, based on the optimal conditions (Modified R medium with 200 mg/L ALA, 20 mg/L FeSO4·7H2O, and 5 g/L vitamin C induced by 0.8 mM isopropylthio-ß-galactoside at 30 °C), the highest reported titer of PCB (28.32 mg/L) was obtained at the fermenter level by feeding glucose and FeSO4·7H2O. The strategies applied in this study will be useful for the synthesis of other natural pigments and PCB or heme derivatives in E. coli.


Assuntos
Escherichia coli , Engenharia Metabólica , Escherichia coli/genética , Escherichia coli/metabolismo , Heme , Ficobilinas , Ficocianina
16.
Photochem Photobiol Sci ; 21(4): 437-446, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35394642

RESUMO

Phycocyanobilin, the primary pigment of both light perception and light-harvesting in cyanobacteria, is synthesized from biliverdin IXα (BV) through intermediate 181, 182-dihydrobiliverdin (181, 182-DHBV) by a phycocyanobilin:ferredoxin oxidoreductase (PcyA). In our previous study, we discovered two PcyA homologs (AmPcyAc and AmPcyAp) derived from Acaryochloris marina MBIC 11017 (A. marina) that exceptionally uses chlorophyll d as the primary photosynthetic pigment, absorbing longer wavelength far-red light than chlorophyll a, the photosynthetic pigment found in most cyanobacteria. Biochemical characterization of the two PcyA homologs identified functional diversification of these two enzymes: AmPcyAc provides 181, 182-DHBV, and PCB to the cyanobacteriochrome (CBCR) photoreceptors, whereas, AmPcyAp specifically provides PCB to the light-harvesting phycobilisome subunit. In this study, we focused on the residues necessary for 181, 182-DHBV supply to the CBCR photoreceptors by AmPcyAc. Based on the SyPcyA structure, we concentrated on the 30 residues that constitute the substrate-binding pocket. Among them, we discovered that Leu151 and Val225 in AmPcyAc were both substituted with isoleucine. During the enzymatic reaction, the SyPcyA variant molecule, possessing V225I and L151I replacements, accumulates the 181, 182-DHBV and supplies it to a CBCR molecule derived from A. marina. It is worth noting that the substitution of Val225 with isoleucine was specifically conserved among the Acaryochloris genus. Collectively, we propose that the specific evolution of PcyA among the Acaryochloris genus may correlate with the acquisition of Chl. d synthetic ability and growth in long-wavelength far-red light environments.


Assuntos
Isoleucina , Oxirredutases , Clorofila , Clorofila A , Ficobilinas/química , Ficocianina
17.
Food Funct ; 13(6): 3294-3307, 2022 Mar 21.
Artigo em Inglês | MEDLINE | ID: mdl-35244658

RESUMO

Phycocyanin is a typical microalgal active compound with antioxidant and anti-inflammatory efficacy, and the pigment moiety phycocyanobilin has been recently proposed as its active structural component. Here, to explore the structural basis for phycocyanin's intestinal protective action, we evaluated the therapeutic effects and mechanism of action of phycocyanin and phycocyanobilin in dextran sodium sulphate (DSS)-induced colitis mice and in Caco-2 and RAW 264.7 cells. Phycocyanobilin was obtained by solvothermal alcoholysis of phycocyanin and characterized by spectroscopy and mass spectrometry methods. Phycocyanin, phycocyanobilin and a positive drug mesalazine were intragastrically administered to C57BL/6 mice daily for 7 days during and after 4-day DSS exposure. Clinical signs and colon histopathology revealed that phycocyanin and phycocyanobilin had an equivalent anti-colitis efficacy that was even superior to mesalazine. Based on biochemical analysis of colonic tight junction proteins, mucus compositions and goblet cells, and colonic and peripheral proinflammatory cytokines, phycocyanin and phycocyanobilin displayed equivalent intestinal epithelial barrier-protecting and anti-inflammatory potential that was evidently superior to that of mesalazine. Flow cytometry analysis of phycocyanobilin fluorescence in Caco-2 cells unveiled a similar uptake efficacy of phycocyanin and phycocyanobilin by intestinal epithelial cells. According to lactic dehydrogenase release, 2',7'-dichlorodihydrofluorescein fluorescence and methylthiazolyldiphenyl-tetrazolium bromide assay in Caco-2 cells, phycocyanin and phycocyanobilin could equally and effectively protect the intestinal epithelial barrier from oxidant-induced disruption. Phycocyanin and phycocyanobilin also showed equivalent anti-inflammatory effects in tumor necrosis factor-α-stimulated Caco-2 cells and in lipopolysaccharides- and tumor necrosis factor-α-activated RAW264.7 cells. Overall, our results demonstrate the phycocyanobilin-dependent anti-colitis role of phycocyanin via antioxidant and anti-inflammatory mechanisms.


Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Antioxidantes/farmacologia , Colite/tratamento farmacológico , Mucosa Intestinal/efeitos dos fármacos , Ficobilinas/farmacologia , Ficocianina/farmacologia , Animais , Anti-Inflamatórios não Esteroides/uso terapêutico , Antioxidantes/uso terapêutico , Células CACO-2 , Colite/fisiopatologia , Células Epiteliais/metabolismo , Humanos , Mucosa Intestinal/metabolismo , Masculino , Mesalamina/farmacologia , Camundongos , Camundongos Endogâmicos C57BL , Ficobilinas/metabolismo , Ficobilinas/uso terapêutico , Ficocianina/metabolismo , Ficocianina/uso terapêutico , Células RAW 264.7
18.
Photosynth Res ; 152(1): 23-42, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35064531

RESUMO

Photosynthetic pigments are an integral and vital part of all photosynthetic machinery and are present in different types and abundances throughout the photosynthetic apparatus. Chlorophyll, carotenoids and phycobilins are the prime photosynthetic pigments which facilitate efficient light absorption in plants, algae, and cyanobacteria. The chlorophyll family plays a vital role in light harvesting by absorbing light at different wavelengths and allowing photosynthetic organisms to adapt to different environments, either in the long-term or during transient changes in light. Carotenoids play diverse roles in photosynthesis, including light capture and as crucial antioxidants to reduce photodamage and photoinhibition. In the marine habitat, phycobilins capture a wide spectrum of light and have allowed cyanobacteria and red algae to colonise deep waters where other frequencies of light are attenuated by the water column. In this review, we discuss the potential strategies that photosynthetic pigments provide, coupled with development of molecular biological techniques, to improve crop yields through enhanced light harvesting, increased photoprotection and improved photosynthetic efficiency.


Assuntos
Cianobactérias , Ficobilinas , Carotenoides/metabolismo , Clorofila , Cianobactérias/metabolismo , Fotossíntese/fisiologia , Plantas/metabolismo
19.
Anal Biochem ; 642: 114557, 2022 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-35092720

RESUMO

Cyanobacteriochromes are the extended family of phytochrome photosensors characterized in cyanobacteria. Alr1966g2C56A is a cyanobacteriochrome mutant of Alr1966g2 in Nostoc sp. PCC 7120 from freshwater. In this paper, we truncated ten residues in the N-terminus and ten residues in the C-terminus of Alr1966g2C56A and obtained truncated Alr1966g2C46A, termed as Alr1966g2C46A-tr. Alr1966g2C46A-tr binded covalently not only phycocyanobilin but also biliverdin via Cys74 of the conserved CH motif, and showed a significant improvement in binding-PCB efficiency in E. coli, compared with that of untruncated Alr1966g2C56A. We also captured a persistent red fluorescence of Alr1966g2C46A-tr-PCB or Alr1966g2C46A-tr-BV expressed in live E. coli. Thus, Alr1966g2C46A-tr was suitable for the stable red fluorescent probe as a starting material.


Assuntos
Biliverdina/química , Cianobactérias/química , Proteínas Luminescentes/análise , Ficobilinas/química , Ficocianina/química , Fitocromo/química , Proteína Vermelha Fluorescente
20.
J Cell Sci ; 134(24)2021 12 15.
Artigo em Inglês | MEDLINE | ID: mdl-34806750

RESUMO

Near-infrared fluorescent protein (iRFP) is a bright and stable fluorescent protein with near-infrared excitation and emission maxima. Unlike the other conventional fluorescent proteins, iRFP requires biliverdin (BV) as a chromophore. Here, we report that phycocyanobilin (PCB) functions as a brighter chromophore for iRFP than BV, and that biosynthesis of PCB allows live-cell imaging with iRFP in the fission yeast Schizosaccharomyces pombe. We initially found that fission yeast cells did not produce BV and therefore did not show any iRFP fluorescence. The brightness of iRFP-PCB was higher than that of iRFP-BV both in vitro and in fission yeast. We introduced SynPCB2.1, a PCB biosynthesis system, into fission yeast, resulting in the brightest iRFP fluorescence. To make iRFP readily available in fission yeast, we developed an endogenous gene tagging system with iRFP and all-in-one integration plasmids carrying the iRFP-fused marker proteins together with SynPCB2.1. These tools not only enable the easy use of multiplexed live-cell imaging in fission yeast with a broader color palette, but also open the door to new opportunities for near-infrared fluorescence imaging in a wider range of living organisms. This article has an associated First Person interview with the first author of the paper.


Assuntos
Schizosaccharomyces , Humanos , Proteínas Luminescentes/genética , Ficobilinas , Ficocianina , Schizosaccharomyces/genética
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA