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Structural characterization and SARS-CoV-2 inhibitory activity of a sulfated polysaccharide from Caulerpa lentillifera.

ABSTRACT: Caulerpa lentillifera (Bryopsidophyceae, Chlorophyta) is an edible seaweed attracting great attention for its expansion of farming scale and increasing consumption in these years. In the present study, a sulfated polysaccharide (CLSP-2) was isolated and separated from C. lentillifera, and its chemical structure was elucidated by a series of chemical and spectroscopic methods. Among these methods, mild acid hydrolysis and photocatalytic degradation were applied to release mono- and oligo-saccharide fragments which were further identified by HPLC-MSⁿ analysis, affording the information of the sugar sequences and the sulfate substitution in CLSP-2. Results indicated that the backbone of CLSP-2 was constructed of →6)-β-Manp-(1→ with sulfated branches at C2, which were comprised of prevalent →3)-β-Galp4S-(1→, →3)-β-Galp2,4S-(1→, and minor Xyl. In addition, the virus neutralization assay revealed that CLSP-2 could effectively protect HeLa cells against SARS-CoV-2 infection with an IC₅₀ of 48.48 μg/mL. Hence, the present study suggests CLSP-2 as a promising agent against SARS-CoV-2.

SUBMITTER: You Y

PROVIDER: S-EPMC8688187 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:The polysaccharides found in Caulerpa lentillifera (sea grape algae) are potentially an important bioactive resource. This study makes use of RSM (response surface methodology) to determine the optimal conditions for the extraction of valuable SGP (sea grape polysaccharides). The findings indicated that a water/raw material ratio of 10:1 mL/g, temperature of 90 °C, and extraction time of 45 min would maximize the yield, with experimentation achieving a yield of 21.576 %. After undergoing purification through DEAE-52 cellulose and Sephacryl S-100 column chromatography, three distinct fractions were obtained, namely SGP11, SGP21, and SGP31, each possessing average molecular weights of 38.24 kDa, 30.13 kDa, and 30.65 kDa, respectively. Following characterization, the fractions were shown to comprise glucose, galacturonic acid, xylose, and mannose, while the sulfate content was in the range of 12.2-21.8 %. Using Fourier transform infrared spectroscopy (FT-IR) it was possible to confirm with absolute certainty the sulfate polysaccharide attributes of SGP11, SGP21, and SGP31. NMR (nuclear magnetic resonance) findings made it clear that SGP11 exhibited α-glycosidic configurations, while the configurations of SGP21 and SGP31 were instead β-glycosidic. The in vitro antioxidant assays which were conducted revealed that each of the fractions was able to demonstrate detectable scavenging activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical cations. All fractions were also found to exhibit the capacity to scavenge NO radicals in a dose-dependent manner. SGP11, SGP21, and SGP31 were also able to display cellular antioxidant activity (CAA) against the human adenocarcinoma colon (Caco-2) cell line when oxidative damage was induced. The concentration levels were found to govern the extent of such activity. Moreover, purified SGP were found to exert strong inhibitory effects upon glycation, with the responses dependent upon dosage, thus confirming the potential for SGP to find a role as a natural resource for the production of polysaccharide-based antioxidant drugs, or products to promote improved health.

Project description:Background: The emergence of artemisinin resistance in South East Asia calls for urgent discovery of new drug compounds that have antiplasmodial activity. Unlike the classical compound screening drug discovery methods, the rational approach involving targeted drug discovery is less cumbersome and therefore key for innovation of new antiplasmodial compounds. Plasmodium falciparum (Pf) utilizes the process of host erythrocyte remodeling using Plasmodium-helical interspersed sub-telomeric domain (PHIST) containing proteins, which are amenable drug targets. The aim of this study is to identify inhibitors of PHIST from sulfated polysaccharides as new antimalarials. Methods: 251 samples from an ongoing study of epidemiology of malaria and drug resistance sensitivity patterns in Kenya were sequenced for PHISTb/RLP1 gene using Sanger sequencing. The sequenced reads were mapped to the reference Pf3D7 protein sequence of PHISTb/RLP1 using CLC Main Workbench. Homology modeling of both reference and mutant protein structures was achieved using the LOMETs tool. The models were refined using ModRefiner for energy minimization. Ramachandran plot was generated by ProCheck to assess the conformation of amino acids in the protein model. Protein binding sites predictions were assessed using FT SITE software. We searched for prospective antimalarials from PubChem. Docking experiments were achieved using AutoDock Vina and analysis results visualized in PyMOL. Results: Sanger sequencing generated 86 complete sequences. Upon mapping of the sequences to the reference, 12 non-synonymous single nucleotide polymorphisms were considered for mutant protein structure analysis. Eleven drug compounds with antiplasmodial activity were identified. Both modeled PHISTb/RLP1 reference and mutant structures had a Ramachandran score of >90% of the amino acids in the favored region. Ten of the drug compounds interacted with amino acid residues in PHISTb and RESA domains, showing potential activity against these proteins. Conclusion: This research identifies inhibitors of exported proteins that can be used in in vitro tests against the Plasmodium parasite.

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Structural characterization and SARS-CoV-2 inhibitory activity of a sulfated polysaccharide from Caulerpa lentillifera.

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