Fucoidan from Ascophyllum nodosum and Undaria pinnatifida attenuate SARS-CoV-2 infection in vitro and in vivo by suppressing ACE2 and alleviating inflammation.

The global healthcare challenge posed by COVID-19 necessitates the continuous exploration for novel antiviral agents. Fucoidans have demonstrated antiviral activity. However, the underlying structure-activity mechanism responsible for the inhibitory activity of fucoidans from Ascophyllum nodosum (FUCA) and Undaria pinnatifida (FUCU) against SARS-CoV-2 remains unclear. FUCA was characterized as a homopolymer with a backbone structure of repeating (1 â†’ 3) and (1 â†’ 4) linked α-l-fucopyranose residues, whereas FUCU was a heteropolysaccharide composed of Fuc1-3Gal1-6 repeats. Furthermore, FUCA demonstrated significantly higher anti-SARS-CoV-2 activity than FUCU (EC50: 48.66 vs 69.52 µg/mL), suggesting the degree of branching rather than sulfate content affected the antiviral activity. Additionally, FUCA exhibited a dose-dependent inhibitory effect on ACE2, surpassing the inhibitory activity of FUCU. In vitro, both FUCA and FUCU treatments downregulated the expression of pro-inflammatory cytokines (IL-6, IFN-α, IFN-γ, and TNF-α) and anti-inflammatory cytokines (IL-10 and IFN-ß) induced by viral infection. In hamsters, FUCA demonstrated greater effectiveness in attenuating lung and gastrointestinal injury and reducing ACE2 expression, compared to FUCU. Analysis of the 16S rRNA gene sequencing revealed that only FUCU partially alleviated the gut microbiota dysbiosis caused by SARS-CoV-2. Consequently, our study provides a scientific basis for considering fucoidans as poteintial prophylactic food components against SARS-CoV-2.

An Extended Approach to Quantify Triacylglycerol in Microalgae by Characteristic Fatty Acids.

Microalgae represent a third generation biofuel feedstock due to their high triacylglycerol (TAG) content under adverse environmental conditions. Microalgal TAG resides in a single cell and serves as a lipid class mixed with complicated compositions. We previously showed that TAG possessed characteristic fatty acids (CFAs) for quantification and was linearly correlated with the relative abundance of CFA within certain limits in microalgae. Here, we defined the application range of the linear correlation between TAG and CFA in the oleaginous microalgae Chlamydomonas reinhardtii and Phaeodactylum tricornutum. In addition, TAG quantification was further expanded to a wide range of levels and the absolute amounts of saturated or monounsaturated CFAs, 16:0 and 18:1n9 of C. reinhardtii and 16:0 and 16:1n7 of P. tricornutum, instead of polyunsaturated CFAs, were verified to be linearly correlated to TAG levels throughout the entire period of nitrogen stress. This approach utilizes a single fatty acid to quantify TAG mixtures, and is rapid, simple and precise, which provides a useful tool for monitoring TAG accumulation of distinct microalgal species and facilitating high-throughput mutant screening for microalgae.

Identification of Characteristic Fatty Acids to Quantify Triacylglycerols in Microalgae.

The fatty acid profiles of lipids from microalgae are unique. Polyunsaturated fatty acids are generally enriched in polar lipids, whereas saturated and monounsaturated fatty acids constitute the majority of fatty acids in triacylglycerols (TAG). Each species has characteristic fatty acids, and their content is positively or negatively correlated with TAGs. The marine oleaginous diatom Phaeodactylum tricornutum was used as the paradigm to determine the quantitative relationship between TAG and characteristic fatty acid content. Fatty acid profiles and TAG content of Phaeodactylum tricornutum were determined in a time course. C16:0/C16:1 and eicosapentaenoic acid (EPA, C20:5n3) were identified as characteristic fatty acids in TAGs and polar lipids, respectively. The percentage of those characteristic fatty acids in total fatty acids had a significant linear relationship with TAG content, and thus, the correlation coefficient presenting r (2) were 0.96, 0.94, and 0.97, respectively. The fatty acid-based method for TAG quantification could also be applied to other microalgae such as Nannochloropsis oceanica in which the r (2) of C16:0 and EPA were 0.94 and 0.97, respectively, and in Chlorella pyrenoidosa r (2)-values for C18:1 and C18:3 with TAG content were 0.91 and 0.99, repectively. This characteristic fatty acid-based method provided a distinct way to quantify TAGs in microalgae, by which TAGs could be measured precisely by immediate transesterification from wet biomass rather than using conventional methods. This procedure simplified the operation and required smaller samples than conventional methods.