Production of gluconic acid from the washed rice waste water using Au/MgO catalyst - A sustainable route.

The majority of the waste produced by the food and agriculture industries is abundant in proteins, carbohydrates, and fats, which can be utilized effectively in other food products or industrial products. Especially, washed rice water (WRW) contains a significant quantity of starch that has been discarded without being utilized properly. In the present investigation, we have successfully upgraded washed rice water into the industrially important intermediate, i.e., gluconic acid, using an Au/MgO catalyst in a single pot reaction. The upgrading strategy was developed in three consecutive phases using two different model reactions: (1) glucose to gluconic acid, (2) hydrolysis of starch into glucose, followed by the oxidation reaction. The results showed that almost 60% gluconic acid was achieved at room temperature with atmospheric pressure. The present investigation highlighted that hydrolysis, followed by oxidation reaction is the most promising route for upgrading WRW to gluconic acid.

Preparation of bovine serum albumin-arabinoxylan cold-set gels by glucono-δ-lactone and salt ions double induction.

In order to investigate the effect of glucono-δ-lactone (GDL) and different salt ions (Na+ and Ca2+) induction on the cold-set gels of bovine serum albumin (BSA)-arabinoxylan (AX), the gel properties and structure of BSA-AX cold-set gels were evaluated by analyzing the gel strength, water-holding capacity, thermal properties, and Fourier Transform Infrared (FTIR) spectra. It was shown that the best gel strength (109.15 g) was obtained when the ratio of BSA to AX was 15:1. The addition of 1 % GDL significantly improved the water-holding capacity, gel strength and thermal stability of the cold-set gels (p < 0.05), and the microstructure was smoother. Low concentrations of Na+ (3 mM) and Ca2+ (6 mM) significantly enhanced the hydrophobic interaction and hydrogen bonding between BSA and AX after acid induction, and the Na+-induced formation of a denser microstructure with a higher water-holding capacity (75.51 %). However, the excess salt ions disrupted the stable network structure of the cold-set gels and reduced their thermal stability and crystalline structure. The results of this study contribute to the understanding of the interactions between BSA and AX induced by GDL and salt ions, and provide a basis for designing hydrogels with different properties.

Effects of secondary cross-linking on the physicochemical properties of sodium alginate-hydrogel and in vitro release of anthocyanins.

To enhance the physicochemical properties and extend the release duration of sodium alginate (SA) hydrogels, this study explored the impact of acidifier type and the number of cross-linking on the physicochemical characteristics and in vitro anthocyanin release from SA hydrogels, utilizing calcium carbonate as the cross-linking agent. The findings revealed that the utilization of gluconolactone (GDL) as an acidifying agent in the preparation of SA hydrogels, as opposed to hydrochloric acid, resulted in a deceleration of the hydrolysis process of calcium carbonate. This deceleration led to the strengthening of hydrogen-bonding interactions and the development of a more compact network structure within the SA hydrogels. Consequently, there was a noticeable enhancement in the hardness, relaxation time, and anthocyanin encapsulation efficiency of the gels. Additionally, the release of anthocyanins in simulated intestinal fluid was delayed. Secondary cross-linking was found to facilitate ionic interactions between SA and Ca2+, further intensifying the denseness of the network structure and enhancing the physicochemical characteristics of the SA hydrogels. Overall, SA hydrogels processed with GDL as the acidifier and subjected to secondary cross-linking exhibited improved physicochemical properties, delayed release effects, and proved to be an efficient system for the delayed release of anthocyanins.

Identification of Tellurium Metabolite in Broccoli Using Complementary Analyses of Inorganic and Organic Mass Spectrometry.

Tellurium (Te) is a chalcogen element like sulfur and selenium. Although it is unclear whether Te is an essential nutrient in organisms, unique Te metabolic pathways have been uncovered. We have previously reported that an unknown Te metabolite (UKTe) was observed in plants exposed to tellurate, a highly toxic Te oxyanion, by liquid chromatography-inductively coupled plasma mass spectrometer (LC-ICP-MS). In the present study, we detected UKTe in tellurate-exposed broccoli (Brassica oleracea var. italica) by LC-ICP-MS and identified it as gluconic acid-3-tellurate (GA-3Te) using electrospray ionization mass spectrometer with quadrupole-Orbitrap detector and tandem MS analysis, the high-sensitivity and high-resolution mass spectrometry for organic compounds. We also found that GA-3Te was produced from one gluconic acid and one tellurate molecule by direct complexation in an aqueous solution. GA-3Te was significantly less toxic than tellurate on plant growth. This study is the first to identify the Te metabolite GA-3Te in plants and will contribute to the investigation of tellurate detoxification pathways. Moreover, gluconic acid, a natural and biodegradable organic compound, is expected to be applicable to eco-friendly remediation strategies for tellurate contamination.

Monitoring sodium caseinate and whey protein isolate interaction with mild preheat treatment using ultrasound spectroscopy and confocal laser scanning microscopy.

Ultrasound spectroscopy and confocal laser scanning microscopy (CLSM) methods were developed to visualize the interaction between sodium caseinate (SC) and whey protein isolate (WPI) with a mild preheat treatment (57°C, 10 min) followed by adding glucono-δ-lactone (GDL). Ultrasonic velocity changes during incubation at 25°C after adding GDL for four kinds of mixtures (no-treated SC plus no-treated WPI, preheated SC plus no-treated WPI, no-treated SC plus preheated WPI and preheated SC plus preheated WPI) were monitored. The results reveal that the mild preheating treatment of the proteins affected the timing of the increase in compressibility of each system. CLSM observation with individualized dyes which have different maxima of excitation and emission wavelengths, showed the preheated SC plus no-treated WPI mixture had a slightly coarse structure and the highest correlation coefficient, suggesting the highest colocalization of the SC and WPI among the four kinds of mixed-protein systems. Furthermore, the scanning electron microscopy (SEM) observation suggests that there are some differences among the gels, namely, preheated WPI leads to the formation of developed three-dimensional gel networks with filamentous structures, whereas SC promotes the formation of cluster-like crowded networks composed of more fine aggregated particles instead of developed filamentous structures. These results demonstrated that although SC is known as a heat-stable protein, pretreated SC could lead to an increase of the collaboration with WPI in the presence of GDL. This finding anticipated the possibility creating a food material with another texture using a milk-protein mixed system.

Enhancing Lactobacillus plantarum delivery: Impact of gluconolactone concentration on high-internal-phase emulsion gels and gastrointestinal viability.

In this study, the impact of Gluconolactone (GDL) concentration on the formation of high-internal-phase emulsion gels (HIPEGs) and the gastrointestinal digestive viability of Lactobacillus plantarum encapsulated within these HIPEGs were demonstrated. Increasing GDL concentrations led to cross-linking of particles at the oil-water interface, thereby stabilizing smaller oil droplets. The addition of GDL to HIPEs results in a significant increase in the secondary structure of SPI, specifically in ß-sheet and ß-turn formations, accompanied by a reduction in α-helix percentage. This alteration enhanced the binding effect of protein on water, leading to changes in intermolecular force. Notably, HIPEGs containing 3.0% GDL demonstrated superior encapsulation efficiency and delivery efficiency, reaching 99.0% and 84.5%, respectively. After 14 d of continuous zebrafishs feeding, the intestinal viable cells count of Lactobacillus plantarum reached 1.18 × 107 CFU/mL. This finding supports the potential use of HIPEGs as a probiotic delivery carrier, effectively enhancing the intestinal colonization rate.

Self-propelled enzyme-controlled IR-mesoporous silica Janus nanomotor for smart delivery.

Here, we report the preparation of a novel Janus nanoparticle with opposite Ir and mesoporous silica nanoparticles through a partial surface masking with toposelective modification method. This nanomaterial was employed to construct an enzyme-powered nanomachine with self-propulsion properties for on-command delivery. The cargo-loaded nanoparticle was provided with a pH-sensitive gate and unit control at the mesoporous face by first attaching boronic acid residues and further immobilization of glucose oxidase through reversible boronic acid esters with the carbohydrate residues of the glycoenzyme. Addition of glucose leads to the enzymatic production of H2O2 and gluconic acid, being the first compound catalytically decomposed at the Ir nanoparticle face producing O2 and causing the nanomachine propulsion. Gluconic acid leads to a pH reduction at the nanomachine microenvironment causing the disruption of the gating mechanism with the subsequent cargo release. This work demonstrates that enzyme-mediated self-propulsion improved release efficiency being this nanomotor successfully employed for the smart release of Doxorubicin in HeLa cancer cells.

Gelling properties of acid-induced tofu (soybean curd): Effects of acid type and nano-fish bone.

The effects of different types of acid coagulants and nano fish bone (NFB) additives on the characteristics of tofu were investigated using texture analyzers, SEM, FT-IR, and other techniques. The breaking force and penetration distance, in descending order, were found in the tofu induced by glucono-d-lactone (GDL) (180.27 g and 0.75 cm), citric acid (152.90 g and 0.74 cm), lactic acid (123.33 g and 0.73 cm), and acetic acid (69.84 g and 0.58 cm), respectively. The syneresis of these tofu samples was in the reverse order (35.00, 35.66, 39.66, and 44.50%). Lightness and whiteness were not significantly different among the different samples. Regardless of the acid type, the soluble calcium content in the soybean milk was significantly increased after adding NFB. As a result, the breaking force and penetration distance of all tofu samples increased significantly, but the syneresis decreased. Compared with tofu coagulated by other acids, GDL tofu formed a more uniform and dense gel network maintained by the highest intermolecular forces (especially hydrophobic interactions). Regarding the secondary structure, the lowest percentage of α-helix (22.72%) and, correspondingly, the highest ß-sheet (48.32%) and random coil (18.81%) were noticed in the GDL tofu. The effects of NFB on the tofu characteristics can be explained by the changes in the gel network, intermolecular forces, and secondary structure, which were in line with the acid type. The characteristics of acid-induced tofu can be most synergistically improved by coagulation with GDL and NFB.

Structural effects of oxidation on sugars: glucose as a precursor of gluconolactone and glucuronolactone.

Although structural information on sugars is wide, experimental studies on the oxidation products of sugars in the gas phase, free from solvent interactions, have been rarely reported. We present an experimental work on the changes in the structure and interactions of two products of glucose oxidation (D-glucono-1,5-lactone (GlcL) and D-glucurono-6,3-lactone (GlcurL)) with respect to their precursor. Features such as intramolecular interactions, ring puckering and tautomerism were observed.

Enabling High Activation of Glucose-6-Phosphate Dehydrogenase Activity Through Liquid Condensate Formation and Compression.

Droplet formation via liquid-liquid phase separation is thought to be involved in the regulation of various biological processes, including enzymatic reactions. We investigated a glycolytic enzymatic reaction, the conversion of glucose-6-phosphate to 6-phospho-D-glucono-1,5-lactone with concomitant reduction of NADP+ to NADPH both in the absence and presence of dynamically controlled liquid droplet formation. Here, the nucleotide serves as substrate as well as the scaffold required for the formation of liquid droplets. To further expand the process parameter space, temperature and pressure dependent measurements were performed. Incorporation of the reactants in the liquid droplet phase led to a boost in enzymatic activity, which was most pronounced at medium-high pressures. The crowded environment of the droplet phase induced a marked increase of the affinity of the enzyme and substrate. An increase in turnover number in the droplet phase at high pressure contributed to a further strong increase in catalytic efficiency. Enzyme systems that are dynamically coupled to liquid condensate formation may be the key to deciphering many biochemical reactions. Expanding the process parameter space by adjusting temperature and pressure conditions can be a means to further increase the efficiency of industrial enzyme utilization and help uncover regulatory mechanisms adopted by extremophiles.

Mechanism of the glucono-δ-lactone induced soymilk gelation: Enthalpy and entropy transformation in the cross-linking of protein molecules.

This study aimed to explore new techniques to regulate the quality of soy products. The glucono-δ-lactone (GDL) induced soymilk gelation process and the gel network structure characteristic were compared as a matter of temperature, and the role of reaction kinetics was discussed. Results showed that there were similarities in the development of G' curves under different temperatures, whereas the gel network structures and the energy requirements of cross-linking reactions were different. In the high-temperature region (70 °C-95 °C), the exposure and binding of reactive groups were promoted. The activation enthalpy (ΔH*) required by protein aggregates decreased and the effect of entropy reduction (-TΔS*) was enhanced, which led to shorten the preaggregation time (tg) and increase the gelation rate (k), resulting in the formation of rough, porous gel network with high stiffness. By contrast, in the low-temperature region (40 °C-70 °C), high enthalpy contributions and low entropy changes were required, then a fine, soft, and tender gel network formed. Besides, a funnel-shaped model of the enthalpy-entropy energy transformation mechanism of soymilk gelation was proposed. The results of this study revealed that adjusting the enthalpy-entropy energy requirements of the protein cross-linking reaction could be utilized to the regulation of the network structure and quality of soymilk gels, which could enrich the reaction kinetics theory and provide innovative ideas for food quality control technology from the perspective of energy requirement and energy input.

Design of glucono-δ-lactone-induced pinto bean protein isolate/κ-carrageenan mixed gels with various microstructures: fabrication, characterization, and release behavior.

BACKGROUND: Protein gels are used for different purposes, such as providing good texture, serving as fat replacers, and enhancing the nutritional and functional characteristics of foods. They can also deliver controlled release agents for sensitive drugs. The objective of this study was to investigate the impact of κ-carrageenan (kcr) concentration (0, 1.5, 3, and 4.5 mg g-1 ) on the morphological and physicochemical properties and release behavior of glucono-δ-lactone (GDL)-induced pinto bean protein aggregate (PBA) gels. RESULTS: When κ-carrageenan concentration increased from 0 to 1.5 and 3 mg.g-1 , the firmness of the samples increased significantly, by 2.04 and 3.7 fold, respectively (P < 0.05). A compact and homogenous network with considerable strength and maximum water-holding capacity (97.52 ± 1.17%) was obtained with the addition of 3 mg g-1 κ-carrageenan to the gel system. Further increasing the κ-carrageenan concentration to 4.5 mg g-1 produced a coarse gel structure with higher storage modulus (G'), firmness (6.30-fold), thermal stability, and entrapment efficiency (85.6%). Depending on the κ-carrageenan concentration, various microstructures from protein continuous phase to κ-carrageenan continuous phase were observed. The release test indicated that 70.25% of the loaded curcumin was released in the simulated gastrointestinal tract for pure PBA gels. In contrast, for binary gels containing 4.5 mg g-1 κ-carrageenan, curcumin was protected in the upper gastrointestinal tract, and 64.45% of loaded curcumin was delivered to the colon. CONCLUSION: Our study showed that κ-carrageenan/PBA gels had high entrapment efficiency and could protect curcumin in the upper gastrointestinal tract. The hydrogels are therefore very valuable for colon-targeting delivery purposes. © 2022 Society of Chemical Industry.

Characterization of 3 phylogenetically distinct membrane-bound d-gluconate dehydrogenases of Gluconobacter spp. and their biotechnological application for efficient 2-keto-d-gluconate production.

We identified a novel flavoprotein-cytochrome c complex d-gluconate dehydrogenase (GADH) encoded by gndXYZ of Gluconobacter oxydans NBRC 3293, which is phylogenetically distinct from previously reported GADHs encoded by gndFGH and gndSLC of Gluconobacter spp. To analyze the biochemical properties of respective GADHs, Gluconobacter japonicus NBRC 3271 mutant strain lacking membranous d-gluconate dehydrogenase activity was constructed. All GADHs (GndFGH, GndSLC, and GndXYZ) were successfully overexpressed in the constructed strain. The optimal pH and KM value at that pH of GndFGH, GndSLC, and GndXYZ were 5, 6, and 4, and 8.82 ± 1.15, 22.9 ± 5.0, and 11.3 ± 1.5 m m, respectively. When the mutants overexpressing respective GADHs were cultured in d-glucose-containing medium, all of them produced 2-keto-d-gluconate, revealing that GndXYZ converts d-gluconate to 2-keto-d-gluconate as well as other GADHs. Among the recombinants, the gndXYZ-overexpressing strain accumulated the highest level of 2-keto-d-gluconate, suggesting its potential for 2-keto-d-gluconate production.

Xylonolactonase from Caulobacter crescentus Is a Mononuclear Nonheme Iron Hydrolase.

Caulobacter crescentus xylonolactonase (Cc XylC, EC 3.1.1.68) catalyzes an intramolecular ester bond hydrolysis over a nonenzymatic acid/base catalysis. Cc XylC is a member of the SMP30 protein family, whose members have previously been reported to be active in the presence of bivalent metal ions, such as Ca2+, Zn2+, and Mg2+. By native mass spectrometry, we studied the binding of several bivalent metal ions to Cc XylC and observed that it binds only one of them, namely, the Fe2+ cation, specifically and with a high affinity (Kd = 0.5 µM), pointing out that Cc XylC is a mononuclear iron protein. We propose that bivalent metal cations also promote the reaction nonenzymatically by stabilizing a short-lived bicyclic intermediate on the lactone isomerization reaction. An analysis of the reaction kinetics showed that Cc XylC complexed with Fe2+ can speed up the hydrolysis of d-xylono-1,4-lactone by 100-fold and that of d-glucono-1,5-lactone by 10-fold as compared to the nonenzymatic reaction. To our knowledge, this is the first discovery of a nonheme mononuclear iron-binding enzyme that catalyzes an ester bond hydrolysis reaction.

Development of a pulse-induced electrochemical biosensor based on gluconamide for Gram-negative bacteria detection.

Pathogenic bacteria can cause the outbreaks of disease and threaten human health, which stimulates the development of advanced detection techniques. Herein, a specific and sensitive electrochemical biosensor for Gram-negative bacteria was established based on the conductive polymer with artificial muscle properties.  The effective recognition was achieved through the specific carbohydrate-carbohydrate interaction between gluconamide and lipopolysaccharide.  The application of impulse voltage enhances the efficiency of recognition and shortens the detection time through the temporary deformation of the electrode surface, with a limit of detection (LOD)  of 1 × 100 CFU/mL and a linear range of 1 × 100 - 1 × 106 CFU/mL for Escherichia coli (E. coli). In addition  to the merits of low cost, high efficiency, and rapidity,  the developed label-free electrochemical biosensor can also be applicable for other Gram-negative bacteria, owning promising potential in the application of portable devices and paving a potential way for the construction of electrochemical biosensors.

N-Terminal Modification of Gly-His-Tagged Proteins with Azidogluconolactone.

Site-specific protein modifications are vital for biopharmaceutical drug development. Gluconoylation is a non-enzymatic, post-translational modification of N-terminal HisTags. We report high-yield, site-selective in vitro α-aminoacylation of peptides, glycoproteins, antibodies, and virus-like particles (VLPs) with azidogluconolactone at pH 7.5 in 1 h. Conjugates slowly hydrolyse, but diol-masking with borate esters inhibits reversibility. In an example, we multimerise azidogluconoylated SARS-CoV-2 receptor-binding domain (RBD) onto VLPs via click-chemistry, to give a COVID-19 vaccine. Compared to yeast antigen, HEK-derived RBD was immunologically superior, likely due to observed differences in glycosylation. We show the benefits of ordered over randomly oriented multimeric antigen display, by demonstrating single-shot seroconversion and best virus-neutralizing antibodies. Azidogluconoylation is simple, fast and robust chemistry, and should accelerate research and development.

Modulation of the clinically accessible gelation time using glucono-d-lactone and pyridoxal 5'-phosphate for long-acting alginate in situ forming gel injectable.

The purpose of this study was to design alginate in situ forming gel (ISFG) injectable with clinically acceptable gelation time and controlled release of hydrophobic drug. Milled or unmilled paliperidone palmitate (PPP) was used. The gelation time was controlled by varying the ratios of glucono-d-lactone (GDL) and pyridoxal 5'-phosphate (PLP) in prefilled alginate solution mixtures (ASMs) containing PPP, CaCO3, GDL and PLP for clinically acceptable injectability. However, the gelation time was varied by the alginate type (M/G ratio), storage condition, and drug solubilizers. This ISFG exhibited 32.15 kPa of the maximal compressive stress without causing pain and stiffness. The ISFG containing conically milled PPP released PPP in a controlled manner without exhibiting any initial burst release for 4 weeks. The current alginate ISFG injectable using new combination of PLP and GDL could be used to deliver long-acting injectable drugs.

Differences in the physicochemical, digestion and microstructural characteristics of soy protein gel acidified with lactic acid bacteria, glucono-δ-lactone and organic acid.

This study evaluated the differences in the physicochemical, digestion and microstructure of soy protein gels acidified with Lactobacillus casei (L. casei), glucono-δ-lactone (GDL) and citric acid. The maximum acidification rate was as follows: citric acid > GDL > L. casei. The gelation points of L. casei-induced gel (LC gel) and GDL-induced gel (GDL gel) occurred at 74 min and 55 min; however, gelation point of citric acid-induced gel (CA gel) was not detected because acidification was too fast. LC gel showed the high gel hardness (20.40 ± 2.23 g) and water holding capacity (84.58 ± 0.59%). At the end of intestinal digestion, the average particle size of the LC gel was the largest, but there was no significant difference between GDL gel and CA gel. The microstructure of the GDL gel was found to be the densest. Acidification rate was the "key step" of acid-induced gels, while both the proteolytic and exopolysaccharide (EPS) production capacity were involved in LC gel.

Enzymatic Management of pH in White Wines.

This paper investigates the potential of the enzymatic management of high pH in white juice and wine using a combination of enzymes-glucose oxidase coupled with catalase. Catazyme® 25 L, a commercially available blend of the two enzymes, was added at different doses (0.2 g/L, 0.6 g/L, and 1g/L) to white grape juice and various parameters (glucose, gluconic acid, pH) were monitored over 24 h of treatment. Treated wines were fermented to dryness without any difficulty and the wines were chemically and sensorially evaluated. At the highest dose (1 g/L), pH was reduced from 3.9 to 3.2, with 20.5 g of gluconic acid produced, while at the lowest dose (0.2 g/L), pH decreased from 4.0 to 3.5 and 8.8 g of gluconic acid was produced. Flash profiling indicated that treated wines were lighter in color than the control and were described using terms such as floral, fruit, citrus, and sour while the control wine was described as being fermented, medicinal, pungent, and oxidized. In conclusion, glucose oxidase coupled with catalase was shown to be effective at significantly reducing juice and wine pH in a short amount of time and with a positive impact on the organoleptic profiles of the treated wines.

Cloning, purification, and characterization of the 6-phosphogluconate dehydrogenase (6 PGDH) from Giardia lamblia.

Giardia lamblia, due to the habitat in which it develops, requires a continuous supply of intermediate compounds that allow it to survive in the host. The pentose phosphate pathway (PPP) provides essential molecules such as NADPH and ribulose-5-phosphate during the oxidative phase of the pathway. One of the key enzymes during this stage is 6-phosphogluconate dehydrogenase (6 PGDH) for generating NADPH. Given the relevance of the enzyme, in the present work, the 6pgdh gene from G. lamblia was amplified and cloned to produce the recombinant protein (Gl-6 PGDH) and characterize it functionally and structurally after the purification of Gl-6 PGDH by affinity chromatography. The results of the characterization showed that the protein has a molecular mass of 54 kDa, with an optimal pH of 7.0 and a temperature of 36-42 °C. The kinetic parameters of Gl-6 PGDH were Km = 49.2 and 139.9 µM (for NADP+ and 6-PG, respectively), Vmax =26.27 µmol*min-1*mg-1, and Kcat = 24.0 s-1. Finally, computational modeling studies were performed to obtain a structural visualization of the Gl-6 PGDH protein. The generation of the model and the characterization assays will allow us to expand our knowledge for future studies of the function of the protein in the metabolism of the parasite.