We exposed Akoya cultured pearls separately to heat (60–120 °C) and artificial light to investigate changes to fluorescence in the visible range and yellowing. We found that for both heat-treated and light-treated pearls, the fluorescence peak shifted from 480 to 430 nm with an increase in fluorescence intensity. This change in intensity was more prominent in heat-treated pearls, with the initial speed of increase rising with treatment temperature; treatment at 100 °C caused the greatest increase in fluorescence intensity. However, aminoguanidine suppressed the heat-induced change in the fluorescence of an ethylenediaminetetraacetic acid–soluble nacreous layer matrix. These results suggest that the heated-induced changes in the fluorescence of Akoya cultured pearls were caused largely by a buildup of fluorescent advanced glycation end products through the Maillard reaction. Although heat treatment led to a large increase in fluorescence intensity of the peak at approximately 430 nm in a deoxygenized environment, hardly any change in fluorescence intensity was observed after light treatment in this environment. Moreover, a new shoulder peak appeared at about 460 nm after light treatment. These results suggest that the Maillard reaction was not a major factor in the light-induced changes in the fluorescence of Akoya cultured pearls.