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Trametes lactinea polysaccharides have a high medicinal value; however, we still know little about the structure and bioactivity of intracellular and extracellular polysaccharides in the mycelial liquid fermentation of T. lactinea. This study analyzed the structures of intracellular (IP-1, IP-2, and IP-3) and extracellular (EP-1 and EP-2) polysaccharide components isolated from T. lactinea liquid fermentation, as well as investigated their antioxidant, antibacterial, and immunomodulatory properties. The results showed that IP-3 was the only component with a triple-helix structure, while the other four components did not possess this structure. IP3 has a higher molecular weight, flavonoid, and total phenolic content compared to other components. Both intracellular and extracellular polysaccharide components exhibited strong scavenging abilities against ABTS and DPPH radicals. The components showed limited antibacterial effects against four types of bacteria (Staphylococcus aureus, Bacillus subtilis, Erwinia carotovora, and Escherichia coli), and were found to be non-toxic to RAW264.7 cells, even promoting cell proliferation. Furthermore, within a specific concentration range, all components enhanced the phagocytic activity of RAW264.7 cells, increased the secretion of NO, TNF-α, and IL-6, and demonstrated concentration-dependent effects, with IP-3 displaying the most potent immunomodulatory activity. This study shows a high potential for the development and utilization of polysaccharides derived from the liquid fermentation of T. lactinea mycelium. Full article

Trametes lactinea is a macrofungus commonly found on broadleaf tree felling piles with rich bioactivity, and its main active substances are polysaccharides, which have a certain medicinal value. Few studies have been conducted on the simultaneous study of intracellular and extracellular polysaccharides of fermented mycelia in liquid culture. In this study, T. lactinea collected from Eucalyptus stumps was subjected to liquid fermentation culturing and the extraction of intracellular and extracellular polysaccharides from the mycelium was optimized using unidirectional and orthogonal test methods. The intracellular and extracellular crude polysaccharides were isolated and purified, and polysaccharide fractions were obtained and assayed for their chemical composition. The main findings of this study are as follows: (1) The optimal conditions for the extraction of intracellular polysaccharides from T. lactinea mycelium were a material–liquid ratio of 1:40, an extraction temperature of 100 °C, a time of 5 h, and four repeats, wherein the polysaccharide extraction rate reached 5.1%. The optimal extraction conditions for the extracellular polysaccharides were a concentration ratio of 5:1, a concentration temperature of 70 °C, an ethanol concentration of 100.0%, and an alcohol settling time of 12 h. The polysaccharides could thus be extracted up to 0.63 mg/mL. (2) The intracellular and extracellular crude polysaccharides of T. lactinea mycelium were successfully isolated and purified, resulting in three homogeneous fractions of intracellular polysaccharides (IP-1, IP-2, and IP-3) and two homogeneous fractions of extracellular polysaccharides (EP-1 and EP-2). (3) The polysaccharide chemical composition resulted in the highest total sugar content of EP-1. IP-3 had the highest uronic acid content. The results of the monosaccharide composition analysis showed that the mycelial intracellular polysaccharides IP-1, IP-2, and IP-3 were all highest in glucose. Both IP-2 and IP-3 contained guluronic acid and glucuronic acid. Mannose was the most abundant extracellular polysaccharide in both EP-1 and EP-2. This study provides theoretical and technical bases for the comprehensive development and utilization of T. lactinea polysaccharides. Full article