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1.
An Acad Bras Cienc ; 96(3): e20230348, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39166650

RESUMO

Cyanobacterial phycocyanin and phycoerythrin are gaining commercial interest due to their nutrition and healthcare values. This research analyzed the biomass accumulation and pigment production of two strains of Leptolyngbya under different combinations of light colors and intensities. The results showed that while Leptolyngbya sp.4 B1 (B1) produced all phycobiliproteins, Leptolyngbya sp.5 F2 (F2) only had phycocyanin and allophycocyanin. Both the color of the light and its light intensity affect the biomass accumulation and phycoerythrin concentration in strain B1. Although white light at medium intensity (50 µmol m-2 s-1) causes greater biomass accumulation (1.66 ± 0.13 gDW L-1), low-intensity (25 µmol m-2 s-1) green light induces lower biomass accumulation with twice the pigment content (87.70 ± 2.46 mg gDW -1), culminating in 71% greater productivity. In contrast, for the F2 strain, light intensity positively influenced biomass and pigment accumulation, being observed 2.25 ± 0.10 gDW L-1 under white light at 100 µmol m-2 s-1 and higher phycocyanin concentration (138.38 ± 3.46 mg gDW -1) under red light at 100 µmol m-2 s-1. These findings provide insights into optimizing the growth conditions by altering the intensity and wavelength of light for future production of phycocyanin and phycoerythrin from local cyanobacteria.


Assuntos
Biomassa , Cianobactérias , Luz , Ficobiliproteínas , Ficobiliproteínas/metabolismo , Cianobactérias/metabolismo , Cianobactérias/efeitos da radiação , Cianobactérias/classificação , Florestas , Ficocianina/metabolismo , Ficocianina/biossíntese , Ficoeritrina/metabolismo , Ficoeritrina/biossíntese
2.
ACS Synth Biol ; 13(8): 2391-2401, 2024 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-39038807

RESUMO

Phycobilisomes (PBSs) are light-harvesting antenna complexes in cyanobacteria that adapt to diverse light environments through the use of phycobiliproteins within the PBS structures. Freshwater cyanobacteria, such as Synechococcus elongatus PCC 7942, thrive under red light because of the presence of phycocyanin (PC) and its chromophore, phycocyanobilin (PCB), in the PBS. Cyanobacteria in shorter-wavelength light environments such as green light, employ phycoerythrin paired with phycoerythrobilin (PEB) along with PC in the PBS. Synthetic biology studies have shown that PEB production can be achieved by expression of the heterologous PEB synthases 15,16-dihydrobiliverdin:ferredoxin oxidoreductase (PebA) and PEB:ferredoxin oxidoreductase (PebB), leading to PEB accumulation and cellular browning. This approach is genetically unstable, and the properties of the resulting PEB-bound PBS complexes remain uncharacterized. In this study, we engineered a novel strain of Synechococcus 7942 PEB1 with finely tuned control of PEB biosynthesis. PEB1 exhibited a reversible change in the color of the culture from green to brown and pink based on PebA and PebB induction levels. High induction led to complete PCB-to-PEB substitution, causing the disassembly of the PBS rod complex. In contrast, low induction levels of PebA and PebB resulted in the formation of a stable chimeric PBS complex with partial PCB-to-PEB substitution. This acclimation enabled efficient light harvesting in the green spectrum and energy transfer to the photosynthetic reaction center. These findings, which improve our understanding of PBS and highlight the structural importance of the bilin composition, provide a foundation for future studies on PBS adaptation in bioengineering, synthetic biology, and renewable energy.


Assuntos
Proteínas de Bactérias , Ficobiliproteínas , Ficobilissomas , Ficocianina , Synechococcus , Synechococcus/metabolismo , Synechococcus/genética , Ficobilissomas/metabolismo , Ficobiliproteínas/metabolismo , Ficobiliproteínas/genética , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/genética , Ficocianina/metabolismo , Ficocianina/genética , Ficobilinas/metabolismo , Ficoeritrina/metabolismo , Ficoeritrina/química , Pigmentos Biliares/metabolismo , Luz , Biologia Sintética/métodos , Cianobactérias/metabolismo , Cianobactérias/genética
3.
Int J Mol Sci ; 25(14)2024 Jul 10.
Artigo em Inglês | MEDLINE | ID: mdl-39062809

RESUMO

The study evaluated the effects of Arthrospira maxima phycobiliproteins (PBPs), rosiglitazone (RSG), and 17ß-estradiol (E) on the differentiation process of 3T3-L1 cells and on their regulation of lipogenic and inflammatory gene expression at different stages of the process. The results showed that phycobiliproteins promoted cell proliferation after 24 h of treatment. Furthermore, for all three treatments, the regulation of the highest number of markers occurred on days 6 and 12 of differentiation, regardless of when the treatment was applied. Phycobiliproteins reduced lipid droplet accumulation on days 3, 6, 10, and 13 of the adipogenic process, while rosiglitazone showed no differences compared to the control. On day 6, both phycobiliproteins and rosiglitazone positively regulated Acc1 mRNA. Meanwhile, all three treatments negatively regulated Pparγ and C/ebpα. Phycobiliproteins and estradiol also negatively regulated Ucp1 and Glut4 mRNAs. Rosiglitazone and estradiol, on the other hand, negatively regulated Ppara and Il-6 mRNAs. By day 12, phycobiliproteins and rosiglitazone upregulated Pparγ mRNA and negatively regulated Tnfα and Il-1ß. Additionally, phycobiliproteins and estradiol positively regulated Il-6 and negatively regulated Ppara, Ucp2, Acc1, and Glut4. Rosiglitazone and estradiol upregulate C/ebpα and Ucp1 mRNAs. The regulation exerted by phycobiliproteins on the mRNA expression of the studied markers was dependent on the phase of cell differentiation. The results of this study highlight that phycobiliproteins have an anti-adipogenic and anti-inflammatory effect by reducing the expression of adipogenic, lipogenic, and inflammatory genes in 3T3-L1 cells at different stages of the differentiation process.


Assuntos
Células 3T3-L1 , Adipócitos , Adipogenia , Diferenciação Celular , Estradiol , Ficobiliproteínas , Rosiglitazona , Animais , Camundongos , Estradiol/farmacologia , Rosiglitazona/farmacologia , Adipócitos/efeitos dos fármacos , Adipócitos/metabolismo , Adipócitos/citologia , Diferenciação Celular/efeitos dos fármacos , Adipogenia/efeitos dos fármacos , Adipogenia/genética , Ficobiliproteínas/farmacologia , Ficobiliproteínas/metabolismo , Ficobiliproteínas/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Lipogênese/efeitos dos fármacos , Lipogênese/genética , PPAR gama/metabolismo , PPAR gama/genética , Proliferação de Células/efeitos dos fármacos , Inflamação/metabolismo , Inflamação/genética , Spirulina
4.
World J Microbiol Biotechnol ; 40(9): 272, 2024 Jul 20.
Artigo em Inglês | MEDLINE | ID: mdl-39030303

RESUMO

Microalgae are a source of a wide variety of commodities, including particularly valuable pigments. The typical pigments present in microalgae are the chlorophylls, carotenoids, and phycobiliproteins. However, other types of pigments, of the family of water-soluble polyphenols, usually encountered in terrestrial plants, have been recently reported in microalgae. Among such microalgal polyphenols, many flavonoids have a yellowish hue, and are used as natural textile dyes. Besides being used as natural colorants, for example in the food or cosmetic industry, microalgal pigments also possess many bioactive properties, making them functional as nutraceutical or pharmaceutical agents. Each type of pigment, with its own chemical structure, fulfills particular biological functions. Considering both eukaryotes and prokaryotes, some species within the four most promising microalgae groups (Cyanobacteria, Rhodophyta, Chlorophyta and Heterokontophyta) are distinguished by their high contents of specific added-value pigments. To further enhance microalgae pigment contents during autotrophic cultivation, a review is made of the main related strategies adopted during the last decade, including light adjustments (quantity and quality, and the duration of the photoperiod cycle), and regard to mineral medium characteristics (salinity, nutrients concentrations, presence of inductive chemicals). In contrast to what is usually observed for growth-related pigments, accumulation of non-photosynthetic pigments (polyphenols and secondary carotenoids) requires particularly stressful conditions. Finally, pigment enrichment is also made possible with two new cutting-edge technologies, via the application of metallic nanoparticles or magnetic fields.


Assuntos
Microalgas , Pigmentos Biológicos , Microalgas/metabolismo , Microalgas/química , Pigmentos Biológicos/química , Carotenoides/química , Carotenoides/metabolismo , Carotenoides/análise , Ficobiliproteínas/química , Ficobiliproteínas/metabolismo , Cianobactérias/metabolismo , Cianobactérias/química , Rodófitas/química , Rodófitas/metabolismo , Clorófitas/química , Clorófitas/metabolismo , Clorofila/análise , Polifenóis/análise , Polifenóis/química , Polifenóis/metabolismo , Meios de Cultura/química
5.
Commun Biol ; 7(1): 560, 2024 May 11.
Artigo em Inglês | MEDLINE | ID: mdl-38734819

RESUMO

Photosynthetic cryptophytes are eukaryotic algae that utilize membrane-embedded chlorophyll a/c binding proteins (CACs) and lumen-localized phycobiliproteins (PBPs) as their light-harvesting antennae. Cryptophytes go through logarithmic and stationary growth phases, and may adjust their light-harvesting capability according to their particular growth state. How cryptophytes change the type/arrangement of the photosynthetic antenna proteins to regulate their light-harvesting remains unknown. Here we solve four structures of cryptophyte photosystem I (PSI) bound with CACs that show the rearrangement of CACs at different growth phases. We identify a cryptophyte-unique protein, PsaQ, which harbors two chlorophyll molecules. PsaQ specifically binds to the lumenal region of PSI during logarithmic growth phase and may assist the association of PBPs with photosystems and energy transfer from PBPs to photosystems.


Assuntos
Criptófitas , Complexo de Proteína do Fotossistema I , Complexo de Proteína do Fotossistema I/metabolismo , Criptófitas/metabolismo , Criptófitas/genética , Complexos de Proteínas Captadores de Luz/metabolismo , Clorofila/metabolismo , Proteínas de Ligação à Clorofila/metabolismo , Proteínas de Ligação à Clorofila/genética , Fotossíntese , Ficobiliproteínas/metabolismo , Ficobiliproteínas/genética
6.
Chembiochem ; 25(11): e202400068, 2024 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-38623786

RESUMO

Far-red and near-infrared fluorescent proteins have regions of maximum transmission in most tissues and can be widely used as fluorescent biomarkers. We report that fluorescent phycobiliproteins originating from the phycobilisome core subunit ApcF2 can covalently bind biliverdin, named BDFPs. To further improve BDFPs, we conducted a series of studies. Firstly, we mutated K53Q and T144A of BDFPs to increase their effective brightness up to 190 % in vivo. Secondly, by homochromatic tandem fusion of high-brightness BDFPs to achieve monomerization, which increases the effective brightness by up to 180 % in vivo, and can effectively improve the labeling effect. By combining the above two approaches, the brightness of the tandem BDFPs was much improved compared with that of the previously reported fluorescent proteins in a similar spectral range. The tandem BDFPs were expressed stably while maintaining fluorescence in mammalian cells and Caenorhabditis elegans. They were also photostable and resistant to high temperature, low pH, and chemical denaturation. The tandem BDFPs advantages were proved in applications as biomarkers for imaging in super-resolution microscopy.


Assuntos
Caenorhabditis elegans , Proteínas Luminescentes , Animais , Proteínas Luminescentes/química , Proteínas Luminescentes/genética , Proteínas Luminescentes/metabolismo , Caenorhabditis elegans/metabolismo , Humanos , Ficobiliproteínas/química , Ficobiliproteínas/metabolismo , Biliverdina/química , Biliverdina/metabolismo , Corantes Fluorescentes/química , Células HEK293
7.
Commun Biol ; 6(1): 1158, 2023 11 13.
Artigo em Inglês | MEDLINE | ID: mdl-37957226

RESUMO

Cryptophyte algae have a unique phycobiliprotein light-harvesting antenna that fills a spectral gap in chlorophyll absorption from photosystems. However, it is unclear how the antenna transfers energy efficiently to these photosystems. We show that the cryptophyte Hemiselmis andersenii expresses an energetically complex antenna comprising three distinct spectrotypes of phycobiliprotein, each composed of two αß protomers but with different quaternary structures arising from a diverse α subunit family. We report crystal structures of the major phycobiliprotein from each spectrotype. Two-thirds of the antenna consists of open quaternary form phycobiliproteins acting as primary photon acceptors. These are supplemented by a newly discovered open-braced form (~15%), where an insertion in the α subunit produces ~10 nm absorbance red-shift. The final components (~15%) are closed forms with a long wavelength spectral feature due to substitution of a single chromophore. This chromophore is present on only one ß subunit where asymmetry is dictated by the corresponding α subunit. This chromophore creates spectral overlap with chlorophyll, thus bridging the energetic gap between the phycobiliprotein antenna and the photosystems. We propose that the macromolecular organization of the cryptophyte antenna consists of bulk open and open-braced forms that transfer excitations to photosystems via this bridging closed form phycobiliprotein.


Assuntos
Criptófitas , Fotossíntese , Ficobiliproteínas/química , Ficobiliproteínas/metabolismo , Clorofila
8.
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
9.
Curr Microbiol ; 80(8): 242, 2023 Jun 10.
Artigo em Inglês | MEDLINE | ID: mdl-37300570

RESUMO

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.


Assuntos
Ficocianina , Synechococcus , Ficocianina/genética , Ficocianina/metabolismo , Ficobiliproteínas/metabolismo , Ficobiliproteínas/farmacologia , Synechococcus/genética , Campos Magnéticos
10.
Chemosphere ; 335: 139107, 2023 09.
Artigo em Inglês | MEDLINE | ID: mdl-37270039

RESUMO

The production of biodiesel has become an important issue in the effort to reduce gas emissions due to the climate change crisis; therefore, algae have widely used to produce biodiesel for energy sustainability. The present study represented an effort to assess the ability of the alga Arthrospira platensis to produce fatty acids involved in biofuel (diesel) by cultivation in Zarrouk media enriched with different municipal wastewater concentrations. Wastewater was used in different concentrations (5, 15, 25, 35 and 100% [control]). Five fatty acids from the alga were determined and included in the present study. These were inoleic acid, palmitic acid, oleic acid, gamma-linolenic acid, and docosahexaenoic acid. Impact of different cultivation conditions were studied in terms of observed changes in growth rate, doubling time, total carbohydrate, total protein, chlorophyll a, carotenoids, phycocyanin, allophycocyanin, and phycobiliproteins. Results showed an increase in the values of growth rate, total protein content, chlorophyll a, and levels of carotenoids at all treatments except for carbohydrate content, which decreased with an increasing concentration of wastewater. The high value of doubling time (11.605 days) was recorded at treatment 5%. Fatty acids yields were increased at treatment 5% and 15%. The highest concentrations of fatty acids were 3.108 mg/g for oleic acid, gamma-linolenic acid (28.401 mg/g), docosahexaenoic acid (41.707 mg/g), palmitic acid (1.305 mg/g), and linoleic acid (0.296 mg/g). Moreover, the range of phycocyanin (0.017-0.084 mg/l), allophycocyanin (0.023-0.095 mg/l), and phycobiliproteins (0.041-0.180 mg/l) were obtained in treatment with 15-100%, respectively. Cultivation with municipal wastewater reduced the values of nitrate, phosphate, and electrical conductivity as well as increased dissolved oxygen. Maximum electrical conductivity was recorded in untreated wastewater with algae, while the highest level of dissolved oxygen was noted at 35% concentration. The use of the household wastewater is more environmentally friendly as an alternative of the traditional cultivation techniques used for long-term for biofuel production.


Assuntos
Microalgas , Spirulina , Águas Residuárias , Ficocianina , Clorofila A/metabolismo , Biocombustíveis , Ácido gama-Linolênico/metabolismo , Ácidos Docosa-Hexaenoicos , Spirulina/metabolismo , Ácidos Graxos/metabolismo , Ficobiliproteínas/metabolismo , Carotenoides/metabolismo , Carboidratos , Biomassa
11.
Proc Natl Acad Sci U S A ; 120(17): e2300770120, 2023 04 25.
Artigo em Inglês | MEDLINE | ID: mdl-37071675

RESUMO

Terrestrial ecosystems and human societies depend on oxygenic photosynthesis, which began to reshape our atmosphere approximately 2.5 billion years ago. The earliest known organisms carrying out oxygenic photosynthesis are the cyanobacteria, which use large complexes of phycobiliproteins as light-harvesting antennae. Phycobiliproteins rely on phycocyanobilin (PCB), a linear tetrapyrrole (bilin) chromophore, as the light-harvesting pigment that transfers absorbed light energy from phycobilisomes to the chlorophyll-based photosynthetic apparatus. Cyanobacteria synthesize PCB from heme in two steps: A heme oxygenase converts heme into biliverdin IXα (BV), and the ferredoxin-dependent bilin reductase (FDBR) PcyA then converts BV into PCB. In the current work, we examine the origins of this pathway. We demonstrate that PcyA evolved from pre-PcyA proteins found in nonphotosynthetic bacteria and that pre-PcyA enzymes are active FDBRs that do not yield PCB. Pre-PcyA genes are associated with two gene clusters. Both clusters encode bilin-binding globin proteins, phycobiliprotein paralogs that we designate as BBAGs (bilin biosynthesis-associated globins). Some cyanobacteria also contain one such gene cluster, including a BBAG, two V4R proteins, and an iron-sulfur protein. Phylogenetic analysis shows that this cluster is descended from those associated with pre-PcyA proteins and that light-harvesting phycobiliproteins are also descended from BBAGs found in other bacteria. We propose that PcyA and phycobiliproteins originated in heterotrophic, nonphotosynthetic bacteria and were subsequently acquired by cyanobacteria.


Assuntos
Cianobactérias , Ficobiliproteínas , Humanos , Filogenia , Ficobiliproteínas/metabolismo , Oxirredutases/metabolismo , Ecossistema , Pigmentos Biliares/química , Cianobactérias/química
12.
Int J Mol Sci ; 24(3)2023 Jan 24.
Artigo em Inglês | MEDLINE | ID: mdl-36768613

RESUMO

Eukaryotic photosynthesis originated in the course of evolution as a result of the uptake of some unstored cyanobacterium and its transformation to chloroplasts by an ancestral heterotrophic eukaryotic cell. The pigment apparatus of Archaeplastida and other algal phyla that emerged later turned out to be arranged in the same way. Pigment-protein complexes of photosystem I (PS I) and photosystem II (PS II) are characterized by uniform structures, while the light-harvesting antennae have undergone a series of changes. The phycobilisome (PBS) antenna present in cyanobacteria was replaced by Chl a/b- or Chl a/c-containing pigment-protein complexes in most groups of photosynthetics. In the form of PBS or phycobiliprotein aggregates, it was inherited by members of Cyanophyta, Cryptophyta, red algae, and photosynthetic amoebae. Supramolecular organization and architectural modifications of phycobiliprotein antennae in various algal phyla in line with the endosymbiotic theory of chloroplast origin are the subject of this review.


Assuntos
Cianobactérias , Ficobilissomas , Ficobilissomas/química , Ficobilissomas/metabolismo , Ficobiliproteínas/metabolismo , Simbiose , Oxigênio/metabolismo , Fotossíntese , Cianobactérias/genética , Cianobactérias/metabolismo , Complexo de Proteína do Fotossistema II/metabolismo , Complexo de Proteína do Fotossistema I/metabolismo , Clorofila/metabolismo
13.
J Am Nutr Assoc ; 42(6): 559-572, 2023 08.
Artigo em Inglês | MEDLINE | ID: mdl-35916491

RESUMO

Spirulina is a blue-green algae which is cultivated not only for its maximum protein content but also due to the presence of other essential nutrients such as carbohydrates and vitamins (A, C and E). It is also a storehouse of minerals including iron, calcium, chromium, copper, magnesium, manganese, phosphorus, potassium, sodium and zinc. Simultaneously, γ- linolenic acid (an essential fatty acid), as well as pigments such as chlorophyll A and phycobiliproteins (C-phycocyanin, allophycocyanin and ß-carotene), is also a major component of its rich nutritional profile. Spirulina is known to have various promising effects on the prevention of cancer, oxidative stress, obesity, diabetes, cardiovascular diseases and anemia. Moreover, it also plays a positive role in treating muscular cramps. The safety recommended dosage of Spirulina is approximately 3-10 g/d for adults and it's biological value (BV) is 75 with a net protein utilization (NPU) of 62. Spirulina does not have pericardium due to which it does not hinder the absorption of iron by chelation with phytates or oxalates. On the contrasting note, it may have some adverse effects due to the toxins (microcystins, ß-methylamino-L-alanine (BMAA)) produced by Spirulina which might contribute to acute poisoning, cancer, liver damage as well as gastrointestinal disturbances. Its long-term consumption may also lead to the pathogenesis of Alzheimer's disease and Parkinson's disease. The current review focuses on the various aspects of spirulina including its cultivation, nutritional composition, extraction techniques, health benefits, adverse effects, industrial scope and market value which could be beneficial for its utilization in the development of value-added products and supplementary foods due to its high content of protein and bioavailability of nutrients.


• Spirulina is a nutrient-dense cyanobacterium which is composed of protein, carbohydrates, vitamins, minerals, essential fatty acids, antioxidants and pigments including chlorophyll A and Phycocyanin.• To avoid the contamination of Spirulina species by other algae, the specific pH maintenance of the media around 9-11 (alkaline) is mandatory.• Positive effects were noticed on the yield and productivity of Spirulina after its biomass was grown in polybags and greenhouse.• Its beneficial effects have been identified in particular reference to obesity, diabetes, hypertension, cardiovascular diseases, anemia, cancer, oxidative stress, arthritis, immunity as well as muscular cramps.• The toxins such as microcystins and hepatotoxins, produced by Spirulina, are accountable to cause acute poisoning, liver damage, gastrointestinal disturbances and cancer.• The safe recommended dosage of Spirulina for adults accounts to approximately 3-10 g/d, with 30 g/d being the maximum limit for consumption.


Assuntos
Spirulina , Clorofila A/metabolismo , Carboidratos , Ficobiliproteínas/metabolismo , Ferro/metabolismo
14.
J Microbiol Biotechnol ; 32(10): 1325-1334, 2022 Oct 28.
Artigo em Inglês | MEDLINE | ID: mdl-36224760

RESUMO

Global warming has accelerated in recent decades due to the continuous consumption of petroleum-based fuels. Cyanobacteria-derived biofuels are a promising carbon-neutral alternative to fossil fuels that may help achieve a cleaner environment. Here, we propose an effective strategy based on the large-scale cultivation of a newly isolated cyanobacterial strain to produce phycobiliprotein and biodiesel, thus demonstrating the potential commercial applicability of the isolated microalgal strain. A native cyanobacterium was isolated from Goryeong, Korea, and identified as Pseudanabaena mucicola GO0704 through 16s RNA analysis. The potential exploitation of P. mucicola GO0704 was explored by analyzing several parameters for mixotrophic culture, and optimal growth was achieved through the addition of sodium acetate (1 g/l) to the BG-11 medium. Next, the cultures were scaled up to a stirred-tank bioreactor in mixotrophic conditions to maximize the productivity of biomass and metabolites. The biomass, phycobiliprotein, and fatty acids concentrations in sodium acetate-treated cells were enhanced, and the highest biodiesel productivity (8.1 mg/l/d) was achieved at 96 h. Finally, the properties of the fuel derived from P. mucicola GO0704 were estimated with converted biodiesels according to the composition of fatty acids. Most of the characteristics of the final product, except for the cloud point, were compliant with international biodiesel standards [ASTM 6761 (US) and EN 14214 (Europe)].


Assuntos
Cianobactérias , Microalgas , Biocombustíveis/análise , Ficobiliproteínas/metabolismo , Acetato de Sódio/metabolismo , Microalgas/metabolismo , Cianobactérias/genética , Biomassa , Ácidos Graxos/metabolismo
15.
Chembiochem ; 23(18): e202200267, 2022 09 16.
Artigo em Inglês | MEDLINE | ID: mdl-35811374

RESUMO

Far-red and near-infrared fluorescent proteins can be used as fluorescence biomarkers in the region of maximal transmission of most tissues and facilitate multiplexing. Recently, we reported the generation and properties of far-red and near-infrared fluorescent phycobiliproteins, termed BeiDou Fluorescent Proteins (BDFPs), which can covalently bind the more readily accessible biliverdin. Far-red BDFPs maximally fluoresce at ∼670 nm, while near-infrared BDFPs fluoresce at ∼710 nm. In this work, we molecularly evolved BDFPs as follows: (a) mutations L58Q, S68R and M81K of BDFPs, which can maximally enhance the effective brightness in vivo by 350 %; (b) minimization and monomerization of far-red BDFPs 2.1, 2.2, 2.3, and near-infrared BDFPs 2.4, 2.5 and 2.6. These newly developed BDFPs are remarkably brighter than the formerly reported far-red and near-infrared fluorescent proteins. Their advantages are demonstrated by biolabeling in mammalian cells using super-resolution microscopy.


Assuntos
Biliverdina , Ficobiliproteínas , Animais , Proteínas de Bactérias/metabolismo , Biomarcadores , Corantes Fluorescentes/metabolismo , Mamíferos/metabolismo , Microscopia de Fluorescência , Ficobiliproteínas/metabolismo
16.
Structure ; 30(4): 564-574.e3, 2022 04 07.
Artigo em Inglês | MEDLINE | ID: mdl-35148828

RESUMO

Chromophore attachment of the light-harvesting apparatus represents one of the most important post-translational modifications in photosynthetic cyanobacteria. Extensive pigment diversity of cyanobacteria critically depends on bilin lyases that covalently attach chemically distinct chromophores to phycobiliproteins. However, how bilin lyases catalyze bilin ligation reactions and how some lyases acquire additional isomerase abilities remain elusive at the molecular level. Here, we report the crystal structure of a representative bilin lyase-isomerase MpeQ. This structure has revealed a "question-mark" protein architecture that unambiguously establishes the active site conserved among the E/F-type bilin lyases. Based on structural, mutational, and modeling data, we demonstrate that stereoselectivity of the active site plays a critical role in conferring the isomerase activity of MpeQ. We further advance a tyrosine-mediated reaction scheme unifying different types of bilin lyases. These results suggest that lyases and isomerase actions of bilin lyases arise from two coupled molecular events of distinct origin.


Assuntos
Cianobactérias , Liases , Pigmentos Biliares/metabolismo , Cianobactérias/metabolismo , Isomerases/genética , Isomerases/metabolismo , Liases/química , Liases/genética , Liases/metabolismo , Ficobiliproteínas/metabolismo
17.
FEBS J ; 289(15): 4646-4656, 2022 08.
Artigo em Inglês | MEDLINE | ID: mdl-35156751

RESUMO

Blue-green algae, also known as cyanobacteria, contain some of the most efficient light-harvesting complexes known. These large, colourful complexes consist of phycobiliproteins which are extremely valuable in the cosmetics, food, nutraceutical and pharmaceutical industries. Additionally, the colourful and fluorescent properties of phycobiliproteins can be modulated by metal ions, making them highly attractive as heavy metal sensors and heavy metal scavengers. Although the overall quenching ability metal ions have on phycobiliproteins is known, the mechanism of heavy metal binding to phycobiliproteins is not fully understood, limiting their widespread quantitative applications. Here, we show using high-resolution native mass spectrometry that phycobiliprotein complexes bind metal ions in different manners. Through monitoring the binding equilibria and metal-binding stoichiometry, we show in particular copper and silver to have drastic, yet different effects on phycobiliprotein structure, both copper and silver modulate the overall complex properties. Together, the data reveals the mechanisms by which metal ions can modulate phycobiliprotein properties which can be used as a basis for the future design of metal-related phycobiliprotein applications.


Assuntos
Cianobactérias , Ficobiliproteínas , Cobre/metabolismo , Cianobactérias/metabolismo , Ficobiliproteínas/química , Ficobiliproteínas/metabolismo , Prata/metabolismo
18.
J Chem Phys ; 156(7): 070901, 2022 Feb 21.
Artigo em Inglês | MEDLINE | ID: mdl-35183095

RESUMO

Single-molecule fluorescence spectroscopy allows direct, real-time observation of dynamic photophysical changes in light harvesting complexes. The Anti-Brownian ELectrokinetic (ABEL) trap is one such single-molecule method with useful advantages. This approach is particularly well-suited to make detailed spectroscopic measurements of pigment-protein complexes in a solution phase because it enables extended-duration single-molecule observation by counteracting Brownian motion. This Perspective summarizes recent contributions by the authors and others that have utilized the unique capabilities of the ABEL trap to advance our understanding of phycobiliproteins and the phycobilisome complex, the primary light-harvesting apparatus of cyanobacteria. Monitoring the rich spectroscopic data from these measurements, which include brightness, fluorescence lifetime, polarization, and emission spectra, among other measurable parameters, has provided direct characterization of pigments and energy transfer pathways in the phycobilisome, spanning scales from single pigments and monomeric phycobiliproteins to higher order oligomers and protein-protein interactions of the phycobilisome complex. Importantly, new photophysical states and photodynamics were observed to modulate the flow of energy through the phycobilisome and suggest a previously unknown complexity in phycobilisome light harvesting and energy transport with a possible link to photoadaptive or photoprotective functions in cyanobacteria. Beyond deepening our collective understanding of natural light-harvesting systems, these and future discoveries may serve as inspiration for engineering improved artificial light-harvesting technologies.


Assuntos
Cianobactérias , Ficobilissomas , Cianobactérias/metabolismo , Transferência de Energia , Complexos de Proteínas Captadores de Luz/metabolismo , Ficobiliproteínas/metabolismo , Ficobilissomas/metabolismo , Espectrometria de Fluorescência/métodos
19.
Environ Sci Pollut Res Int ; 29(12): 17441-17455, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-34664174

RESUMO

Cadmium (Cd) is known for its many toxic effects on male population such as hypogonadism and fertility difficulties, which are oftenly associated with oxidative stress. As beneficial food, Spirulina(Sp) has been proved efficient against the heavy metal toxicity. This capacity can be associated with its phycobiliproteins (PBP). In this study, the capability of PBP and Sp to treat Cd-induced oxidative damage on the testes and spermatozoa was considered. CD-1 strain mice were orally treated with either Sp or PBP for 10 days prior to single-dose Cd challenge. Sperm quality determinations and testicle histology analysis were performed. Testosterone on serum was measured using enzyme-linked immunosorbent assay (ELISA). Oxidative damage was determined. Antioxidant enzyme activity was analyzed by measuring the activity of super oxide dismutase (SOD), catalase (Cat), and glutathione peroxidase (GpX). The motility and viability of sperm decrease with Cd and improve with PBP and Sp, as the acrosomal reaction (AR) is diminished by PBPs. Testosterone levels decrease due to Cd, and only Sp maintains elevated levels. Cd increases the production of malondialdehyde in the spermatozoa, but not in testes; this production of malondialdehyde in the spermatozoa decreases in the presence of PBP. ROS only decreases with Cd, FBP, and Sp at high concentrations. Advanced oxidative protein products (AOPP) decrease with Cd and PBPs. Cat and GpX increase their activity with Cd and are altered by FBP. Cd produces vascular alterations testes. Within the seminiferous tubule, it produces areas of necrosis and apoptosis, which improve with PBPs and Sp. PBPs have a strong antioxidant activity as they show protective properties against Cd oxidative-induced toxicity on testes and sperm.


Assuntos
Cádmio , Spirulina , Animais , Antioxidantes/metabolismo , Antioxidantes/farmacologia , Cádmio/metabolismo , Catalase/metabolismo , Masculino , Camundongos , Estresse Oxidativo , Ficobiliproteínas/metabolismo , Ficobiliproteínas/farmacologia , Extratos Vegetais/farmacologia , Espermatozoides , Spirulina/metabolismo , Superóxido Dismutase/metabolismo , Testículo
20.
Mol Phylogenet Evol ; 155: 106991, 2021 02.
Artigo em Inglês | MEDLINE | ID: mdl-33098986

RESUMO

Cyanobacteria are often reported as abundant components of the sponge microbiome; however their diversity below the phylum level is still underestimated. Aiming to broaden our knowledge of sponge-cyanobacteria association, we isolated cyanobacterial strains from Aegean Sea sponges in previous research, which revealed high degree of novel cyanobacterial diversity. Herein, we aim to further characterize sponge-associated cyanobacteria and re-evaluate their classification based on an extensive polyphasic approach, i.e. a combination of molecular, morphological and ecological data. This approach resulted in the description of five new genera (Rhodoploca, Cymatolege, Metis, Aegeococcus, and Thalassoporum) and seven new species (R. sivonenia, C. spiroidea, C. isodiametrica, M. fasciculata, A. anagnostidisi, A. thureti, T. komareki) inside the order Synechococcales, and a new pleurocapsalean species (Xenococcus spongiosum). X. spongiosum is a baeocyte-producing species that shares some morphological features with other Xenococcus species, but has distinct phylogenetic and ecological identity. Rhodoploca, Cymatolege, Metis and Thalassoporum are novel well supported linages of filamentous cyanobacteria that possess distinct characters compared to their sister taxa. Aegeococcus is a novel monophyletic linage of Synechococcus-like cyanobacteria exhibiting a unique ecology, as sponge-dweller. The considerable number of novel taxa characterized in this study highlights the importance of employing polyphasic culture-dependent approaches in order to reveal the true cyanobacterial diversity associated with sponges.


Assuntos
Cianobactérias/classificação , Poríferos/microbiologia , Animais , Sequência de Bases , Cianobactérias/genética , Cianobactérias/ultraestrutura , DNA Espaçador Ribossômico/genética , Conformação de Ácido Nucleico , Ficobiliproteínas/metabolismo , Filogenia , RNA Ribossômico 16S/química , RNA Ribossômico 16S/genética
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