Decaglycerol laurates have been widely used as emulsifiers in food, medicine, and cosmetic industries for many years. Currently, they are synthesized using alkaline catalysts under stringent conditions. Here, decaglycerol laurates were prepared through a green lipase-catalyzed process, employing the transesterification of methyl laurate with decaglycerol by the immobilized lipase (Novozym 435). Single-factor experiments and orthogonal test were used for reaction optimization. The optimum conditions were obtained as follows: reaction temperature of 65°C, a laurate/decaglycerol molar ratio of 2 : 1, an oscillating speed of 180 rpm, an enzyme dosage (based on amount of methyl laurate) of 8 wt.%, initial water content (based on the total substrate mass) of 5.0 wt.%, and reaction time of 4.5 h with 84.4% conversion of methyl laurate. The transesterification products were identified by electrospray ionization mass spectrometer. In addition, the surface activity of decaglycerol laurates was also characterized, and the surface tension of water was reduced to 33 mN·m−1 at a concentration magnitude of 10−5 g/mL.