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Furuki, Takao; Maeda, Shuichi; Imajo, Satoshi; Hiroi, Tetsuya; Amaya, Tsutomu; Hirokawa, Takahiko; Itô, Kazuo; Nozawa, H

doi:10.1023/a:1025118516888

Cited by 80 publications

(15 citation statements)

References 19 publications

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“…For phycocyanin extraction of cyanobacteria living on feather moss, samples were oven-dried at 35°C for 4 h and milled with a blender. Then, 0.15-0.20 g of moss sample were placed in 50 ml sterile tubes with 15 ml of sterilized sodium phosphate buffer (0.025 M, pH 7; Sarada et al, 1999;Furuki et al, 2003;Horváth et al, 2013) prepared by mixing 2.99 g of Na 2 HPO 4 • 2H 2 O and 1.91 g of NaH 2 PO 4 • 2H 2 O in 1 L of Milli-Q water. Phycocyanin extraction procedure was then performed similarly for cyanobacteria cells and moss samples.…”

Section: Phycocyanin Extraction and Quantification Proceduresmentioning

confidence: 99%

Quantification of Moss-Associated Cyanobacteria Using Phycocyanin Pigment Extraction

2021

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In the boreal forest, cyanobacteria can establish associations with feather moss and realize the biological nitrogen fixation (BNF) reaction, consisting in the reduction of atmospheric dinitrogen into bioavailable ammonium. In this ecosystem, moss-associated cyanobacteria are the main contributors to BNF by contributing up to 50% of new N input. Current environmental changes driven by anthropogenic activities will likely affect cyanobacteria activity (i.e., BNF) and populations inhabiting mosses, leading to potential important consequences for the boreal forest. Several methods are available to efficiently measure BNF activity, but quantifying cyanobacteria biomass associated with moss is challenging because of the difficulty to separate bacteria colonies from the host plant. Attempts to separate cyanobacteria by shaking or sonicating in water were shown to be poorly efficient and repeatable. The techniques commonly used, microscopic counting and quantitative PCR (qPCR) are laborious and time-consuming. In aquatic and marine ecosystems, phycocyanin (PC), a photosynthesis pigment produced by cyanobacteria, is commonly used to monitor cyanobacteria biomass. In this study, we tested if PC extraction and quantification can be used to estimate cyanobacteria quantity inhabiting moss. We report that phycocyanin can be easily extracted from moss by freeze/thaw disturbance of cyanobacteria cells and can be quickly and efficiently measured by spectrofluorometry. We also report that phycocyanin extraction is efficient (high recovery), repeatable (relative SD < 13%) and that no significant matrix effects were observed. As for aquatic systems, the main limitation of cyanobacteria quantification using phycocyanin is the difference of cellular phycocyanin content between cyanobacteria strains, suggesting that quantification can be impacted by cyanobacteria community composition. Nonetheless, we conclude that phycocyanin extraction and quantification is an easy, rapid, and efficient tool to estimate moss-associated cyanobacteria number.

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Section: Phycocyanin Extraction and Quantification Proceduresmentioning

confidence: 99%

Quantification of Moss-Associated Cyanobacteria Using Phycocyanin Pigment Extraction

2021

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“…The differentiated sedimentation of molecules, according to their densities and under the influence of gravity (g-force), is the basic principle of particles separation by centrifugation. In laboratory scale, the g-force of ultracentrifugation can reach values of up to 200,000× g, being able, for example, to remove impurities (e.g., chlorophyll, cell debris) in an extract of C-phycocyanin [201] and to estimate molecular weight of enzymes and proteins from microalgae [202]. However, for this last purpose ultracentrifugation is already outdated.…”

Section: Ultracentrifugationmentioning

confidence: 99%

Microalgae Biomolecules: Extraction, Separation and Purification Methods

et al. 2020

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Several microalgae species have been exploited due to their great biotechnological potential for the production of a range of biomolecules that can be applied in a large variety of industrial sectors. However, the major challenge of biotechnological processes is to make them economically viable, through the production of commercially valuable compounds. Most of these compounds are accumulated inside the cells, requiring efficient technologies for their extraction, recovery and purification. Recent improvements approaching physicochemical treatments (e.g., supercritical fluid extraction, ultrasound-assisted extraction, pulsed electric fields, among others) and processes without solvents are seeking to establish sustainable and scalable technologies to obtain target products from microalgae with high efficiency and purity. This article reviews the currently available approaches reported in literature, highlighting some examples covering recent granted patents for the microalgae’s components extraction, recovery and purification, at small and large scales, in accordance with the worldwide trend of transition to bio-based products.

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“…Chlorophyll content was determined spectrophotometrically, after extraction with DMSO and dark incubation at 65˚C, using formulas and absorbance wavelengths for chl a & b [43] and for chl c [44]. Following the procedures of Chaffin [45] and equations of Furuki [46] and Sampath and Neefus [47], phycocyanin content was determined after sonication (to break cells) for 15 min at 4˚C in 0.1 M phosphate buffer (pH 6.8) (Bransonic #1510). Samples were then incubated for 1 h at 4˚C, and centrifuged for 10 min at 3800 g. Phycocyanin concentration of samples was determined by fluorescence (10-AU fluorometer with P/N 10 -305 filters, Turner Design).…”

Section: Pigment Contentmentioning

confidence: 99%

Interactive Effects of Temperature, Nitrogen, and Zooplankton on Growth and Protein and Carbohydrate Content of Cyanobacteria from Western Lake Erie

Bista¹,

Heckathorn²,

Bridgeman³

et al. 2014

JWARP

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Harmful algal blooms (HABs) in freshwater ecosystems, especially of cyanobacterial species, are becoming more frequent and expanding geographically, including in Lake Erie in North America. HABs are the result of complex and synergistic environmental factors, though N or P eutrophication is a leading cause. With global mean temperatures expected to increase an additional 2˚C -5˚C by 2100, cyanobacterial blooms are predicted to increase even more, given their typically-high temperature optimum for growth. We investigated how increases in temperature and nitrogen, singly or in combination, affect the growth, food quality, and herbivory of Lake Erie cyanobacteria. Algal community samples collected from Lake Erie, and isolated non-N-fixing (Microcystis aeruginosa) and N-fixing (Anabaena flos-aquae) cyanobacterial species, were cultured at 20˚C, 25˚C, or 30˚C, and at 5, 50, 150, or 250 μM N, and then analyzed for growth and (for isolates) content of total protein and non-structural carbohydrates (NSC). Temperature and N both affected algal growth, and there were temperature × N interactions, which were sometimes affected by presence/absence of zooplankton. For example, cyanobacteria (but not green algae) growth increased with both temperature and N, especially from 25˚C to 30˚C, but N and herbivore presence increased cyanobacterial growth primarily only at 30˚C. In general, temperature and N had little consistent effect on NSC, but increasing temperature and N tended to increase protein content in Microcystis and Anabaena (temperature effects mostly at higher N levels). In Anabaena, increases in N did not increase growth or protein at 20˚C or 25˚C, but did increase both at 30˚C, indicating that N fixation * Corresponding author. D. Bista et al.1140 is damaged at high temperatures and that high NO3 can overcome this damage. These results indicate that future global warming and continued eutrophication will increase cyanobacterial growth, as well influence algal herbivory and competition between N-fixing and non-N-fixing cyanobacteria.

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Cited by 80 publications

References 19 publications

Quantification of Moss-Associated Cyanobacteria Using Phycocyanin Pigment Extraction

Quantification of Moss-Associated Cyanobacteria Using Phycocyanin Pigment Extraction

Microalgae Biomolecules: Extraction, Separation and Purification Methods

Interactive Effects of Temperature, Nitrogen, and Zooplankton on Growth and Protein and Carbohydrate Content of Cyanobacteria from Western Lake Erie

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