Design of whey protein nanostructures for incorporation and release of nutraceutical compounds in food

Whey proteins are widely used as nutritional and functional ingredients in formulated foods because they are relatively inexpensive, generally recognized as safe (GRAS) ingredient, and possess important biological, physical, and chemical functionalities. Denaturation and aggregation behavior of these proteins is of particular relevance toward manufacture of novel nanostructures with a number of potential uses. When these processes are properly engineered and controlled, whey proteins may be formed into nanohydrogels, nanofibrils, or nanotubes and be used as carrier of bioactive compounds. This review intends to discuss the latest understandings of nanoscale phenomena of whey protein denaturation and aggregation that may contribute for the design of protein nanostructures. Whey protein aggregation and gelation pathways under different processing and environmental conditions such as microwave heating, high voltage, and moderate electrical fields, high pressure, temperature, pH, and ionic strength were critically assessed. Moreover, several potential applications of nanohydrogels, nanofibrils, and nanotubes for controlled release of nutraceutical compounds (e.g. probiotics, vitamins, antioxidants, and peptides) were also included. Controlling the size of protein networks at nanoscale through application of different processing and environmental conditions can open perspectives for development of nanostructures with new or improved functionalities for incorporation and release of nutraceuticals in food matrices.     

Description of the kinetic enzymatic browning in banana (Musa cavendish) slices using non-uniform color information from digital images

A novel methodology “fractal browning indicator” (FBI) is presented, that describes the enzymatic browning kinetic based on the use of irregular color patterns from banana slice images. It uses the fractal Fourier texture image value in a selected area, to calculate a fractal dimension (FD), which represents the complexity of color distribution. During the procedure, colors from digital images were first transformed to L^*a^*b^* space color using a transformation function (quadratic model), in order to derivate three color channels, lightness (L^*), redness (a^*), and yellowness (b^*). In the results, lightness and yellowness parameters decreased during the browning kinetic, when their respective FD values increased, indicating major color distribution complexity in a selected area analyzed during the kinetic. The redness color (a^*) did not show any statistical variation. The empirical power law model was suitable to correlate enzymatic browning kinetic data both for FBI and for the traditional method (when an L^* mean was used). However, enzymatic browning rates using the FBI method, were between 8.5 and 35 times higher than rates calculated with the traditional method.     

Response surface models for effects of temperature, pH, and previous growth pH on growth kinetics of Salmonella Typhimurium in brain heart infusion broth

Response surface models were developed for effects of temperature (15 to 40 degrees C), pH (5.2 to 7.4), and previous growth pH (5.7 to 8.6) on lag time (lambda) and specific growth rate (mu) of Salmonella Typhimurium in brain heart infusion broth (BHIB). Seventy-five growth curves for model development and 30 growth curves for model validation were fit to a two-phase linear growth model to obtain direct estimates of lambda and mu of Salmonella Typhimurium in BHIB. Response surface models for natural logarithm transformations of lambda and mu as a function of temperature, pH, and previous growth pH were obtained by regression analysis. Previous growth pH did not alter (P > 0.05) or interact with temperature or pH to alter subsequent growth kinetics of Salmonella Typhimurium. However, lambda and mu of Salmonella Typhimurium in BHIB were affected (P < 0.05) by linear and quadratic effects of temperature and pH. The models were validated against data not used in their development. Mean absolute relative error of predictions (model accuracy) was 7.8% for lambda and 6.6% for mu. Median relative error of predictions (model bias) was -1.8% for lambda and -2.8% for mu. Results of the current study indicated that the models developed accurately predicted growth kinetics of Salmonella Typhimurium in BHIB within the matrix of factors modeled and that the range of previous growth pH (5.7 to 8.6) investigated did not alter the subsequent growth kinetics of Salmonella Typhimurium in BHIB.     

A filtration-based real-time PCR method for the quantitative detection of viable Salmonella enterica and Listeria monocytogenes in food samples

We developed a novel filtration-based method that can eliminate dead or severally damaged Salmonella enterica and Listeria monocytogenes in food samples. This new method can recover all viable bacteria in less than 30 min, and can be coupled with a subsequent bacterial DNA extraction and real-time PCR. No statically significant differences (p < 0.01) were found between real-time PCR results obtained separately from S. enterica and L. monocytogenes when different ratios of living and dead cells were used. The analytical sensitivity in both cases was 1 genome equivalent (GE), and the quantification was linear (R² > 0.9969) over a 5-log dynamic range with PCR efficiencies >0.9754. When compared with the standard microbiological methods for the detection of these foodborne pathogens, the relative accuracy was excellent ranging from 95.72% to 104.48%. Finally, we applied the pre-treatment method to the direct detection of viable forms of these foodborne pathogens in food samples using yogurt as a model, the results being similar to those obtained using pure cultures.     

Impact of ultrafiltration and nanofiltration of an industrial fish protein hydrolysate on its bioactive properties

BACKGROUND: Numerous studies have demonstrated that in vitro controlled enzymatic hydrolysis of fish and shellfish proteins leads to bioactive peptides. Ultrafiltration (UF) and/or nanofiltration (NF) can be used to refine hydrolysates and also to fractionate them in order to obtain a peptide population enriched in selected sizes. This study was designed to highlight the impact of controlled UF and NF on the stability of biological activities of an industrial fish protein hydrolysate (FPH) and to understand whether fractionation could improve its content in bioactive peptides. RESULTS: The starting fish protein hydrolysate exhibited a balanced amino acid composition, a reproducible molecular weight (MW) profile, and a low sodium chloride content, allowing the study of its biological activity. Successive fractionation on UF and NF membranes allowed concentration of peptides of selected sizes, without, however, carrying out sharp separations, some MW classes being found in several fractions. Peptides containing Pro, Hyp, Asp and Glu were concentrated in the UF and NF retentates compared to the unfractionated hydrolysate and UF permeate, respectively. Gastrin/cholecystokinin‐like peptides were present in the starting FPH, UF and NF fractions, but fractionation did not increase their concentration. In contrast, quantification of calcitonin gene‐related peptide (CGRP)‐like peptides demonstrated an increase in CGRP‐like activities in the UF permeate, relative to the starting FPH. The starting hydrolysate also showed a potent antioxidant and radical scavenging activity, and a moderate angiotensin‐converting enzyme (ACE)‐1 inhibitory activity, which were not increased by UF and NF fractionation. CONCLUSION: Fractionation of an FPH using membrane separation, with a molecular weight cut‐off adapted to the peptide composition, may provide an effective means to concentrate CGRP‐like peptides and peptides enriched in selected amino acids. The peptide size distribution observed after UF and NF fractionation demonstrates that it is misleading to characterize the fractions obtained by membrane filtration according to the MW cut‐off of the membrane only, as is currently done in the literature. Copyright © 2010 Society of Chemical Industry     

Revalorisation of vine trimming wastes using Lactobacillus acidophilus and Debaryomyces hansenii

BACKGROUND: Mild acid treatments of vine‐shoot trimmings result in the hydrolysis of hemicellulosic sugars that can be utilised by Lactobacillus acidophilus CECT‐4179 (ATCC 832) and Debaryomyces hansenii NRRL Y‐7426 as carbon sources to obtain food additives. Since the high content of glucose in these hydrolysates reduces the effective bioconversion of xylose into xylitol by D. hansenii, the use of Lactobacillus acidophilus, one of the main probiotic species, allows this problem to be solved by the selective consumption of glucose. In order to use both sugars (glucose and xylose), hemicellulosic vine‐shoot trimming hydrolysates can be sequentially fermented by both micro‐organisms. RESULTS: It was found that, in the first step, L. acidophilus generated almost exclusively lactic acid (32.7 g of lactic acid L^?1, Q_LA = 1.363 g L^?1 h^?1, Y_LA/S = 0.72 g g^?1) by homofermentative degradation of sugars (mainly glucose), and in the second step, the remaining hemicellulosic sugars were transformed primarily into xylitol by Debaryomyces hansenii (31.3 g of xylitol L^?1, Q_Xylitol = 0.708 g L^?1 h^?1, Y_Xylitol/S = 0.66 g g^?1). Furthermore, L. acidophilus proved to be a strong cell‐bounded biosurfactant producer. Cell extracts were able to reduce the surface tension (ST) of PBS in 18 mN m^?1 units. Lactobacillus acidophilus cells showed no difference in viability before or after PBS extraction of biosurfactants, achieving values of 0.9 × 10⁹ colony‐forming units (CFU) mL^?1 in both cases. CONCLUSIONS: These results have made a serious contribution to the re‐evaluation of a useless and pollutant residue, producing a wide range of natural food additives. Copyright © 2008 Society of Chemical Industry     

Thermomechanical properties of biodegradable films based on blends of gelatin and poly(vinyl alcohol)

The aim of this work was to study the effect of the poly(vinyl alcohol) (PVA) concentration on the thermal and viscoelastic properties of films based on blends of gelatin and PVA using differential scanning calorimetry (DSC) and dynamic-mechanical analysis (DMA). One glass transition was observed between 43 and 49 °C on the DSC curves obtained in the first scanning of the blended films, followed by fusion of the crystalline portion between 116 and 134 °C. However, the DMA results showed that only the films with 10% PVA had a single peak in the tan δ spectrum. However, when the PVA concentration was increased the dynamic mechanical spectra showed two peaks on the tan δ curves, indicating two T_gs. Despite this phase separation behavior the Gordon and Taylor model was successfully applied to correlate T_g as a function of film composition, thus determining k=7.47. In the DMA frequency tests, the DMA spectra showed that the storage modulus values decreased with increasing temperature. The master curves for the PVA–gelatin films were obtained applying the TTS principle (T_r=100 °C). The WLF model was thus applied allowing for the determination of the constants C₁ and C₂. The values of these constants increased with increasing PVA concentrations in the blend: C₁=49–66 and C₂=463–480. These values were used to calculate the fractional free volume of the films at the T_g and the thermal expansion coefficient of the films above the T_g.