In vitro oral processing of raw tomato: Novel insights into the role of endogenous fruit enzymes.

During consumption of fruits, the breakdown of the fruit tissue due to oral processing (chewing, mixing with saliva) may activate or increase the rate of endogenous enzyme activities via the disruption of the cell wall, cellular decompartmentalization, and particle size reduction allowing the enzymes to reach their substrates. The aim of this study was to investigate the activity of one such endogenous fruit enzyme (pectin methylesterase [PME] [E.C. 3.1.1.11]) during in vitro oral processing of raw tomatoes and associated changes in viscosity and microstructure. Oral processing of tomatoes purees was examined in the presence of artificial saliva (AS) at 37°C. in vitro oral processing was followed using immunofluorescence microscopy, apparent viscosity measurements, spectrophotometric, and titrimetric techniques. The results demonstrated that PME had slight but significant activity in the tomato fruit during in vitro oral processing generating methanol as a function of oral processing time, which was further evidenced using immunolabeling techniques to detect methylated pectin epitopes. A significant shear-thinning behavior of the tomato puree was observed due to dilution and/or endogenous fruit enzyme activity. These results suggest that activity of other fruit enzymes, such as polygalacturonase, which catalyzed the depolymerization of unmethylated pectin chains, might have resulted in a decrease in viscosity, which compensated for the increased potential for gel formation (if any) caused by PME. These interesting insights into the role of endogenous fruit enzymes might pave the way to the understanding of fruit viscosity modification occurring in the mouth and help in rational design of new fruit-based products.

Novel cell wall architecture of isoxaben-habituated Arabidopsis suspension-cultured cells: global transcript profiling and cellular analysis.

The herbicide isoxaben is a highly specific and potent inhibitor of cellulose synthesis in plants. Nevertheless, suspension-cultured cells can be habituated to grow in high concentrations of isoxaben, and apparently compensate for the disruption of cellulose synthesis by the modulation of other cell wall components. We have habituated Arabidopsis cells to isoxaben and characterized the cellular and genetic consequences. Near whole-genome transcript profiling implicated novel genes in cell wall assembly and extended our understanding of the activity of known cell wall-related genes including glycosyltransferases involved in cellulose and pectin biosynthesis. Habituation does not appear to be mediated by stress response processes, nor by functional redundancy within the cellulose synthase (AtCesA) family. Uniquely, amongst the cellulose synthase superfamily, AtCslD5 was highly upregulated and may play a role in the biosynthesis of the novel walls of habituated cells. In silico analysis of differentially expressed genes with unknown functions identified a putative glycosyltransferase and collagen-like putative cell wall protein.