The catalytic and kinetic characterization of Bacillus subtilis MK775302 milk clotting enzyme: comparison with calf rennet as a coagulant in white soft cheese manufacture.

BACKGROUND: Calf rennet is considered the traditional source of milk clotting enzyme (MCE). However, increasing cheese consumption with decreasing the calf rennet supply had encouraged the quest for new rennet alternatives. The purpose of this study is to acquire more information about the catalytic and kinetic properties of partially purified Bacillus subtilis MK775302 MCE and to assess the role of enzyme in cheese manufacture. RESULTS: B. subtilis MK775302 MCE was partially purified by 50% acetone precipitation with 5.6-fold purification. The optimum temperature and pH of the partially purified MCE were 70 °C and 5.0, respectively. The activation energy was calculated as 47.7 kJ/mol. The calculated Km and Vmax values were 36 mg/ml and 833 U/ml, respectively. The enzyme retained full activity at NaCl concentration of 2%. Compared to the commercial calf rennet, the ultra-filtrated white soft cheese produced from the partially purified B. subtilis MK775302 MCE exhibited higher total acidity, higher volatile fatty acids, and improved sensorial properties. CONCLUSIONS: The partially purified MCE obtained in this study is a promising milk coagulant that can replace calf rennet at a commercial scale to produce better-quality cheese with improved texture and flavor.

Microencapsulation of Plant Phenolic Extracts Using Complex Coacervation Incorporated in Ultrafiltered Cheese Against AlCl3-Induced Neuroinflammation in Rats.

Plant-derived phenolic compounds have numerous biological effects, including antioxidant, anti-inflammatory, and neuroprotective effects. However, their application is limited because they are degraded under environmental conditions. The aim of this study was to microencapsulate plant phenolic extracts using a complex coacervation method to mitigate this problem. Red beet (RB), broccoli (BR), and spinach leaf (SL) phenolic extracts were encapsulated by complex coacervation. The characteristics of complex coacervates [zeta potential, encapsulation efficiency (EE), FTIR, and morphology] were evaluated. The RB, BR, and SL complex coacervates were incorporated into an ultrafiltered (UF) cheese system. The chemical properties, pH, texture profile, microstructure, and sensory properties of UF cheese with coacervates were determined. In total, 54 male Sprague-Dawley rats were used, among which 48 rats were administered an oral dose of AlCl3 (100 mg/kg body weight/d). Nutritional and biochemical parameters, including malondialdehyde, superoxide dismutase, catalase, reduced glutathione, nitric oxide, acetylcholinesterase, butyrylcholinesterase, dopamine, 5-hydroxytryptamine, brain-derived neurotrophic factor, and glial fibrillary acidic protein, were assessed. The RB, BR, and SL phenolic extracts were successfully encapsulated. The RB, BR, and SL complex coacervates had no impact on the chemical composition of UF cheese. The structure of the RB, BR, and SL complex coacervates in UF cheese was the most stable. The hardness of UF cheese was progressively enhanced by using the RB, BR, and SL complex coacervates. The sensory characteristics of the UF cheese samples achieved good scores and were viable for inclusion in food systems. Additionally, these microcapsules improved metabolic strategies and neurobehavioral systems and enhanced the protein biosynthesis of rat brains. Both forms failed to induce any severe side effects in any experimental group. It can be concluded that the microencapsulation of plant phenolic extracts using a complex coacervation technique protected rats against AlCl3-induced neuroinflammation. This finding might be of interest to food producers and researchers aiming to deliver natural bioactive compounds in the most acceptable manner (i.e., food).

Synthesis and evaluation of eco-friendly carboxymethyl cellulose/polyvinyl alcohol/CuO bionanocomposites and their use in coating processed cheese.

In the present study, we formulated and characterized CMC/PVA/CuO bionanocomposites to evaluate their use in coating processed cheese. Copper oxide nanoparticles (CuO-NPs) were prepared and added to a mixed solution of carboxymethyl cellulose (CMC)/polyvinyl alcohol (PVA) using compositions of 0.3, 0.6 and 0.9% (w/v). The CMC/PVA/CuO bionanocomposites were prepared by a solution casting method and used for coating processed cheese. The fabricated bionanocomposite films and CuO-NPs were characterized by TEM, SEM, EDEX, XRD, DLS, and FT-IR analysis. Inclusion of CuO-NPs decreased the gas transmission rate (GTR) and water vapor transmission rate (WVTR) of the prepared film. Also, the bionanocomposite suspensions exhibited high but variable inhibitory effects against several pathogenic bacteria and fungi. The impact of coating of processed cheese surfaces with the prepared bionanocomposite films on microbiological, physicochemical, textural and sensory properties of the processed cheese were assessed during 6 months of cold storage. Coating cheese with film containing CuO-NPs eliminated mould growth on the cheese surface and decreased significantly (P < 0.05) the total bacterial count of the cheese. Furthermore, coating of cheese decreased the moisture losses and retarded the increase in the cheese hardness during storage. The highest acceptability at the end of the storage period was given for processed cheese coated with the bionanocomposite containing 0.9% CuO-NPs. Thus, the obtained CMC/PVA/CuO bionanocomposite films could be a promising candidate for cheese packaging applications.

Development of bionanocomposite materials and its use in coating of Ras cheese.

The aim of the present study was to prepare and characterize bionanocomposite materials, and to evaluate its use in the coating of Ras cheese. The bionanocomposite materials were made from mixture of chitosan/polyvinyl alcohol with loading of titanium dioxide nanoparticles (TiO2-NPs) from (0.5-2%). The prepared nanoparticles as well as the bionanocomposites were evaluated using, XRD, SEM, TEM, FT-IR and final contact angle. Furthermore, the mechanical properties and water vapor transmission rate (WVTR) of the fabricated bionanocomposites were evaluated. The impacts of coating Ras cheese with the prepared bionanocomposite on weight losses and microbiological, chemical, and physical characteristics of the Ras cheese were assessed during ripening in comparison to the uncoated cheese. Coating of cheese decreased the weight and moisture losses but didn't affect the normal ripening changes in the microbiological, chemical and textural properties of Ras cheese. Coating cheese with film containing 2% TiO2-NPs eliminated mold growth on the cheese surface.