Transcriptome sequencing and metabolome analysis reveal the molecular mechanism of Salvia miltiorrhiza in response to drought stress.

Salvia miltiorrhiza is commonly used as a Chinese herbal medicine to treat different cardiovascular and cerebrovascular illnesses due to its active ingredients. Environmental conditions, especially drought stress, can affect the yield and quality of S. miltiorrhiza. However, moderate drought stress could improve the quality of S. miltiorrhiza without significantly reducing the yield, and the mechanism of this initial drought resistance is still unclear. In our study, transcriptome and metabolome analyses of S. miltiorrhiza under different drought treatment groups (CK, A, B, and C groups) were conducted to reveal the basis for its drought tolerance. We discovered that the leaves of S. miltiorrhiza under different drought treatment groups had no obvious shrinkage, and the malondialdehyde (MDA) contents as well as superoxide dismutase (SOD) and peroxidase (POD) activities dramatically increased, indicating that our drought treatment methods were moderate, and the leaves of S. miltiorrhiza began to initiate drought resistance. The morphology of root tissue had no significant change under different drought treatment groups, and the contents of four tanshinones significantly enhanced. In all, 5213, 6611, and 5241 differentially expressed genes (DEGs) were shared in the A, B, and C groups compared with the CK group, respectively. The results of KEGG and co-expression analysis showed that the DEGs involved in plant-pathogen interactions, the MAPK signaling pathway, phenylpropanoid biosynthesis, flavonoid biosynthesis, and plant hormone signal transduction responded to drought stress and were strongly correlated with tanshinone biosynthesis. Furthermore, the results of metabolism analysis indicated that 67, 72, and 92 differentially accumulated metabolites (DAMs), including fumarate, ferulic acid, xanthohumol, and phytocassanes, which were primarily involved in phenylpropanoid biosynthesis, flavonoid biosynthesis, and diterpenoid biosynthesis pathways, were detected in these groups. These discoveries provide valuable information on the molecular mechanisms by which S. miltiorrhiza responds to drought stress and will facilitate the development of drought-resistant and high-quality S. miltiorrhiza production.

Effect of bamboo shoot dietary fiber on gel properties, microstructure and water distribution of pork meat batters.

OBJECTIVE: To develop healthier comminuted meat products to meet consumer demand, the gel properties, rheological properties, microstructure and water distribution of pork meat batters formulated with various amounts of bamboo shoot dietary fiber (BSDF) were investigated. METHODS: Different levels of BSDF (0% to 4%) were added to pork batters, and the pH, color, water-holding capacity, texture and rheological properties of pork batters were determined. Then, pork batters were analyzed for their microstructure and water distribution using scanning electron microscopy (SEM) and low-field nuclear magnetic resonance (LF-NMR). RESULTS: Compared with the control, BSDF addition into meat batters showed a significant reduction in L*-value and a significant increase in b*-value (p<0.05). BSDF addition of up to 4% reduced the pH value of pork batters by approximately 0.15 units; however, the cooking loss and expressible water loss decreased significantly (p<0.05) with the increased addition of BSDF. The hardness and gel strength were noticeably enhanced (p<0.05) as the content of BSDF increased. The rheological results showed that BSDF added into pork batters produced higher storage modulus (G') and loss modulus (G″) values. The SEM images suggested that the addition of BSDF could promote pork batters to form a more uniform and compact microstructure. The proportion of immobilized water increased significantly (p<0.05), while the population of free water was decreased (p<0.05), indicating that BSDF improved the water-holding capability of pork batters by decreasing the fraction of free water. CONCLUSION: BSDF could improve the gel properties, rheological properties and water distribution of pork meat batters and decrease the proportion of free water, suggesting that BSDF has great potential as an effective binder in comminuted meat products.

Comparative study of thermal gelation properties and molecular forces of actomyosin extracted from normal and pale, soft and exudative-like chicken breast meat.

OBJECTIVE: The objectives of this study were to investigate the thermal gelation properties and molecular forces of actomyosin extracted from two classes of chicken breast meat qualities (normal and pale, soft and exudative [PSE]-like) during heating process to further improve the understanding of the variations of functional properties between normal and PSE-like chicken breast meat. METHODS: Actomyosin was extracted from normal and PSE-like chicken breast meat and the gel strength, water-holding capacity (WHC), protein loss, particle size and distribution, dynamic rheology and protein thermal stability were determined, then turbidity, active sulfhydryl group contents, hydrophobicity and molecular forces during thermal-induced gelling formation were comparatively studied. RESULTS: Sodium dodecyl sulphate-polyacrylamide gel electrophoresis showed that protein profiles of actomyosin extracted from normal and PSE-like meat were not significantly different (p>0.05). Compared with normal actomyosin, PSE-like actomyosin had lower gel strength, WHC, particle size, less protein content involved in thermal gelation forming (p<0.05), and reduced onset temperature (To), thermal transition temperature (Td), storage modulus (G') and loss modulus (G″). The turbidity, reactive sulfhydryl group of PSE-like actomyosin were higher when heated from 40°C to 60°C. Further heating to 80°C had lower transition from reactive sulfhydryl group into a disulfide bond and surface hydrophobicity. Molecular forces showed that hydrophobic interaction was the main force for heat-induced gel formation while both ionic and hydrogen bonds were different significantly between normal and PSE-like actomyosin (p<0.05). CONCLUSION: These changes in chemical groups and inter-molecular bonds affected protein-protein interaction and protein-water interaction and contributed to the inferior thermal gelation properties of PSE-like meat.

Effect of vegetable oils on the thermal gel properties of PSE-like chicken breast meat protein isolate-based emulsion gels.

To enhance the gel properties of PSE (pale, soft, and exudative)-like chicken meat protein isolate (PPI), the effect of peanut, corn, soybean, and sunflower oils on the gel properties of PPI emulsion gels was investigated. Vegetable oils improved emulsion stability and gel strength and enhanced viscosity and elasticity. The gel strength of the PPI-sunflower oil emulsion gel increased by 163.30 %. The thermal denaturation temperature and enthalpy values were increased. They decreased the particle size of PPI emulsion (P < 0.05) and changed the three-dimensional network structure of PPI emulsion gels from reticular to sheet with a smooth surface and pore-reduced lamellar. They elevated the content of immobile water PPI emulsion gels, decreased the α-helix and ß-turn, and increased the ß-sheet and random coil. Vegetable oil improved the gel properties of PPI in the following order: sunflower oil > soybean oil > corn oil ≈ peanut oil > control group.

Unraveling the effect of combined heat and high-pressure homogenization treatment on the improvement of chickpea protein solubility from the perspectives of colloidal state change and structural characteristic modification.

Chickpea protein (CP) is a promising plant protein ingredient, but the poor solubility has limited its broad application. In this study, heating followed by high-pressure homogenization (HPH) was used to improve the solubility of CP. The results showed that combined heat (80℃, 30 min) and HPH (80 MPa, 2 cycles) treatment exhibited an additive effect in improving the solubility of CP. This improvement could be attributed to the dissociation and the rearrangement of large insoluble protein aggregates into small-sized soluble protein aggregates, the increased exposure of hydrophobic residues and reactive sulfhydryl groups, the transformation of α-helices to ß-sheets and ß-turns. Moreover, the 11S subunits of CP could form reinforced disulfide covalent cross-links under heating + HPH, which may provide steric hindrance preventing the reassembly of large protein bodies. This work proposes an interesting approach to enhance the physicochemical properties of CP for tailoring techno-functional plant protein ingredients in food formulations.

The crucial role of Y109 and R162 as catalytic residues of nanoKAZ: insights from molecular docking, molecular dynamics simulation, and quantum chemical investigations.

CONTEXT: nanoKAZ is a compact luciferase that exhibits intense blue light emission when it catalyzes the substrate Furimazine (FMZ) as a luciferin, making it an excellent candidate as a reporter protein. However, the specific catalytic residues and mechanism of nanoKAZ have not been revealed. Recently, the structure of nanoKAZ was determined, and it was observed that the luminescent properties changed when FMZ analogs with naphthalene replacing benzene were used. It is speculated that the substituted naphthalene may influence the interaction between the catalytic residues and luciferins, thereby affecting the energy of the emitted light signal. METHOD: Therefore, the primary objective of this study is to analyze and compare the molecular recognition between nanoKAZ and FMZ along with its four activity-altered naphthalene analogs, with aiming to identify the catalytic residues. Molecular docking was employed to construct all nanoKAZ-luciferin models, followed by a 500 ns molecular dynamics simulation. The simulation trajectory was subjected to MM/PBSA analysis to identify crucial residues that contribute significantly to luciferin binding. In the result, two polar residues Y109, and R162 were identified as active residues as their notable contributions to the binding energy. Subsequently, an oxygen molecule was introduced into the local region of the nanoKAZ-FMZ complex and followed with quantum chemical calculations (semiempirical and DFT methods were used) to investigate the catalysis details. The results illustrated the involvement of Y109 and R162 in the oxygenation of FMZ, leading to the formation of dioxetanone, which has been suggested as an important intermediate in the oxidation process among various luciferins sharing the same functional group as FMZ.

Evaluation of chickpea protein isolate as a partial replacement for phosphate in pork meat batters: Techno-functional properties and molecular characteristic modifications.

The effects of chickpea protein isolate (CPI, 0.5-2 %, w/w) on the techno-functional properties of 50 % reduced-phosphate pork meat batters (RPMBs) were explored. The results showed that 1.5-2 % CPI significantly decreased the cooking loss but significantly increased the emulsion stability, hardness, gumminess, chewiness and yellowness (b*) of RPMBs (P < 0.05). CPI altered molecular characteristics of RPMBs, as demonstrated by the increased storage modulus (G'), the conversion of free water into immobilized water, the reduced intensities of the aliphatic residue Raman bands, the decreased α-helical structure and the formation of well-organized gel networks with evenly distributed small fat globules. Principal component analysis and Pearson's correlation analysis indicated that CPI-induced changes in RPMB techno-functional properties were closely related to molecular characteristics. Hierarchical cluster analysis suggested that RPMBs supplemented with 1.5-2 % CPI were highly similar in techno-functional properties to the high-phosphate group. Therefore, CPI may potentially be used to develop reduced-phosphate meat products.

Use of basic amino acids to improve gel properties of PSE-like chicken meat proteins isolated via ultrasound-assisted alkaline extraction.

To improve the gel quality of pale, soft, and exudative (PSE)-like chicken protein isolate (PPI) obtained via ultrasound-assisted alkaline extraction (UAE), l-lysine (l-Lys), l-arginine (l-Arg), or l-histidine (l-His) were used and the effects on the thermal gelling characteristics of PPI were studied. Compared with the nonbasic amino acid addition group, the addition of l-His/l-Arg/l-Lys significantly increased the solubility and absolute zeta potential of PPI, whereas reduced the particle size and turbidity (p < 0.05). They enhanced the gel strength and textural properties of PPI (p < 0.05) and reduced the cooking loss of PPI in the following order: l-Lys > l-Arg > l-His. The solubility, gel strength, and hardness of PPI with l-Lys were increased by 18.6%, 44.6%, and 57.6%, respectively, and cooking loss was decreased by 18.1%. Low-field nuclear magnetic resonance and magnetic resonance imaging revealed that basic amino acids addition decreased the water mobility in PPI gels with increasing immobile water content. Scanning electron microscopy revealed that the addition of basic amino acids promoted the formation of a more uniform and tight network microstructure in PPI gels. The α-helix content was decreased, whereas the ß-sheet content was increased in PPI gels after basic amino acid addition. Therefore, addition of basic amino acids, especially l-Lys, enhances the gel properties of PPI. PRACTICAL APPLICATION: This study revealed that adding basic amino acids effectively improved the gel properties of PPI obtained via UAE method, with l-Lys exerting the best improvement effect. Our findings highlight the application value of PSE-like meat by the improvement of gel characteristics of PPI, providing a theoretical reference for the processing and utilization of PPI.

Effects of combined chickpea protein isolate and chitosan on the improvement of technological quality in phosphate-free pork meat emulsions: Its relation to modifications on protein thermal and structural properties.

The effects of combined chickpea protein isolate (CPI, 1%, w/w) and chitosan (CHI, 1%, w/w) on the technological, thermal, and structural properties of phosphate-free pork meat emulsions (PPMEs) were investigated. The results showed that CPI + CHI significantly improved the emulsion stability (P < 0.05), synergistically elevated the hardness and chewiness, and did not negatively impact the color attributes, which endowed the PPMEs with similar or even better technological performances compared to the high-phosphate control. These alterations were related to the reduced myosin enthalpy values, the rearrangement of free water into immobilized water, the synergistic reduction in α-helical structure and increase in ß-sheet structure, the increased trans-gauche-trans SS conformation intensity of the Raman bands, and the formation of interactive protein gel networks where small-sized fat particles were evenly dispersed in the protein matrix. Therefore, combined CPI and CHI shows promise as a phosphate replacer for meat products.

Effect of boiling on texture of scallop adductor muscles and its mechanism based on label-free quantitative proteomic technique.

The mechanism behind textural changes in scallop adductor muscle during boiling was investigated through proteomic analysis, determination of water holding capacity (WHC) and oxidative indices, as well as observation with scanning electron microscopy and multiphoton nonlinear optical microscopy. The hardness and shear force showed the trend of first rising and then falling in 45 min-boiling time. The results suggested that short-time boiling caused the oxidation, denaturation and aggregation of proteins, resulting in the transverse contraction of myofibers and lateral cross-linked aggregation of muscle fibers and a rise in WHC, which led to the increase in hardness and shear force. While long-time boiling caused the progressive degradation of structural proteins such as fibrillin-1, collagen alpha-2(I) chain, myosin heavy chain, basement membrane-specific heparan sulfate proteoglycan core protein, and paramyosin, resulting in a loose myofibril network and the decrease in WHC, which led to the decrease in hardness and shear force.

Effect of different sous-vide cooking conditions on textural properties, protein physiochemical properties and microstructure of scallop (Argopecten irradians) adductor muscle.

Among 26 sous-vide cooking conditions of scallop adductor muscle (SAM), 65 °C-5.5 h, 70 °C-1.5 h and 100 °C-5 min were selected by the differential scanning calorimetry analysis. After sous-vide cooking, the shear force, hardness, springiness, cohesiveness, chewiness and recoverability of SAM increased significantly compared to fresh sample. The cooking also changed the secondary structures of the proteins in SAM with the rising ß-sheet and descending α-helix, and the chemical interactions with the rising hydrophobic interactions and disulfide bonds but the descending ionic bonds and hydrogen bonds. These caused the longitudinal shrinkage and transverse aggregation of muscle fibers, and the aggregation and cross-linking between myofibrils which led to the squeeze of immobile water from myofibril network structure. This indicated that the denaturation, oxidation, aggregation and cross-linking of proteins caused by heat treatment changed the microstructure and water distribution, which contributed to the increased textural indicators of sous-vide cooked SAM.

Effects of Boiling Processing on Texture of Scallop Adductor Muscle and Its Mechanism.

The objective of this study was to reveal the effects of boiling processing on the texture of scallop adductor muscle (SAM) and its mechanism. Compared to the fresh sample, all the texture indicators, including the hardness, chewiness, springiness, resilience, cohesiveness, and shear force of 30-s- and 3-min-boiled SAMs increased time-dependently (p < 0.05). As the boiling time increased further to 15 min, the shear force and cohesiveness still increased significantly (p < 0.05), and the resilience and hardness were maintained (p > 0.05), but the springiness and chewiness decreased significantly (p < 0.05). The overall increase in the texture indicators of the boiled SAMs was due to the boiling-induced protein denaturation, aggregation, and increased hydrophobicity, resulting in the longitudinal contraction and lateral expansion of myofibrils, the longitudinal contraction and lateral cross-linked aggregation of muscle fibers, and the loss of free water. However, the decreasing springiness and chewiness of the 15-min-boiled SAMs was due to the significant degradation of proteins (especially collagen), resulting in the destruction of the connective tissue between the muscle fiber clusters. Both from a subjective sensory point of view and from the objective point of view of protein denaturation and degradation, 3-min-boiled SAMs are recommended. The quality improvement of thermally processed products by controlled, moderate cooking is of practical value from the perspective of food consumption.

Formation and disappearance of aldehydes during simulated gastrointestinal digestion of fried clams.

The formation and disappearance of aldehydes during simulated gastrointestinal digestion (SD) of fried clams was investigated in order to shed light on the underlying mechanism. Results from the thiobarbituric acid reactive substance (TBARS) and fluorometric assays using a specific aldehyde probe indicated that the SD (with lipase) of fried clams initially reduced (at the gastric stage), but subsequently increased (mainly at the intestinal stage) the contents of total aldehydes. Meanwhile, eight specific aldehydes including propanal, acrolein, trans-2-pentenal, hexanal, trans,trans-2,4-octadienal, trans,trans-2,4-decadienal, 4-hydroxy-hexenal and 4-hydroxy-nonenal in the digested meal were determined by using a high-performance liquid chromatography-tandem electrospray ionization mass spectrometry (HPLC-ESI-MS/MS) method. Results indicated that the changes in the trend of the contents of the eight aforementioned aldehydes were similar to those of total aldehydes during SD (with lipase) of fried clams. However, a similar SD process without lipase time-dependently reduced the contents of total and individual aldehydes. Moreover, lipid classes and free fatty acids (FFAs) in the digested meal were determined to reveal the degree of hydrolysis of lipids during the SD process. Results indicated that the SD (with lipase) of fried clams significantly hydrolyzed triacylglycerols (TAG) and polar lipids (PL) and produced FFAs, but the SD process without lipase resulted in negligible lipid hydrolysis. Thus, our results demonstrated a positive correlation between lipid hydrolysis and aldehyde generation during the SD of fried clams. Alternatively, unsaturated FFAs instead of TAG and PL could have served as the main precursors for aldehyde generation due to their high oxidative susceptibility.

Non-thermal plasmas chemistry as a tool for environmental pollutants abatement.

Over the past several decades, interest in environmental pollutants abatement has greatly increased. This interest is derived from growing concern about environmental pollution and the serious deterioration of many ecosystems as a result of environmental pollution. NTPs chemistry is a proven and effective tool both for decomposing a range of pollutants and for cleaning contaminated surfaces. NTPs chemistry has crucial advantages such as high energy efficiency, higher treatment effectiveness, effective treatment in more confined spaces, and near zero-emissions. When NTPs chemistry is combined with the use of certain catalysts, synergetic pollution abatement results may be achieved; however, the mechanism by which the synergy occurs is still unclear. The purpose of this paper is to provide a basic understanding of NTPs chemistry, including the commonly employed chemical mechanisms, examples of NTPs chemistry applications, and an opinion on the future for NTPs chemistry in the field of environmental pollutants abatement.

Effect of gellan gum on functional properties of low-fat chicken meat batters.

The effects of substituting pork back-fat with addition of gellan gum and water on gel characteristics, rheological property, water mobility, and distribution of chicken meat batters were studied. The addition of gellan gum significantly affected the cooking yield, color, texture, and rheological property of chicken meat batters (p < .05). The cooking yield and textural properties were not significantly different (p > .05) when gellan gum was added with 0, 0.2, and 0.4%. When the level of gellan gum reached 0.6%, the cooked chicken meat batters had significantly lower L* value, cooking yield, hardness, springiness, cohesiveness, and chewiness values (p < .05). The initial storage modulus (G') of chicken batters was significantly decreased (p < .05) with increasing gellan gum. During the heating, an increase in denaturation temperature of the myosin head and tail were observed in chicken batters with added gellan gum. The G' of chicken batters with 0, 0.2, and 0.4% gellan gum were not significant difference (p > .05) at 80C, but the G' of chicken batters with 0.6% gellan gum was significantly decreased (p < .05). LF-NMR revealed that T21 and T22 were significantly increased (p < .05) with increasing the water and gellan gum content while P21 and P22 were not significantly different (p > .05) in the chicken batters with 0, 0.2, and 0.4% gellan gum. A significant decrease was observed (p < .05) in the chicken batter with 0.6% gellan gum. Overall, added 0.2 and 0.4% gellan gum to chicken meat batters could reduce pork back-fat. PRACTICAL APPLICATIONS: Emulsion-type meat products contain a high level of fat, saturated fatty acids, and cholesterol, which have a potential negative effect on consumers health. However, animal fat plays an important role in the textural properties and water-holding capacity of meat products. One of great challenges in meat processing is how to find a fat substitute to produce low-fat meat products which have overall acceptable quality. This study showed that the addition of gellan gum influenced the textural properties of chicken batters and changed the dynamic rheological behavior. The addition of 0.2 and 0.4% gellan gum kept a stable cooking yield and textural properties of cooked chicken batters and also reduced the pork back-fat. LF-NMR could reflect the cooking yield and gel properties of the chicken batters. Gellan gum may be used to replace pork back-fat to produce low-fat chicken meat batters having overall acceptability in meat industry for meeting consumer demand.

A rapid method for the determination of dopamine in porcine muscle by pre-column derivatization and HPLC with fluorescence detection.

A rapid method has been developed based on the sample preparation procedure named as QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe), combined with reversed-phase high performance liquid chromatography with fluorescence detector and C18 column after pre-column derivatization using o-phthalaldehyde and 2-mercaptoethanol to determine dopamine in porcine muscle. Methanol and deionized water (0.1% acetic acid, v/v) with a ratio of 60:40 was used as mobile phase. The flow rate was 0.8 mL/min and dopamine was eluted within 15 min. The linearity range was 0.003-8 µg/mL with r=0.9992. The detection limit for dopamine was 4 µg/kg and the quantification limit was 9 µg/kg. Recovery studies were carried out at 0.1, 0.5 and 1.0 mg/kg fortification levels and the average recoveries obtained ranged from 90.4% to 98.2% with relative standard deviations between 3.5% and 8.1%. The method was found to be suitable for detection of dopamine in animal product tissues at the maximum residue level.

Effect of chickpea protein modified with combined heating and high-pressure homogenization on enhancing the gelation of reduced phosphate myofibrillar protein.

The effects of chickpea protein (CP) modified by heating and/or high-pressure homogenization (HPH) on the gelling properties of myofibrillar protein under reduced phosphate conditions (5 mM sodium triphosphate, STPP) were investigated. The results showed that heating and HPH dual-modified CP could decrease the cooking loss by 29.57 %, elevate the water holding capacity by 17.08 %, and increase the gel strength by 126.88 %, which conferred myofibrillar protein with gelation performance comparable with, or even surpassing, that of the high-phosphate (10 mM STPP) control. This gelation behavior improvement could be attributed to enhanced myosin tail-tail interactions, decreased myosin thermal stability, elevated trans-gauche-trans disulfide conformation, strengthened hydrophobic interactions and hydrogen bonding, the uncoiling of α-helical structures, the formation of well-networked myofibrillar protein gel, and the disulfide linkages between the myosin heavy chain, actin, and CP subunits. Therefore, the dual-modified CP could be a promising phosphate alternative to develop healthier meat products.