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TEMPO oxidized cellulose nanofibers (T-CNF) were prepared from cellulose pulp which is extracted from bagasse. Soy protein hydrolysate (SPH) was grafted on T-CNF via amidation of carboxylic groups. Biomineralization was, then, assessed via calcium phosphates (CaP) precipitation in twice-simulated body fluid until formation of a new bioactive material. Protein was efficiently grafted without alteration of morphology and nanofibrils packing as reported by Fourier Transform infrared analysis /X Ray Diffraction /Scanning and Transmission Electron Microscopy / Atomic Force Microscopy. Highly crystalline calcium phosphate deposits - ca. 22.1% - were detected, with a Ca/P ratio equal to 1.63, in agreement with native bone apatite composition. In vitro response of human Mesenchymal Stem Cells confirmed the biocompatibility. No significant differences in terms of cell adhesion were recognized while a significant increase in cell proliferation was detected until 7 days. The presence of calcium phosphates tends to cover the nanofibrillar pattern, inducing the inhibition of cell proliferation and promoting the ex-novo precipitation of mineral phases. All the results suggest a promising use of these biomaterials in the repair and/or the regeneration of hard tissues such as bone., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The state of dispersion and the interactions between a polymer and a filler in a nanocomposite crucially define its properties and performance. The affinity of polylactide (PLA) with vegetable and animal proteins (casein, gelatin, soy protein isolate, and hydrolysate) is investigated and their role as eco-friendly dispersants and compatibilizers of cellulose nanofibrils (CNF) is elucidated. The affinity of the proteins with PLA is determined by using sensograms acquired by electroacoustic (quartz crystal microgravimetry) and optical (surface plasmon resonance) techniques. The surface energy of PLA increases upon protein adsorption while the opposite effect is observed for CNF, under identical experimental conditions. A significant improvement in the thermodynamic work of adhesion for PLA/CNF systems is predicted by application of the denatured proteins at low concentrations (∼20% and ∼15% enhancement with soy protein and casein at pH 3 and pH 8, respectively). We offer a robust method to screen denatured proteins and to tailor the wettability and material compatibility in the synthesis of bionanocomposites based on CNF and PLA.

Biocatalytic hydrolysis reactions were designed for preparation of bioactive hydrolysate of duck egg white protein (DEWP) employing two enzymes in one pot. Firstly, the fresh DEWP was thermal treated at 95 °C, for 40 min at pH 10, to effectively deactivate enzyme inhibitors thus facilitating the following two-step enzymatic hydrolysis. Compared with single-enzyme processes, the two-step enzymatic procedures showed much higher reaction efficiency. The first enzymatic step (in the presence of Alcalase or hydrolase SEEP) allowed to hydrolyze DEWP with degree of hydrolysis (DH) of 8.8-10% and soluble peptide yield (SEP) of 60.5-70.2% in a short period (4 h). The second enzymatic step (in the presence of Trypsin or Alcalase) gave a further degradation of DEWP with DH and SEP being more than 26.2% and 90.4%, respectively. The final hydrolysates exhibited high antioxidant activity in an evident DH dependent manner. The hydrolysates achieved by sequential addition of the proteinase SEEP and Alcalase at DH value 21% gave the highest antioxidant activity, which was mainly ascribed to the changes in the amino acid compositions that the contents of some key amino acids and total hydrophobic amino acids were significantly improved by the enzymatic hydrolysis.