This study aimed to determine the potential of sugar fatty acid esters (SFAE) derived from crude palm oil (CPO) and palm oil hydrolysates as cytotoxic and anti-acne agents. The influence of structural variations among different saccharides on the SFAE properties was also investigated. Three types of saccharides were tested in this study; D-mannitol (a sugar alcohol), D-mannose (an aldose) and D-fructose (a ketose). SFAEs were prepared enzymatically by reacting fatty acids and saccharides using the low-cost liquid lipase Eversa^® Transform 2.0 at 5% (w/w) concentration. Thin layer chromatography (TLC) analysis, fourier transform infrared spectroscopy (FTIR) and nuclear magnetic resonance (NMR) spectra, emulsification properties, conversion rate and degree of esterification indicated the formation of SFAE. Some SFAE exhibited strong cytotoxic and antibacterial activities. SFAE of palmitic acid-mannitol and SFAE of stearic acid-fructose showed the highest cytotoxic activity against MCF-7 cells, with an IC50 value of 29 ppm. The SFAE of palm oil hydrolysate exhibited stronger cytotoxic activity than the SFAE of CPO hydrolysate. Additionally, the SFAE from CPO hydrolysate and D-mannitol exhibited the strongest antibacterial activity, achieving 100% inhibition against Cutibacterium acnes at 0.9 mg/mL. These findings highlight the potential of SFAE as a promising raw materials for the development of health and cosmetic products.