Biosurfactants based on carbohydrates are of special interest because of their production from renewable resources, nontoxicity, biocompatibility, and environmental friendliness. Dodecyl polyglucosides, a type of nonionic surfactant synthesized with methyl polyglucosides obtained from the eco-friendly subcritical methanolysis of renewable resource of starch, is no doubt good for ecosystem. With the subcritical methanolysis of starch, the methyl polyglucosides were obtained without any catalyst. Under the reaction condition of the weight ratio of methanol to starch of 7.5, temperature of 220 °C, and reaction time of 2 h, the yield of methyl polyglucosides was 85%. Dodecyl polyglucosides were synthesized by transglycosidation with methyl polyglucosides, and the green nonionic surfactant has excellent surface activity. The critical micelle concentration and hydrophilic lipophilic balance are 0.022 wt % and 12, respectively.