Molluscan shell evolution with review of shell calcification hypothesis.

Biomineralization research on mollusc shells has mostly focused on nacre formation. Chitin, silk fibroin protein, and acidic macromolecules are important components for shell formation. Although the principle concept behind shell calcification was developed many years ago, the individual components have not been well scrutinized. Besides that, Mollusca are the second largest invertebrate phylum, but comprehensive biochemical research involving a comparison of different taxa is still rare. This study reconsiders the above three components with adding some biochemical data of aculiferans. The presence of chitin in polyplacophorans sclerites was confirmed by IR and pyrolysis GC/MS. DMMB staining data inferred that sulphated groups present in aplacophoran cuticle but not in polyplacophorans cuticle. These insight suggested importance of comparison between acuriferans and conchiferans.

Pyrolysis GC/MS and IR spectroscopy in chitin analysis of molluscan shells.

Chitin is an insoluble component in the shells of several molluscan species. It is thought to play important roles, in biomineralization and shell structure. To date, however, reports are scarce and sometimes contradictory, and suffer from methodological problems. Only in a single cephalopod species has the chitin been identified as beta-chitin. We present data on chitin occurrence in 22 species of shell-bearing Mollusca (Conchifera) and Polyplacophora, including the first evidence for scaphopods, based on pyrolysis gas chromatography, mass spectrometry (GC-MS), and infrared spectroscopy (IR). Pyrolysis GC-MS detected chitin in every tested member of the Conchifera. IR spectroscopy before and after chitinase treatment revealed at least three distinct patterns of peak changes. The contents of the insoluble shell organics included not only chitin and proteins, but also insoluble polysaccharides, e.g., glucan. We conclude that chitin was present in the last common ancestor of the Conchifera and that its abundance in the shell matrix depends on the differentiation of the shell.