Reducing bulk materials to layers or dots results in profound alterations in their physiochemical and optoelectronic properties, leading to a wide array of applications, spanning from device manufacturing to biomedicine. In this regard, the preparation of sulfur nanomaterials has garnered significant attention due to their low toxicity. Traditional methods for sulfur nanomaterial synthesis often involve harsh reaction conditions, leaving a gap for convenient approaches to create nanomaterials, such as nanosheets (NSs) and nanodots (NDs). Herein, the mechanical exfoliation of bulk sulfur using a surfactant thiol ligand with probe sonication is reported, making a unique contribution to existing methods. In the reported method, the thiol group binds to sulfur surfaces, facilitating exfoliation and stabilization, while the hydrophilic ends provide functional groups for exfoliated nanomaterials. Exfoliation can yield either nanosheets or nanodots, depending on the thiol ligand and exfoliation time. This approach offers the opportunity to exfoliate bulk sulfur using bioactive thiol ligands. With this goal in mind, bulk sulfur was exfoliated with 4-mercaptophenylboronic acid (BA) to target Gram-positive bacteria. This innovative exfoliation strategy of bulk sulfur using thiol ligands holds immense promise for synthesizing functionalized sulfur nanomaterials with wide-ranging applications, particularly in biomedicine.