Lentinula edodes is one of the most popular edible macrofungi, the first recognized medicinal macrofungus with many therapeutic potentials, like anti-inflammatory, antimicrobial, antifungal, and antiviral activities. Also, this mushroom was used to synthesize eco-friendly nanoparticles to treat microbial infections. The present chapter discusses the availability of different molecules in extracts from L. edodes, which could be used to prevent and treat many bacterial infections, including antibiotic-resistant strains in humans. In addition, it is also used as an antiviral agent, for the modulation of the immune system, and for the lowering of the sugar and triglyceride levels in obese and diabetic patients. Taken together, these findings suggest L. edodes’s usefulness as a natural source of bactericidal candidates for human drug applications. The goal of the current chapter is to discuss some medicinal uses against pathogenic microbes.