Effect of ultrasound treatment on thawing process of frozen tofu prepared with different salt coagulants.

This study investigated the effects of ultrasound-assisted water thawing (UWT) at different power levels (0, 100, 150, 200, and 250 W) on the thawing rate and gel properties of frozen tofu made using three different salt coagulants (CaCl2, CaSO4, and MgCl2). Tofu produced with CaCl2 and CaSO4 elicited gel structures with dense and homogeneous networks, while that with MgCl2 had rough pores and irregular networks. UWT treatment significantly decreased thawing time by 30.9-53.5% compared to the control. Water holding capacity and scanning electron microscopy analyses demonstrated that UWT-100, UWT-150, and UWT-200 should be used to increase the amount of fixed water for CaCl2, CaSO4, and MgCl2. These findings suggest that appropriate ultrasonic treatment could improve the water retention capacity of the tofu network and make the gel network structure more compact. Additionally, protein structural analysis showed a decrease in the exposure of hydrophobic groups and reduced protein denaturation when tofu prepared with all the coagulants were thawed with UWT energies of 100-200 W ultrasonication. These findings offer theoretical support for improving the frozen tofu thawing process while ensuring optimal final product quality.

Effect of polysaccharides on the rheological behaviour of soy-wheat protein aggregation and conformational changes during high-moisture extrusion.

BACKGROUND: Due to the extrusion black box effect, polysaccharides determine the formation of meat-like fibrous structures by modulating the flow behaviour and structural changes of plant proteins under high-moisture extrusion conditions. However, there is limited knowledge on the mechanism of resolution. This study simulated the rheological properties of soy protein-wheat protein under 57% moisture conditions with addition of 4% sodium alginate (SA), 2% xanthan gum (XG), and 2% maltodextrin (MD). The effect of these polysaccharides on the aggregation behaviour and conformation of raw protein during high-moisture extrusion was investigated. RESULTS: It was revealed that the three polysaccharides were effective in increasing the interaction between proteins and between proteins and water. Among them, 4% SA elicited a significantly stronger storage modulus (gelation behaviour) compared to the control. Analysis of different zones of extrudates by protein electrophoresis, particle size, and turbidity showed that SA-4% was able to form more high molecular protein aggregates (> 245 kDa) and promoted crosslinking of low molecular subunits (< 48 kDa), resulting in moderately sized protein aggregated particles. Fluorescence and ultraviolet spectra showed the transformation of protein tertiary structures in different extrusion zones, confirming that the key extrusion zone for protein conformational transformation by polysaccharides is the die-cooling zone. Furthermore, stretching of polypeptide chains and accelerated protein rearrangement facilitated the formation of more fibrillar structures. CONCLUSION: Theoretical support for polysaccharide modulation of plant protein quality in high moisture extruded products is provided by this study. © 2023 Society of Chemical Industry.

Effect of high-pressure homogenization on Ca2+ -induced gel formation of soybean 11 S globulin.

BACKGROUND: High-pressure homogenization (HPH) is commonly used as a non-thermal processing technique for soybean and soy protein products, and the preparation of soy protein gel products often requires the synergistic effect of HPH and heat treatment. The dissociative association behavior of 11 S is the key to the protein gel formation state. In this study, therefore, 11 S thermal gels were prepared by high-pressure homogenization and co-induction (90 °C, 30 min) (adding Ca2+ to promote gel formation before heat treatment), and the effects of different high-pressure homogenization pressures (0-100 MPa) and co-treatment on the dissociative association behavior of 11 S protein, gel properties, and microstructure of 11 S gels were investigated. RESULTS: The results showed that HPH at higher pressures led to the breaking of disulfide bonds of aggregates and disrupted non-covalent interactions in protein aggregates, leading to collisions between protein aggregates and the reduction of large protein aggregates. High-pressure homogenization treatment at 60 MPa improved the gel properties of 11 S more. The HPH combined with heating changed the binary and tertiary structure of 11 S soy globulin and enhanced the hydrophobic interaction between 11 S molecules, thus improving the gel properties of 11 S. The change in intermolecular forces reflected the positive effect of HPH treatment on the formation of denser and more homogeneous protein gels. CONCLUSION: In conclusion, high-pressure homogenization combined with heating can improve the properties of 11 S gels by changing the structure of 11 S protein, providing data and theoretical support for soy protein processing and its further applications. © 2022 Society of Chemical Industry.

Effect of maltodextrin on the oxidative stability of ultrasonically induced soybean oil bodies microcapsules.

Introduction: Encapsulation of soybean oil bodies (OBs) using maltodextrin (MD) can improve their stability in different environmental stresses and enhance the transport and storage performance of OBs. Methods: In this study, the effects of different MD addition ratios [OBs: MD = 1:0, 1:0.5, 1:1, 1:1.5, and 1:2 (v/v)] on the physicochemical properties and oxidative stability of freeze-dried soybean OBs microcapsules were investigated. The effect of ultrasonic power (150-250 W) on the encapsulation effect and structural properties of oil body-maltodextrin (OB-MD) microcapsules were studied. Results: The addition of MD to OBs decreased the surface oil content and improved the encapsulation efficiency and oxidative stability of OBs. Scanning electron microscopy images revealed that the sonication promoted the adsorption of MD on the surface of OBs, forming a rugged spherical structure. The oil-body-maltodextrin (OB-MD) microcapsules showed a narrower particle size distribution and a lower-potential absolute value at an MD addition ratio of 1:1.5 and ultrasonic power of 250 W (32.1 mV). At this time, MD-encapsulated OBs particles had the highest encapsulation efficiency of 85.3%. Ultrasonic treatment improved encapsulation efficiency of OBs and increased wettability and emulsifying properties of MD. The encapsulation of OBs by MD was improved, and its oxidative stability was enhanced by ultrasound treatment, showing a lower hydrogen peroxide value (3.35 meq peroxide/kg) and thiobarbituric acid value (1.65 µmol/kg). Discussion: This study showed that the encapsulation of soybean OBs by MD improved the stability of OBs microcapsules and decreased the degree of lipid oxidation during storage. Ultrasonic pretreatment further improved the encapsulation efficiency of MD on soybean OBs, and significantly enhanced its physicochemical properties and oxidative stability.

Structural Analysis and Study of Gel Properties of Thermally-Induced Soybean Isolate-Potato Protein Gel System.

Heat-induced composite gel systems consisting of different soybean protein isolate (SPI) and potato protein (PP) mixtures were studied to elucidate their "backbone" and property changes. This was achieved by comparing the ratio of non-network proteins, protein subunit composition, and aggregation of different gel samples. It was revealed that SPI was the "gel network backbone" and PP played the role of "filler" in the SPI-PP composite gel system. Compared with the composite gels at the same ratio, springiness and WHC decrease with PP addition. For hardness, PP addition showed a less linear trend. At the SPI-PP = 2/1 composite gel, hardness was more than doubled, while springiness and WHC did not decrease too much and increased the inter-protein binding. The hydrophobic interactions and electrostatic interactions and hydrogen bonding of the SPI gel system were enhanced. The scanning electron microscopy results showed that the SPI-based gel system was able to form a more compact and compatible gel network. This study demonstrates the use of PP as a potential filler that can effectively improve the gelling properties of SPI, thus providing a theoretical basis for the study of functional plant protein foods.

The regulation of 2,3-butanediol synthesis in Klebsiella pneumoniae as revealed by gene over-expressions and metabolic flux analysis.

A variety of microorganism species are able naturally to produce 2,3-butanediol (2,3-BDO), although only a few of them are suitable for consideration as having potential for mass production purposes. Klebsiella pneumoniae (K. pneumoniae) is one such strain which has been widely studied and used industrially to produce 2,3-BDO. In the central carbon metabolism of K. pneumoniae, the 2,3-BDO synthesis pathway is dominated by three essential enzymes, namely acetolactate decarboxylase, acetolactate synthase, and butanediol dehydrogenase, which are encoded by the budA, budB, and budC genes, respectively. The mechanisms of the three enzymes have been characterized with regard to their function and roles in 2,3-BDO synthesis and cell growth (Blomqvist et al. in J Bacteriol 175(5):1392-1404, 1993), while a few studies have focused on the cooperative mechanisms of the three enzymes and their mutual interactions. Therefore, the K. pneumoniae KCTC2242::ΔwabG wild-type strain was utilized to reconstruct seven new mutants by single, double, and triple overexpression of the three enzymes key to this study. Subsequently, continuous cultures were performed to obtain steady-state metabolism in the organisms and experimental data were analyzed by metabolic flux analysis (MFA) to determine the regulation mechanisms. The MFA results showed that the seven overexpressed mutants all exhibited enhanced 2,3-BDO production, and the strain overexpressing the budBA gene produced the highest yield. While the enzyme encoded by the budA gene produced branched-chain amino acids which were favorable for cell growth, the budB gene enzyme rapidly enhanced the conversion of acetolactate to acetoin in an oxygen-dependent manner, and the budC gene enzyme catalyzed the reversible conversion of acetoin to 2,3-BDO and regulated the intracellular NAD(+)/NADH balance.

Effect of pH on the metabolic flux of Klebsiella oxytoca producing 2,3-butanediol in continuous cultures at different dilution rates.

The efficiency of the bioconversion process and the achievable end-product concentration decides the economic feasibility of microbial 2,3-butanediol (2,3-BDO) production. In 2,3-BDO production, optimization of culture condition is required for cell growth and metabolism. Also, the pH is an important factor that influences microbial performance. For different microorganisms and substrates, it has been shown that the distribution of the metabolites in 2,3-BDO fermentation is greatly affected by pH, and the optimum pH for 2,3-BDO production seems dependently linked to the particular strain and the substrate employed. Quantification analysis of intracellular metabolites and metabolic flux analysis (MFA) were used to investigate the effect of pH on the Klebsiella oxytoca producing 2,3-BDO and other organic acids. The main objectives of MFA are the estimation of intracellular metabolic fluxes and the identification of rate-limiting step and the key enzymes. This study was conducted under continuous aerobic conditions at different dilution rates (0.1, 0.2, and 0.3 h(-1)) and different pH values (pH 5.5 and 7.0) for the steady-state experimental data. In order to obtain the flux distribution, the extracellular specific rates were calculated from the experimental data using the metabolic network model of K. oxytoca. Intracellular metabolite concentration profiles were generated using ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry.

Enhanced 2,3-butanediol production in recombinant Klebsiella pneumoniae via overexpression of synthesis-related genes.

2,3-Butanediol (2,3-BD) is a major metabolite produced by Klebsiella pneumoniae KCTC2242, which is a important chemical with wide applications. Three genes important for 2,3-BD biosynthesis acetolactate decarboxylase (budA), acetolactate synthase (budB), and alcohol dehydrogenase (budC) were identified in K. pneumoniae genomic DNA. With the goal of enhancing 2,3-BD production, these genes were cloned into pUC18K expression vectors containing the lacZ promoter and the kanamycin resistance gene to generate plasmids pSB1-7. The plasmids were then introduced into K. pneumoniae using electroporation. All strains were incubated in flask experiments and 2,3-BD production was increased by 60% in recombinant bacteria harboring pSB04 (budA and budB genes), compared with the parental strain K. pneumoniae KCTC2242. The maximum 2,3-BD production level achieved through fedbatch fermentation with K. pneumoniae SGJSB04 was 101.53 g/l over 40 h with a productivity of 2.54 g/l.h. These results suggest that overexpression of 2,3-BD synthesisrelated genes can enhance 2,3-BD production in K. pneumoniae by fermentation.

Identification of factors regulating Escherichia coli 2,3-butanediol production by continuous culture and metabolic flux analysis.

2,3-Butanediol (2,3-BDO) is an organic compound with a wide range of industrial applications. Although Escherichia coli is often used for the production of organic compounds, the wild-type E. coli does not contain two essential genes in the 2,3-BDO biosynthesis pathway, and cannot ferment 2,3-BDO. Therefore, a 2,3-BDO biosynthesis mutant strain of Escherichia coli was constructed and cultured. To determine the optimum culture factors for 2,3-BDO production, experiments were conducted under different culture environments ranging from strongly acidic to neutral pH. The extracellular metabolite profiles were obtained using high-performance liquid chromatography (HPLC), and the intracellular metabolite profiles were analyzed by ultra-performance liquid chromatography and quadruple time-of-flight mass spectrometry (UPLC/ Q-TOF-MS). Metabolic flux analysis (MFA) was used to integrate these profiles. The metabolite profiles showed that 2,3-BDO production favors an acidic environment (pH 5), whereas cell mass favors a neutral environment. Furthermore, when the pH of the culture fell below 5, both the cell growth and 2,3-BDO production were inhibited.

Dynamic modeling of lactic acid fermentation metabolism with Lactococcus lactis.

A dynamic model of lactic acid fermentation using Lactococcus lactis was constructed, and a metabolic flux analysis (MFA) and metabolic control analysis (MCA) were performed to reveal an intensive metabolic understanding of lactic acid bacteria (LAB). The parameter estimation was conducted with COPASI software to construct a more accurate metabolic model. The experimental data used in the parameter estimation were obtained from an LC-MS/ MS analysis and time-course simulation study. The MFA results were a reasonable explanation of the experimental data. Through the parameter estimation, the metabolic system of lactic acid bacteria can be thoroughly understood through comparisons with the original parameters. The coefficients derived from the MCA indicated that the reaction rate of L-lactate dehydrogenase was activated by fructose 1,6-bisphosphate and pyruvate, and pyruvate appeared to be a stronger activator of L-lactate dehydrogenase than fructose 1,6-bisphosphate. Additionally, pyruvate acted as an inhibitor to pyruvate kinase and the phosphotransferase system. Glucose 6-phosphate and phosphoenolpyruvate showed activation effects on pyruvate kinase. Hexose transporter was the strongest effector on the flux through L-lactate dehydrogenase. The concentration control coefficient (CCC) showed similar results to the flux control coefficient (FCC).