Pearl Science Laboratory

We found filamentous structures resembling conchocelis of red algae within the nacreous layer of the Akoya pearl oyster Pinctada fucata shell. Microscopic analyses revealed that these filamentous structures had an autofluorescence and varied in length, occasionally branched, and exhibited diameters ranging from 2 to 5μm-consistent with previously reported sizes for Bangiales conchocelis. We extracted DNA from organic matrix sheets obtained by decalcifying the shell and performed PCR with primers targeting red algae from the family Bangiales to identify these structures. Sequencing analysis confirmed a 94% match with that of Bangia fuscopurpurea, establishing the presence of Bangia conchocelis within the shell. Conchocelis distributed predominantly near the anterior ear across all examined shells. In 3-year-old oysters, conchocelis filament densities reached approximately 250 filaments per 1 mm2, with a broader distribution area than a 2-year-old oyster, where densities were lower, peaking at 35 filaments per 1 mm2. Conchocelis distribution was consistent between the left and right shells of the identical oyster. These findings suggest that conchocelis density and distribution expand with oyster growth, providing insights into the age-related dynamics of conchocelis localization in Akoya pearl oyster shells.

The gold and cream colors of cultured Akoya pearls, as well as natural yellow nacre of pearl oyster shells, are thought to arise from intrinsic yellow pigments. While the isolation of the yellow pigments has been attempted using a large amount of gold pearls, the substance concerned is still unknown. We report here on the purification and characterization of yellow pigments from the nacre of Akoya pearl oyster shells. Two yellow components, YC1 and YC2, were isolated from the HCl-methanol (HCl-MeOH) extract from nacreous organic matrices obtained by decalcification of the shells with ethylenediaminetetraacetic acid (EDTA). Energy-dispersive X-ray and infrared spectroscopy analyses suggested that YC1 and YC2 precipitated under basic conditions are composed of Fe-containing inorganic and polyamide-containing organic compounds, respectively. YC1 solubilized under acidic conditions exhibited positive reactions to KSCN and K4[Fe(CN)6] reagents, showing the same ultraviolet-visible absorption spectrum as those of Fe(III)-containing compounds. In addition, X-ray absorption fine structure analysis supported the compound in the form of Fe(III). The total amount of Fe was approximately 2.6 times higher in the yellow than white nacre, and most Fe was fractionated into the EDTA-decalcifying and HCl-MeOH extracts. These results suggest that Fe(III) coordinated to EDTA-soluble and insoluble matrix compounds are mainly associated with yellow color development not only in the Akoya pearl oyster shells but also in the cultured Akoya pearls.

The nacreous layer of shells and pearls is composed of aragonite crystals arranged in an organic matrix. The organic matrix contains chitin and several proteins that regulate the formation of the nacreous layer. Owing to their strong interactions in the organic matrix, the current method for extraction of insoluble proteins from the pre-powdered nacreous layer involves heating to high temperatures in the presence of a detergent (e.g., sodium dodecyl sulfate, SDS) and reductant (e.g., dithiothreitol, DTT), which is likely to induce protein degradation. Therefore, we have developed an electroextraction method to isolate proteins from the organic matrix of a nacreous organic sheet, that was obtained following the decalcification of shells in their original shape. Our electroextraction method employs milder conditions without heating or detergent. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) patterns of the electro-extracted proteins (EEPs) under non-reduced and reduced conditions revealed that this method yielded a greater number of different proteins compared with the conventional extraction method and the isolated EEPs retained their disulfide bonds. Our method is able to easily extract insoluble proteins from the nacreous layer under mild conditions and will undoubtedly aid future analyses into the functions of the nacreous layer proteins.

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.