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The cell-associated dextransucrase produced by sucrose-grown cells of cariogenic Streptococcus mutans K1-R is derived from soluble dextransucrase. Synthesis of insoluble dextran by soluble dextransucrase gives rise to two dextransucrase fractions bound to the insoluble polysaccharide; a reversibly bound enzyme, which can be eluted in solutions of clinical dextran, and an irreversibly bound enzyme, which cannot be solubilized in this manner. Both of these fractions of dextransucrase are also present on sucrose-grown cells. During the synthesis of insoluble dextran by sucrose-grown cells, dextransucrase is progressively converted from soluble enzyme, first to the reversibly bound fraction and then to the irreversibly bound fraction, and is finally inactivated as insoluble dextran accumulates. The two cell-associated dextransucrase fractions therefore represent two stages in the insolubilization and inactivation of their precursor, soluble dextransucrase. As a result of this process of inactivation, the yield of dextransucrase from cells cultured on sucrose is markedly decreased by high concentrations of sucrose in the culture medium.