The production yield of Coenzyme Q(10) (CoQ(10),) from the sucrose consumed by Agrobacterium tumefaciens KCCM 10413 decreased, and high levels of exopolysaccharide (EPS) accumulated after switching from batch culture to fed-batch culture. Therefore, we examined the effect of sucrose concentration on the fermentation profile by A. tumefaciens. In the continuous fed-batch culture with the sucrose concentration maintained constantly at 10, 20, 3 0, and 40 g l(-1), the dry cell weight (DCW), specific CoQ(10) content, CoQ(10) production, and the production yield of CoQ(10) from the sucrose consumed increased, whereas EPS production decreased as maintained sucrose concentration decreased. The pH-stat fed-batch culture system was adapted for CoQ(10) production to minimize the concentration of the carbon source and osmotic stress from sucrose. Using the pH-stat fed-batch culture system, the DCW, specific CoQ(10) content, CoQ(10) production, and the product yield of CoQ(10) from the sucrose consumed increased by 22.6, 13.7, 39.3, and 39.3%, respectively, whereas EPS production decreased by 30.7% compared to those of fed-batch culture in the previous report (Ha SJ, Kim SY, Seo JH, Oh DK, Lee JK, Appl Microbiol Biotechnol, 74:974980, 2007). The pH-stat fed-batch culture system was scaled up to a pilot scale (300 1), and the CoQ(10) production results obtained (626.5 mg l(-1) of CoQ(10) and 9.25 mg g DCW-1 of specific CoQ(10) content) were similar to those obtained at the laboratory scale. Thus, an efficient and highly competitive process for microbial CoQ(10) production is available.