In this study, two new glycosylation derivatives of β-mangostin synthesized by enzymatic reactions were reported, together with the cytotoxic activity in various cancer cell lines including epidermal carcinoma (KB), breast cancer cell line (MCF-7), hepatocellular carcinoma (HepG2), pulmonary carcinoma (A549), and anti-microbial as well as acetylcholinesterase (AChE) and α-glucosidase inhibitory activities of β-mangostin and its novel glycosyl derivatives. Here, ACHE is linked to hepatocellular carcinoma.