A novel enzymatic synthesis method of cyclodextrin (CD) from low-mol-wt maltose using cyclomaltodextrin glucanotransferase (CGTase) from Bacillus macerans has been developed in various water-organic solvent systems. A beta-CD was synthesized in a two-phase system consisting of water and cyclohexane. However, no CDs could be synthesized in an aqueous buffer solution. A maximal yield of beta-CD has been obtained at a cyclohexane content volume of 44%. This synthesis has been obtained only at low temperatures, i.e., 7 degrees C, and did not take place at 50 degrees C. In addition, various organic solvents have been used for the enzymatic synthesis of CD from maltose. Consequently, beta-CD could be synthesized in various water-organic solvent systems, e.g., cyclohexane, benzene, xylene, and chloroform, but no enzymatic reaction occurred using aliphatic n-hydrocarbon solvents such as hexane, dodecane, and hexadecane. Furthermore, alpha- and beta-CD could be synthesized in water mixture solutions using organic solvents having an alcoholic group (e.g., ethanol, propanol, butanol, and pentanol) in a wide range of the reaction temperatures, typically 7-50 degrees C. In this temperature range, alpha- and beta-CD were also formed and the maximal yield from maltose to beta-CD of approx 13% was reached in 60 h.