Search Results (20)

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

Understanding characteristic post-source decay (PSD) fragmentation patterns in tandem mass spectrometry (MS/MS) is important for the identification of target molecules. In this study, we explored the characteristic PSD patterns associated with O-linked glycopeptides and their cyclization using the MALDI-TOF/TOF MS analysis of linear and cyclic antifreeze glycoproteins. We performed a comparative analysis of the proton and sodium adduct ions of the peptide backbones of antifreeze glycoproteins, which have a simple repeating sequence, shedding light on the characteristics of the fragmentation of the threonine side chain and that of its cyclized form. Furthermore, the presence or absence of a glycan on the threonine side chain and its substitution with serine caused changes in its fragmentation. These findings are expected to contribute to the prediction of three-dimensional peptide structures and the search for physiologically active O-linked glycopeptides and cyclic (glyco)peptides. 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

Antifreeze peptides have become effective antifreeze agents for frozen products, but their low quantity of active ingredients and high cost limit large-scale application. This study used the glycosylation of fish collagen peptides with glucosamine hydrochloride catalyzed by transglutaminase to obtain a transglutaminase-catalyzed glycosylation product (TGP) and investigate its antifreeze effect on tilapia. Compared with the blank group, the freshness (pH value of 6.31, TVB-N value of 21.7 mg/100 g, whiteness of 46.28), textural properties (especially hardness and elasticity), and rheological properties of the TGP groups were significantly improved. In addition, the protein structures of the samples were investigated using UV absorption and fluorescence spectroscopy. The results showed that the tertiary structure of the TGP groups changed to form a dense polymer. Therefore, this approach can reduce the denaturation and decomposition of muscle fibers and proteins in fish meat more effectively and has a better protective effect on muscle structure and protein aggregation, improving the stability of fish meat. This study reveals an innovative method for generating antifreeze peptides by enzymatic glycosylation, and glycosylated fish collagen peptide products can be used as new and effective green antifreeze agents in frozen foods. Full article

Aquatic products are gaining popularity due to their delicacy and high nutrient value. However, they are perishable, with a short shelf-life. Frozen storage is associated with adverse effects, leading to protein oxidation and degradation, thereby altering the protein’s structural integrity and subsequently influencing the palatability of protein-based food products. To address these challenges, novel antifreeze peptides have gained significant attention. Antifreeze peptides are a class of small molecular weight proteins or protein hydrolysates that offer protection to organisms in frozen or sub-frozen environments. They offer distinct advantages over conventional commercial antifreeze agents and natural antifreeze proteins. This review provides an overview of the current state of research on antifreeze agents, elucidates their characteristics and mechanisms, and examines their applications in aquatic products. Furthermore, the article offers insights into the prospective development and application prospects of antifreeze peptides. Full article

Antifreeze peptides are a class of small molecule protein hydrolysates that protect frozen products from cold damage under freezing or subcooling conditions. In this study, three different Pseudosciaena crocea (P. crocea) peptides were from pepsin, trypsin, and neutral protease enzymatic hydrolysis. It aimed to elect the P. crocea peptides with better activity through molecular weight, antioxidant activity, and amino acid analysis, as well as to compare the cryoprotective effects with a commercial cryoprotectant. The results showed that the untreated fillets were prone to be oxidized, and the water-holding capacity after freeze-thaw cycle decreased. However, the treatment of the trypsin hydrolysate of P. crocea protein significantly promoted the water-holding capacity level and reduced the loss of Ca²⁺-ATP enzyme activity and the structural integrity damage of myofibrillar protein in surimi. Moreover, compared with 4% sucrose-added fillets, trypsin hydrolysate treatment enhanced the umami of frozen fillets and reduced the unnecessary sweetness. Therefore, the trypsin hydrolysate of P. crocea protein could be used as a natural cryoprotectant for aquatic products. Hence, this study provides technical support for its use as a food additive to improve the quality of aquatic products after thawing and provides a theoretical basis and experimental foundation for the in-depth research and application of antifreeze peptides. Full article

The purpose of this study is to obtain new antifreeze peptides (AFPs) that are natural, safe, and high activity from Ctenopharyngodon idella scales. The optimal hydrolysis conditions were investigated, and chromatography-based isolation was conducted using thermal hysteresis activity (THA) as an index. Molecular dynamic simulation (MDs) was explored to reveal the antifreeze mechanism of the AFPs. The results showed that the optimal hydrolysis conditions were 4000 U/g papain enzyme for 60 °C at pH 5.0 and substrate concentration (1:10) for 3 h, as unveiled by single-factor experiment results. The AFPs documented a THA of 2.7 °C when the T_h was 1.3 °C. Hydrophilic peptide, named GCFSC-AFPs, with a THA of 5.09 °C when the T_h was 1.1 °C was obtained after a series isolation of gel filtration, ion exchange, and reversed-phase HPLC chromatography. The AFPs had a molecular weight of 1107.54~1554.72 Da with three main peptides in the amino acid sequence of VGPAGPSGPSGPQ, RGSPGERGESGPAGPSG, and VGPAGPSGPSGPQG, respectively. The survival rate of yeast with GCFSC-AFPs reached 84.4% following one week of exposure at −20 °C. MDs indicated that GCFSC-AFPs interfered with the ice-water interaction and thus inhibited the ice crystallization process. Our data suggested that the GCFSC-AFPs were a novel and potential antifreeze agent in the food industry. Full article

Antifreeze proteins (AFPs) from the model crop, Brachypodium distachyon, allow freeze survival and attenuate pathogen-mediated ice nucleation. Intriguingly, Brachypodium AFP genes encode two proteins, an autonomous AFP and a leucine-rich repeat (LRR). We present structural models which indicate that ice-binding motifs on the ~13 kDa AFPs can “spoil” nucleating arrays on the ~120 kDa bacterial ice nucleating proteins used to form ice at high sub-zero temperatures. These models are consistent with the experimentally demonstrated decreases in ice nucleating activity by lysates from wildtype compared to transgenic Brachypodium lines. Additionally, the expression of Brachypodium LRRs in transgenic Arabidopsis inhibited an immune response to pathogen flagella peptides (flg22). Structural models suggested that this was due to the affinity of the LRR domains to flg22. Overall, it is remarkable that the Brachypodium genes play multiple distinctive roles in connecting freeze survival and anti-pathogenic systems via their encoded proteins’ ability to adsorb to ice as well as to attenuate bacterial ice nucleation and the host immune response. Full article

Defatted Antarctic krill powder is the main by-product in the manufacturing of krill oil. Exploring a high value-added approach for utilizing this protein-rich material has received much attention in research and industry. Given this, the preparation and primary characterization of antifreeze peptides from defatted Antarctic krill (AKAPs) were carried out in this study. The cryoprotective effect of AKAPs on Lactobacillus rhamnosus ATCC7469 was also investigated. The results showed that Protamex was the optimum protease for AKAP preparation from defatted Antarctic krill. AKAPs were found to be rich in short peptides, with the MW ranging from 600 to 2000 Da (69.2%). An amino acid composition analysis showed that AKAPs were rich in glutamic acid (18.71%), aspartic acid (12.19%), leucine (7.87%), and lysine (7.61%). After freezing, the relative survival rate of Lactobacillus rhamnosus in the 1.0 mg/mL AKAP-treated group (96.83%) was significantly higher than in the saline group (24.12%) (p < 0.05). AKAPs also retarded the loss of acidifying activity of L. rhamnosus after freezing. AKAPs showed even better cryoprotective activity than three commercial cryoprotectants (sucrose, skim milk, and glycerol). In addition, AKAPs significantly alleviated the decrease in β-galactosidase and lactic dehydrogenase activities of L. rhamnosus (p < 0.05). Furthermore, AKAPs effectively protected the integrity of L. rhamnosus cell membranes from freezing damage and alleviated the leakage of intracellular substances. These findings demonstrate that AKAPs can be a potential cryoprotectant for preserving L. rhamnosus, providing a new way to use defatted Antarctic krill. Full article

Antifreeze peptides can protect cell membranes and maintain the cell viability of probiotics under cold stress. Given this, antifreeze peptides were prepared from tilapia processing byproducts of tilapia skin by enzymolysis using the response surface methodology (RSM) method. The cryoprotective effects on Lacticaseibacillus rhamnosus ATCC7469 were investigated. Trypsin was selected as the protease for tilapia skin hydrolysis. The optimal hydrolysis conditions consisted of the amount of enzyme (2200 U/g), solid–liquid ratio (1:10, w/v), reaction temperature (49 °C), and reaction time (6.8 h), and the relative survival rate of L. rhamnosus reached 98.32%. Molecular weight (M_w) distribution and peptide sequences of the antifreeze peptides prepared from tilapia skin (APT) under the optimal conditions were analyzed. APT significantly reduced the leakage of extracellular proteins and protected β-galactosidase and lactate dehydrogenase activities of L. rhamnosus. Compared with the saline group, scanning electron microscopy (SEM) observation showed that cells had a more normal, smooth, and entire surface under the protection of APT. These findings indicate that APT can be a new cryoprotectant in preserving probiotics. Full article

Antifreeze proteins (AFPs) or thermal hysteresis (TH) proteins are biomolecular gifts of nature to sustain life in extremely cold environments. This family of peptides, glycopeptides and proteins produced by diverse organisms including bacteria, yeast, insects and fish act by non-colligatively depressing the freezing temperature of the water below its melting point in a process termed thermal hysteresis which is then responsible for ice crystal equilibrium and inhibition of ice recrystallisation; the major cause of cell dehydration, membrane rupture and subsequent cryodamage. Scientists on the other hand have been exploring various substances as cryoprotectants. Some of the cryoprotectants in use include trehalose, dimethyl sulfoxide (DMSO), ethylene glycol (EG), sucrose, propylene glycol (PG) and glycerol but their extensive application is limited mostly by toxicity, thus fueling the quest for better cryoprotectants. Hence, extracting or synthesizing antifreeze protein and testing their cryoprotective activity has become a popular topic among researchers. Research concerning AFPs encompasses lots of effort ranging from understanding their sources and mechanism of action, extraction and purification/synthesis to structural elucidation with the aim of achieving better outcomes in cryopreservation. This review explores the potential clinical application of AFPs in the cryopreservation of different cells, tissues and organs. Here, we discuss novel approaches, identify research gaps and propose future research directions in the application of AFPs based on recent studies with the aim of achieving successful clinical and commercial use of AFPs in the future. Full article

The intracellular homeostasis of lyophilized lactic acid bacteria (LAB) is destroyed by extreme cold stress, resulting in decreased stability. This study aimed to verify the validity of collagen as a potential protective agent for improving microbial stability deteriorated by freezing. The collagen types used in this study were low molecular weight collagen (LC) of less than 1000 Da and low molecular weight collagen-peptide (LCP) of less than 300 Da. By the accelerated stability test according to the addition of each collagen type, a 3% LCP displaying a protective effect on the viability of various LAB strains (Lactoplantibacillus plantarum MG989, Lactococcus lactis MG5125, Enterococcus faecium MG5232, Bifidobacterium animalis ssp. lactis MG741, and Streptococcus thermophilus MG5140) was finally selected. It was evaluated whether LCP enhances bacterial stability, survivability in the gastrointestinal (GI) tract, and heat resistance. LCP significantly improved the viability of all strains in the GI tract compared to sucrose and skim milk, which are conventional protective agents. Based on morphological observations, LCP was uniformly coated on the cell surface, resulting in protective effects against multiple external stress stimuli. Such findings indicate the applicability of LCP as an unprecedented protective agent, which can improve the stability of various probiotics with antifreeze effects. Full article

Beetle hyperactive antifreeze protein (AFP) has a unique ability to maintain a supercooling state of its body fluids, however, less is known about its origination. Here, we found that a popular stag beetle Dorcus hopei binodulosus (Dhb) synthesizes at least 6 isoforms of hyperactive AFP (DhbAFP). Cold-acclimated Dhb larvae tolerated −5 °C chilled storage for 24 h and fully recovered after warming, suggesting that DhbAFP facilitates overwintering of this beetle. A DhbAFP isoform (~10 kDa) appeared to consist of 6−8 tandem repeats of a 12-residue consensus sequence (TCTxSxNCxxAx), which exhibited 3 °C of high freezing point depression and the ability of binding to an entire surface of a single ice crystal. Significantly, these properties as well as DNA sequences including the untranslated region, signal peptide region, and an AFP-encoding region of Dhb are highly similar to those identified for a known hyperactive AFP (TmAFP) from the beetle Tenebrio molitor (Tm). Progenitor of Dhb and Tm was branched off approximately 300 million years ago, so no known evolution mechanism hardly explains the retainment of the DNA sequence for such a lo­ng divergence period. Existence of unrevealed gene transfer mechanism will be hypothesized between these two phylogenetically distant beetles to acquire this type of hyperactive AFP. Full article

Antifreeze proteins (AFPs) are specific proteins, glycopeptides, and peptides made by different organisms to allow cells to survive in sub-zero conditions. AFPs function by reducing the water’s freezing point and avoiding ice crystals’ growth in the frozen stage. Their capability in modifying ice growth leads to the stabilization of ice crystals within a given temperature range and the inhibition of ice recrystallization that decreases the drip loss during thawing. This review presents the potential applications of AFPs from different sources and types. AFPs can be found in diverse sources such as fish, yeast, plants, bacteria, and insects. Various sources reveal different α-helices and β-sheets structures. Recently, analysis of AFPs has been conducted through bioinformatics tools to analyze their functions within proper time. AFPs can be used widely in various aspects of application and have significant industrial functions, encompassing the enhancement of foods’ freezing and liquefying properties, protection of frost plants, enhancement of ice cream’s texture, cryosurgery, and cryopreservation of cells and tissues. In conclusion, these applications and physical properties of AFPs can be further explored to meet other industrial players. Designing the peptide-based AFP can also be done to subsequently improve its function. Full article