Recent advances on enhancing 3D printing quality of protein-based inks: A review.

3D printing is an additive manufacturing technology that locates constructed models with computer-controlled printing equipment. To achieve high-quality printing, the requirements on rheological properties of raw materials are extremely restrictive. Given the special structure and high modifiability under external physicochemical factors, the rheological properties of proteins can be easily adjusted to suitable properties for 3D printing. Although protein has great potential as a printing material, there are many challenges in the actual printing process. This review summarizes the technical considerations for protein-based ink 3D printing. The physicochemical factors used to enhance the printing adaptability of protein inks are discussed. The post-processing methods for improving the quality of 3D structures are described, and the application and problems of fourth dimension (4D) printing are illustrated. The prospects of 3D printing in protein manufacturing are presented to support its application in food and cultured meat. The native structure and physicochemical factors of proteins are closely related to their rheological properties, which directly link with their adaptability for 3D printing. Printing parameters include extrusion pressure, printing speed, printing temperature, nozzle diameter, filling mode, and density, which significantly affect the precision and stability of the 3D structure. Post-processing can improve the stability and quality of 3D structures. 4D design can enrich the sensory quality of the structure. 3D-printed protein products can meet consumer needs for nutritional or cultured meat alternatives.

Solubilization of fish myofibrillar proteins in NaCl and KCl solutions: A DIA-based proteomics analysis.

Understanding the basic solubilization of fish myofibrillar proteins (MPs) in common monovalent chloride solutions is crucial for muscle food processing. In this study, the differential proteomic profiles of MPs during extraction and solubilization in NaCl and KCl solutions were investigated by using advanced four-dimensional data-independent acquisition (4D DIA) quantitative proteomics for the first time. Compared to routine biochemical analysis, this could provide insights into the solubilization of muscle proteins. We ensure the consistency of the effective ionic strength of NaCl and KCl buffers by adjusting the conductivity. The results showed that NaCl extractor mainly facilitated the solubilization of cytoskeletal proteins, biochemical enzymes, and stromal proteins compared to KCl, such as tubulin, myosin-9, collagen, plectin, protein phosphatase, and cathepsin D. However, no significant difference was observed in the extraction of major sarcomeric proteins, including myosin, actin, troponin C, myosin-binding protein C, M-Protein, α-actinin-3, and tropomyosin.

Efficient Polytelluride Anchoring for Ultralong-Life Potassium Storage: Combined Physical Barrier and Chemisorption in Nanogrid-in-Nanofiber.

Metal tellurides (MTes) are highly attractive as promising anodes for high-performance potassium-ion batteries. The capacity attenuation of most reported MTe anodes is attributed to their poor electrical conductivity and large volume variation. The evolution mechanisms, dissolution properties, and corresponding manipulation strategies of intermediates (K-polytellurides, K-pTex) are rarely mentioned. Herein, we propose a novel structural engineering strategy to confine ultrafine CoTe2 nanodots in hierarchical nanogrid-in-nanofiber carbon substrates (CoTe2@NC@NSPCNFs) for smooth immobilization of K-pTex and highly reversible conversion of CoTe2 by manipulating the intense electrochemical reaction process. Various in situ/ex situ techniques and density functional theory calculations have been performed to clarify the formation, transformation, and dissolution of K-pTex (K5Te3 and K2Te), as well as verifying the robust physical barrier and the strong chemisorption of K5Te3 and K2Te on S, N co-doped dual-type carbon substrates. Additionally, the hierarchical nanogrid-in-nanofiber nanostructure increases the chemical anchoring sites for K-pTex, provides sufficient volume buffer space, and constructs highly interconnected conductive microcircuits, further propelling the battery reaction to new heights (3500 cycles at 2.0 A g-1). Furthermore, the full cells further demonstrate the potential for practical applications. This work provides new insights into manipulating K-pTex in the design of ultralong-cycling MTe anodes for advanced PIBs.

Postmortem Muscle Proteome Characteristics of Silver Carp (Hypophthalmichthys molitrix): Insights from Full-Length Transcriptome and Deep 4D Label-Free Proteomic.

The coverage of the protein database directly determines the results of shotgun proteomics. In this study, PacBio single-molecule real-time sequencing technology was performed on postmortem silver carp muscle transcripts. A total of 42.43 Gb clean data, 35,834 nonredundant transcripts, and 15,413 unigenes were obtained. In total, 99.32% of the unigenes were successfully annotated and assigned specific functions. PacBio long-read isoform sequencing (Iso-Seq) analysis can provide more accurate protein information with a higher proportion of complete coding sequences and longer lengths. Subsequently, 2671 proteins were identified in deep 4D proteomics informed by a full-length transcriptomics technique, which has been shown to improve the identification of low-abundance muscle proteins and potential protein isoforms. The feature of the sarcomeric protein profile and information on more than 30 major proteins in the white dorsal muscle of silver carp were reported here for the first time. Overall, this study provides valuable transcriptome data resources and the comprehensive muscle protein information detected to date for further study into the processing characteristic of early postmortem fish muscle, as well as a spectral library for data-independent acquisition and data processing. This batch of muscle-specific dependent acquisition data is available via PRIDE with identifier PXD043702.

Insight into Ionic Strength-Induced Solubilization of Myofibrillar Proteins from Silver Carp (Hypophthalmichthys molitrix): Structural Changes and 4D Label-Free Proteomics Analysis.

In this study, changes in the physical, structural, and assembly characteristics of silver carp myofibrillar proteins (MPs) at different ionic strength (I) values were investigated. Moreover, the differential proteomic profile of soluble MPs was analyzed using 4D proteomics based on timsTOF Pro mass spectrometry. Solubility of MPs significantly increased at high I (>0.3), and the increase in I enhanced the apparent viscosity, fluorescence intensity, surface hydrophobicity, and α-helix content of MPs solution. Particle size and sodium dodecyl sulfate-polyacrylamide gel electrophoresis patterns also supported the solubility profiles. Transmission electron microscopy and atomic force microscopy observations revealed the morphological assembly and disassembly of MPs under different I conditions. Finally, proteomic analysis revealed the evolution law of salt-induced solubilization of MPs and the critical molecular characteristics in different I environments. The number of differentially abundant proteins (DAPs) decreased with the increase of I, and most DAPs related to the muscle filament sliding, contraction and assembly, actinin binding, and actin filament binding. The soluble abundance of myosin and some structural proteins was dependent on I, and structural proteins in the Z-disk and M-band might contribute to the solubilization of myosin. Our findings provide insightful information about the impact of common I on the solubility pattern of MPs from freshwater fish.

Microwave-assisted depolymerization of chitin and chitosan extracted from crayfish shells waste: A sustainable approach based on graphene oxide catalysis.

This study targeted the sustainable utilization of chitin and chitosan from crayfish shell waste, and further depolymerization of the recovered products in one step through synergy between microwaves and graphene oxide, aiming for the monosaccharides, 5-hydroxymethylfurfural and other high-value products. The results indicated that graphene oxide was more effective than graphene in enhancing the microwave absorption properties of the system, which is contrary to the parameters of their dielectric properties. The heating rate was increased by 0.37 K/s and 0.26 K/s when graphene oxide was introduced into the chitin and chitosan depolymerization systems, respectively, at a microwave power of 5 W/g. The mechanism underlying the impact of graphene oxide on chitin and chitosan under a microwave field was proposed by analyzing the variations in the depolymerization products of chitin and chitosan systems under different reaction conditions, including holding time, catalyst content, solvent content, and reaction temperature. Furthermore, the recovered graphene oxide exhibited delamination upon redispersion in water, which was not observed in the initial samples. The infrared spectra and scanning electron microscopy results suggest that the catalytic reaction is associated with oxygen-containing functional groups. This study demonstrated the synergistic effect of microwaves and graphene oxide on the depolymerization of chitin and chitosan, and the ability to achieve rapid one-step depolymerization in an acid/alkali-free solvent, which provides a green and promising development for the degradation of carbohydrate macromolecules in crustacean solid waste.

Advances in flavor peptides with sodium-reducing ability: A review.

Condiments (such as sodium chloride and glutamate sodium) cause consumers to ingest too much sodium and may lead to a variety of diseases, thus decreasing their quality of life. Recently, a salt reduction strategy using flavor peptides has been established. However, the development of this strategy has not been well adopted by the food industry. There is an acute need to screen for peptides with salty and umami taste, and to understand their taste characteristic and taste mechanism. This review provides a thorough analysis of the literature on flavor peptides with sodium-reducing ability, involving their preparation, taste characteristic, taste mechanism and applications in the food industry. Flavor peptides come from a wide range of sources and can be sourced abundantly from natural foods. Flavor peptides with salty and umami tastes are mainly composed of umami amino acids. Differences in amino acid sequences, spatial structures and food matrices will cause different tastes in flavor peptides, mostly attributed to the interaction between peptides and taste receptors. In addition to being used in condiments, flavor peptides have also anti-hypertensive, anti-inflammatory and anti-oxidant abilities, offering the potential to be used as functional ingredients, thus making their future in the food industry extremely promising.

Investigation on the quality regulating mechanism of antifreeze peptides on frozen surimi: From macro to micro.

Freeze denaturation of protein caused by ice crystals is the main motivation for the quality deterioration of surimi during circulation and storage. This investigation aimed to cryoprotect surimi by adding antifreeze peptides from Takifugu obscurus skin (TsAFP) which can inhibit ice recrystallization, and to elucidate regulating mechanism. The comprehensive results showed that 4% TsAFP, half dosage of commercial cryoprotectant, had good cryoprotection on surimi by reducing the moisture variation and maintaining protein solubility of surimi at macro level, as well as inhibiting the degeneration and structure changes of myofibrillar proteins at micro level. Meanwhile, TsAFP could directly bind to the structural cavity of myosin, inhibit protein freezing-induced oxidation, maintain the spatial structure of myosin and water retention ability to preserve the surimi quality. This study helped better comprehend the protective mechanisms of antifreeze peptides in frozen surimi and was expected to provide a promising cryoprotectant for low-sweetness and low-calorie surimi.

New insight into the mechanism by which antifreeze peptides regulate the physiological function of Streptococcus thermophilus subjected to freezing stress.

INTRODUCTION: Antifreeze peptides regulate the physiological functions of frozen cells and even their apoptosis; however, the mechanisms by which antifreeze peptides regulate these processes remain unclear, although the interactions between cell membranes and ice are well known to be important in this process. OBJECTIVES: Our study aims to investigate how antifreeze peptides regulate cell physiological functions during the freezing process. METHODS: We investigated the cryoprotective effect of rsfAFP on the physiological functions of S. thermophilus under freezing stress by measuring cellular metabolism activity, intracellular enzyme activity, cell membrane characterization, and cell apoptosis. The mechanism by which rsfAFP impacts S. thermophilus physiological functions under freezing stress was investigated using multispectral techniques and cryo-TEM. RESULTS: We show that a recombinant antifreeze peptide (rsfAFP) interacts with the extracellular capsular polysaccharides and peptidoglycan of Streptococcus thermophilus and ice to cover the outer layer of the membrane, forming a dense protective layer that regulates the molecular structure of extracellular ice crystals, which results in reduced extracellular membrane damage, depressed apoptosis and increased intracellular metabolic activity. This interaction mechanism was indicated by the fact that S. thermophilus better maintained its permeability barrier, membrane fluidity, membrane structural integrity, and cytoplasmic membrane potential during freezing stress with rsfAFP treatment. CONCLUSION: These results provide new insights into the mechanism by which rsfAFP regulates frozen cellphysiological functionsand apoptosis under freezing stress.

Microwave heating process of moderate-minced surimi based on multiphase porous media model.

Moderately processed surimi products exhibit better nutrient retention and enhanced gels, and the great potential of microwaves application and changes in the way of chopping meat has been reported by previous research. In this study, a systematic analysis of the novel surimi product was made to explore the heat and mass transfer characteristics. A porous media model combining electromagnetic heat and hygroscopic expansion was developed to evaluate this process, and its accuracy has been verified experimentally. It was found that the dielectric characterization of multiphase mixture system has great influence on the results, the complex refractive index mixture equation was used due to its lowest root-mean-square error value. In addition, the effect of moderate processing on microwave heating was examined in terms of porosity changes. However, nonuniform temperature distributions were found in the higher porous samples, especially when the porosity is greater than 0.81. Moreover, the developed model was coupled with the evaluation for gel properties and the results showed the significant effect of moderate crushing on the gel quality during the microwave heating process.

Analysis of the shape retention ability of antifreeze peptide-based surimi 3D structures: Potential in freezing and thawing cycles.

The effects of freeze-thaw (F-T) cycles on the shape retention of antifreeze peptides-based surimi ink (ASI) 3D structures were analyzed. The results showed that the ASI 3D structure has good shape retention ability, and the width, height, weight, and water holding capacity were 22.42 mm, 21.07 mm, 9.99 g, and 68.30 % even after F-T 4 times, respectively. The average area and equivalent diameter of ice crystals in ASI 3D structures only expand from 0.001 mm2 and 0.040 mm to 0.015 mm2 and 0.139 mm, respectively. The α-helix and ß-sheet of myofibrillar protein in ASI 3D structure were slightly decreased by 44.16 ± 0.98 % to 33.33 ± 0.92 % and increased by 18.28 ± 4.45 % to 24.43 ± 1.60 %, respectively. The chemical bond and protein interaction have changed to some extent. AFPs can prevent denaturation and juice loss of surimi 3D structures after F-T. The results provide theoretical guidance for maintaining the shape retention of frozen 3D food structures.

Identification of Novel Antifreeze Peptides from Takifugu obscurus Skin and Molecular Mechanism in Inhibiting Ice Crystal Growth.

Foodborne hydrolyzed antifreeze peptides have been widely used in the food industry and the biomedical field. However, the components of hydrolyzed peptides are complex and the molecular mechanism remains unclear. This study focused on identification and mechanism analysis of novel antifreeze peptides from Takifugu obscurus skin by traditional methods and computer-assisted techniques. Results showed that three peptides (EGPRAGGAPG, GDAGPSGPAGPTG, and GEAGPAGPAG) possessed cryoprotection via reducing the freezing point and inhibiting ice crystal growth. Molecular docking confirmed that the cryoprotective property was related to peptide structure, especially α-helix, and hydrogen bond sites. Moreover, the antifreeze peptides were double-faces, which controlled ice crystals while affecting the arrangement of surrounding water molecules, thus exhibiting a strong antifreeze activity. This investigation deepens the comprehension of the mechanism of antifreeze peptides at molecular scale, and the novel efficient antifreeze peptides can be developed in antifreeze materials design and applied in food industry.

Effects of preheating-induced denaturation treatments on the printability and instant curing property of soy protein during microwave 3D printing.

Soybean protein pastes were preheated for different times (20, 25 and 30 min) with l-cysteine to improve the printability and self-gelation properties and to explore the relationship between the degree of denaturation and the feasibility of microwave 3D printing. As the pre-denaturation degree increased, ordered secondary structures in proteins gradually decreased, and tertiary structures unfolded. Soybean protein printability, moulding quality increased, and soybean protein, which is not suitable for printing in its natural state, achieved self-gelling after 3D printing. Soybean protein pastes preheated for 25 min with l-cysteine achieved a complete hollow sphere structure compared to other formulations, while pastes preheated for 30 min formed a coarser structure, and blockage occurred during printing. The covalent bond of ε- (γ-glutamine) formed by the synergistic effect of transglutaminase (TGase) and microwaves was the main force driving the gel network structure formation. The most suitable pre-denaturation degree of soybean protein was quantitatively characterized as 57 %.

Effects and mechanism of antifreeze peptides from silver carp scales on the freeze-thaw stability of frozen surimi.

The objective of this work was to investigate the cryoprotective effects of antifreeze peptides obtained from silver carp scales (ScAFPs) on the freeze-thaw stability of surimi, and to explore the action mechanisms of ScAFPs on frozen surimi. The comprehensive analysis of ice crystal size, myofibril protein oxidation, water retention, surimi gel properties, and rheological properties of surimi after different freeze-thaw cycles were investigated. Results showed that frozen surimi treated with ScAFPs exhibited a higher Ca2+-ATPase activity, salt-soluble protein concentration and sulfhydryl group content, while lower surface hydrophobicity, carbonyl content and disulfide bond content. Moreover, the gel properties and water holding capacity of surimi and surimi gel were improved significantly by regulating the size of ice crystals during freeze-thaw process. These findings indicate that ScAFPs could serviced as a new food ingredient with anti-freezing function for frozen products.

Changes in physicochemical properties of silver carp (Hypophthalmichthys molitrix) surimi during chilled storage: The roles of spoilage bacteria.

Chilled surimi has become increasingly popular owing to its superior texture and freshness. In this study, changes in the microbiota and gel properties during chilled surimi storage, and the contributions of dominant bacteria to the physicochemical properties of chilled surimi were investigated. The results showed that Pseudomonas gessardii, Aeromonas media, and Acinetobacter johnsonii were the dominant bacteria during chilled surimi storage. P. gessardii was the key bacteria that degraded protein in the process of surimi spoilage, which led to high total volatile base nitrogen (TVB-N), trichloroacetic acid (TCA)-soluble peptides as well as poor gel properties. Both P. gessardii and A. media were high putrescine producers, whereas only A. media produced cadaverine. In this study, spoilage microorganisms in chilled surimi were investigated for the first time, and it was found that P. gessardii had the greatest influence on surimi quality, which provides a research basis for in-depth study on the mechanism of microbial spoilage and the preservation of chilled surimi.

Snow flea antifreeze peptide for cryopreservation of lactic acid bacteria.

Cryogenic machining is one of the most commonly used techniques for processing and preserving in food industry, and traditional antifreeze agents cannot regulate the mechanical stress damage caused by ice crystals formed during recrystallization or thawing. In this study, we successfully developed an express system of a novel recombinant snow flea antifreeze peptide (rsfAFP), which has significant ice recrystallization inhibition ability, thermal hysteresis activity and alters ice nucleation, thus regulating extracellular ice crystal morphology and recrystallization. We showed that rsfAFP improved the survival rate, acid-producing ability, freezing stability, and cellular metabolism activity of Streptococcus thermophilus. We further showed that rsfAFP interacts with the membrane and ice crystals to cover the outer layer of cells, forming a dense protective layer that maintains the physiological functions of S. thermophilus under freezing stress. These findings provide the scientific basis for using rsfAFP as an effective antifreeze agent for lactic acid bacteria cryopreservation or other frozen food.

Investigation of the cryoprotective mechanism and effect on quality characteristics of surimi during freezing storage by antifreeze peptides.

Freezing technology is important for storage of animal products such as surimi. However, mechanical damage caused by ice crystals would lead to quality deterioration. This study aims to investigate the protective effect of antifreeze peptides (AFPs) on the quality of surimi during freezing storage and its possible mechanism. We found that AFPs exhibited a strong inhibition of ice crystal recrystallization, and the molecular weight ranged from 180 to 3000 Da. AFPs can prevent the degeneration of myofibrillar protein by reducing the loss of Ca2+-ATPase activity, slowing oxidation of sulfhydryl groups to disulfide bonds, and maintaining surface hydrophobicity and solubility of myofibrillar protein. Moreover, AFPs can reduce the influence of freezing stress on water mobility, thereby protecting the surimi from losing immobilized water and bound water during frozen storage. These findings indicate that AFPs could potentially serve as a food ingredient with antifreeze functional for the storage of surimi products.

Redox Proteomic Analysis Reveals Microwave-Induced Oxidation Modifications of Myofibrillar Proteins from Silver Carp (Hypophthalmichthys molitrix).

To provide an insight into the oxidation behavior of cysteines in myofibrillar proteins (MPs) during microwave heating (MW), a quantitative redox proteomic analysis based on the isobaric iodoacetyl tandem mass tag technology was applied in this study. MPs from silver carp muscles were subjected to MW and water bath heating (WB) with the same time-temperature profiles to eliminate the thermal differences caused by an uneven energy input. Altogether, 422 proteins were found to be differentially expressed after thermal treatments as compared to that with no heat treatment. However, MW triggered a larger number of proteins and cysteine sites for oxidation. Myosin heavy chain, myosin-binding protein C, nebulin, α-actinin-3-like, and titin were found to be highly susceptible to oxidation under microwave irradiation. Notably, MW caused such modifications at cysteine site 9 in the head of myosin, revealing the enhancement mechanism of MP gelation by excess cysteine cross-linking during microwave processing. Furthermore, Gene Ontology and functional enrichment analyses suggested that the two thermal treatments resulted in some differences in ion binding, muscle cell development, and protein-containing complex assembly. Overall, this study is the first to report the redox proteomic changes caused by MW and WB treatments, thus providing a further understanding of the microwave-induced oxidative modifications of MPs.

Glycated peptides obtained from cultured crocodile meat hydrolysates via Maillard reaction and the anti-aging effects on Drosophila in vivo.

With the aging problems increasing, the discovery of anti-aging compounds has become a popular research direction. Accumulation of free radicals and the consequent oxidative stress are the chief culprit of aging. Given this, cultured crocodile meat peptides-Maillard reaction product (CMP-MRP) with remarkable antioxidant activity was obtained via Maillard reaction of cultured crocodile meat hydrolysates and xylose. The antioxidant activity in vitro and anti-aging activity in vivo of CMP-MRP were investigated. Results indicated that the lifespan and the athletic ability of Drosophila were significantly improved after the administration of CMP-MRP in natural aging, H2O2- and paraquat-induced models. Furthermore, the antioxidant enzyme activities of Drosophila treated with CMP-MRP were enhanced while the levels of malondialdehyde (MDA) and protein carbonyl (PCO) were reduced in three Drosophila models. With the supplement of 5 mg/mL CMP-MRP in natural aging Drosophila model, the maximum lifespan increased from 61 days to 73 days, athletic ability raised by 95.45%, MDA and PCO reduced by 52.72% and 47.43%, respectively. Taken together, CMP-MRP exhibited outstanding antioxidant and anti-aging capacities in Drosophila models, suggesting that CMP-MRP possesses great potential in the health food and biomedicine fields as a food-derived anti-aging agent.