Type and Amount of Lipids Influence the Molecular and Textural Properties of a Soy Soft Pretzel

Altering baked goods by the addition of nutrient-rich ingredients, such as soy and ground almonds, affects the water and lipid distribution of the product and, subsequently, its final quality. Here, we studied how three lipid sources, shortening, canola oil, and ground almonds, affected texture and water distribution in a baked soy pretzel and the molecular mobility in the dough. Pretzel crumb from all formulations exhibited 40–43% moisture with a little more than half present as “freezable” water. Firmness and chewiness decreased with increased shortening and canola oil, whereas firmness and chewiness increased with additional almonds. In contrast, neither springiness nor cohesiveness was affected by the lipid quantity or source. Finally, magnetic resonance imaging of the soy pretzel dough revealed two or three populations of dough components that have distinct molecular mobilities. With increased lipid content, the mobility of each population increased in magnitude and heterogeneity. Interestingly, almonds had the smallest effect on the molecular mobility of the dough but had the largest effect on textural properties. These results provide quantitative insight into the mechanisms by which the lipid source can influence molecular properties that have textural implications for bakery products

Homotropic Cooperativity from the Activation Pathway of the Allosteric Ligand-Responsive Regulatory <i>trp</i> RNA-Binding Attenuation Protein

The <i>trp</i> RNA-binding attenuation protein (TRAP) assembles into an 11-fold symmetric ring that regulates transcription and translation of <i>trp</i>-mRNA in bacilli via heterotropic allosteric activation by the amino acid tryptophan (Trp). Whereas nuclear magnetic resonance studies have revealed that Trp-induced activation coincides with both microsecond to millisecond rigidification and local structural changes in TRAP, the pathway of binding of the 11 Trp ligands to the TRAP ring remains unclear. Moreover, because each of 11 bound Trp molecules is completely surrounded by protein, its release requires flexibility of Trp-bound (holo) TRAP. Here, we used stopped-flow fluorescence to study the kinetics of Trp binding by <i>Bacillus stearothermophilus</i> TRAP over a range of temperatures and observed well-separated kinetic steps. These data were analyzed using nonlinear least-squares fitting of several two- and three-step models. We found that a model with two binding steps best describes the data, although the structural equivalence of the binding sites in TRAP implies a fundamental change in the time-dependent structure of the TRAP rings upon Trp binding. Application of the two-binding step model reveals that Trp binding is much slower than the diffusion limit, suggesting a gating mechanism that depends on the dynamics of apo TRAP. These data also reveal that dissociation of Trp from the second binding mode is much slower than after the first Trp binding mode, revealing insight into the mechanism for positive homotropic allostery, or cooperativity. Temperature-dependent analyses reveal that both binding modes imbue increases in bondedness and order toward a more compressed active state. These results provide insight into mechanisms of cooperative TRAP activation and underscore the importance of protein dynamics for ligand binding, ligand release, protein activation, and allostery