This study was carried out to investigate the effects of agitation, aeration and scale-up on the mycelial growth, exo-polysaccharide (EPS) production, and mycelial morphology in the liquid culture of Ganoderma lucidum. A correlation between roughness and operating variables was also studied to scale-up the liquid culture of G. lucidum in a jar fermenter. When the agitation speed or aeration rate increased, the morphological form was changed from rough pellet to smooth pellet form. Increase of the agitation and aeration reduced the mycelial roughness. On the other hand, in the case of pellet size, it was not affected by aeration. The higher EPS production was obtained at approximately 17% of roughness and mycelial pellet size of 3~5 mm. The morphology at each fermenter was closely correlated with kLa value, and it was found that similarity of morphology would be used as a criteria of scale-up for liquid culture of G. lucidum.