Fuguo Liu

Utilization of interfacial engineering to improve physicochemical stability of β-carotene emulsions: Multilayer coatings formed using protein and protein–polyphenol conjugates

The impact of lactoferrin (LF)-chlorogenic acid (CA) and (-)-Epigallocatechin-3-gallate (EGCG) conjugates on the physicochemical properties of β-carotene emulsions was investigated. Formation of lactoferrin-polyphenol conjugates, which was confirmed by SDS-PAGE, caused changes in the structure and nature of lactoferrin. Based on layer-by-layer electrostatic deposition, β-carotene bilayer emulsions were prepared by lactoferrin and lactoferrin-polyphenol conjugates at pH 7.0. The physicochemical properties of primary and secondary emulsions were evaluated and the results suggested that LF-polyphenol conjugates-stabilized primary and secondary emulsions exhibited better emulsifying properties and improved physical stability of β-carotene bilayer emulsions under freeze-thaw, ionic strength and thermal treatments. In addition, the lactoferrin-polyphenol conjugates could effectively enhance chemical stability of β-carotene in oil-in-water emulsions against heat treatment and ultraviolet light exposure, and the least degradation of β-carotene occurred in LF-EGCG conjugate-stabilized primary emulsion. The interfacial engineering technology utilized in this study may lead to the formation of emulsions with improved physicochemical and functional performance.

Structure and antimicrobial mechanism of ɛ-polylysine–chitosan conjugates through Maillard reaction

The aim of the study was to testify the formation and antimicrobial activity of ɛ-polylysine-chitosan conjugate through Maillard reaction. The results of UV absorbance, browning index and fluorescence changes of Maillard reaction products (MRPs) between ɛ-polylysine and chitosan indicated there existed Maillard reaction between ɛ-polylysine and chitosan and the formation of their conjugate. The conjugate showed strong antibacterial activity against Escherichia coli, Staphylococcus aureus, Bacillus subtilis and beer yeast. Morphologies of E. coli and S. aureus treated with the conjugate were observed by transmission electron microscopy (TEM). The results revealed that the conjugate of ɛ-polylysine and chitosan increased the permeability of the outer membrane (OM) and inner membrane (IM) and ultimately disrupted bacterial cell membranes, with the release of cellular cytoplasm.

Structural characterization and functional evaluation of lactoferrin–polyphenol conjugates formed by free-radical graft copolymerization

Covalent modifications of lactoferrin (LF) with epigallocatechin gallate (EGCG), chlorogenic acid (CA) and gallic acid (GA) were performed by adopting a free-radical grafting procedure in aqueous media. The resulting LF–polyphenol conjugates were characterized in terms of structural and functional properties. Results showed that the covalent binding amount into the LF molecule of EGCG, CA and GA was 68, 58 and 17 nmol mg−1, respectively. Covalent insertion of polyphenols into the LF molecule was verified by SDS-PAGE and MALDI-TOF-MS analysis, and in particular the molecular weight was increased from 84 011 Da (LF) to 85 906 Da (LF–CA conjugate). The circular dichroism and Fourier transform infrared spectroscopy analyses revealed that the content of α-helix increased and the contents of the remaining structures decreased, while the differential scanning calorimetry data indicated that the thermal stability of LF–polyphenol conjugates was enhanced after the modification. In addition, the antioxidant activity of LF–polyphenol conjugates was 0.23- to 2.10-fold (ABTS˙+ scavenging assay), and 0.04- to 2.19-fold (reducing power assay) higher than that of the control LF. Moreover, the covalent modification obviously changed the solubility and emulsifying properties of LF. The emulsifying properties of the LF–CA conjugate were better than those of the LF–EGCG and LF–GA conjugates.

Molecular interaction between (−)-epigallocatechin-3-gallate and bovine lactoferrin using multi-spectroscopic method and isothermal titration calorimetry

The molecular interaction between bovine lactoferrin (LF) and (-)-epigallocatechin-3-gallate (EGCG) was investigated in an aqueous solution at pH6.0. The presence of EGCG did not change the size and turbidity of LF-EGCG complex in an aqueous solution until the LF/EGCG molar ratio was over a critical value of 1:25. The fluorescence spectra revealed that both tryptophanyl and tyrosyl groups of LF were associated with the interaction with EGCG. The infrared spectra of freeze-dried LF-EGCG complexes showed that they were different from those of LF and EGCG alone, FTIR and far-UV CD results indicated that EGCG induced a progressive increase in the proportion of α-helix structure at the cost of β-sheet structure of LF. The near-UV CD data testified that LF tertiary conformation was altered in the presence of EGCG. Isothermal titration calorimetry (ITC) analysis implied that EGCG was spontaneously bound to LF by a two-stage mechanism, about 31 EGCG molecules were integrated with 1 molecule LF and hydrogen bonds were always involved in the assembly process.

Food‐Grade Covalent Complexes and Their Application as Nutraceutical Delivery Systems: A Review

Food proteins, polysaccharides, and polyphenols are 3 major food constituents with distinctly different functional attributes. Many proteins and polysaccharides are capable of stabilizing emulsions and foams, thickening solutions, and forming gels, although they differ considerably in their abilities to provide these functional attributes. Many plant polyphenols exhibit beneficial physiological functions, such as antitumor, antioxidant, antibacterial, and antiviral properties. Proteins, polysaccharides, and polyphenols can form complexes with each other, which leads to changes in the functional and nutritional properties of the combined systems. Recently, there has been considerable interest in understanding and utilizing covalent interactions between polyphenols and biopolymers (proteins and polysaccharides). The binary or tertiary conjugates formed may be designed to have physicochemical properties and functional attributes that cannot be achieved using the individual components. This article provides a review of the formation, characterization, and utilization of conjugates prepared using proteins, polysaccharides, and polyphenols. It also discusses the relationship between the structural properties and functionality of the conjugates, and it highlights the bioavailability of bioactive compounds loaded in conjugate-based delivery systems. In addition, it highlights the main challenges to be considered when preparing and analyzing conjugates. This article provides an improved understanding of the chemical reactions that occur between major food ingredients and how they can be utilized to develop biopolymer-based delivery systems with enhanced functional attributes.

Physicochemical characterisation of β-carotene emulsion stabilised by covalent complexes of α-lactalbumin with (−)-epigallocatechin gallate or chlorogenic acid

In this study the impact of covalent complexes of α-lactalbumin (α-La) with (-)-epigallocatechin gallate (EGCG) or chlorogenic acid (CA) was investigated on the physicochemical properties of β-carotene oil-in-water emulsions. EGCG, or CA, was covalently linked to α-La at pH 8.0, as evidenced by increased total phenolic content and declined fluorescence intensity. Compared with those stabilised by α-La alone and α-La-CA or EGCG mixture, the emulsion stabilised by the α-La-EGCG covalent complex exhibited the least changes in particle size and transmission profiles, using a novel centrifugal sedimentation technique, indicating an improvement in the physical stability. The least degradation of β-carotene occurred in the emulsion stabilised with the α-La-EGCG covalent complex when stored at 25 °C. These results implied that protein-polyphenol covalent complexes were able to enhance the physical stability of β-carotene emulsion and inhibit the degradation of β-carotene in oil-in-water emulsion, and the effect was influenced by the types of the phenolic compounds.

Physicochemical properties of β-carotene emulsions stabilized by chlorogenic acid–lactoferrin–glucose/polydextrose conjugates

In this study, the influence of chlorogenic acid (CA)-lactoferrin (LF)-glucose (Glc) conjugate and CA-LF-polydextrose (PD) conjugate on the physicochemical characteristics of β-carotene emulsions was investigated. Novel emulsifiers were formed during Maillard reaction between CA-LF conjugate and Glc/PD. The physicochemical properties of β-carotene emulsions were characterized by droplet size, ζ-potential, rheological behavior, transmission changes during centrifugal sedimentation and β-carotene degradation. Results showed that the covalent attachment of Glc or PD to CA-LF conjugate effectively increased the hydrophilicity of the oil droplets surfaces and strengthened the steric repulsion between the oil droplets. Glucose was better than polydextrose for the conjugation with CA-LF conjugate to stabilize β-carotene emulsions. In comparison with LF and CA-LF-Glc/PD mixtures, CA-LF-Glc/PD ternary conjugates exhibited better emulsifying properties and improved physical stability of β-carotene emulsions during the freeze-thaw treatment. In addition, CA-LF-Glc/PD conjugates significantly enhanced chemical stability of β-carotene in the emulsions against ultraviolet light exposure.

Fabrication mechanism and structural characteristics of the ternary aggregates by lactoferrin, pectin, and (-)-epigallocatechin gallate using multispectroscopic methods.

The ternary aggregates were fabricated by lactoferrin (LF), pectin (high methylated pectin (HMP)/low methylated pectin (LMP)), and (-)-epigallocatechin gallate (EGCG) through three different fabrication methods at pH 5.0. The turbidity, particle size, and ζ-potential of ternary aggregates were influenced by the types of pectin, the concentration of EGCG, and fabrication methods. The fluorescence intensity of LF decreased with an increase in EGCG concentration for all ternary aggregates. Far-UV circular dichroism results indicated that EGCG could alter the secondary structure of LF with an increase in the proportion of β-sheet structure at the cost of unordered coil structure. According to near-UV circular dichroism results, EGCG could also modulate the tertiary structure of LF at the presence of pectin. In addition, EGCG could increase the viscoelasticity of the ternary aggregates with HMP, leading to better stability of the ternary aggregates. An opposite result was observed for the ternary aggregates with LMP. These findings should provide an insight into the fabrication mechanism and applications of ternary aggregates formed by protein, polysaccharide, and polyphenol in the food, pharmaceutical, and cosmetic industries.

Structural characterization, formation mechanism and stability of curcumin in zein-lecithin composite nanoparticles fabricated by antisolvent co-precipitation

Curcumin (Cur) exhibits a range of bioactive properties, but its application is restrained due to its poor water solubility and sensitivity to environmental stresses. In this study, zein-lecithin composite nanoparticles were fabricated by antisolvent co-precipitation technique for delivery of Cur. The result showed that the encapsulation efficiency of Cur was significantly enhanced from 42.03% in zein nanoparticles to 99.83% in zein-lecithin composite nanoparticles. The Cur entrapped in the nanoparticles was in an amorphous state confirmed by differential scanning calorimetry and X-ray diffraction. Fourier transform infrared analysis revealed that hydrogen bonding, electrostatic interaction and hydrophobic attraction were the main interactions among zein, lecithin, and Cur. Compared with single zein and lecithin nanoparticles, zein-lecithin composite nanoparticles significantly improved the stability of Cur against thermal treatment, UV irradiation and high ionic strength. Therefore, zein-lecithin composite nanoparticles could be a potential delivery system for water-insoluble bioactive compounds with enhanced encapsulation efficiency and chemical stability.

Glycosylation improves the functional characteristics of chlorogenic acid–lactoferrin conjugate

In the present study, a chlorogenic acid (CA)–lactoferrin (LF) conjugate prepared via alkali treatment was glycoslated with glucose (Glc) or polydextrose (PD) by the Maillard reaction. Formation of the covalent CA–LF–Glc/PD ternary conjugates was confirmed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), Fourier transform infrared (FTIR) spectroscopy, differential scanning calorimetry (DSC) and fluorescence analyses. The results showed that the grafting ratio between the CA–LF conjugate and Glc/PD was 40.50% for the CA–LF–Glc ternary conjugate and 11.72% for the CA–LF–PD ternary conjugate. Conjugating CA and Glc/PD onto LF changed the conformation of the protein, leading to a reduction in the α-helix content and an increase in the unordered structure. The thermal stability of the CA–LF conjugate was remarkably improved by the Maillard-type conjugation. According to AFM and DLS results, the ternary conjugates showed coarser structures and bigger particle sizes than their mixtures. The reducing power was increased from 206.91 μmol Trolox g−1 of CA–LF conjugate to 255.76 and 273.25 μmol Trolox g−1 sample, respectively, in CA–LF–Glc and CA–LF–PD ternary conjugates, indicating the glycosylation was an effective way to improve the antioxidant activity of the CA–LF conjugate. Moreover, the ternary conjugates were employed to encapsulate β-carotene as a model biologically active macromolecule, and they could enhance the physicochemical stability of β-carotene emulsions. This work presented a simple and general approach to the preparation of polyphenol–protein–polysaccharide conjugates that could be potentially employed as food-grade biomacromolecules in the food and pharmaceutical industries.