Low Salt Conditions Research Articles

A unique structure of bacteriophage T4 gene 32 protein with double-stranded DNA in low-salt conditions is distinguished by antibodies.

Bacteriophage T4 gene 32 protein (gp32) preferentially binds to single-stranded DNA (ssDNA) to facilitate DNA replication but shows weak binding to double-stranded DNA (dsDNA). Polyclonal and monoclonal antibodies against gp32 were raised, and an enzyme-linked immunosorbent assay was used to evaluate their reactivities against gp32. The reactivity of the monoclonal antibody MGP45 was diminished in the presence of 5ng/ml dsDNA, suggesting a conformational change that reduces epitope availability. Notably, the same concentration of ssDNA had little effect; instead, 500ng/ml ssDNA was required to elicit the same degree of inhibition. A decrease in MGP45 reactivity with gp32 was observed in the presence of NaCl at concentrations less than 100mM under neutral conditions. These changes in antibody reactivity reflect differences the gp32 conformation, which may underlie its different affinities for ssDNA and dsDNA.

Transcriptome Analysis Reveals the Key Genes and Pathways for Growth, Ion Transport, and Oxidative Stress in Post-Larval Black Tiger Shrimp (Penaeus monodon) Under Acute Low Salt Stress.

As an abiotic stress factor, salinity significantly affects the physiological activities of crustaceans. In this study, transcriptome sequencing was used to evaluate the mechanism of ion transport and the physiological response of black tiger shrimp (Penaeus monodon) under low salt stress. Four hundred post larval (PL) stage P. monodon were distributed in eight experimental tanks and exposed to 3 or 5 ppt salt concentrations for 96h. Low salinity significantly reduced the survival rate of shrimp but simultaneously activated the activity of ion transporter enzymes Na+/K+-ATPase (NKA) and Ca2+/Mg2+-ATPase), the expression of NKA, galectin 10, and cytochrompe c peroxidase genes, and the activity and expression of antioxidant-related genes (superoxide dismutase, catalase, heat shock protein 60). Low salt stress activated the urea cycle but significantly inhibited glutathione metabolization-related indicators (glutamate dehydrogenase, glutaminase, glutamic acid). RNA-seq analysis identified 221 differentially expressed genes (78 up-regulated and 143 down-regulated). Quantitative real-time PCR and RNA-seq results of 11 of them were consistent, illustrating the validity of the transcriptomic predictions. Gene set enrichment analysis results showed that calcium ion transmembrane transport, calmodulin binding, the stress-activated protein kinase signaling cascade, and regulation of the cytosolic calcium ion concentration process were significantly enriched. These results showed that low salt stress activated the calcium-dominated ion transport pathway and promoted molting growth of P. monodon. They also indicate that there is potential for larval rearing shrimp under low salt conditions.

Systematic free energy insights into the enhanced dispersibility of myofibrillar protein in low-salt solutions through ultrasound-assisted enzymatic deamidation.

This work aimed to investigate the effects of ultrasound assisted enzymatic deamidation by protein-glutaminase (PG) on the dispersion of myofibrillar protein (MP) in low-salt solutions. The solubility, structural characteristics, transmission electron microscopy, asymmetric-flow field-flow fractionation, steady shear rheological property and multiple light scattering of MP deamidated by PG (MP-PG) and MP pretreated with ultrasound followed by PG deamidation (MP-U-PG) were determined. Molecular docking and molecular dynamics (MD) simulations were used to estimate the interaction between PG and MP. Under ultrasound assistance, the MP deamidated for 16h (MP-U-PG16) showed the highest solubility (80.1%) in low-salt conditions, which is attributed to its highest absolute zeta potential and smallest particle size. Although secondary structure analysis showed that MP-PG and MP-U-PG had an increased α-helix ratio and a decreased β-sheet ratio, ultrasonic treatment had a significantly influence on the MD results. The results manifested that hydrogen bond was the primary forces driving the binding between PG and MP, and the hydrogen bond and hydrophobic interaction were the dominant forces responsible the binding between PG and MP pretreated with ultrasound. According to the energy landscapes theory, ultrasound could overcome the energy barriers through external force input and find the best pathway to achieve the final lowest energy state. Our research contributed to the improvement of the colloidal dispersibility of MPs under low-salt conditions and the regulation of protein interaction by ultrasound assistance.

Thermo-reversible gelation and enhanced umami perception of myofibrillar proteins induced by protein-glutaminase-mediated deamidation.

In this study, the absolute electrostatic charge of myofibrillar protein (MP) was substantially increased by protein-glutaminase (PG) treatment, which was a critical step for achieving the dissociation and solubility of MP under low salt condition. The PG-treated MP exhibited the capacity to form thermo-reversible gels that could be melted through heating and subsequently reformed into a stable gel structure upon refrigeration. The results of SDS-PAGE further revealed that the levels of soluble monomeric myosin and actin in the supernatant of deamidated MP (DMP) gels were markedly elevated, and confirmed the increased formation of intermolecular disulfide bond between myosin and actin. Additionally, moderate deamidation was beneficial for the improvements of MP gel properties, especially in terms of water-holding capacity and springiness. Electronic tongue and correlation analysis indicated that the umami perception of DMP was significantly enhanced because of the conversion of glutamine (Gln) to glutamate (Glu) residues that induced by PG deamidation.

Self-Diffusion of Star and Linear Polyelectrolytes in Salt-Free and Salt Solutions.

This work explored solution properties of linear and star poly(methacrylic acids) with four, six, and eight arms (LPMAA, 4PMAA, PMAA, and 8PMAA, respectively) of matched molecular weights in a wide range of pH, salt, and polymer concentrations. Experimental measurements of self-diffusion were performed by fluorescence correlation spectroscopy (FCS), and the results were interpreted using the scaling theory of polyelectrolyte solutions. While all PMAAs were pH sensitive and showed an increase in hydrodynamic radius (R h) with pH in the dilute regime, the R h of star polymers (measured at basic pH values) was significantly smaller for the star polyacids due to their more compact structure. Fully ionized star PMAAs were also found to be less sensitive to changes in salt concentration and type of the counterion compared to linear PMAA. While R h of fully ionized linear PMAA decreased in the series Li+ > Na+ > K+ > Cs+ in agreement with the Hofmeister series, R h of star PMAAs was virtually independent of type of the counterion for eight-arm PMAA. However, molecular architecture strongly affected interactions of counterions with PMAAs. In particular, 7Li NMR revealed that the spin-lattice relaxation time T 1 of Li+ ions in low-salt solutions of eight-arm PMAA was ∼2-fold smaller than that in the solution of linear PMAA, suggesting slower Li+-ion dynamics within star polymers. An increase in concentration of monovalent chloride salts, c s, above that of the PMAA monomer unit concentration (c m) resulted in shrinking of both linear and star molecules, with the hydrodynamic size R h scaling as R h ∝ c s -0.11±0.01. Self-diffusion of linear and star polyelectrolytes was then studied in a wide range of polyelectrolyte concentrations (10-3 mol/L < c m < 0.5 mol/L) in low-salt (<10-4 mol/L of added salt) and high-salt (1 mol/L) solutions. In both the low-salt and high-salt regimes, diffusion coefficient D was lower for PMAAs with a larger number of arms at a fixed c m. In addition, in both cases, D plateaued at low polymer concentrations and decreased at higher polymer concentrations. However, while in the high-salt conditions, the concentration dependence of D reflected transitions between the dilute to semidilute solution regimes as expected for neutral chains in good and theta solvents, analysis of the diffusion data in the low-salt conditions using the scaling theory revealed a different origin of the concentration dependence of D. Specifically, in the low-salt solutions, both linear and star PMAAs exhibited unentangled (Rouse-like) dynamics in the entire range of polyelectrolyte concentrations.

Based on metabolomics analysis: metabolic mechanism of intestinal tract of Scylla paramamosain under low-salt saline-alkali water aquaculture environment

BackgroundIn recent years, the total production of mud crab Scylla paramamosain has been declining, and the breeding areas are faced with land shortage and shortage of breeding production, which needs to be solved urgently. S. paramamosain can survive and grow in a wide range of salinities is an excellent variety suitable for saline-alkali water aquaculture. As a species with high economic value and strong adaptability to the environment, its cultivation under low salt conditions can not only improve the utilization efficiency of saline-alkali land, but also provide new possibilities for the sustainable development of aquaculture.ResultsA total of 248 different metabolites were identified by LC/GC-MS in the intestinal tract of S. paramamosain. These different metabolites were mainly concentrated in ‘Lipids and lips-like molecules’. Among them, 112 metabolites are upregulated, and among these upregulated metabolites are mainly ‘Fatty Acyls’ and ‘Glycerophospholipids’. The upregulation of these metabolites indicates an increase in lipid storage of S. paramamosain, which may increase the resistance of S. paramamosain to adverse environmental stress. Among them, 136 metabolic differentiates were down-regulated, mainly ‘Carboxylic acids and derivatives’. The down-regulation of these organic acids may indicate that organic acids are used as energy sources for the immune response to long-term environmental stress.ConclusionUnder long-term chloride type low-salt saline-alkali water stress, S. paramamosain will shift to another homeostasis for development.

Glycosylated peptides isolated from cheese whey have antifreezing activity.

The glycomacropeptide (GMP) present in the cheese whey byproduct can be an excellent antifreezing agent due to its unique molecular structure. The objective of this study was to concentrate this peptide and investigate its ice recrystallization inhibition (IRI) ability. Heat denaturation of the non-GMP proteins and preparative liquid chromatography were used to create fraction 1 (F1) and fraction 2 (F2) and these were tested using the splat assay and a modified sucrose sandwich assay to investigate their IRI activity. Both F1 and F2 showed moderate IRI activity, but this activity was strongest under low salt and low pH conditions as indicated by the regression trends. F1 demonstrated an ice shaping ability similar to that of antifreeze glycoproteins. Molecular dynamic simulations of GMP, de-glycosylated GMP, and alpha-lactalbumin were performed and confirmed substantial differences of GMP when compared to the others in its high degree of conformational diversity and flexibility, as shown by the combination of radius of gyration, solvent accessible surface area, secondary structure, and hydrogen-bonding measurements together. Thus, GMP was shown for the first time as an effective IRI agent and its concentrate can be feasibly obtained from dairy processing byproduct.

The improved gel properties of myofibrillar protein under low salt condition by ultrasound-assisted sodium tripolyphosphate

Excessive salt intake is associated with increased risks of cardiovascular disease, while directly reducing salt content significantly decreased the quality of meat. The objective of this study was to investigate the synergistic effects of sodium tripolyphosphate (STP) and ultrasound treatment on the gel properties of MP under low-salt conditions (0.3 M NaCl). The results of FTIR spectra showed that P3O105− group of STP bound to -NH2 or -OH group of MP form C-N-P or C-O-P bond, indicating the STP was successfully introduced to MP. The addition of STP significantly increased the absolute value of Zeta-potential, suggesting that the presence of STP increased the electrostatic interaction of MP-MP. Importantly, STP combined with ultrasound treatment under low salt condition (STP-U0.3) significantly increased solubility and decreased particle size of MP. Besides, STP-U0.3 treatment also promoted the exposure of hydrophobic groups and improved the rheological behavior of MP, resulting in the highest gel strength (37.78 ± 0.71 g) and the lowest cooking loss (26.73 ± 0.90 %) especially in 10 mM STP combined 100 W ultrasound treatment. These results corresponded by the gradually increases of α-helix content and the decrease of tryptophan fluorescence intensity. Furthermore, results of SEM illustrated that STP-U0.3 treatment contributed to formation of more homogeneous and dense gel network of MP gel. The above results displayed that the STP-U treatment under 0.3 M NaCl resulted in an equivalent effect to control group of 0.6 M NaCl, indicating that the combined application of STP and ultrasound has a promising potential in the low-salt meat processing industry.

Structure Prediction of Large RNAs with AlphaFold3 Highlights its Capabilities and Limitations

DeepMind’s AlphaFold3 webserver offers exciting new opportunities to make structural predictions of heterogeneous macromolecular systems. Here we attempt to apply AlphaFold3 to large RNA molecules whose 3D atomic structures are unknown but whose physical dimensions have been studied experimentally. One difficulty that we encounter is that models returned by AlphaFold3 often contain severe steric clashes and, less frequently, clear breaks in the phosphodiester backbone, with the probability of both events increasing with the length of the RNA. Restricting attention to those RNAs for which non-clashing models can be obtained, we find that hydrodynamic radii computed from the AlphaFold3 models are much larger than those reported experimentally under low salt conditions but are in better agreement with those reported in the presence of polyvalent cations. For two RNAs whose shapes have been imaged experimentally, the computed anisotropies of the AlphaFold3-predicted structures are too low, indicating that they are excessively spherical; extending this analysis to larger RNAs shows that they become progressively more spherical with increasing length. Overall, the results suggest that AlphaFold3 is capable of producing plausible models for RNAs up to ∼2000 nucleotides in length, but that thousands of predictions may be required to obtain models free of geometric problems.

Having their cake and eat it too: Effects of different cations in reduced-salt myofibrillar protein microgel pickering emulsion under high-intensity ultrasound

Health concerns have increased the demand for low-salt emulsified meat products emulsions; however, salt-soluble myofibrillar proteins (MP) are less than optimal emulsifiers under low-salt conditions. In this study, four different cations (Na+, K+, Ca2+, and Mg2+) were added to a Pickering emulsion and high-intensity ultrasound (HIU, 20 kHz, 600 W) was used to determine the quality of the final emulsion. The divalent cation group was found to significantly (P < 0.05) improve the emulsification performance of the emulsion; in particular, the emulsion activity index and emulsion stability index of Mg2+-H group increased to 16.24 ± 1.22 m2/g and 89.66 ± 3.01%, respectively. Measurements of the particle size and zeta potential confirmed that the emulsion particle size of the Mg2+-H group decreased, and the electrostatic repulsion force increased. Based on the surface hydrophobicity and endogenous tryptophan spectra, the particle structure of the MP microgel was determined to be a more stable emulsifier. What's more, the divalent cations promoted the droplets to become evenly dispersed under high-intensity ultrasound. The micrograph confirmed this result: the divalent cation group emulsion was more uniform and stable than the monovalent cation group emulsion. Divalent cations are more suitable than monovalent cations for improving the quality of reduced-salt Pickering emulsions.

An effective means to improve the flavor quality of traditional fermented sour meat: The salt reduction strategy

The flavor formation of fermented meat products is significantly influenced by salt. In this study, the flavor analysis methods of electronic tongue and nose, gas chromatography-mass spectrometry, and liquid chromatography-mass spectrometry were utilized to explore the effect of salt content on the formation of sour meat flavor. The results demonstrated a significant decrease in pH value with the reduction of salt content (P < 0.05), while there was a significant rise in thiobarbituric acid reactive substances (TBARS) value associated with the decline in salt content (P < 0.05). Alcohols, aldehydes, and esters such as 1-octen-3-ol, 3-methyl-1-butanol, hexanal, heptanal, ethyl octanoate, ethyl heptanoate, etc. were considered to have significant contributions to the overall flavor of sour meat. The rapid accumulation of these aroma compounds under low-salt conditions was regarded as one of the key factors in the overall flavor enhancement of sour meat products. Additionally, the low-salt fermentation also promoted the accumulation of free amino acids and free fatty acids in sour meat products, thereby facilitating the formation of more complex aroma compounds. This study offers new technological ways to improve the conventional method of fermenting sour meat and offers theoretical justification for studies on salt reduction in conventionally fermented meat products.

Effects of Flammulina velutipes powder on the emulsion properties of chicken myofibrillar protein under low-salt conditions.

This study aimed to investigate the influence of varying concentrations of Flammulina velutipes powder (FVP) (0, 4, 8, 12, 16, and 20 g L-1, based on FVP weight per liter of emulsion) on the stability, rheological properties, microstructure, and interfacial protein content of chicken myofibrillar protein (MP) emulsions under low-salt conditions. Visual assessment and the Turbiscan stability index revealed that the stability of MP emulsions improved with increasing FVP concentration. The greatest stability was achieved when the FVP concentration was 16 gL-1. The incorporation of FVP enhanced the elasticity and viscosity of the emulsions by forming a dense three-dimensional network structure. The droplet size of the emulsions initially decreased and then increased with increasing FVP concentration. The addition of FVP increased the amount of protein absorbed by the emulsion layer. Flammulina velutipes powder is promising as a stabilizer that could improve the emulsifying and functional properties of low-salt myofibrillar protein emulsions. © 2024 Society of Chemical Industry.

Investigation on environmental factors contributing to bispecific antibody stability and the reversal of self-associated aggregates

Bispecific antibodies (bsAbs) hold promises for enhanced therapeutic potential surpassing that of their parental monoclonal antibodies. However, bsAbs pose great challenges in their manufacturing, and one of the common reasons is their susceptibility to aggregation. Building on previous studies demonstrating the functionality and potential manufacturability of Fab-scFv format bsAb, this investigation delved into the impact of environmental factors—such as pH, buffer types, ionic strength, protein concentrations, and temperatures—on its stability and the reversal of its self-associated aggregates. Mildly acidic, low-salt conditions were found optimal, ensuring bsAb stability for 30 days even at elevated temperature of 40 °C. Furthermore, these conditions facilitated the reversal of its self-associated aggregates to monomers during the initial 7-day incubation period. Our findings underscore the robustness and resilience of Fab-scFv format bsAb, further confirming its potential manufacturability despite its current absence as commercial products.Graphical

Effects of Different Salinity Stress on the Transcriptomic Responses of Freshwater Crayfish (Procambarus clarkii, Girard, 1852).

Salinization of freshwater ecosystems is a pressing global issue. Changes in salinity can exert severe pressure on aquatic animals and jeopardize their survival. Procambarus clarkii is a valuable freshwater aquaculture species that exhibits some degree of salinity tolerance, making it an excellent research model for freshwater aquaculture species facing salinity stress. In the present study, crayfish were exposed to acute low salt (6 ppt) and high salt (18 ppt) conditions. The organisms were continuously monitored at 6, 24, and 72 h using RNA-Seq to investigate the mechanisms of salt stress resistance. Transcriptome analysis revealed that the crayfish responded to salinity stress with numerous differentially expressed genes, and most of different expression genes was observed in high salinity group for 24h. GO and KEGG enrichment analyses indicated that metabolic pathways were the primary response pathways in crayfish under salinity stress. This suggests that crayfish may use metabolic pathways to compensate for energy loss caused by osmotic stress. Furthermore, gene expression analysis revealed the differential expression of immune and antioxidant-related pathway genes under salinity stress, implying that salinity stress induces immune disorders in crayfish. More genes related to cell proliferation, differentiation, and apoptosis, such as the Foxo, Wnt, Hippo, and Notch signaling pathways, responded to high-salinity stress. This suggests that regulating the cellular replication cycle and accelerating apoptosis may be necessary for crayfish to cope with high-salinity stress. Additionally, we identified 36 solute carrier family (SLC) genes related to ion transport, depicting possible ion exchange mechanisms in crayfish under salinity stress. These findings aimed to establish a foundation for understanding crustacean responses to salinity stress and their osmoregulatory mechanisms.

Effect of temperature and sodium bicarbonate combined on aggregation, rheology and conformation of low-salt chicken myofibrillar protein

To study the impact of temperature (4–50 °C) and sodium bicarbonate (0.4%) on the solubility, protein aggregation and conformation of low-salt (1 mmol/L sodium chloride) chicken myofibrillar protein, the changes in turbidity, particle size, protein secondary and tertiary structures were investigated. There was no significant difference in pH when it did not exceed 30 °C, then a significant increase (p < 0.05) when it was above 30 °C. The solubility, total and reactive sulfhydryl groups, and apparent viscosity increased first and then decreased (p < 0.05), and reached the maximum values at 30 °C; whereas the turbidity, particle size, α-helix, and random coil contents showed opposite trends. Additionally, the fluorescence peak experienced a noticeable redshift with rising temperatures; β-turn content significantly increased but β-sheet content decreased significantly. Finally, combining sodium bicarbonate with a temperature of around 30 °C could enhance chicken myofibrillar protein solubility and change the protein structure under a low-salt condition.

IcmF2 of the type VI secretion system 2 plays a role in biofilm formation of Vibrio parahaemolyticus.

Vibrio parahaemolyticus possesses two distinct type VI secretion systems (T6SS), namely T6SS1 and T6SS2. T6SS1 is predominantly responsible for adhesion to Caco-2 and HeLa cells and for the antibacterial activity of V. parahaemolyticus, while T6SS2 mainly contributes to HeLa cell adhesion. However, it remains unclear whether the T6SS systems have other physiological roles in V. parahaemolyticus. In this study, we demonstrated that the deletion of icmF2, a structural gene of T6SS2, reduced the biofilm formation capacity of V. parahaemolyticus under low salt conditions, which was also influenced by the incubation time. Nonetheless, the deletion of icmF2 did not affect the biofilm formation capacity in marine-like growth conditions, nor did it impact the flagella-driven swimming and swarming motility of V. parahaemolyticus. IcmF2 was found to promote the production of the main components of the biofilm matrix, including extracellular DNA (eDNA) and extracellular proteins, and cyclic di-GMP (c-di-GMP) in V. parahaemolyticus. Additionally, IcmF2 positively influenced the transcription of cpsA, mfpA, and several genes involved in c-di-GMP metabolism, including scrJ, scrL, vopY, tpdA, gefA, and scrG. Conversely, the transcription of scrA was negatively impacted by IcmF2. Therefore, IcmF2-dependent biofilm formation was mediated through its effects on the production of eDNA, extracellular proteins, and c-di-GMP, as well as its impact on the transcription of cpsA, mfpA, and genes associated with c-di-GMP metabolism. This study confirmed new physiological roles for IcmF2 in promoting biofilm formation and c-di-GMP production in V. parahaemolyticus.

Genome-wide identification of ATG genes and their expression profiles under biotic and abiotic stresses in Fenneropenaeus chinensis

BackgroundAutophagy is a conserved catabolic process in eukaryotes that contributes to cell survival in response to multiple stresses and is important for organism fitness. Extensive research has shown that autophagy plays a pivotal role in both viral infection and replication processes. Despite the increasing research dedicated to autophagy, investigations into shrimp autophagy are relatively scarce.ResultsBased on three different methods, a total of 20 members of the ATGs were identified from F. chinensis, all of which contained an autophagy domain. These genes were divided into 18 subfamilies based on their different C-terminal domains, and were found to be located on 16 chromosomes. Quantitative real-time PCR (qRT-PCR) results showed that ATG genes were extensively distributed in all the tested tissues, with the highest expression levels were detected in muscle and eyestalk. To clarify the comprehensive roles of ATG genes upon biotic and abiotic stresses, we examined their expression patterns. The expression levels of multiple ATGs showed an initial increase followed by a decrease, with the highest expression levels observed at 6 h and/or 24 h after WSSV injection. The expression levels of three genes (ATG1, ATG3, and ATG4B) gradually increased until 60 h after injection. Under low-salt conditions, 12 ATG genes were significantly induced, and their transcription abundance peaked at 96 h after treatment.ConclusionsThese results suggested that ATG genes may have significant roles in responding to various environmental stressors. Overall, this study provides a thorough characterization and expression analysis of ATG genes in F. chinensis, laying a strong foundation for further functional studies and promising potential in innate immunity.

Potential mechanisms and effects of ultrasound treatment combined with pre- and post-addition of κ-carrageenan on the gelling properties and rheological behavior of myofibrillar proteins under low-salt condition

This study investigated the effect of ultrasound (US) combined with pre- and post-addition of κ-carrageenan (KC) on the gelling properties, structural characteristics and rheological behavior of myofibrillar proteins (MP) under low-salt conditions. The results showed that US combined with either pre- or post-addition of KC rendered higher gel strength and water holding capacity (WHC) of MP gels than those treated with US alone and added with KC alone (P < 0.05). US combined with pre-addition of KC facilitated the binding between MP and KC, which enhanced the gel strength and WHC of the mixed MP gels and significantly improved the rheological behavior of MP. This was also confirmed by the highest surface hydrophobicity, disulfide bonds and β-sheet content of the MP gels with US combined with pre-addition of KC. Moreover, microstructural results reflected a denser structure for the pre-addition of KC in combination with US. However, US combined with post-addition of KC resulted in limited MP unfolding and relatively weak hydrophobic interactions in the composite gels, which were less effective in improving the gel properties of the MP gels. This study provides potential strategies for enhancing the gelling properties of low-salt meat products via application of US and KC.

Rational engineering of Halomonas salifodinae to enhance hydroxyectoine production under lower-salt conditions

Hydroxyectoine is an important compatible solute that holds potential for development into a high-value chemical with broad applications. However, the traditional high-salt fermentation for hydroxyectoine production presents challenges in treating the high-salt wastewater. Here, we report the rational engineering of Halomonas salifodinae to improve the bioproduction of hydroxyectoine under lower-salt conditions. The comparative transcriptomic analysis suggested that the increased expression of ectD gene encoding ectoine hydroxylase (EctD) and the decreased expressions of genes responsible for tricarboxylic acid (TCA) cycle contributed to the increased hydroxyectoine production in H. salifodinae IM328 grown under high-salt conditions. By blocking the degradation pathway of ectoine and hydroxyectoine, enhancing the expression of ectD, and increasing the supply of 2-oxoglutarate, the engineered H. salifodinae strain HS328-YNP15 (ΔdoeA::PUP119-ectD p-gdh) produced 8.3-fold higher hydroxyectoine production than the wild-type strain and finally achieved a hydroxyectoine titer of 4.9 g/L in fed-batch fermentation without any detailed process optimization. This study shows the potential to integrate hydroxyectoine production into open unsterile fermentation process that operates under low-salinity and high-alkalinity conditions, paving the way for next-generation industrial biotechnology.Key points• Hydroxyectoine production in H. salifodinae correlates with the salinity of medium• Transcriptomic analysis reveals the limiting factors for hydroxyectoine production• The engineered strain produced 8.3-fold more hydroxyectoine than the wild typeGraphical

Underlying the mechanisms of incorporation of κ-carrageenan on the formation of low-salt myofibrillar protein gels during heating process: Perspective on the dynamic changes of protein structures and molecular interactions

This study aimed to systematacially investigate the effects of κ-carrageenan (KC) concentration on the formation of myofibrillar protein (MP) gels under low-salt condition based on the dynamic changes of protein structures and molecular interactions during heating process. The results indicated that incorporating KC markedly improved the water holding capacity (WHC) and gel strength of MP gels (P < 0.05). Those properties were most improved by adding 0.4% KC with WHC of 71.44% and gel strength of 1.62 N. During the heating process, when the temperature was above 50 °C, KC addition significantly increased the turbidity and particle size and decreased solubility of MP in a concentration-dependent manner, as well as exhibiting higher surface hydrophobicity at 0.2% KC concentration (P < 0.05), indicating that KC accelerated the rate of aggregation of MP during heating process. The dynamic rheological behaviors indicated that the addition of KC promoted the early unfolding and aggregation of MP at a relatively lower temperature, which was verified by the fluorescence denaturation mechanism results. Moreover, the addition of KC significantly promoted the heat-induced conformational transition of MP from α-helix companied with β-sheet and β-turn, which was due to the interaction between MP and KC accelerating the formation of MP gels during the heating process. Intermolecular forces results revealed that the aggregation of myosin head and cross-linking of myosin tail of MP added KC was enhanced by ionic bonds, hydrophobic interactions, and disulfide bonds, which facilitated the better gel performance. Our present results provided a comprehensive understanding of gelation mechanism of MP-KC mixed gels under low-salt condition, with implications for the practical application of KC in meat processing.