A Novel Intelligent Indicator Film: Preparation, Characterization, and Application.

The development of intelligent indicator film that can detect changes in food quality is a new trend in the food packaging field. The WPNFs-PU-ACN/Gly film was prepared based on whey protein isolate nanofibers (WPNFs). Anthocyanin (ACN) and glycerol (Gly) were used as the color indicator and the plasticizer, respectively, while pullulan (PU) was added to enhance mechanical properties of WPNFs-PU-ACN/Gly edible film. In the study, the addition of ACN improved the hydrophobicity and oxidation resistance of the indicator film; with an increase in pH, the color of the indicator film shifted from dark pink to grey, and its surface was uniform and smooth. Therefore, the WPNFs-PU-ACN/Gly edible film would be suitable for sensing the pH of salmon, which changes with deterioration, as the color change of ACN was completely consistent with fish pH. Furthermore, the color change after being exposed to grey was evaluated in conjunction with hardness, chewiness, and resilience of salmon as an indication. This shows that intelligent indicator film made of WPNFs, PU, ACN, and Gly could contribute to the development of safe food.

B- and N-doped carbon dots by one-step microwave hydrothermal synthesis: tracking yeast status and imaging mechanism.

BACKGROUND: Carbon dots (CDs) are widely used in cell imaging due to their excellent optical properties, biocompatibility and low toxicity. At present, most of the research on CDs focuses on biomedical application, while there are few studies on the application of microbial imaging. RESULTS: In this study, B- and N-doped carbon dots (BN-CDs) were prepared from citric acid, ethylenediamine, and boric acid by microwave hydrothermal method. Based on BN-CDs labeling yeast, the dead or living of yeast cell could be quickly identified, and their growth status could also be clearly observed. In order to further observe the morphology of yeast cell under different lethal methods, six methods were used to kill the cells and then used BN-CDs to label the cells for imaging. More remarkably, imaging of yeast cell with ultrasound and antibiotics was significantly different from other imaging due to the overflow of cell contents. In addition, the endocytosis mechanism of BN-CDs was investigated. The cellular uptake of BN-CDs is dose, time and partially energy-dependent along with the involvement of passive diffusion. The main mechanism of endocytosis is caveolae-mediated. CONCLUSION: BN-CDs can be used for long-term stable imaging of yeast, and the study provides basic research for applying CDs to microbiol imaging.

Direct separation and purification of α-lactalbumin from cow milk whey by aqueous two-phase flotation of thermo-sensitive polymer/phosphate.

BACKGROUND: α-lactalbumin (α-La) is of great interest to the industry as a result of its excellent functional properties and nutritional value. Aqueous two-phase flotation (ATPF) of thermo-sensitive polymer poly (ethylene glycol-ran-propylene glycol) monobutyl ether (UCON) and KH2 PO4 was applied to directly separate and purify α-La from milk whey, which was purposed to simplify the production process and reduced cost of production. RESULTS: The effect of ATPF composition and operating parameters on the flotation efficiency (E) and purity of α-La were investigated. The optimal conditions included 2 min of premixing time, 30 mL min-1 flow velocity and 20 min of flotation time, whereas the composition conditions comprised 35.0 mL 0.18 g mL-1 phosphate solution (containing 10% (cow milk whey/salt solution, v/v) cow milk whey, 50 ppm defoamer and 2 g NaCl) and 5.0 mL of 40% (w/w) UCON solution. Under the optimal conditions, E of α-La was 95.67 ± 1.04% and purity of α-La was 98.78 ± 1.19%. UCON was recovered by a thermally-induced phase separation and reused in next ATPF process without reducing E of α-La. Purified α-La was characterized by several key technologies. The results indicated that α-La in cow milk whey could be directly separated and purified by the ATPF and the purity was satisfactory. Moreover, it was suggested there was no obvious structure difference between the α-La separated by ATPF and the α-La standard. CONCLUSION: The present study enabled the recycling of UCON, providing an effective, economically viable and environmentally friendly approach for the separation and purification of protein. © 2021 Society of Chemical Industry.

Effect of Antioxidant and Antimicrobial Coating based on Whey Protein Nanofibrils with TiO2 Nanotubes on the Quality and Shelf Life of Chilled Meat.

Whey protein nanofibrils (WPNFs) can be used in edible films and coatings (EFCs) because of its favorable functional properties, which rely on its well-ordered ß-sheet structures, high hydrophobicity, homogeneous structure, and antioxidant activity. In the present study, WPNF-based edible coatings with glycerol (Gly) as plastic and titanium dioxide nanotubes (TNTs) as antimicrobial agents were studied. TNTs not only showed greater antibacterial activity than titanium dioxide nanoparticles (TNPs), but also increased interactions with WPNFs. The WPNF/TNT film had a smooth and continuous surface and was homogeneous with good mechanical properties. WPNF/TNT edible coatings (ECs) can help improve lipid peroxidation and antioxidant activity, limit microbial growth, reduce weight loss, and extend the shelf life of chilled beef. Given that the WPNF/TNT film components are low cost and show high antioxidant and antimicrobial activity, these optimized films have potential applications for various food products, including raw and chilled meat.

Separation and Enrichment of Lectin from Zihua Snap-Bean (Phaseolus vulgaris) Seeds by PEG 600-Ammonium Sulfate Aqueous Two-Phase System.

A fast and efficient method based on a polyethylene glycol (PEG) 600/(NH4)2SO4 aqueous two-phase system for extracting lectin from Zihua snap-bean (Phaseolus vulgaris) seeds was established. According to a Box-Behnken design (BBD), involving four factors at three levels each subjected to analysis of variance (ANOVA) and response surface analysis, the protein recovery and the purification factor of lectin in the top phase were used as the response values of the variance analysis to acquire the multivariate quadratic regression model. SDS-PAGE electrophoresis and the hemagglutination test were used to detect the distribution of lectin in the aqueous two-phase system (ATPS). The obtained data indicated that lectin was preferentially partitioned into the PEG-rich phase, and the ATPS, composed of 15% (NH4)2SO4 (w/w), 18% PEG 600 (w/w), 0.4 g/5 g NaCl and 1 mL crude extract, showed good selectivity for lectin when the pH value was 7.5. Under the optimal conditions, most of the lectin was assigned to the top phase in the ATPS, and the hemagglutination activity of the purified lectin in the top phase was 3.08 times that of the crude extract. Consequently, the PEG 600/(NH4)2SO4 aqueous two-phase system was an effective method for separating and enriching lectin directly from the crude extract of Zihua snap-bean seeds.

Nitrogen-Doped Carbon Quantum Dots as Fluorescent Probes for Sensitive and Selective Detection of Nitrite.

Nitrites are the upstream precursors of the carcinogenic nitrosamines, which are widely found in the natural environment and many food products. It is important to develop a simple and sensitive sensor for detecting nitrites. In this work, a fluorescence probe based on nitrogen-doped carbon quantum dots (N-CQDs) was developed for the sensitive and selective determination of nitrites. At pH 2, the fluorescence of N-CQDs can be selectively quenched by nitrite due to the fact N-nitroso compounds can be formed in the reaction of amide groups with nitrous acid, which results in fluorescence static quenching. Under optimal conditions, fluorescence intensity quenching upon addition of nitrite gives a satisfactory linear relationship covering the linear range of 0.2-20 µM, and the limit of detection (LOD) is 40 nM. Moreover, this method has been successfully applied to the determination of nitrites in tap water, which indicates its great potential for monitoring of nitrites in environmental samples.

Extraction and purification of wheat-esterase using aqueous two-phase systems of ionic liquid and salt.

To explore a new and simple rapid extraction and purification technique for wheat-esterase, an ionic liquids (ILs)-based aqueous two-phase system (ATPS) was developed for the purification of wheat-esterase from wheat extracts. Effects of various process parameters such as the concentrations of [Bmim]BF4, the types and concentrations of phase-forming salt, the system pH and the temperature on partitioning of wheat-esterase were evaluated. The obtained data indicated that wheat-esterase was preferentially partitioned into the ILs-rich phase and the ATPS composed of 20 % [Bmim]BF4 (w/w) and 25 % (w/w) NaH2PO4(pH = 4.8) showed good selectivity on wheat-esterase. Under the optimum conditions, wheat-esterase was purified with an acceptable yield (88.93 %), but produced wheat-esterase was 4.23 times as pure. It was obvious that temperature shows little influence on the purification between 10 and 50 °C. Sephadex G-150FF revealed that the band intensity of contaminating proteins in ATPS fraction almost disappeared. Therefore, ILs-based ATPS was an effective method for partitioning and recovery of wheat-esterase from wheat crude extracts.

One-step high efficiency separation of prolyl endopeptidase from Aspergillus niger and its application.

Prolyl endopeptidase from Aspergillus niger (An-PEP) is an enzyme that recognizes C-terminal peptide bonds of amino acid chains and cleaves them by hydrolysis. An aqueous two-phase system (ATPS) was used to separate An-PEP from fermentation broth. Through single factor experiments, the ATPS containing 16 % (w/w) PEG2000 and 15 % (w/w) (NH4)2SO4 at pH 6.0 obtained the recovery of 79.74 ± 0.16 % and the purification coefficient of 7.64 ± 0.08. It was then used to produce soy protein isolate peptide (SPIP) by hydrolysis of soy protein isolate (SPI), and SPIP-Ferrous chelate (SPIP-Fe) was prepared with SPIP and Fe2+. The chelation conditions were optimized by RSM, as the chelation time was 30 min, chelation temperature was 25 °C, SPIP mass to VC mass was two to one and pH was 6.0. The obtained chelation rate was 82.56 ± 2.30 %. The change in the structures and functional features of SPIP before and after chelation were investigated. The FTIR and UV-Vis results indicated that the chelation of Fe2+ and SPIP depended mainly on the formation of amide bonds. The fluorescence, SEM and amino acid composition analysis results indicated that Fe2+ could induce and stabilize the surface conformation and change the amino acid distribution on the surfaces of SPIP. The chelation of SPIP and Fe2+ resulted in the enhancement of radical scavenging activities and ACE inhibitory activities. This work provided a new perspective for the further development of peptide-Fe chelates for iron supplement.

Structural characteristics and biological activity of lactic acid bacteria exopolysaccharides separated by ethanol/(NH4)2SO4 ATPS.

Exopolysaccharides (EPS) from lactic acid bacteria (LAB) as edible and safe bioproducts with health benefits have become an interesting topic. In this study, aqueous two-phase system (ATPS) was established using ethanol and (NH4)2SO4 as phase-forming substances to separate and purify LAB EPS from Lactobacillus plantarum 1.0665. The operating conditions were optimized by a single factor and response surface method (RSM). The results indicated that an effectively selective separation of LAB EPS was achieved by the ATPS consisted of 28 % (w/w) ethanol and 18 % (w/w) (NH4)2SO4 at pH 4.0. Under optimized conditions, the partition coefficient (K) and recovery rate (Y) were well matched with the predicted value of 3.83 ± 0.019 and 74.66 ± 1.05 %. The physicochemical properties of purified LAB EPS were characterized by various technologies. According to the results, LAB EPS was a complex polysaccharide with a triple helix structure mainly composed of mannose, glucose and galactose in the molar ratio of 1.00: 0.32: 0.14, and it proved that the ethanol/(NH4)2SO4 system had good selectivity for LAB EPS. In addition, LAB EPS displayed excellent antioxidant activity, antihypertension activity, anti-gout capacity and hypoglycemic activity in vitro analysis. The results suggested that LAB EPS could be a dietary supplement applied in functional foods.

Efficient imaging based on P - and N-codoped carbon dots for tracking division and viability assessment of lactic acid bacteria.

Assessment of lactic acid bacteria (LAB) activity plays a key role in the fermented food industry. Fluorescence imaging method based on dye is facile to detect LAB viability. However, it is difficult to obtain stable fluorescence, non-toxic and low-cost dyes. In this study, we prepare P- and N-doped carbon dots (PN-CDs) via microwave-assisted hydrothermal synthesis. The properties of high quantum yield (60.36%) and excitation dependence allowed for multicolor imaging of LAB (Lactobacillus plantarum [L.p] and Streptococcus thermophilus [S.t]). The abundant functional groups and positive charges (+2.34 mV) on the surface of PN-CDs facilitated their quickly integrated into cell wall of live LAB with obvious fluorescence or into dead cells. As a result, PN-CDs can not only be used to rapidly and efficiently monitor bacterial viability (one minute), but can also be used to visualize LAB division using fluorescence imaging. Importantly, the PN-CDs have potential to rapidly detect LAB activity in LAB-fermented juices.

Enhanced viability of probiotics encapsulated within synthetic/natural biopolymers by the addition of gum arabic via electrohydrodynamic processing.

To enhance the probiotics' viability, novel vehicles consisting of synthetic/natural biopolymers, i.e., polyvinyl alcohol (PVOH), polyvinylpyrrolidone, whey protein concentrate and maltodextrin, encapsulated with L. plantarum KLDS 1.0328 and gum arabic (GA) as a prebiotic were fabricated by electrohydrodynamic techniques. Inclusion of cells into composites caused an increase in conductivity and viscosity. Morphological analysis showed that cells were distributed along the electrospun nanofibres or distributed randomly in the electrosprayed microcapsules. Both intramolecular and intermolecular hydrogen bond interactions exist between biopolymers and cells. Thermal analysis revealed that the degradation temperatures (>300 °C) of various encapsulation systems have potential applications in heat-treatment foods. Additionally, cells especially immobilized in PVOH/GA electrospun nanofibres showed the highest viability compared with free cells after exposure to simulated gastrointestinal stress. Furthermore, cells retained their antimicrobial ability after rehydration of the composite matrices. Therefore, electrohydrodynamic techniques have great potential in encapsulating probiotics.

Ultrasonic-microwave synergistic supramolecular solvent liquid-liquid microextraction of trace biogenic amines in fish and beer based on solidification of floating organic droplet.

It is important to detect the presence of biogenic amines (BAs) as indicators of food freshness. The purpose of this study was to develop a novel ultrasonic-microwave synergistic supramolecular solvent liquid-liquid microextraction based on solidification of floating organic droplet (UMS-SUPRAS-SFO-LLME) combined with high-performance liquid chromatography for the determination of BAs. The physical properties and microstructure of SUPRAS based on 1-dodecanol and tetrahydrofuran were studied, and the extraction conditions such as the SUPRAS volume, the UMS process, and the centrifugal conditions were optimized. The results for the extraction kinetics and thermodynamics showed that UMS-SUPRAS-SFO-LLME is a spontaneous, endothermic diffusion process. The linear ranges of this method are 0.1-2.0 × 105 ng·mL-1 (R2 > 0.994), the limits of detection are 4.0 × 10-3-6.0 × 10-2 ng·mL-1, and the recoveries were 96.28-103.15%. Compared with existing analysis methods, UMS-SUPRAS-SFO-LLME is a sensitive, green and economical sample pretreatment method for analyzing the enrichment of BAs in beer and fish.

Covalent and Noncovalent Complexation of Phosvitin and Gallic Acid: Effects on Protein Functionality and In Vitro Digestion Properties.

To investigate the effects of different binding modes on the structure, function, and digestive properties of the phosvitin (Pv) and gallic acid (GA) complex, Pv was covalently and noncovalently combined with different concentrations of GA (0.5, 1.5, and 2.5 mM). The structural characterization of the two Pv-GA complexes was performed by Fourier transform infrared, circular dichroism, and LC-MS/MS to investigate the covalent and noncovalent binding of Pv and GA. In addition, the microstructure of the two Pv-GA complexes was investigated by super-resolution microscopy and transmission electron microscopy. The particle size and zeta potential results showed that the addition of GA increased the particle size and the absolute potential of Pv. The determination of protein digestibility, polyphenol content, SH and S-S group levels, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and antioxidant capacity of the digests indicated that noncovalent complexes had greater antioxidant and protective effects on polyphenols. Molecular docking revealed that GA was conjugated with Pv through hydrogen bond interactions.

Effective separation of prolyl endopeptidase from Aspergillus Niger by aqueous two phase system and its characterization and application.

Aspergillus niger prolyl endopeptidase (An-PEP) has become a research focus because of its advantages in specifically cleaving the C-terminal peptide bond of proline residues, especially it was an industrial food-grade acidic PEP. Aqueous two-phase system (ATPS) was first applied for separating An-PEP from fermentation broth. Via response surface method (RSM) experiment, an effectively separation of An-PEP was achieved by ATPS containing27% (w/w) ethanol and 14.5% (w/w) (NH4)2SO4 at pH 6.0 with the recovery of 90.29 ± 0.23% and purification coefficient of 15.35 ± 0.30. The purified An-PEP was characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), fourier transform infrared (FTIR) and fluorescence spectrometry. The optimum temperature and pH of An-PEP were 40 °C and 4.5-5.0, respectively. An-PEP was activated and stabilized by Ca2+ but inhibited by Fe3+. The enzymatic application of purified An-PEP was evaluated by hydrolyzing egg white protein (EWP) to prepare bioactive peptides. The obtained hydrolysates had good scavenging ability of OH and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) free radicals, angiotensin converting enzyme (ACE) inhibitory activity and anti-gout activity. This research realized a low-cost, high-efficiency and simple separation technology of An-PEP and provided a broader idea for the preparation of bioactive peptides and the application of An-PEP.

Development of Antioxidant and Stable Conjugated Linoleic Acid Pickering Emulsion with Protein Nanofibers by Microwave-Assisted Self-Assembly.

Whey protein isolate nanofibrils (WPNFs) can be used as a novel stabilizer in the Pickering emulsion system to improve the water solubility, stability and bioavailability of lipophilic bioactive ingredients. In this study, conjugated linoleic acid (CLA) and WPNFs were used to prepare a stable Pickering emulsion. We used a transmission electron microscope, low-temperature scanning electron micrographs and other methods to evaluate the micromorphology, surface hydrophobicity and structural units of the obtained WPNFs. Compared with whey protein isolate/CLA Pickering emulsion, the WPNFs/CLA Pickering emulsion has greater ability to remove 2,2-Diphenyl-1-picrylhydrazyl and 2,2'-amino-di(2-ethyl-benzothiazoline sulphonic acid-6) ammonium salt free radicals. Furthermore, the WPNFs/CLA Pickering emulsion has a more stable effect in terms of droplet size and zeta potential over a wider range of ionic strength and temperature conditions. These findings indicate that Pickering emulsion stabilized by WPNFs is more suitable as a carrier of CLA, as it increases the solubility of CLA and has better active applications in biology and food.

Targeting Delivery System for Lactobacillus Plantarum Based on Functionalized Electrospun Nanofibers.

With the increased interest in information on gut microbes, people are realizing the benefits of probiotics to health, and new technologies to improve the viability of probiotics are still explored. However, most probiotics have poor resistance to adverse environments. In order to improve the viability of lactic acid bacteria, polylactic acid (PLA) nanofibers were prepared by coaxial electrospinning. The electrospinning voltage was 16 kV, and the distance between spinneret and collector was 15 cm. The feed rates of the shell and core solutions were 1.0 and 0.25 mL/h, respectively. The lactic acid bacteria were encapsulated in the coaxial electrospun nanofibers with PLA and fructooligosaccharides (FOS) as the shell materials. Scanning electron microscopy, transmission electron microscopy, and laser scanning confocal microscopy showed that lactic acid bacteria were encapsulated in the coaxial electrospun nanofibers successfully. The water contact angle test indicated that coaxial electrospun nanofiber films had good hydrophobicity. An in vitro simulated digestion test exhibited that the survival rate of lactic acid bacteria encapsulated in coaxial electrospun nanofiber films was more than 72%. This study proved that the viability of probiotics can be improved through encapsulation within coaxial electrospun PLA nanofibers and provided a novel approach for encapsulating bioactive substances.

Preparation and Characterization of Coating Based on Protein Nanofibers and Polyphenol and Application for Salted Duck Egg Yolks.

Salted duck egg yolk (SDEY) is one of the traditional pickled egg products in Asian countries, which suffers from the weight loss and deterioration of texture characteristics during storage. To better maintain the texture of SDEY, an edible coating based on whey protein isolate nanofibers (WPNFs) with glycerol (Gly) as a plasticizer and incorporating carvacrol (CA) as an antimicrobial agent was developed. Whey protein isolate (WPI, 5%) was used to self-assemble into WPNFs at 80 °C for 10 h. The particle size, zeta-potential and microstructure of WPNFs-CA emulsion were investigated to evaluate the distribution. Results proved that WPNFs-CA emulsion had smaller particle size and better distribution than WPI-CA emulsion. WPNFs-CA/Gly edible coating was then prepared based on WPNFs-CA emulsion. The WPNFs-CA/Gly edible coating exhibited higher antibacterial activity while the WPNFs-CA/Gly film had smooth and continuous surfaces and better transmittance compared with other samples. Furthermore, weight losses and textural properties changes of SDEYs with WPNFs-CA/Gly coating were evaluated. Results proved that salted duck egg yolks with WPNFs-CA/Gly coating exhibited lower weight losses. Textural properties were significantly improved by the WPNFs-CA/Gly coating on SDEYs than those uncoated samples. It was noted that the egg yolks coated with the WPNFs-CA/Gly coating had the lowest hardness increase rate (18.22%). Hence, WPNF-based coatings may have a good development prospect in the food industry.

Fabrication and Characterization of a Microemulsion Stabilized by Integrated Phosvitin and Gallic Acid.

The purpose of this work was to conjugate phosvitin (Pv) with gallic acid (GA) to explore a new emulsifier that had both good emulsifying properties and antioxidant activity. The Pv-GA complex was prepared at a GA concentration of 1.5 mg/mL with pH 9.0. The Pv-GA complex obtained was identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and characterized with infrared, ultraviolet, and fluorescence spectra. The emulsifying activity and stability of the Pv-GA complex were slightly improved, and antioxidant activities was significantly enhanced. Furthermore, the Pv-GA complex was used to load conjugated linoleic acid (CLA) for microemulsion preparation. Results showed that the Pv-GA complex could increase the viscosity and lipid antioxidant capacity of Pv-GA/CLA microemulsion. The Pv-GA/CLA microemulsion had remarkable emulsifying activity, emulsifying stability, pH, and thermal stability and poor salt stability.

Carbon Dots Derived from the Maillard Reaction for pH Sensors and Cr (VI) Detection.

The Maillard reaction involves a series of complex reactions; fluorescent compounds have been considered as vital intermediate products of the reaction. In this article, carbon dots (CDs) based on the Maillard reaction (MR-CDs) were prepared with L-tryptophan and D-glucose, and they had excellent photoluminescence stability. MR-CDs showed stable pH-dependence behavior and exhibited an excellent linear response to pH in the range of 4.0-7.5 and 7.5-13.0, respectively. Under the masking effect of sodium fluoride for Fe(III), MR-CDs showed excellent selectivity and sensitivity for Cr (VI). The linear range of Cr(VI) was 0.2-50 µM and the limit of detection was 20 nM. (S/N ≥ 3). Furthermore, MR-CDs were used to detect Cr(VI) in tap water samples. The recoveries were between 95.8% and 98.94%, and RSDs were less than 3.17%.

Efficient Imaging of Saccharomyces cerevisiae Based on B- and N-Doped Carbon Dots.

The estimation of yeast viability with B- and N-doped carbon dots (BN-CDs) was investigated in this paper. BN-CDs with a fluorescent quantum yield of 65.47% were prepared by a one-step hydrothermal method. The size distribution of BN-CDs was relatively narrow, with the majority falling within 7.5-8.5 nm, and they were mainly composed of carbon, oxygen, nitrogen, and boron. BN-CDs were shown to have strong and stable fluorescence. They exhibited excitation-independent photoluminescence property, which could avoid the autofluorescence and limitation of the excitation source. Dead and live yeast cells were distinguished well by BN-CD staining in a short time, and there was no strict requirement for light protection. The application of BN-CDs in beer brewing can solve the problem of estimation of yeast viability.