C-phycocyanin extraction from two freshwater cyanobacteria by freeze thaw and pulsed electric field techniques to improve extraction efficiency and purity

Chittapun, Supenya; Jonjaroen, Veasarach; Khumrangsee, Katsaya; Charoenrat, Theppanya

doi:10.1016/j.algal.2020.101789

Cited by 84 publications

(40 citation statements)

References 39 publications

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“…However, phycocyanin extraction also has some limitations. Differences in cellular phycocyanin concentration between cyanobacteria strains have been reported in pure cultures (this study, Rippka et al, 1979;Santiago-Santos et al, 2004;Chittapun et al, 2020). This can be explained by variable phycocyanin production per cell, as well as differences in extraction efficiencies due to cell wall thickness and biofilm production, differing among cyanobacteria strains (Chittapun et al, 2020).…”

Section: Advantages and Limits Of The Methodsmentioning

confidence: 59%

“…Differences in cellular phycocyanin concentration between cyanobacteria strains have been reported in pure cultures (this study, Rippka et al, 1979;Santiago-Santos et al, 2004;Chittapun et al, 2020). This can be explained by variable phycocyanin production per cell, as well as differences in extraction efficiencies due to cell wall thickness and biofilm production, differing among cyanobacteria strains (Chittapun et al, 2020). The composition of cyanobacteria communities colonizing moss can vary with environmental conditions, moss species and time of sampling during the growth season (Ininbergs et al, 2011;Rousk et al, 2013;Warshan et al, 2016).…”

Section: Advantages and Limits Of The Methodsmentioning

confidence: 68%

“…Cyanobacteria cellular lyse efficiency (cellular apparent recovery/standard apparent recovery, C/B, Figure 2) achieved 86.7% for P. crista-castrensis and 73.6% for P. schreberi, which confirms that cell wall breaking was not optimal after the phycocyanin extraction procedure. Chittapun et al (2020) also showed that several species of cyanobacteria had specific cell wall structures which necessitated to using different disruption techniques for each species to obtain an optimal phycocyanin efficiency. Besides the cell wall disruption efficiency, the lower apparent recoveries of cellular phycocyanin could also be explained by the presence of biofilm in cyanobacteria culture, preventing accurate cell counting and pipetting.…”

Section: Phycocyanin Extraction Methods Characterization and Validationmentioning

confidence: 99%

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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: Advantages and Limits Of The Methodsmentioning

confidence: 59%

Section: Advantages and Limits Of The Methodsmentioning

confidence: 68%

Section: Phycocyanin Extraction Methods Characterization and Validationmentioning

confidence: 99%

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Quantification of Moss-Associated Cyanobacteria Using Phycocyanin Pigment Extraction

2021

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“…Several studies have demonstrated that electroporation by PEF improve the extraction of intracellular components such as proteins, lipids, and pigments from microorganisms has been reported ( Ganeva et al, 2003 ; Chittapun et al, 2020 ; Leonhardt et al, 2020 ). Very recently, it has been proposed that PEF not only permeabilized the cytoplasmatic membrane to enable the release of intracellular biomolecules but also trigger some enzymatic process that accelerates the autolysis and the subsequent release of intracellular components ( Martínez et al, 2018 , 2019 ; Silve et al, 2018 ; Scherer et al, 2019 ).…”

Section: Introductionmentioning

confidence: 99%

Enzymatic Processes Triggered by PEF for Astaxanthin Extraction From Xanthophyllomyces dendrorhous

Aguilar-Machado

Delso

Martínez

et al. 2020

Front. Bioeng. Biotechnol.

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The aim of this study was to evaluate the potential of pulsed electric fields (PEF) to improve the extraction of the lipid-soluble astaxanthin from fresh biomass of a wild-type (CECT 11028) and mutant (ATCC 74219) Xanthophyllomyces dendrorhous strain using ethanol as solvent. Inactivation and propidium uptake studies revealed that inactivation is a good index for estimated the proportion of irreversible permeabilized cells when inactivation is higher than 70% in the two strains. Ethanol was ineffective for extracting carotenoids from the PEF-treated cells (20 kV/cm, 135 µs) of the two strains. However, after aqueous incubation of PEF-treated X. dendrorhous ATCC 74219 cells for 12 h, up to 2.4 ± 0.05 mg/g dried weight (d.w.) of carotenoids were extracted in ethanol. From total carotenoid extracted, around 84% corresponded to all-trans astaxanthin. The detection and quantification of esterase activity in the supernatant and the relationship between the percentage of esterase activity quantified and the amount of carotenoids extracted indicate that the extraction of astaxanthin was mediated by enzymatic esterase activity triggered by PEF during incubation. On the other hand, the formation of a large lipid globule into the cytoplasm of PEF-treated X. dendrorhous CECT 11028 cells during aqueous incubation prevented carotenoid extraction. The process developed in this investigation represents a more sustainable and greener method that those previously used for extracting astaxanthin from yeast.

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“…Phycobiliproteins extraction conditions that used in previous studies. References [ 50 ] is cited in the supplementary materials.…”

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confidence: 99%

Optimization of the Freezing-Thawing Method for Extracting Phycobiliproteins from Arthrospira sp.

et al. 2020

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The freezing–thawing method had been reported to be the best phycobiliprotein extraction technique. However, optimum parameters of this extraction method for Arthrospira sp. (one of the major phycobiliprotein sources) still remained unclear. Hence, this study aimed to optimize the freezing–thawing parameters of phycobiliprotein extraction in Arthrospira sp. (UPMC-A0087). The optimization of the freezing–thawing method was conducted using different solvents, biomass/solvent ratios, temperatures, time intervals and freezing–thawing cycles. The extracted phycobiliproteins were quantified using a spectrophotometric assay. Double distilled water (pH 7) with a 0.50% w/v biomass/solvent ratio was the most efficient solvent in extracting high concentrations and purity of phycobiliproteins from Arthrospira sp. In addition, the combination of freezing at −80 °C (2 h) and thawing at 25 °C (24 h) appeared to be the optimum temperature and extraction time to obtain the highest amount of phycobiliproteins. A minimum of one cycle of freezing and thawing was sufficient for extracting high concentrations of phycobiliproteins. The findings from this study could reduce the cost and labor needed for extracting high quality phycobiliproteins. It also allowed the harvesting of large amounts of valuable phycobiliproteins.

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C-phycocyanin extraction from two freshwater cyanobacteria by freeze thaw and pulsed electric field techniques to improve extraction efficiency and purity

Cited by 84 publications

References 39 publications

Quantification of Moss-Associated Cyanobacteria Using Phycocyanin Pigment Extraction

Quantification of Moss-Associated Cyanobacteria Using Phycocyanin Pigment Extraction

Enzymatic Processes Triggered by PEF for Astaxanthin Extraction From Xanthophyllomyces dendrorhous

Optimization of the Freezing-Thawing Method for Extracting Phycobiliproteins from Arthrospira sp.

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