Search Results (89)

This study uniquely explores the impact of a novel natural antifreeze peptide derived from grass carp (GCAFP) on the freeze–thaw characteristics and structural stability of wet gluten protein, providing new insights into the development of natural cryoprotectants for frozen foods. The effects of GCAFP on the physicochemical and structural properties of gluten protein were investigated using differential scanning calorimetry (DSC), nuclear magnetic resonance imaging (NMR), rheology, and scanning electron microscopy (SEM). The results showed that the addition of 0.5% GCAFP significantly reduced the freezing temperature (T_f, from −8.50 ± 1.31 °C to −10.75 ± 2.49 °C) and expanded the melting temperature range (T_m,δ, from 3.60 ± 1.40 °C to 5.65 ± 0.12 °C), indicating improved freezing stability. After five weeks of frozen storage, the ice crystal melting enthalpy (ΔH_m) of gluten protein in the GCAFP group increased by only 20.17 J/g, compared with 27.23 J/g in the control, representing a 6.35% reduction (p < 0.05). Similarly, after five freeze–thaw cycles, the freezable water fraction (F_w) and ΔH_m were reduced by 5.19% and 1.55%, respectively, demonstrating that GCAFP inhibited water migration and ice recrystallization. Low-field NMR revealed that GCAFP maintained a higher proportion of bound water (T21) and decreased free water (T23), confirming its role in restricting water mobility. Rheological analysis showed that GCAFP preserved the viscoelasticity of gluten protein, maintaining higher storage (G′) and loss (G″) moduli than the control after five freeze–thaw cycles, thus mitigating the decline in network elasticity. Structural characterization indicated that GCAFP stabilized the α-helix and β-sheet contents, reduced glutenin macropolymer depolymerization from 24.85% to 18.95%, and strengthened hydrogen bonding within the protein matrix. Overall, GCAFP effectively protected wet gluten protein against ice crystal damage by maintaining water distribution, viscoelasticity, and secondary structure integrity, highlighting its potential as a natural antifreeze ingredient for frozen food applications. Full article

There has been little research on freezing-induced damage in high-moisture dairy products, specifically sour cream and yogurt. This work aimed to investigate, as a proof-of-concept, if antifreeze additives may prevent quality decrease in high-moisture dairy products due to freeze-induced damage. Whey protein isolate and soy protein isolate were complexed with locust bean gum and lambda carrageenan, in both unhydrolyzed and hydrolyzed forms, and their antifreeze activity was evaluated in a model system as well as in sour cream and yogurt. The biomolecules were also tested individually as controls to determine any synergistic effects. Protein and polysaccharide complexes were found to have ice recrystallization inhibition activity in the model systems by reducing the ice crystal size significantly (35–64%) compared to the negative control at both pH 4.5 and 7.0. However, the complexes failed to prevent freeze-induced damage in the dairy system and all treatments resulted in decreased firmness, cohesiveness, and consistency along with increased graininess, possibly due to the complex interacting with different food components that may have interfered with the antifreeze activity of the tested compounds. Full article

Porcine embryo cryopreservation remains challenging due to high lipid content, oxidative stress, and ice recrystallization that compromise post-thaw survival and developmental competence. We evaluated an integrated vitrification approach combining antioxidants (berberine, melatonin), iron oxide (Fe₃O₄) nanoparticles, and antifreeze protein I (AFP I) with post-thaw interventions (glutathione and zona pellucida digestion) to synergistically improve cryosurvival and developmental competence of porcine parthenogenetic embryos. In vitro-matured parthenogenetic embryos were vitrified on Cryotop using a protocol that included berberine and melatonin in embryo culture, Fe₃O₄ nanoparticles and AFP I in cryoprotectant solutions, and post-warming treatment with glutathione and a brief zona pellucida digestion. Survival, hatching, adenosine triphosphate (ATP) content, reactive oxygen species (ROS) levels, cytoskeletal integrity, and the expression of BAX, BCL2, OCT4, and SOX2 genes were measured. Both the dual antioxidant (berberine + melatonin) and nanoparticle + AFP interventions produced greater improvements than individual additives. Fully integrating all components yielded the highest post-thaw viability, with ~94% survival and ~90% hatching, values statistically equivalent to those of fresh embryos. Treated embryos also showed significantly higher ATP levels, lower ROS accumulation (approaching levels in fresh embryos), and preserved microtubule structure (~91% normal). Vitrification alone upregulated BAX and downregulated BCL2, OCT4, and SOX2, whereas the integrated protocol restored their expression levels to near control levels. This multi-component antioxidant, nanoparticle, antifreeze strategy synergistically enhances the cryotolerance and developmental competence of vitrified porcine embryos by mitigating oxidative stress and cryoinjury. Post-thaw viability and molecular markers were restored to near-fresh conditions, demonstrating a promising approach to improve embryo cryopreservation outcomes in swine and potentially other species. Full article

Aqueous batteries, which replace flammable organic electrolytes with water, offer advantages such as intrinsic safety, low cost, and environmental friendliness, making them well-suited to the energy storage needs driven by the increasing proliferation of renewable energy. However, their widespread adoption is hampered by the narrow electrochemical stability window of water, dendrite growth on metal anodes, and various parasitic interfacial reactions. This review proposes a unified three-part framework for biomimetic electrolytes—SEI-mimetic, antifreeze-protein-mimetic, and ion-channel-mimetic—corresponding to three mechanistic strands—water activity regulation, interfacial mechanics, and sub-nanometer transport—to organize and compare various strategies. This paper systematically reviews and evaluates the latest advances in biomimetic electrolytes. It discusses these three biomimetic concepts and their applications in different battery chemistries (monovalent and multivalent metal systems, as well as aqueous redox-flow batteries). It also proposes a roadmap and engineering thresholds for both basic research and commercialization. Full article

Freezing is a crucial technology for preserving food quality and extending shelf life. However, frozen storage often leads to protein oxidation, degradation, and cellular structural damage, compromising food palatability. To address these challenges, antifreeze substances have emerged as a promising solution. This review comprehensively summarizes the current research on antifreeze substances, including natural compounds and artificial analogs. For natural antifreeze substances, the mechanisms of antifreeze proteins (AFPs), antifreeze peptides (AFPPs), antifreeze polysaccharides (AFPLs), and antifreeze phosphates (AFPSs) are elucidated. Additionally, the preparation of artificial synthesis analogs and the application of antifreeze substances are discussed. By presenting their properties and research advancements, this review aims to provide a reference for the practical utilization of antifreeze substances in food-freezing applications. Full article

Developing safe and efficient cryoprotectants is critical for effective cryopreservation in biomedical applications. Inspired by natural antifreeze proteins (AFPs), a series of linear and star-shaped peptoids featuring isopropanol side chains to mimic the amphiphilic characteristics of threonine were prepared. The effects of chain length and molecular topology on antifreeze properties were systematically investigated. Both ice recrystallization inhibition (IRI) and ice crystal growth suppression improved with increasing chain length, and star-shaped peptoids exhibited superior performance. Notably, the star-shaped peptoid S-(A₆)₃ demonstrated excellent antifreeze activity and low cytotoxicity, highlighting its promise as a novel, non-toxic alternative to conventional cryoprotectants like DMSO. These findings provide valuable insight into the structure-property relationship of peptoids for cryopreservation applications. Full article

This review provides an overview of terrestrial extremophiles, highlighting their adaptive strengths and strategies for coping with environmental challenges through the use of specialized proteins. It also explores why their unique lifestyle and ability to adapt to extreme conditions have become a major focus of research, as well as the main benefits and advancements in the study of these organisms in recent decades. This review aims to present an objective summary of the knowledge acquired and its translation into applied science and biotechnological applications. Full article

The antifreeze and antioxidant capacities of tilapia (Oreochromis mossambicus) gelatin hydrolysates were investigated, after glycosylation with saccharides of varying molecular weights, to enhance their functional properties to widen its commercial application in frozen aquatic products. Glycosylation was conducted by mixing gelatin hydrolysates with ribose, glucose, maltose, and dextran (20 kDa) at a 1:1 mass ratio; the glycosylation products had a pH of 10 and were incubated at 80 °C for 1 h. The results showed that the glycosylation degree ranked as: ribose > glucose > maltose > dextran. The mass spectrometry analysis showed that 17, 32, and 5 glycosylation sites were identified for ribose, glucose, and maltose, respectively, suggesting a molecular weight-dependent effect. Spectroscopic analyses, including ultraviolet and infrared spectroscopy, revealed that the gelatin hydrolysate structure was expanded, with chromophores in hydrophilic environments; a blue shift in the amide A and II bands confirmed that the amino group was involved. Fluorescence spectroscopy showed conformational changes with a red shift at 303.4 nm and a reduction in intensity. Antifreeze activity, such as catalase freezing protection and shrimp surimi protein stability, and antioxidant activity, including radical scavenging and metal ion chelation, were significantly improved. Ribose exhibited the strongest effects, followed by maltose and glucose. These results demonstrate the potential of glycosylation to improve gelatin hydrolysates for functional applications. Full article

Cryopreservation can adversely affect sperm motility, structural integrity, and fertilization ability. This study investigated the effects of MitoQ and antifreeze protein III (AFP III) on frozen–thawed semen from eight adult dogs using a Tris–fructose extender. Ejaculates were divided and diluted with a standard Tris–fructose–egg yolk extender containing MitoQ (200 nM/mL) and AFP III (0.75, 1.0, 2.0 µg/mL), individually or combined. Post-thaw, samples were evaluated for motility, viability, membrane and acrosome integrity, lipid peroxidation, apoptosis indicators, mitochondrial function, and reactive oxygen species (ROS-H₂O₂). The results showed significant (p < 0.05) improvements in motility rate, progressive motility, VAP, VSL, VCL, ALH, and BCF with MitoQ or AFP alone. AFP III (0.75, 1.0 µg/mL) showed higher values than controls (p > 0.05), while MitoQ alone showed no significant effect. Viability and acrosome integrity improved with AFP III. Membrane integrity and lipid peroxidation were better in 0.75 and 1.0 µg/mL AFP III groups. ROS-H₂O₂ levels and mitochondrial membrane potential were unaffected except at 1.0 µg/mL AFP III. The phosphatidylserine translocation assay showed no significant differences in dead sperm between controls and individual treatments, but significant differences occurred with combined MitoQ/AFP III. In conclusion, AFP III and MitoQ in diluents protect canine sperm cells from cryodamage. Full article

Sliver carp is a nutritious and abundant species in China, but its low market value stems from its thin meat, small bones and strong odor. Processing it into surimi enhances its economic value, though surimi typically has low gel strength and is prone to deterioration. Recently, three-dimensional (3D) printing has gained attention as an innovative additive manufacturing technique for personalization and process simplification requiring high-performance materials. This study intended to develop an optimized surimi formula for 3D printing with dynamic high-pressure microfluidization (DHPM)-modified pea protein isolate (PPI) and microcrystalline cellulose (MCC). Firstly, the effect of DHPM on PPI properties was evaluated, followed by the optimization of the surimi gel formula (72.093% water content, 3.203% PPI, 1.728% MCC, 1% salt, 1% collagen peptide and 20.976% sliver carp paste) and 3D printing parameters (2000 mm/min at 25 °C with a 1.5 mm nozzle). Rheological comparisons between the optimized surimi, surimi with commercial antifreeze and surimi with only PPI or MCC indicated that the optimized formulation exhibited clearer 3D printing outlines and reduced stickiness due to a higher recovery and lower loss modulus. These results demonstrated that DHPM-treated PPI and MCC enhanced the 3D printability of silver carp surimi gel, providing a new idea for a surimi product and supporting its potential applications in food 3D printing. Full article

Antifreeze agents play a critical role in various fields including tissue engineering, gene therapy, therapeutic protein production, and transplantation. Commonly used antifreeze agents such as DMSO and other organic substances are known to have cytotoxic effects. Antifreeze proteins sourced from cold-adapted organisms offer a promising solution by inhibiting ice crystal formation; however, their effectiveness is hindered by a dynamic ice-shaping (DIS) effect and thermal hysteresis (TH) properties. In response to these limitations, antifreeze peptides (AFPs) have been developed as alternatives to antifreeze proteins, providing similar antifreeze properties without the associated drawbacks. This review explores the methods for acquiring AFPs, with a particular emphasis on chemical synthesis. It aims to offer valuable insights and practical implications to drive the realm of sub-zero storage. Full article

The Chinese rare minnow (Gobiocypris rarus), an important model fish in China, faces endangerment in the wild. Sperm cryopreservation facilitates the development of new strains and germplasm conservation, but the quality of its cryopreserved sperm remains low. This study evaluates the protective effects of different concentrations of antifreeze proteins (AFP I and AFP III) on the cryopreservation of Chinese rare minnow sperm. Cryopreserved sperm showed significant declines in progressive motility, curvilinear velocity (VCL), average path velocity (VAP), and lifespan compared to fresh sperm, except for straight-line velocity (VSL). The cryomedium containing 10 μg/mL AFP I improved these parameters to their highest levels. However, no significant difference was found in progressive motility and kinetic parameters between cryopreserved sperm with and without AFPs. Cryopreserved sperm with 10 μg/mL AFP I showed the highest plasma membrane integrity, mitochondrial activity, and DNA integrity, significantly better than without AFPs; importantly, the fertilization rate of cryopreserved sperm with 10 μg/mL AFP I was not significantly different from that of fresh sperm. These results indicate that the addition of 10 μg/mL AFP I to the cryomedium for Chinese rare minnow sperm does not improve kinetic parameters but significantly enhances sperm quality, aiding in its new strain development and germplasm conservation. Full article

Cryopreservation is a crucial technique for the long-term ex situ conservation of plant genetic resources, particularly in the context of global biodiversity decline. This process entails freezing biological material at ultra-low temperatures using liquid nitrogen, which effectively halts metabolic activities and preserves plant tissues over extended periods. Over the past seven decades, a plethora of techniques for cryopreserving plant materials have been developed. These include slow freezing, vitrification, encapsulation dehydration, encapsulation–vitrification, droplet vitrification, cryo-plates, and cryo-mesh techniques. A key challenge in the advancement of cryopreservation lies in our ability to understand the molecular processes underlying plant freezing tolerance. These mechanisms include cold acclimatization, the activation of cold-responsive genes through pathways such as the ICE–CBF–COR cascade, and the protective roles of transcription factors, non-coding RNAs, and epigenetic modifications. Furthermore, specialized proteins, such as antifreeze proteins (AFPs) and late embryogenesis abundant (LEA) proteins, play crucial roles in protecting plant cells during freezing and thawing. Despite its potential, cryopreservation faces significant challenges, particularly in standardizing protocols for a wide range of plant species, especially those from tropical and subtropical regions. This review highlights the importance of ongoing research and the integration of omics technologies to improve cryopreservation techniques, ensuring their effectiveness across diverse plant species and contributing to global efforts regarding biodiversity conservation. Full article

The effects of MRS, whey protein and blueberry alone, and mixed fermentation on the survival rate of lactic acid bacteria under various freeze-drying conditions were investigated. The surface structure of the freeze-dried powders was also investigated to explore the anti-freezing protection mechanism of mixed whey protein and blueberry fermentation on the bacteria. It was found that the mixed fermentation medium of blueberry and whey protein has a protective effect on the freeze-drying bacteria and is better than the traditional MRS and whey protein medium. The optimal concentration of blueberry juice addition was 9%. The survival rate of the pre-freezing temperature at −80 °C was higher than at −20 °C after the pre-freezing and freeze-drying processes. The freeze-drying thickness of 0.3 cm could improve the survival rate of the bacteria. The Fourier transform infrared spectroscopy results indicated the interaction between the whey protein, anthocyanins, and the surface composition of the lactic acid bacteria. Full article

In this study, ultrafiltration fractions (<3 k Da, LMH; >3 k Da, HMH) and solid-phase extraction fractions (hydrophilic hydrolysate, HIH; hydrophobic hydrolysate, HOH) from trypsin hydrolysate purified from croceine croaker (Pseudosciaena crocea) isolate were obtained to investigate the cryoprotective effects of the different fractions, achieved by means of maceration of turbot fish meat after three freeze-thaw cycles. Alterations in the texture, color, moisture loss, myofibrillar protein oxidation stability and conformation, and microstructure of the fish were analyzed after freezing and thawing. The results demonstrate that HIH maximized the retention of fish texture, reduced moisture loss, minimized the oxidation and aggregation of myofibrillar proteins, and stabilized the secondary and tertiary structures of myofibrillar proteins compared to the control group. In conclusion, the HIH component in the trypsin hydrolysates of croceine croaker significantly contributes to minimizing freeze damage in fish meat and acts as an anti-freezing agent with high industrial application potential. Full article