Yuqian Liu

Urea-Driven Epigallocatechin Gallate (EGCG) Permeation into the Ferritin Cage, an Innovative Method for Fabrication of Protein–Polyphenol Co-assemblies

The 8 nm diameter cavity endows the ferritin cage with a natural space to encapsulate food components. In this work, urea was explored as a novel medium to facilitate the formation of ferritin-polyphenol co-assemblies. Results indicated that urea (20 mM) could expand the 4-fold channel size of apo-red bean ferritin (apoRBF) with an increased initial iron release rate υ0 (0.22 ± 0.02 μM min-1) and decreased α-helix content (5.6%). Moreover, urea (20 mM) could facilitate the permeation of EGCG into the apoRBF without destroying the ferritin structure and thus form ferritin-EGCG co-assemblies (FECs) with an encapsulation ratio and loading capacity of 17.6 and 2.1% (w/w), respectively. TEM exhibited that FECs maintained a spherical morphology with a 12 nm diameter in size. Fluorescence analysis showed that urea intervention could improve the binding constant K [(1.22 ± 0.8) × 104 M-1] of EGCG to apoRBF. Furthermore, the EGCG thermal stability was significantly improved (20-60 °C) compared with free EGCG. Additionally, this urea-involved method was applicable for chlorogenic acid and anthocyanin encapsulation by the apoRBF cage. Thus, urea shows potential as a novel potential medium to encapsulate and stabilize bioactive polyphenols for food usage based on the ferritin protein cage structure.

Channel directed rutin nano-encapsulation in phytoferritin induced by guanidine hydrochloride

Phytoferritin cage has a nano-sized cavity to encapsulate bioactive molecules. In this work, a novel approach is presented that guanidine hydrochloride (GuHCl) (2mM) can expand the channel of apo-soybean seed ferritin (apoSSF) and promote the encapsulation of rutin molecules in apoSSF at pH 7.0 without the disassembly of the protein cage. Upon removal of GuHCl from SSF, a rutin encapsulation ratio of 10.1% was obtained; and the prepared rutin-loaded ferritin nanoparticles were homogeneously distributed, showing a shell-like morphology with a size of 12nm. By virtue of this interesting method, core molecules can be encapsulated within the ferritin cage in a benign condition without extreme pH changes, which is beneficial for the stability and bioactivity of the pH-sensitive molecules in food encapsulation and delivery of functional molecules.

Thermal Stability Improvement of Rice Bran Albumin Protein Incorporated with Epigallocatechin Gallate

Rice bran albumin protein (RAP) is sensitive to thermal changes and tends to degrade when exposed to high-temperature processing. In this work, RAP-epigallocatechin-3-gallate (EGCG) complex (RAPE) was prepared and the thermal stability was evaluated. Fluorescence results showed that EGCG could interact with RAP with a binding number n of 0.0885:1 (EGCG:RAP, w/w) and a binding constant K of 1.02 (± 0.002) ×104 /M, suggesting both hydrogen bonding and van der Waals forces played an important role. FTIR analysis demonstrated that EGCG could induce secondary structural changes in RAP above a ratio of 1.6:1 (EGCG:RAP, w/w). Interestingly, the secondary structure changes of RAPE at different temperatures (25, 50, 60, 70, and 80 °C) were inhibited compared with that for RAP, suggesting RAPE was more resistant and stable to the heat treatment. In addition, a dense porous structure of RAPE was achieved due to the EGCG binding after thermal treatment. Furthermore, the Tpeak temperature of RAPE increased significantly from 64.58 to 74.16 °C and the enthalpy also increased from 85.53 to 138.52 J/g with a mass ratio increasing from 0 to 3.2 (EGCG:RAP, w/w), demonstrating the thermal stability of RAPE. In addition, the valine, methionine, and lysine content in RAPE were significantly higher than RAP following 80 °C treatment for 20 min (P < 0.05), exhibiting enhanced amino acid profiles, which might be due to EGCG-RAP interactions and microenvironment changes around relevant amino acids. These findings demonstrate that EGCG has the potential to improve the thermal stability of sensitive proteins and is beneficial for usage in the food industry.

Alcalase Enzymolysis of Red Bean (adzuki) Ferritin Achieves Nanoencapsulation of Food Nutrients in a Mild Condition

Classical methods to fabricate ferritin-nutrients shell-core nanoparticles usually apply extremely acid/alkaline pH transition, which may cause the activity loss of nutrients or the formation of insoluble aggregates. In this work, we prepared an extension peptide (EP) deleted red bean (adzuki) ferritin (apoRBFΔEP) by Alcalase 3.0T enzymolysis. Such enzymolysis could delete the EP domain and remain the typical shell-like structure of the ferritin. Meanwhile, the α-helix content of apoRBFΔEP was decreased by 5.5%, and the transition temperature (Tm) was decreased by 4.1 °C. Interestingly, the apoRBFΔEP can be disassembled into subunits under a benign condition at pH 4.0 and is assembled to form an intact cage protein when the pH was increased to 6.7. By using this novel route, the epigallocatechin gallate (EGCG) molecules were successfully encapsulated into the apoRBFΔEP cage with an encapsulation ratio of 11.6% (w/w), which was comparable with that by the traditional pH 2.0 transition. The newly prepared EGCG-loaded apoRBFΔEP exhibited a similarly protective effect on the EGCG upon simulated gastrointestinal tract and thermal treatment as compared with the control. In addition, the EGCG-loaded apoRBFΔEP could significantly relieve the ferritin association induced by pH transition, which was superior to traditional method. The thinking of this work will be especially suitable for encapsulating pH-sensitive molecules based on ferritin in a benign condition.

Nano-encapsulation of epigallocatechin gallate in the ferritin-chitosan double shells: Simulated digestion and absorption evaluation

In this work, a double shell material chitosan (CS)-recombinant soybean seed H-2 ferritin (H-2F) was fabricated to encapsulate epigallocatechin gallate (EGCG) molecules. EGCG-loaded H-2F complex (EHF) was firstly prepared by taking advantages of the reversible self-assembly of the ferritin, and the EHF-CS composite (EHFC) was fabricated by electrostatic interactions with binding number n of (4.1 ± 0.11) and binding constant K of ((5.3 ± 0.2) × 105 M-1), respectively. It was calculated that about 12.6 of EGCG molecules can be encapsulated in one H-2F ferritin cage with an encapsulation efficiency of 9.69% (w/w). SDS-PAGE analysis indicated that the CS binding to H-2F could inhibit ferritin degradation by pepsin and trypsin; the stability of EGCG molecules in EHFC was also significantly improved in simulated gastrointestinal tract. In addition, the chitosan-ferritin double shells were beneficial for the transport of EGCG across the Caco-2 monolayer model based on ferritin uptake. This work demonstrates a novel method to promote stabilization and absorption of food bioactive molecules.

Effect of atmospheric cold plasma on structure, activity, and reversible assembly of the phytoferritin

Ferritin is characterized by a shell-like structure and a reversible self-assembly property. In this study, atmospheric cold plasma (ACP) was applied to red bean seed ferritin (RBF) to prepare an ACP-treated RBF (ACPF). Results indicated that the ACP treatment retained the shell-like structure of ferritin but reduced the α-helix/β-sheet contents and thermal stability. Iron oxidative deposition and release activities were also markedly changed. The ACPF could be disassembled at pH 4.0 and then assembled into an intact ferritin cage when pH was increased to 7.0, which was a more benign transition condition than that of the traditional method (pH 2.0/7.0 transition). By using this assembly routine, curcumin was successfully encapsulated within the ACPF with a size distribution of 12 nm. Moreover, the encapsulation ratio of curcumin in the ACPF reached 12.7% (w/w). This finding can be used to expand the application of ACP and improve the functionalization of the ferritin.

Impact on the nutritional attributes of rice bran following various stabilization procedures

ABSTRACT Rice bran, a valuable byproduct of the rice milling process, has limitations in food industrial applications due to its instability during storage. This review summaries the methodology for stabilization and its impact on the nutritional properties of rice bran. A variety of treatments have been used and these include heat treatment, low-temperature storage, biological and chemical approaches and these will be discussed in terms of their ability to destroy/inhibit enzyme activity and improve storage performance of rice bran. More importantly, changes in the nutritional value of rice bran in terms of vitamins, polyphenols, tocopherols, flavonoids, free fatty acids caused by stabilization of rice bran will also be discussed. This review highlights the importance of appropriate design of processes for stabilization and controlling storage conditions to ensure quality of the rice bran and enhancing levels of phytochemicals in the bran for novel applications in functional foods.

Thermally Induced Encapsulation of Food Nutrients into Phytoferritin through the Flexible Channels without Additives

The cavity of phytoferritin provides a nanospace to encapsulate and deliver food nutrient molecules. However, tranditional methods to prepare the ferritin-nutrient complexes must undergo acid/alkaline conditions or apply additives. In this work, we provide a novel guideline that thermal treatment at 60 °C can expand ferritin channels by uncoiling the surrounding α-helix. Upon reduction of the temperature to 20 °C, food nutrient rutin can be encapsulated in apo-soybean seed ferritin (apoSSF) at pH 7.0 through channels without disassembly of the protein cage and with no addition of additives. Results indicated that one apoSSF could encapsulate about 10.5 molecules of rutin, with an encapsulation ratio of 8.08% (w/w). In addition, the resulting rutin-loaded SSF complexes were monodispersed in a size of 12 nm in aqueous solution. This work provides a novel pathway for the encapsulation of food nutrient molecules into the nanocavity of ferritin under a neutral pH condition induced by thermal treatment.

Microelectric Current Treatment Enhanced Biodegradation of Pumpkin Lignocelluloses by Trichoderma reesei RUT-C30

A homemade microcurrent reactor was used to treat the fermentation of Trichoderma reesei. Results indicated that the yield of saccharides for T. reesei RUT-C30 cultivated in pumpkin lignocellulose broth reaches 38.86% (w/w) when a microcurrent treatment (20 mA, at the 48th hour for 60 min) was carried out, which is significantly higher than the control group (p < 0.05). Additionally, activities of endoglucanase, cellobiohydrolase, xylanase, and pectinase were significantly increased in days 3-7. Furthermore, the fungal growth was facilitated by microelectric treatment, showing a 0.57-fold increase of spore numbers at the sixth day of cultivation. Besides, the monosaccharide composition, including glucose (1.03 mg/mL), xylose (0.12 mg/mL), arabinose (0.31 mg/mL), and fructose (0.13 mg/mL), extracted from the reactor was higher than that without the current treatment. In this work, we improved the biodegradation of lignocellulosic wastes by applying a microcurrent to lignocellulose-degrading fungal cultures and provided a new idea for the lignocellulose material pretreatment and bioconversion.

Ferritin glycosylated by chitosan as a novel EGCG nano-carrier: Structure, stability, and absorption analysis

Ferritin is a shell-like carrier protein with an 8nm diameter cavity which endows a natural space to encapsulate food and drug components. In this work, phytoferritin was unprecedentedly glycosylated by chitosan to fabricate ferritin-chitosan Maillard reaction products (FCMPs) (grafting degree of 26.17%, 24h, 55°C). Results indicated that the amide I and II bands of ferritin were altered due to the chitosan grafting, whereas the ferritin spherical structure were retained. Simulated digestion analysis showed that the FCMPs were more resistant to pepsin and trypsin digestion as compared with ferritin alone. Furthermore, FCMPs were employed as carrier to encapsulate epigallocatechin gallate (EGCG) molecules with an encapsulation ratio of 12.87% (w/w), and the resulting FCMPs-EGCG complexes showed a slow release of EGCG in simulated gastrointestinal tract. Interestingly, different types of food components displayed different effects in EGCG release behavior from the FCMPs, wherein proanthocyanidin, milk and soy protein inhibited the EGCG release. In addition, the absorption of EGCG encapsulated in FCMPs in Caco-2 monolayer model was significantly improved as compared with free EGCG. This work provides a novel nano-vehicle for fabricating core-shell systems in food and drug delivery domain.