Application of experimental design as a statistical approach to recover bioactive peptides from different food sources.

Bioactive peptides (BAPs) derived from samples of animals and plants have been widely recommended and consumed for their beneficial properties to human health and to control several diseases. This work presents the applications of experimental designs (DoE) used to perform factor screening and/or optimization focused on finding the ideal hydrolysis condition to obtain BAPs with specific biological activities. The collection and discussion of articles revealed that Box Behnken Desing and Central Composite Design were the most used. The main parameters evaluated were pH, time, temperature and enzyme/substrate ratio. Among vegetable protein sources, soy was the most used in the generation of BAPs, and among animal proteins, milk and shrimp stood out as the most explored sources. The degree of hydrolysis and antioxidant activity were the most investigated responses in obtaining BAPs. This review brings new information that helps researchers apply these DoE to obtain high-quality BAPs with the desired biological activities.

Structural characterization of the human DjC20/HscB cochaperone in solution.

J-domain proteins (JDPs) form a very large molecular chaperone family involved in proteostasis processes, such as protein folding, trafficking through membranes and degradation/disaggregation. JDPs are Hsp70 co-chaperones capable of stimulating ATPase activity as well as selecting and presenting client proteins to Hsp70. In mitochondria, human DjC20/HscB (a type III JDP that possesses only the conserved J-domain in some region of the protein) is involved in [FeS] protein biogenesis and assists human mitochondrial Hsp70 (HSPA9). Human DjC20 possesses a zinc-finger domain in its N-terminus, which closely contacts the J-domain and appears to be essential for its function. Here, we investigated the hDjC20 structure in solution as well as the importance of Zn+2 for its stability. The recombinant hDjC20 was pure, folded and capable of stimulating HSPA9 ATPase activity. It behaved as a slightly elongated monomer, as attested by small-angle X-ray scattering and SEC-MALS. The presence of Zn2+ in the hDjC20 samples was verified, a stoichiometry of 1:1 was observed, and its removal by high concentrations of EDTA and DTPA was unfeasible. However, thermal and chemical denaturation in the presence of EDTA led to a reduction in protein stability, suggesting a synergistic action between the chelating agent and denaturators that facilitate protein unfolding depending on metal removal. These data suggest that the affinity of Zn+2 for the protein is very high, evidencing its importance for the hDjC20 structure.

Solubility of Cd, Cr, Cu, Ni, and Pb and Its Correlation with Total Polyphenols and Soluble Melanoidins in Hot Infusions of Green and Roasted Mate.

The total and soluble contents of three potentially toxic elements (PTEs) (Cd, Cr, and Pb) and two essential elements (Cu and Ni) in commercial green and roasted mate (Ilex paraguariensis St. Hil.) and their hot infusions were accessed by inductively coupled plasma mass spectrometry. Total polyphenolic (TP) contents in the infusions were accessed by Folin-Ciocalteu method, and the presence of soluble melanoidins (SM) was evaluated by measuring absorbances at 420 nm. The soluble element contents in hot infusions of green mate were present in the following increasing order: Pb < Cd < Cr < Cu, and Ni. For hot infusions of roasted mate, the increasing order was: Cu < Pb < Cd < Cr < Ni. Except for Cr, hot infusions of roasted mate supply lower contents of the soluble PTEs than hot infusions of green mate. Cadmium was the only of evaluated trace element found in hot infusions of green and roasted mate as well as in some of their mate samples above the maximum allowed contents established by Brazilian legislation. Despite this, polyphenol and melanoidin contents in green and roasted mate infusions may reduce Cd bioavailability. The soluble trace elements that correlated directly with total polyphenols in hot infusions of green or roasted mate are indirectly correlated with the presence of soluble melanoidins. Moreover, this is the first study to evaluate the correlations of soluble forms of Cd, Cr, Cu, Ni, and Pb with the presence of SM and TP contents in hot infusions of green and roasted mate.

Multi-elemental determination in meat samples using multi-isotope calibration strategy by ICP-MS.

The multi-isotope calibration (MICal) is a fast and accurate calibration strategy for multi-elemental determination that uses only two solutions, (1) 50% (v v-1) sample plus 50% (v v-1) analytical blank solution, and (2) 50% (v v-1) sample plus 50% (v v-1) standard solution (containing the analytes). The calibration curves were built as a function of the intensity signals of solution 1 (y-axis) versus solution 2 (x-axis), employing the various isotopes of each monitored mass/charge. The concentration of the analytes was calculated from the slope and the concentrations of the standards added to solution 2. The validation of the method was evaluated using standard reference materials of bovine liver and bovine muscle. The recoveries varied from 87 to 113% for MICal, which were similar to the conventional standard additions calibration procedure (83 to 120%).

A compact miniaturized continuous flow system for the determination of urea content in milk.

A multicommutation-based flow system with photometric detection was developed, employing an analytical microsystem constructed with low temperature co-fired ceramics (LTCC) technology, a solid-phase reactor containing particles of Canavalia ensiformis DC (urease source) immobilized with glutaraldehyde, and a mini-photometer coupled directly to the microsystem which monolithically integrates a continuous flow cell. The determination of urea in milk was based on the hydrolysis of urea in the solid-phase reactor and the ammonium ions produced were monitored using the Berthelot reaction. The analytical curve was linear in the urea concentration range from 1.0 x 10(-4) to 5.0 x 10(-3) mol L(-1) with a limit of detection of 8.0 x 10(-6) mol L(-1). The relative standard deviation (RSD) for a 2.0 x 10(-3) mol L(-1) urea solution was lower than 0.4% (n = 10) and the sample throughput was 13 h(-1). To check the reproducibility of the flow system, calibration curves were obtained with freshly prepared solutions on different days and the RSD obtained was 4.7% (n = 6). Accuracy was assessed by comparing the results of the proposed method with those from the official procedure and the data are in close agreement, at a 95% confidence level.