Analytical Protocols in Phycobiliproteins Analysis

Nikolić, Milan; Minić, Simeon; Mačvanin, Mirjana; Stanić-Vučinić, Dragana; Veličković, Tanja Ćirković

doi:10.1007/978-3-030-50971-2_8

Cited by 4 publications

(8 citation statements)

References 109 publications

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“…The supernatant was collected for measuring the amount of phycobilisome. The absorbance of the supernatant was measured using a UV–vis spectrophotometer at 620 and 650 nm for phycocyanin (PC) and allophycocyanin (APC), respectively …”

Section: Methodsmentioning

confidence: 99%

“…The absorbance of the supernatant was measured using a UV−vis spectrophotometer at 620 and 650 nm for phycocyanin (PC) and allophycocyanin (APC), respectively. 23 Intracellular NADH and NADPH concentrations were measured using NADH and NADPH Assay Kits with WST-8 (Beyotime Biome, Shanghai, China) according to the manufacturer's instructions. Pretreatments for the samples analyzed for NADH and NADPH were the same as the assay for phycobilisome.…”

Section: Cellular Growth Ofmentioning

confidence: 99%

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Toward Understanding the Ecophysiology of Freshwater and Marine Cyanobacteria under Different Salinities

Chen

Huang

et al. 2023

ACS EST Water

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The mechanism by which cyanobacteria have adapted from low to high salinity and ultimately diversified in marine environments remains poorly understood. To understand how freshwater and marine cyanobacteria respond to high salinity, we selected a marine cyanobacteria, Synechococcus sp. PCC7002, and freshwater cyanobacteria, Synechococcus elongatus PCC7942. Photosynthetic growth of PCC7002 and PCC7942 were enhanced and inhibited by the high salinity, respectively, which were also supported by the quantification of reducing equivalents and ATP in these two species. Growth under the high salinity reduced the photosynthetic activities for PCC7942, resulting in more oxidative stress, which was opposite of the effect of high salinity on PCC7002. Transcriptomic analysis demonstrated that those key genes involved in synthetic and energy metabolisms as well as certain processes for salinity acclimation and salinity signal transduction were significantly downregulated in PCC7942 and upregulated in PCC7002 under the high salinity, suggesting a potential functional regulatory mechanism from perception, transduction, and acclimation of cyanobacteria to high salinities. The findings of this study improve the understanding of cyanobacterial ecophysiology under different salinities and shed light on the evolutionary acclimation of cyanobacteria from freshwater to marine environments.

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Section: Methodsmentioning

confidence: 99%

Section: Cellular Growth Ofmentioning

confidence: 99%

Toward Understanding the Ecophysiology of Freshwater and Marine Cyanobacteria under Different Salinities

Chen

Huang

et al. 2023

ACS EST Water

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“…C-PC was purified according to the published procedure [ 45 ]. The concentration of C-PC was calculated using the following equation [ 46 ]: C-PC ( mg / mL ) = (A 615 − 0.474 × A 652 )/5.34 where A 615 and A 652 are absorbances of isolated C-PC at 615 and 652 nm. The molar concentration of C-PC was calculated by dividing the protein mass concentration by the molar mass of trimeric C-PC (~110 kDa).…”

Section: Methodsmentioning

confidence: 99%

Investigation of the Potential of Selected Food-Derived Antioxidants to Bind and Stabilise the Bioactive Blue Protein C-Phycocyanin from Cyanobacteria Spirulina

Gligorijević,

Jovanović,

Cvijetić

et al. 2023

IJMS

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Blue C-phycocyanin (C-PC), the major Spirulina protein with innumerable health-promoting benefits, is an attractive colourant and food supplement. A crucial obstacle to its more extensive use is its relatively low stability. This study aimed to screen various food-derived ligands for their ability to bind and stabilise C-PC, utilising spectroscopic techniques and molecular docking. Among twelve examined ligands, the protein fluorescence quenching revealed that only quercetin, coenzyme Q10 and resveratrol had a moderate affinity to C-PC (Ka of 2.2 to 3.7 × 105 M–1). Docking revealed these three ligands bind more strongly to the C-PC hexamer than the trimer, with the binding sites located at the interface of two (αβ)3 trimers. UV/VIS absorption spectroscopy demonstrated the changes in the C-PC absorption spectra in a complex with quercetin and resveratrol compared to the spectra of free protein and ligands. Selected ligands did not affect the secondary structure content, but they induced changes in the tertiary protein structure in the CD study. A fluorescence-based thermal stability assay demonstrated quercetin and coenzyme Q10 increased the C-PC melting point by nearly 5 °C. Our study identified food-derived ligands that interact with C-PC and improve its thermal stability, indicating their potential as stabilising agents for C-PC in the food industry.

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Section: Methodsmentioning

confidence: 99%

“…The content of light-harvesting complexes was determined by measuring the absorbances of the supernatants at 565, 620, and 650 nm, respectively, for phycoerythrin (PE), phycocyanin (PC), and phycocyanin (APC) with a UV−vis spectrophotometer (UV-2700, Shimadzu Co., Japan) according to the standard protocols for phycobiliprotein analysis. 18 NADPH, ATP, and ATPase were determined by a colorimetry method. 19−21 Specifically, a pellet of Microcystis aeruginosa cells was collected by centrifugation at 10500g and 4 °C for 5 min (the same for all the centrifugation processes that were conducted in this study).…”

Section: ■ Experimental Sectionmentioning

confidence: 99%

Acetylacetone Interferes with Carbon and Nitrogen Metabolism of Microcystis aeruginosa by Cutting Off the Electron Flow to Ferredoxin

Yilimulati

Zhou

Shevela

et al. 2022

Environ. Sci. Technol.

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The regulation of photosynthetic machinery with a nonoxidative approach is a powerful but challenging strategy for the selective inhibition of bloom-forming cyanobacteria. Acetylacetone (AA) was recently found to be a target-selective cyanocide for Microcystis aeruginosa, but the cause and effect in the studied system are still unclear. By recording of the chemical fingerprints of the cells at two treatment intervals (12 and 72 h with 0.1 mM AA) with omics assays, the molecular mechanism of AA in inactivating Microcystis aeruginosa was elucidated. The results clearly reveal the effect of AA on ferredoxin and the consequent effects on the physiological and biochemical processes of Microcystis aeruginosa. In addition to its role as an electron acceptor of photosystem I, ferredoxin plays pivotal roles in the assimilation of nitrogen in cyanobacterial cells. The effect of AA on ferredoxin and on nonheme iron of photosystem II first cut off the photosynthetic electron transfer flow and then interrupted the synthesis of adenosine triphosphate (ATP) and reduced nicotinamide adenine dinucleotide phosphate (NADPH), which ultimately might affect carbon fixation and nitrogen assimilation metabolisms. The results here provide missing pieces in the current knowledge on the selective inhibition of cyanobacteria, which should shed light on the better control of harmful blooms.

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Analytical Protocols in Phycobiliproteins Analysis

Cited by 4 publications

References 109 publications

Toward Understanding the Ecophysiology of Freshwater and Marine Cyanobacteria under Different Salinities

Toward Understanding the Ecophysiology of Freshwater and Marine Cyanobacteria under Different Salinities

Investigation of the Potential of Selected Food-Derived Antioxidants to Bind and Stabilise the Bioactive Blue Protein C-Phycocyanin from Cyanobacteria Spirulina

Acetylacetone Interferes with Carbon and Nitrogen Metabolism of Microcystis aeruginosa by Cutting Off the Electron Flow to Ferredoxin

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