ABSTRACT
Aiming at valorizing the ricotta cheese exhausted whey (RCEW), one of the most abundant by-products from the dairy industry, a biotechnological protocol to obtain bioactive peptides with angiotensin-I-converting enzyme (ACE)-inhibitory activity was set up. The approach was based on the combination of membrane filtration and fermentation. A Lactobacillus helveticus strain selected to be used as starter for the fermentation of the ultrafiltration protein-rich retentate (R-UF) obtained from RCEW. The fermented R-UF was characterized by a high anti-ACE activity. Peptides responsible for the bioactivity were purified and identified through nano-LC-ESI-MS/MS. The sequences identified in the purified active fractions of the fermented R-UF showed partial or complete overlapping with previously reported κ-casein antihypertensive fragments. The fermented R-UF was spray-dried and used to enrich ricotta cheese at different fortification level (1 and 5% w/w). An integrated approach including the assessment of the microbiological, chemical, functional, textural, and sensory properties was used to characterize the fortified products. A significantly higher anti-ACE activity was found in the ricotta cheese fortified with fermented R-UF as compared to the control and to the samples obtained with the unfermented R-UF fraction at the same levels of fortification. In particular, a 100 g portion of the ricotta cheese produced at 5% fortification level contained circa 30 mg of bioactive peptides. The fortification led to a moderate acidification, increased hardness and chewiness, and decreased the milk odor and taste of the ricotta cheese as compared to the control, while flavor persistence and sapidity improved.
ABSTRACT
Designing bioactive materials, with controlled metal ion release, exerting a significant biological action and associated to low toxicity for humans, is nowadays one of the most important challenges for our community. The most looked-for nanoantimicrobials are capable of releasing metal species with defined kinetic profiles, either by slowing down or inhibiting bacterial growth and pathogenic microorganism diffusion. In this study, laser ablation synthesis in solution (LASiS) has been used to produce bioactive Ag-based nanocolloids, in isopropyl alcohol, which can be used as water-insoluble nano-reservoirs in composite materials like poly(3-hydroxybutyrate-co-3-hydroxyvalerate). Infrared spectroscopy was used to evaluate the chemical state of pristine polymer and final composite material, thus providing useful information about synthesis processes, as well as storage and processing conditions. Transmission electron microscopy was exploited to study the morphology of nano-colloids, along with UV-Vis for bulk chemical characterization, highlighting the presence of spheroidal particles with average diameter around 12 nm. Electro-thermal atomic absorption spectroscopy was used to investigate metal ion release from Ag-modified products, showing a maximum release around 60 ppb, which ensures an efficient antimicrobial activity, being much lower than what recommended by health institutions. Analytical spectroscopy results were matched with bioactivity tests carried out on target microorganisms of food spoilage.
ABSTRACT
In the last decade, the dairy industry underwent a rapid expansion due to the increasing demand of milk-based products, resulting in high quantity of wastewater, i.e., whey and ricotta cheese exhausted whey (RCEW). Although containing high content of nutritional compounds, dairy by-products are still disposed as waste rather being reintroduced in a new production chain, hence leading to environmental and economic issues. This study proposes a new biotechnological approach based on the combination of membrane filtration and fermentation to produce poly-hydroxyalkanoates (PHA), biodegradable bioplastics candidate as an alternative to petroleum-derived plastics. The protocol, exploiting the metabolic capability Haloferax mediterranei to synthesize PHA from RCEW carbon sources, was set up under laboratory and pilot scale conditions. A multi-step fractionation was used to recover a RCEW fraction containing 12.6% (w/v) of lactose, then subjected to an enzymatic treatment aimed at releasing glucose and galactose. Fermentation conditions (culture medium for the microorganism propagation, inoculum size, time, and temperature of incubation) were selected according to the maximization of polymer synthesis, under in-flasks experiments. The PHA production was then tested using a bioreactor system, under stable and monitored pH, temperature, and stirring conditions. The amount of the polymer recovered corresponded to 1.18 g/L. The differential scanning calorimetry (DSC) analysis revealed the poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) as the polymer synthesized, with a relatively high presence of hydroxyvalerate (HV). Identity and purity of the polymer were confirmed by attenuated total reflectance-Fourier transform infrared (ATR-FTIR) and X-ray photoelectron (XPS) spectroscopy analyses. By combining the fractionation of RCEW, one of the most abundant by-products from the agri-food industry, and the use of the halophile Hfx mediterranei, the production of PHBV with high purity and low crystallinity has successfully been optimized. The process, tested up to pilot scale conditions, may be further implemented (e.g., through fed-batch systems) and used for large-scale production of bioplastics, reducing the economical and environmental issues related the RCEW disposal.
ABSTRACT
A straight synthetic route to fabricate hybrid nanocomposite films of well-dispersed CdS nanocrystals (NCs) in poly[2-methoxy-5-(2'-ethyl-hexyloxy)-1,4-phenylene vinylene] (MEH-PPV) is reported. A soluble cadmium complex [Cd(SBz)2]2·MI, obtained by incorporating a Lewis base (1-methylimidazole, MI) on the cadmium bis(benzyl)thiol, is used as starting reagent in an in situ thermolytic process. CdS NCs with spherical shape nucleate and grow well below 200°C in a relatively short time (30 min). Photoluminescence spectroscopy measurements performed on CdS/MEH-PPV nanocomposites show that CdS photoluminescence peaks are totally quenched inside MEH-PPV, if compared to CdS/PMMA nanocomposites, as expected due to overlapping of the polymer absorption and CdS emission spectra. The CdS NCs are well-dispersed in size and homogeneously distributed within MEH-PPV matrix as proved by transmission electron microscopy. Nanocomposites with different precursor/polymer weight ratios were prepared in the range from 1:4 to 4:1. Highly dense materials, without NCs clustering, were obtained for a weight/weight ratio of 2:3 between precursor and polymer, making these nanocomposites particularly suitable for optoelectronic and solar energy conversion applications.
ABSTRACT
The in vitro effects of polyalkyl-imide hydrogel (PAI) containing 96% pyrogen-free water on the viability, apoptosis, cell shape and metabolic activities of murine 3T3 and human Detroit 555 fibroblasts were investigated. Analysis of the viscous-elastic properties and the ultrastructure of PAI, performed by rheometer and AFM respectively, showed that the material has the typical characteristics of hydrogel, including a three-dimensional configuration of molecules arranged in a regular network with many discrete caveolae where most of the water is captured. Hydrogel biocompatibility was found to be high for both cell lines, with some differences. Cell viability decreased more in 3T3 cells than Detroit 550 fibroblasts when cultured in the presence of 100 mg/ml hydrogel but not at concentrations of 25 and 50 mg/ml hydrogel. The period of incubation with PAI (24 and 48 h) only partially affected cell viability. Apoptosis, most likely due to cells' inability to adhere to the polymer, was the only type of cell death observed. Fibroblasts grown in the presence of polymer were always metabolically active since they continued to synthesize collagen. In conclusion, PAI hydrogel, even at high concentrations, was biocompatible for both fibroblasts, but in particular for human cells, thus encouraging its use as a dermal filler.
