For the production of ethanol from seaweed as the source material, thermal acid hydrolysis and enzymatic saccharification were carried out for monosugars production of 25.5 g/l galactose and 7.6 g/l glucose using Gelidium amansii. The fermentation was performed with Pichia stipitis KCTC 7228 or Saccharomyces cerevisiae KCCM 1129. When wild P. stipitis and S. cerevisiae were used, the ethanol productions of 11.2 g/l and 6.9 g/l were produced, respectively. The ethanol productions of 16.6 g/l and 14.6 g/l were produced using P. stipitis and S. cerevisiae acclimated to high concentration of galactose, respectively. The yields of ethanol fermentation increased to 0.5 and 0.44 from 0.34 and 0.21 using acclimated P. stipitis and S. cerevisiae, respectively. Therefore, acclimation of yeasts to a specific sugar such as galactose reduced the glucose-induced repression on the transport of galactose.