Few reports about the biosynthesis of alpha-arbutin using amylosucrase are available. In this work, an amylosucrase from Cellulomonas carboniz T26, CcAS, was identified for highly efficient synthesis of alpha-arbutin using 20 mM sucrose and 5 mM hydroquinone as a glucosyl donor and acceptor, respectively. The optimum conditions for transglucosylation are a pH of 7.0 and a temperature of 40 degrees C. Under these conditions, 40%-44.7% of molar production yield of alpha-arbutin can be obtained at low concentration of hydroquinone (5 mM) without addition of ascorbic acid after 2 h, which saves time and raw material compared with the reported amylosucrase DGAS. In spite of oxidation of hydroquinone at high concentration of 23.6 mM, 16.1% of molar production yield of alpha-arbutin is acquired using CcAS without addition of ascorbic acid, while only 1.3% is obtained using DGAS. This work presents that higher molar production yield of alpha-arbutin at low concentrations of hydroquinone can be obtained using amylosucrase, because hydroquinone at high concentrations is easily oxidized, which inhibits the synthesis of alpha-arbutin. Simultaneously, CcAS likely further glycosylates the synthesized alpha-arbutin after a 2-h reaction, which is not reported using DGAS. CcAS exhibits a potential in industrialized application.