Antioxidant synergistic anti-inflammatory effect in the MAPK/NF-κB pathway of peptide KGEYNK (KK-6) from giant salamander (Andrias davidianus).

BACKGROUND: Giant salamander protein peptide is a peptide with rich functional properties. Giant salamander protein peptide KGEYNK (KK-6) is a peptide with both antioxidant and anti-inflammatory properties. The antioxidant and anti-inflammatory mechanisms of KK-6 are still unclear. When we studied the functional mechanism of KK-6, we found that the antioxidant property of KK-6 has a synergistic and promoting effect on anti-inflammatory properties. RESULTS: KK-6 enhances cellular resistance to LPS via the MAPK/NF-κB signaling pathway, leading to increased levels of inflammatory factors: interleukin-1ß (764.81 ng mL-1), interleukin-6 (1.06 ng mL-1) and tumor necrosis factor-α (4440.45 ng mL-1). KK-6 demonstrates potent antioxidant properties by activating the Nrf2 signaling pathway, resulting in elevated levels of antioxidant enzymes (glutathione peroxidase: 0.03 µg mL-1; superoxide dismutase: 0.589 µg mL-1) and a reduction in the concentration of the oxidative product malondialdehyde (967.05 µg mL-1). CONCLUSION: Our findings highlight the great potential of KK-6, a peptide extracted from giant salamander protein, as a remedy for intestinal inflammation. Through its dual role as an antioxidant and anti-inflammatory agent, KK-6 offers a promising avenue for alleviating inflammation-related damage and oxidative stress. This study lays the foundation for further exploration of giant salamander products and highlights their importance in health and novel food development. © 2024 Society of Chemical Industry.

Effect of Ca2+ on 3D printing accuracy and antioxidant efficacy of nano starch-lutein surimi: mechanical properties, chemical bonds and release rate.

BACKGROUND: Nano starch-lutein (NS-L) can be used in three-dimensional (3D) printed functional surimi. However, the lutein release and printing effect are not ideal. The purpose of this study was to facilitate the function and printing properties of surimi by adding the combination of calcium ion (Ca2+ ) and NS-L. RESULTS: Printing properties, lutein release and antioxidation of printed Ca2+ -NS-L-surimi were determined. The NS-L-surimi with 20 mM kg-1 Ca2+ had the best printing effects (fine accuracy, 99 ± 1%). Compared to NS-L-surimi, the structure became denser after adding Ca2+ , the gel strength, hardness, elasticity, yield stress (τ), water holding capacity of Ca2+ -NS-L-surimi increased by about 17 ± 4%, 3 ± 1%, 9 ± 2%, 20 ± 4%, 40 ± 5% respectively. These enhanced mechanical strength and self-supporting ability to resist binding deformation and improve printing accuracy. Moreover, salt dissolution and increased hydrophobic force by Ca2+ stimulated protein stretching and aggregation, leading to enhancement of gel formation. Decreased printing effects of NS-L-surimi with excessive Ca2+ (> 20 mM kg-1 ) caused by excessive gel strength and τ, leading to strong extrusion force and low extrudability. Additionally, Ca2+ -NS-L-surimi had higher digestibility and lutein release rate (increased from 55 ± 2% to 73 ± 3%), because Ca2+ made NS-L-surimi structure porous, which promoted contact of enzyme-protein. Furthermore, weakened ionic bonds reduced electron binding bondage that combined with released lutein to provide more electrons for enhancing antioxidation. CONCLUSION: Collectively, 20 mM kg-1 Ca2+ could better promote printing process and function exertion of NS-L-surimi, facilitating the application of 3D printed functional surimi. © 2023 Society of Chemical Industry.

Excited-state dynamics of all-trans protonated retinal Schiff base in CRABPII-based rhodopsin mimics.

The rhodopsin mimic is a chemically synthetized complex with retinyl Schiff base (RSB) formed between protein and the retinal chromophore that can mimic the natural rhodopsin-like protein. The artificial rhodopsin mimic is more stable and designable than the natural protein and hence has wider uses in photon detection devices. The mimic structure RSB, like the case in the actual rhodopsin-like protein, undergoes isomerization and protonation throughout the photoreaction process. As a result, understanding the dynamics of the RSB in the photoreaction process is critical. In this study, the ultrafast transient absorption spectra of three mutants of the cellular retinoic acid-binding protein II-based rhodopsin mimic at acidic environment were recorded, from which the related excited-state dynamics of the all-trans protonated RSB (AT-PRSB) were investigated. The transient fluorescence spectra measurements are used to validate some of the dynamic features. We find that the excited-state dynamics of AT-PRSB in three mutants share a similar pattern that differs significantly from the dynamics of 15-cis PRSB of the rhodopsin mimic in neutral solution. By comparing the dynamics across the three mutants, we discovered that the aromatic residues near the ß-ionone ring structure of the retinal may help stabilize the AT-PRSB and hence slow down its isomerization rate. The experimental results provide implications on designing a rhodopsin-like protein with significant infrared fluorescence, which can be particularly useful in the applications in biosensing or bioimaging in deeper tissues.

Ultralow-threshold green fluorescent protein laser based on high Q microbubble resonators.

Biological lasers have attracted vast attention because of their potential medical application prospects, especially the low threshold biological laser that can be used for ultrasensitive biological detection while leaving the luminous gain medium undamaged by the pump light. By coupling the low concentration green fluorescent protein (GFP) solution with a high Q whispering gallery mode microbubble resonator, we managed to fabricate a miniature GFP laser with the lowest threshold and highest Q value compared to any known type of the GFP laser. The threshold energy is as low as 380 fJ, two orders of magnitude lower than any type of GFP laser at present. The Q value of the optical cavity in this biological laser is 5.3 × 107, two orders higher than the highest Q value of GFP lasers. We further confirmed the long-term stability of the working characteristics of GFP laser. It can work well nearly a month in temperature 3-4°C. Finally, we measured the effects of different concentrations of fluorescent protein on laser threshold. The data show that this biological laser can be used for highly sensitive detection of GFP concentration, which is particularly useful when the GFP is used as tracers.

Purification and identification of a novel hypotensive and antioxidant peptide from porcine plasma.

BACKGROUND: Pig plasma contains a large amount of protein. Porcine plasma polypeptide can be prepared by the enzymatic hydrolysis of porcine plasma protein. The present study investigated the function, structure, and mechanisms of porcine plasma peptides. RESULTS: The results showed that WVRQAPGKGL had a major ability to scavenge hydroxyl radical scavenging activity (HRSA) (35.25%), 2,2'-azino-bis (3-ethylbenzothiazo line-6-sulfonic acid) diammonium salt radical scavenging activity (ABTS RSA) (93.09%) and 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity (DPPH RSA) (25.72%), as well as in angiotensin converting enzyme (ACE) inhibition (91.64%). WVRQAPGKGL could inactivate ACE by binding to Zn2+ because of the presence of carboxyl in WVRQAPGKGL. The ACE inhibition, HRSA, and DPPH of synthetic WVRQAPGKGL were improved by 12.70%, 16.06%, and 117.11% respectively after in vitro digestion. It (0.1 mg mL-1 ) also increased superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) by 59.78%, 69.05%, and 59.06%, and decreased reactive oxygen species (ROS) and malondialdehyde (MDA) by 22.08% and 50.59%, respectively, to protect HepG2 cells induced by H2 O2 . Furthermore, in a spontaneously hypertensive rat (SHR) model, the systolic blood pressure (SBP) and diastolic blood pressure (DBP) of the peptide group (30 mg kg-1 ) both decreased by about 33.33% in comparison with captopril. CONCLUSION: A new difunctional (antioxidant and hypotensive) peptide, WVRQAPGKGL, derived from porcine plasma hydrolyzate was isolated by gel filtration and reverse phase chromatography, and identified by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS)-1 . The difunctional peptide WVRQAPGKGL from porcine plasma could therefore be used in formulating functional foods or pharmaceuticals. © 2022 Society of Chemical Industry.

Excited-State Dynamics of a CRABPII-Based Microbial Rhodopsin Mimic.

Microbial rhodopsin, a pivotal photoreceptor protein, has garnered widespread application in diverse fields such as optogenetics, biotechnology, biodevices, etc. However, current microbial rhodopsins are all transmembrane proteins, which both complicates the investigation on the photoreaction mechanism and limits their further applications. Therefore, a specific mimic for microbial rhodopsin can not only provide a better model for understanding the mechanism but also can extend the applications. The human protein CRABPII turns out to be a good template for design mimics on rhodopsin due to the convenience in synthesis and the stability after mutations. Recently, Geiger et al. designed a new CRABPII-based mimic M1-L121E on microbial rhodopsin with the 13-cis, syn (13C) isomerization after irradiation. However, it still remains a question as to how similar it is compared with the natural microbial rhodopsin, in particular, in the aspect of the photoreaction dynamics. In this article, we investigate the excited-state dynamics of this mimic by measuring its transient absorption spectra. Our results reveal that there are two components in the solution of mimic M1-L121E at pH 8, known as protonated Schiff base (PSB) and unprotonated Schiff base (USB) states. In both states, the photoreaction process from 13-cis, syn(13C) to all-trans,anti (AT) is faster than that from the inverse direction. In addition, the photoreaction process in the PSB state is faster than that in the USB state. We compared the isomerization time of the PSB state to that of microbial rhodopsin. Our findings indicate that M1-L121E exhibits behaviors similar to those of microbial rhodopsins in the general pattern of PSB isomerization, where the isomerization from 13C to AT is much faster than its inverse direction. However, our results also reveal significant differences in the excited-state dynamics of the mimic relative to the native microbial rhodopsin, including the slower PSB isomerization rates as well as the unusual USB photoreaction dynamics at pH = 8. By elucidating the distinctive characteristics of mimics M1-L121E, this study enhances our understanding of microbial rhodopsin mimics and their potential applications.

The highly stable indicator film incorporating roselle anthocyanin co-pigmented with oxalic acid: Preparation, characterization and freshness monitoring application.

A novel stable PVA/HPMC/roselle anthocyanin (RAE) indicator film co-pigmented with oxalic acid (OA) was prepared, its properties, application effects and stability enhancement mechanism were investigated correspondingly. The structural characterization revealed that more stable network was formed due to the co-pigmentation facilitated generation of molecular interactions. Meanwhile, the co-pigmentation improved film mechanical and hydrophobic properties compared to both PVA/HPMC/RAE newly prepared (PHRN) or stored (PHRS) film, expressing as higher tensile strength values (12.25% and 14.44% higher than PHRN and PHRS), lower water solubility (7.22% and 10.09% lower than PHRN and PHRS) and water vapor permeability values (33.20% and 21.05% lower than PHRN and PHRS) of PVA/HPMC/RAE/OA newly prepared (PHON) or stored (PHOS) film. Compared with the PHRS film, the PHOS film still presented more distinguishable color variations when being applied to monitor shrimp freshness, owing to the stabilization behaviors of co-pigmentation in anthocyanin conformation. Hence, the co-pigmentation was an effective strategy to enhance film stability, physical and pH-responsive properties after long term storage, leading to better film monitoring effects when applied in real-time freshness monitoring.

Enhanced fresh-keeping capacity of printed surimi by Ca2+-nano starch-lutein and its nondestructive freshness monitoring based on 4D printed anthocyanin.

To solve undiscernible freshness changes of printed functional surimi while maintaining printed shape, 4D printable color-changing material were prepared. Firstly, based on results of printing properties and fresh-keeping of Ca2+-NS-L-surimi, it showed better printing effects (enhanced mechanical strength) and good preservation (inhibition of amino acids decomposition, bacterial growth). However, freshness changes of printed Ca2+-NS-L-surimi were not distinguished directly. To avoid that, 4D printable color-changing material-anthocyanin-hydroxypropyl methyl cellulose-xanthan gum-carrageenan (AHXK) was prepared for indicating freshness through discoloration. Printing results showed AHX with 5 % K had the most suitable mechanical strength (appropriate gel strength, texture, rheology) for printing. Based on that, AHXK had stable color (ΔE fluctuation <5) and was sensitive to pH and ammonia (obvious discoloration; ΔE > 10). Actual freshness monitoring results (co-printing of AHXK-surimi) exhibited significant discolorations, especially for HXK with 0.75 % A. It became green during refrigeration of 3-5 d (keeping fresh, ΔE < 4), brighter green at 7 d (decreased freshness, ΔE > 6), turned yellow at 9 d (spoilage, ΔE > 16), which were distinguished significantly with naked eyes rather than traditional freshness determining. In conclusion, printed AHXK-functional surimi exhibited good printing, preservation and nondestructive freshness monitoring, facilitating application of 3D printed functional surimi.

Identification of a novel hypotensive peptide from porcine plasma hydrolysate by in vitro digestion and rat model.

We separated a novel functional peptide IFPPKPKDTL from porcine plasma hydrolysate by chromatography, HPLC, and identified by Q Exactive LC-MS/MS. Results showed that IFPPKPKDTL had a significant ability of ACE inhibition (76.6%) likely due to the presence of hydrophobic, aromatic, and acidic amino acids that can inactivate ACE by binding Zn2+, providing a hydrogen atom to maintain the link between ACE and the peptide. Furthermore, the ACE inhibition of synthetic IFPPKPKDTL was improved by 15.6% after in vitro digestion. Additionally, the systolic blood pressure and diastolic blood pressure of spontaneously hypertensive rats gavaged by the peptide (30 mg/kg). Thereby, ACE inhibitory peptide IFPPKPKDTL from porcine plasma was stable and has potential functional value.

Identification of novel antioxidant peptide from porcine plasma hydrolysate and its effect in in vitro digestion/HepG2 cells model.

A novel antioxidant peptide EDEQKFWGK from porcine plasma hydrolysate (PPH) was separated by chromatography, HPLC, and identified by LC-MS/MS. Results showed that EDEQKFWGK had better antioxidant ability (Hydroxyl RAS 32.19%, ABTS RAS 92.93% and DPPH RAS 26.76%) compared with glutathione (30.11%, 82.01%, 26.44%) due to the presence of hydrophobic, aromatic acids (F, W) and acidic amino acids (E, D), decreasing ROS by providing hydrogen atom and chelating metal ions. Furthermore, the antioxidant properties of synthetic EDEQKFWGK still significant despite in vitro digestion because of the production of smaller active peptide. Additionally, it could increase SOD, CAT, GSH-Px to resist oxidative damage in HepG2 cells by inhibiting ROS (O2- , OH·), forming complexes to prevent OH· from destroying DNA and binding to ARE to promote antioxidase expression. Thereby, the novel peptide EDEQKFWGK from porcine plasma had much stable antioxidant properties and hade great potential in formulating functional foods. PRACTICAL APPLICATIONS: This research isolated a novel antioxidant peptide. Moreover, the antioxidant effects of peptide were confirmed under the in vitro digestion model and oxidative damage HepG2 cells model. The results showed the antioxidant peptide could play better effect after digestion and protect the cells from oxidative damage. These data could expand the sequence data of antioxidant peptides and promote the high-value utilization of PPH.

Differentiation of three commercial tuna species through Q-Exactive Orbitrap mass spectrometry based lipidomics and chemometrics.

Mislabeling and adulteration of tuna are common due to the diminishing of morphological characteristics during processing. The tuna authenticity is now being focused in the seafood supply chain. In this study, the lipid profiles of 3 commercial tuna species (skipjack tuna, bigeye tuna and yellowfin tuna) were investigated via ultra-high performance liquid chromatography coupled with Q-Exactive Orbitrap mass spectrometry (UPLC-Q-Exactive Orbitrap MS). A total of 439 lipid species were identified and semi-quantitated by MS-DIAL. Further biomarkers discovery was carried out by chemometrics, leading to 27 lipids being identified as potential lipid biomarkers. Comparisons to reference standards revealed that lipid biomarkers were effective for discrimination of different tuna species. Interestingly, the proposed lipid biomarkers were all glycerophospholipids, implying that they might be the focus of future study.

Chitosan-based films with antioxidant of bamboo leaves and ZnO nanoparticles for application in active food packaging.

The function of chitosan film was reinforced by ZnO nanoparticles and antioxidant of bamboo leaves (AOB) for food packaging application. The results of structural characterization indicated the good compatibility among chitosan, ZnO nanoparticles and AOB. The chitosan film had the best mechanical strength and the highest light transmittance. The addition of AOB remarkably reduced the UV light transmittance and significantly enhanced the antioxidant activity of the films. Meanwhile, AOB and ZnO nanoparticles synergistically enhanced the antibacterial activity against E. coli and S. aureus. Our results suggested that the chitosan/ZnO/AOB films could be applied as potential active packaging materials in food industry to extend the shelf-life of packaged food.

Determination of the iron bioavailability, conformation, and rheology of iron-binding proteins from Tegillarca granosa.

The increased interest in achieving, solely through diet, the same effect on iron levels with supplementation, leads to numerous studies on iron absorption of iron binding proteins (IBPs). The characteristics of IBPs from Tegillarca granosa (T. granosa) and its iron utilization were determined to analyze their relationship. The results showed in T. granosa, Fe(ӀӀ) was main iron form in hemoglobin (TH) and that Fe(ӀӀ) and Fe(ӀӀӀ) coexisted in ferritin (TF). After in vitro digestion, TH was easier to be digested than TF, bovine hemoglobin, and bovine ferritin. In caco-2 cells model, iron bioavailability of TH also was the best, which related to TH's superior fluid properties, higher ratios of α-helix to ß-sheet and amide I to amide II. These suggest TH could be used as a good source of organic iron and provide references for application of T. granosa in human nutrition. PRACTICAL APPLICATIONS: This research investigated the iron bioavailability and structural properties of iron-binding proteins from Tegillarca granosa (T. granosa). Moreover, the effects of iron absorption in bovine hemoglobin and ferritin were compared with those from T. granosa. The results showed the hemoglobin in T. granosa had better iron bioavailability and it could be a good source of iron. These data could provide a basic instruction of the application of T. granosa in functional food production.