Ganoderma has served as a valuable food supplement and medicinal ingredient with outstanding active compounds that are essential for human protection against chronic diseases. Modern pharmacology studies have proven that Ganoderma β-d-glucan exhibits versatile biological activities, such as immunomodulatory, antitumor, antioxidant, and antiviral properties, as well as gut microbiota regulation. As a promising polysaccharide, β-d-glucan is widely used in the prevention and treatment of various diseases. In recent years, the extraction, purification, structural characterization, and pharmacological activities of polysaccharides from the fruiting bodies, mycelia, spores, and fermentation broth of Ganoderma species have received wide attention from scholars globally. Unfortunately, comprehensive studies on the preparation, structure and bioactivity, toxicology, and utilization of β-d-glucans from Ganoderma species still need to be further explored, which may result in limitations in future sustainable industrial applications of β-d-glucans. Thus, this review summarizes the research progress in recent years on the physicochemical properties, structural characteristics, and bioactivity mechanisms of Ganoderma β-d-glucan, as well as its toxicological assessment and applications. This review is intended to provide a theoretical basis and reference for the development and application of β-d-glucan in the fields of pharmaceuticals, functional foods, and cosmetics.