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Solid Matrix-Supported Supercritical CO₂ Enhances Extraction of γ-Linolenic Acid from the Cyanobacterium Arthrospira (Spirulina) platensis and Bioactivity Evaluation of the Molecule in Zebrafish.

ABSTRACT: Marine cyanobacteria represent a large untapped source of functional glycolipids enriched with polyunsaturated fatty acids (PUFAs) for human health. However, advanced methods for scalable isolation of diverse species containing high-purity PUFA-rich glycolipids will have to be developed and their possible pharmaceutical and nutraceutical functions identified. This paper introduces a novel solid matrix-supported supercritical CO₂ extraction method for scalable isolation of the PUFA γ-linolenic acid (GLA)-enriched glycolipids from the cyanobacterium Arthrospira (Spirulina) platensis, which has been the most widely used among microalgae in the nutraceutical and pharmaceutical industries. Of various porous materials studied, diatomite was the best to facilitate extraction of GLA-rich glycolipids, resulting in an extraction efficiency of 98%. Gamma-linolenic acid made up 35% of total fatty acids (TFAs) in the extracts, which was considerably greater than that obtained with ethanol (26%), Bligh and Dyer (24%), and in situ transesterification (24%) methods, respectively. Lipidomics analysis revealed that GLA was exclusively associated with galactolipids. Pharmaceutical functions of GLA-rich galactolipids were investigated on a zebrafish caudal fin regeneration model. The results suggested that GLA extracted from A. platensis possessed anti-oxidative, anti-inflammatory, and anti-allergic activities, which acted in a concerted manner to promote post-injury regeneration of zebrafish.

SUBMITTER: Yang X

PROVIDER: S-EPMC6520994 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Background:The extensive usage of zinc oxide nanoparticles (ZnO NPs) in industrial and consumer products raises the risk of releasing their residues into the aquatic environment. The presence of ZnO NPs in the aquatic environment could potentially cause cytotoxic effects on aquatic organisms. Thus, investigating the cytotoxic effects of ZnO NPs on microalgae, which form the base for the food web of aquatic biota, is essential to gain information regarding the ecotoxicological effects of metallic oxide nanoparticles in the aquatic ecosystem. Therefore, the present study has investigated in detail the assorted cytotoxic effects of ZnO NPs on S. platensis using various concentrations of ZnO NPs (10-200 mg/L) from 6 to 96 h to explore the dose- and time-dependent cytotoxic effects. Methods:The cytotoxic effects were all assessed through quantification of loss in cell viability, reduction in biomass and decrease in photosynthetic pigments such as chlorophyll-a, carotenoids and phycocyanin. The surface interactions of nanoparticles and the subsequent morphological alterations on algal cells were examined by optical and scanning electron microscopy (SEM). The intracellular alterations of algal cells were studied using transmission electron microscopy. Furthermore, Fourier transformed infrared (FTIR) spectrum was obtained to investigate the involvement of algal surface biomolecules in surface binding of ZnO NPs on algal cells. Results:The treatment of ZnO NPs on S. platensis exhibited a typical concentration- and time-dependent cytotoxicity. Results showed a significant (p < 0.05) cytotoxicity from 24 h onwards for all tested concentrations of ZnO NPs. The maximum cytotoxicity on algal cells was achieved at 96 h of exposure to ZnO NPs. In comparison with control, the algal cells that interacted with 200 mg/L of ZnO NPs for 96 h showed 87.3 ± 1% loss in cell viability, 76.1 ± 1.7% reduction in algal biomass, 92.5 ± 2.2%, 76.2 ± 2.2% and 74.1 ± 3.4% decrease in chlorophyll-a, carotenoids and phycocyanin contents respectively. Our study confirmed the cytotoxicity of ZnO NPs through the algal growth inhibition with 72 h EC10 and EC50 values of 1.29 and 31.56 mg/L, respectively. The microscopic examinations of the algal cells that interacted with ZnO NPs showed severe cell membrane and intracellular damage. The SEM EDX spectrum of ZnO NPs treated algal biomass evidenced the surface accumulation of zinc in the biomass. Finally, the FTIR spectrum confirmed the involvement of amino, hydroxyl and carboxylic groups of algal cell wall in the surface interaction of ZnO NPs on the algal cells. Discussion:The results showed that the treatment of ZnO NPs on S. platensis triggered substantial cytotoxicity and caused cell death. Hence, S. platensis could be potentially used as a bioindicator for testing toxicity of ZnO NPs in aquatic environment.

Project description:Arthrospira (Spirulina) platensis (ASP) is a representative filamentous, non-N2-fixing cyanobacterium that has great potential to enhance the food supply and possesses several valuable physiological features. ASP tolerates high and low temperatures along with highly alkaline and salty environments, and can strongly resist oxidation and irradiation. Based on genomic sequencing of ASP, we compared the protein expression profiles of this organism under different temperature conditions (15°C, 35°Cand 45°C) using 2-DE and peptide mass fingerprinting techniques. A total of 122 proteins having a significant differential expression response to temperature were retrieved. Of the positively expressed proteins, the homologies of 116 ASP proteins were found in Arthrospira (81 proteins in Arthrospira platensis str. Paraca and 35 in Arthrospira maxima CS-328). The other 6 proteins have high homology with other microorganisms. We classified the 122 differentially expressed positive proteins into 14 functions using the COG database, and characterized their respective KEGG metabolism pathways. The results demonstrated that these differentially expressed proteins are mainly involved in post-translational modification (protein turnover, chaperones), energy metabolism (photosynthesis, respiratory electron transport), translation (ribosomal structure and biogenesis) and carbohydrate transport and metabolism. Others proteins were related to amino acid transport and metabolism, cell envelope biogenesis, coenzyme metabolism and signal transduction mechanisms. Results implied that these proteins can perform predictable roles in rendering ASP resistance against low and high temperatures. Subsequently, we determined the transcription level of 38 genes in vivo in response to temperature and identified them by qRT-PCR. We found that the 26 differentially expressed proteins, representing 68.4% of the total target genes, maintained consistency between transcription and translation levels. The remaining 12 genes showed inconsistent protein expression with transcription level and accounted for 31.6% of the total target genes.

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Solid Matrix-Supported Supercritical CO₂ Enhances Extraction of γ-Linolenic Acid from the Cyanobacterium Arthrospira (Spirulina) platensis and Bioactivity Evaluation of the Molecule in Zebrafish.

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