Hot alkali-extracted antioxidative crude polysaccharide from a novel mushroom enhances immune response via TLR-mediated NF-κB activation: A strategy for full utilization of a neglected tribal food.

Mushroom polysaccharides have been revered for decades as incredibly versatile compounds suitable for application in biomedical, pharmaceutical, food, and cosmetic industries. In that essence Russula alatoreticula, a traditionally appraised unique macrofungus, has earlier been explored to contain functional macromolecules elicited by hydrothermal and cold alkali systems. Remnants of that multistep extraction could further be utilized not only to isolate high-value components, but also to reduce waste material. With this in mind, we decided to recycle leftover residue of the tribal delicacy, rather than undervalue, by following heated NaOH reflux. Intriguingly, the effort resulted productive isolation of carbohydrate fraction (RualaHap) enriched in ß-glucan that showed excellent antioxidant activity in terms of radical scavenging, chelating ability, and reducing power with EC50 of 207-5000 µg/ml. Besides strong immune boosting prowess was also witnessed as exposure of RAW 264.7 cells to the metabolites influenced macrophage viability and phagocytosis in a time-dependent manner. Induction of intracellular NO and ROS generation was also enumerated along with aggrandized production of pseudopods representing hallmark of monocyte stimulation. Moreover, the treatment drastically triggered level of TLR-2, TLR-4, NF-κB, Iκ-Bα, iNOS, COX-2, TNF-α, IFN-γ, and IL-10 maintaining balance between M1 and M2 cytokines via TLR coordinated NF-κB pathway. Conclusively the research offers a roadmap for better utilization of a species boasted in traditional practice aiding future implementation of RualaHap as ingredients of nutraceuticals or functional foods. PRACTICAL APPLICATIONS: At present, macrofungal polysaccharides are regarded as an ideal resource for a range of safe uses encompassing direct contact or ingestion by human. In this context, RualaHap isolated from the wild edible mushroom, R. alatoreticula, could have a broad application prospect, particularly in development of natural therapeutics to ameliorate radical-induced ailment and immune-deficiency condition. The biopolymers could thus be utilized in functional diet or complementary medicine to improve health that in turn might raise local cost of the abandoned taxon boosting rural empowerment.