Application of taste modifying protein, neoculin and miraculin for food preperation

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Neoculin and miraculin are taste modifying proteins which convert sourness to sweetness. They are occurring in tropical fruits, Curculigo latiforia and Synsepalum dulcificum, respectively. Each native protein is difficult to obtain constantly. We succeeded to produce recombinant neoculin and miraculin with Aspergillus oryzae that was designated to GRAS. To verify the safety of neoculin, viability of Caco-2 cells exposed to neoculin was investigated. The viability of cells was not changed with neoculin exposure. DNA microarray analysis also revealed that neoculin did not affect the gene expression of Caco-2 cells. Neoculin is received by human sweet taste receptor, T1R2-T1R3. The interaction sites of neoculin to T1R2-T1R3 were analyzed using HEK293T cells that express T1R2-T1R3. As a result, neoculin was received at the N-terminal extra cellar domain in T1R3.