In this study we tried to optimize the enzyme reaction conditions for the synthesis of highly branched isomaltodextrin (Mw > 2.5 kDa) using two dextransucrases from L. mesenteroides B-742CB and B-512FMCM that are dextransucrase constitutive mutants. As the concentration of sucrose or the ratio of maltose to sucrose increased, the amount of dextran decreased and the number and the amount of acceptor-products (of sucrose or maltose) increased. With high sucrose concentration (over 34%), there was more branched isomaltodextrin (as acceptor products) than dextran. When the ratio of sucrose to maltose was 2.5, there produced 86.7% of isomaltodextrin were produced. The Mw of dextrans, however, was over 2${ imes}$10(sup)6 and there was no significant amounts of branched clinical dextran or high molecular weight oligosaccharides. With the combined activities of B-742CB dextransucrase and B-512FMCM dextransucrase we could synthesize high molecular weight branched isomaltodextrin (Mw>2.5 kDa). The high molecular weight dextran was composed of high branches as B-742CB dextran.