Effects of heat treatment parameters on liquid whole egg proteins.

The aim of this study was to analyse the effect of heat treatment parameters on liquid whole egg (LWE) proteins by using ultraviolet-visible (UV-VIS) spectroscopy and capillary electrophoresis (CE). Heat treatment (at 60-68°C for 1-5min) was applied to LWE. Treated LWE was centrifuged and supernatant was taken for measurement of UV-VIS spectroscopy and CE. The change in UV absorbance showed loss of protein solubility depending on heat treatments parameters. Electropherograms of samples demonstrated the effect of treatment parameters on composition of LWE proteins. It was found that conalbumin and lysozyme were influenced by the treatment, while ovalbumin and ovomucoid were not affected. CE combined with principal component analysis (PCA) was used for classification of samples untreated or treated and treated at different treatment parameters. The results of the study revealed that the extent of heat treatment in LWE samples could be determined with PCA of the CE measurements.

Monitoring multiple components in vinegar fermentation using Raman spectroscopy.

In this study, the utility of Raman spectroscopy (RS) with chemometric methods for quantification of multiple components in the fermentation process was investigated. Vinegar, the product of a two stage fermentation, was used as a model and glucose and fructose consumption, ethanol production and consumption and acetic acid production were followed using RS and the partial least squares (PLS) method. Calibration of the PLS method was performed using model solutions. The prediction capability of the method was then investigated with both model and real samples. HPLC was used as a reference method. The results from comparing RS-PLS and HPLC with each other showed good correlations were obtained between predicted and actual sample values for glucose (R(2)=0.973), fructose (R(2)=0.988), ethanol (R(2)=0.996) and acetic acid (R(2)=0.983). In conclusion, a combination of RS with chemometric methods can be applied to monitor multiple components of the fermentation process from start to finish with a single measurement in a short time.

Thermodynamic and structural analysis of interactions between peptide ligands and SEB.

Staphylococcal enterotoxin B (SEB) is an exotoxin produced by Staphylococcus aureus and commonly associated with food poisoning. In this study, SEB-binding peptides were identified by screening a phage displayed peptide library. The binding of peptides to SEB was tested with isothermal titration calorimetry (ITC) and of the five selected peptides, three showed affinity to SEB, with one measured to have the highest affinity constant (10(5) M(-1)). ITC revealed that the interaction of peptide ligands with SEB was driven entropically and the binding was dominated by hydrophobic interactions. Circular dichroism (CD) measurements and molecular dynamics (MD) simulations, together, give a structural insight into the interaction of peptides with SEB. While SEB binding peptides showed random coil structure before binding, after complex formation they had more ordered structures. The peptide with highest affinity to SEB showed stable conformation during MD simulation. Taken together, our approach about thermodynamic and structural characterization of peptide ligands can be used to develop aptamers, with high affinity and selectivity, for biosensor applications.

Analyzing and monitoring of phage-bacteria interaction using CE.

The utilization of CE for monitoring bacteria-phage interaction was investigated in this study. Streptococcus thermophilus and Lactobacillus bulgaricus strains and their phages were used as model bacteria and phages for the purpose of validation in this study. CE with heterogeneous polymer polyethylene oxide was utilized for the separation of intact bacteria and investigation of phage-bacteria interaction. An intact phage detection was carried out with CZE by adding SDS in the running buffer. Calibration graphs of bacteria and phages were obtained with R(2) values of 0.963 and 0.937, respectively. S. thermophilus strain was infected with its virulent phage B3-X18 for investigation of phage-bacteria interaction. It was observed in capillary electropherogram that the culture was lysed depending on the multiplicity of infection value and it showed to be completely lysed when the multiplicity of infection value was 10. The interaction of S. thermophilus strain with L. bulgaricus phage was also investigated by using a CE and a microbiological method and it was observed that the L. bulgaricus phage attached itself to the cell wall of S. thermophilus strain without damaging the cell.

Selection of staphylococcal enterotoxin B (SEB)-binding peptide using phage display technology.

In this study, peptides were selected to recognize staphylococcal enterotoxin B (SEB) which cause food intoxication and can be used as a biological war agent. By using commercial M13 phage library, single plaque isolation of 38 phages was done and binding affinities were investigated with phage-ELISA. The specificities of the selected phage clones showing high affinity to SEB were checked by using different protein molecules which can be found in food samples. Furthermore, the affinities of three selected phage clones were determined by using surface plasmon resonance (SPR) sensors. Sequence analysis was realized for three peptides showing high binding affinity to SEB and WWRPLTPESPPA, MNLHDYHRLFWY, and QHPQINQTLYRM amino acid sequences were obtained. The peptide sequence with highest affinity to SEB was synthesized with solid phase peptide synthesis technique and thermodynamic constants of the peptide-SEB interaction were determined by using isothermal titration calorimetry (ITC) and compared with those of antibody-SEB interaction. The binding constant of the peptide was determined as 4.2+/-0.7 x 10(5)M(-1) which indicates a strong binding close to that of antibody.