Repulsive Electrostatic Interactions Research Articles

Formation and Transport of Cr(III)-NOM-Fe Colloids upon Reaction of Cr(VI) with NOM-Fe(II) Colloids at Anoxic–Oxic Interfaces

Natural organic matter-iron (NOM-Fe) colloids are ubiquitous at anoxic-oxic interfaces of subsurface environments. Fe(II) or NOM can chemically reduce Cr(VI) to Cr(III), and the formation of Cr(III)-NOM-Fe colloids can control the fate and transport of Cr. We explored the formation and transport of Cr(III)-humic acid (HA)-Fe colloids upon reaction of Cr(VI) with HA-Fe(II) colloids over a range of environmentally relevant conditions. Cr(VI) was completely reduced by HA-Fe(II) complexes under anoxic conditions, and the formation of Cr(III)-HA-Fe colloids depended on HA concentration (or molar C/Fe ratio) and redox conditions. No colloids formed at HA concentrations below 3.5 mg C/L (C/Fe ratio below 1.6), but Cr(III)-HA-Fe colloids formed at higher HA concentrations. In column experiments, Cr(III)-HA-Fe(III) colloids formed under oxic conditions were readily transported through sand-packed porous media. Colloidal stability measurements further suggest that Cr(III)-HA-Fe colloids are highly stable and persist for at least 20 days without substantial change in particle size. This stability is attributed to the enrichment of free HA adsorbed on the Cr(III)-HA-Fe colloid surfaces, intensifying the electrostatic and/or steric repulsion interactions between particles. The new insights provided here are important for evaluating the long-term fate and transport of Cr in organic-rich redox transition zones.

Electrostatic Interactions Govern Extreme Nascent Protein Ejection Times from Ribosomes and Can Delay Ribosome Recycling

The ejection of nascent proteins out of the ribosome exit tunnel, after their covalent bond to transfer-RNA has been broken, has not been experimentally studied due to challenges in sample preparation. Here, we investigate this process using a combination of multiscale modeling, ribosome profiling, and gene ontology analyses. Simulating the ejection of a representative set of 122 E. coli proteins we find a greater than 1000-fold variation in ejection times. Nascent proteins enriched in negatively charged residues near their C-terminus eject the fastest, while nascent chains enriched in positively charged residues tend to eject much more slowly. More work is required to pull slowly ejecting proteins out of the exit tunnel than quickly ejecting proteins, according to all-atom simulations. An energetic decomposition reveals, for slowly ejecting proteins, that this is due to the strong attractive electrostatic interactions between the nascent chain and the negatively charged ribosomal-RNA lining the exit tunnel, and for quickly ejecting proteins, it is due to their repulsive electrostatic interactions with the exit tunnel. Ribosome profiling data from E. coli reveals that the presence of slowly ejecting sequences correlates with ribosomes spending more time at stop codons, indicating that the ejection process might delay ribosome recycling. Proteins that have the highest positive charge density at their C-terminus are overwhelmingly ribosomal proteins, suggesting the possibility that this sequence feature may aid in the cotranslational assembly of ribosomes by delaying the release of nascent ribosomal proteins into the cytosol. Thus, nascent chain ejection times from the ribosome can vary greatly between proteins due to differential electrostatic interactions, can influence ribosome recycling, and could be particularly relevant to the synthesis and cotranslational behavior of some proteins.

A computational study of XCCl3 (X = NC, F, Cl, Br) interacting with model ions

A computational study of model ionic complexes of XCCl3 (X = NC, F, Cl, Br) and Y (Y = Li+, Na+, K+, F−, Cl−, Br−) was undertaken. Both anionic and cationic XCCl3⋯Y dyads were found to be stable, despite repulsive electrostatic interactions in the cationic complexes. The deformation of XCCl3 and electronic charge shifts are important in stabilizing these complexes. Hydrogen bonding enhances the binding in anionic FH⋯NCCCl3⋯Y triads, but destabilizes the intermolecular interaction if Y is cationic. Complexes of NCCCl3 and LiF were also predicted, including an unusual insertion complex Li+⋯NCCCl3⋯F−.

Release of colloidal biochar during transient chemical conditions: The humic acid effect

Our understanding of colloidal biochar (CB) transport and release is largely unknown in environments with transient chemical conditions, e.g., ionic strength (IS), pH, and especially humic acid (HA). In this study, column experiments were conducted to investigate CB transport and retention in the presence and absence of HA, and CB release under transient IS and pH conditions in saturated sand. Step reductions in solution IS from 25 to 0.01 mM produced significant release peaks of CB due to a reduction in the depth of the primary minima on rough surfaces with small energy barriers. In contrast, step increases of solution pH from 4 to 10 only slightly increased CB release presumably due to the strong buffering capacity of CB. The CB retention was diminished by HA during the deposition phase. However, the release of CB with transients in IS and pH was not influenced much when deposition occurred in the presence of HA. These observations indicate that HA increased the energy barrier during deposition but did not have a large influence on the depth of the interacting minimum during transient release. Potential explanations for these effects of HA on CB retention and transient release include enhanced repulsive electrostatic interactions and/or altering of surface roughness properties. Our findings indicated that the release of retained CB is sensitive to transient IS conditions, but less dependent on pH increases and CB deposition in the presence of HA. This information is needed to quantify potential benefits and/or adverse risks of mobile CB in natural environments.

A novel mutation A212T in chloroplast Protoporphyrinogen oxidase (PPO1) confers resistance to PPO inhibitor Oxadiazon in Eleusine indica.

Protoporphyrinogen oxidase (PPO) with two isoforms, chloroplast-targeted (PPO1) and mitochondrial-targeted (PPO2), catalyzes a step in the biosynthesis of chlorophyll and heme. PPO1 and PPO2 are herbicide target sites of PPO-inhibiting herbicides. Target-site mutations conferring resistance to PPO inhibitors have all thus far been in PPO2. Oxadiazon is a unique PPO inhibitor utilized for preemergence Eleusine indica control. In this research, we evaluated the response of two previously confirmed oxadiazon-resistant and susceptible E. indica biotypes to other PPO inhibitors and identified the resistance mechanism in two oxadiazon-resistant E. indica biotypes. Two E. indica biotypes were resistant to oxadiazon, but not to other structurally unrelated PPO inhibitors, such as lactofen, flumioxazin and sulfentrazone. A novel mutation A212T was identified in the chloroplast-targeted PPO1, conferring resistance to oxadiazon in a heterologous expression system. Computational structural modeling provided a mechanistic explanation for reduced herbicide binding to the variant protein: the presence of a methyl group of threonine 212 changes the PPO1 active site and produces repulsive electrostatic interactions that repel oxadiazon from the binding pocket. The novel A212T mutation in PPO1 conferring resistance specifically to PPO inhibitor oxadiazon was characterized. This is the first evidence of the direct role of PPO1 in the PPO mode of action, and the first evidence of evolved resistance in PPO1. © 2019 Society of Chemical Industry.

Influence of dissolved organic matter on the removal of 12 organic micropollutants from wastewater effluent by powdered activated carbon adsorption

The presence of dissolved organic matter (DOM) in wastewater effluents is recognized as the main factor limiting the adsorption of organic micropollutants (OMPs) onto activated carbon. The degree of the negative effect that DOM, depending on its quality, exerts on OMPs adsorption is still unclear. The influence of the interactions between DOM and OMPs on their removal is also not fully understood. Adsorption isotherms and conventional batch tests were performed in ultra-pure water and in wastewater effluent to study the influence of DOM on the adsorption of 12 OMPs onto powdered activated carbon. Best fit of adsorption pseudo-isotherms was obtained with the Freundlich equation and showed, as expected, that OMPs adsorption was higher in ultra-pure water than in wastewater effluent due to the presence of DOM leading to pore blockage and competition for adsorption sites. LC-OCD analysis revealed that biopolymers and hydrophobic molecules were the most adsorbed fractions while humic acids were not removed after a contact time of either 30 min or 72 h. The presence of DOM had a negative impact on the removal of all OMPs after 30 min of adsorption, but similar removals to ultra-pure water were obtained for 6 OMPs after 72 h of adsorption. This demonstrated that competition between DOM and OMPs for adsorption sites was not a major mechanism as compared to pore blockage, which only slowed down the adsorption and did not prevent it. The charge of OMPs had a clear impact: the adsorption of negatively charged compounds was reduced in the presence of wastewater effluent due to repulsive electrostatic interactions with the adsorbed DOM and the PAC surface. On the other hand, the removal of positively charged compounds was improved. A 24 h pre-equilibrium between OMPs and DOM improved their removal onto PAC, which suggest that OMPs and DOM interacted in solution which decreased the negative effects caused by the presence of DOM, e.g. through co-adsorption of an OMP-DOM complex.

Understanding the Carbon-Bio Interface: Influence of Surface Chemistry and Buffer Composition on the Adsorption of Phospholipid Liposomes at Carbon Surfaces.

Surface active phospholipids are present in fluids of biological relevance, and their adsorption may condition and determine the response of carbon and nanocarbon surfaces when they are immersed in physiological media. In this work, the adsorption and assembly of liposomes at carbon interfaces were investigated to understand the effect of surface termination on the extent and mode of assembly of lipid aggregates. Liposomes of natural lipids were prepared from a mixture of phosphatidylcholine (PC) and phosphatidylserine (PS), and their hydrodynamic size and surface zeta potential were studied as a function of pH. Adsorption was investigated at graphitic amorphous carbon surfaces (a-C) and at these surfaces after oxidative treatments (a-C:O). Infrared surface spectroscopy experiments show that PC/PS liposomes adsorb at a-C surfaces exclusively, independently of pH, while no adsorption is observed at a-C:O materials. Nanogravimetry and fluorescence imaging experiments in solution indicate that adsorption at a-C occurs as supported intact vesicles. Interestingly, PC/PS adsorption at oxidized surfaces was observed only in the presence of a dication such as Ca2+, a behavior that was attributed to screening of surface-liposome repulsive electrostatic interactions. Vesicle rupture experiments show that lipids adsorb as monolayers on graphitic surfaces, whereas adsorbate structures correspond to bilayers in the case of oxidized carbons. These results therefore demonstrate a strong dependence of adsorbate structure on both carbon chemistry and buffer composition. These findings have important implications for the design of carbon nanoparticles, carbon electrodes, or carbon coatings for applications in biology and medicine.

Anomalous Salt Dependence Reveals an Interplay of Attractive and Repulsive Electrostatic Interactions in α-synuclein Fibril Formation.

α-Synuclein (α-syn) is an intrinsically disordered protein with a highly asymmetric charge distribution, whose aggregation is linked to Parkinson's disease. The effect of ionic strength was investigated at mildly acidic pH (5.5) in the presence of catalytic surfaces in the form of α-syn seeds or anionic lipid vesicles using thioflavin T fluorescence measurements. Similar trends were observed with both surfaces: increasing ionic strength reduced the rate of α-syn aggregation although the surfaces as well as α-syn have a net negative charge at pH 5.5. This anomalous salt dependence implies that short-range attractive electrostatic interactions are critical for secondary nucleation as well as heterogeneous primary nucleation. Such interactions were confirmed in Monte Carlo simulations of α-syn monomers interacting with surface-grafted C-terminal tails, and found to be weakened in the presence of salt. Thus, nucleation of α-syn aggregation depends critically on an attractive electrostatic component that is screened by salt to the extent that it outweighs the screening of the long-range repulsion between negatively charged monomers and negative surfaces. Interactions between the positively charged N-termini of α-syn monomers on the one hand, and the negatively C-termini of α-syn on fibrils or vesicles surfaces on the other hand, are thus critical for nucleation.

Expanding the structural diversity of peptide assemblies by coassembling dipeptides with diphenylalanine.

Molecular self-assembly is a bottom-up approach to fabricate novel supramolecular structures. While the structural diversity obtained by the use of a single type of building block is limited, coassembly of different peptides has recently evolved as an extended strategy to expand the diversity of peptide nanoarchitectures. Here we systematically investigate the coassembly of diphenylalanine (FF) with each one of the 399 non-FF dipeptides by micro-second molecular dynamics simulations. Our simulations show that dipeptides, by coassembling with FF, display a greatly enhanced aggregation propensity and a significantly expanded structural diversity. Regular-shaped vesicles, single- or multi-cavity assemblies, and planar sheets are formed by coassembly of FF with different types of non-FF dipeptides, which are rarely observed in self-assemblies of non-FF dipeptides. Interaction analyses reveal that the formation of these varied structures is attributed to a delicate balance between aromatic stacking, hydrophobic, and electrostatic repulsion interactions. This study provides structural and mechanistic insights into the coassembly of FF and non-FF dipeptides, thus offering a possible way to achieve a controllable design of bionanomaterials through FF-involved dipeptide coassembly.

Influence of ns laser texturing of AISI 316L surfaces for reducing bacterial adhesion

Nanosecond pulsed laser texturing has been performed on stainless steel with the objective of developing surface treatments to reduce bacterial adhesion on mechanical components in food handling machinery. The adhesion of Escherichia coli (E. coli) on four distinct textures has been investigated with standardised protocols for measurement of antibacterial performance. Surface morphology has been studied in detail for each texture to ascertain the presence of hierarchical structures and determine the role of topography in reducing bacterial adhesion. Despite the absence of sub-micrometric features comparable with bacterial size, this work highlights the crucial role that nanosecond pulsed laser irradiation plays in promoting a thin layer of iron oxide that reduces E. coli adhesion through local repulsive electrostatic interactions.

A facile non-solvent induced phase separation process for preparation of highly porous polybenzimidazole separator for lithium metal battery application

The drawbacks of low porosity, inferior electrolyte wettability, low thermal dimensional stability and permissive lithium dendrite growth of the conventional microporous polyolefin-based separators hinder their widely application in the high power density and safe Lithium ion batteries. Herein, highly porous polybenzimidazole-based separator is prepared by a facile non-solvent induced phase separation process (NIPS) using water, ethanol, chloroform and ethyl acetate as the coagulation bath solvent, respectively. It was found that the ethanol is suitable to fabricate uniform morphology macroporous separator with the porosity of 92%, electrolyte uptake of 594 wt.%, and strong mechanical strength of 15.9 MPa. In addition, the experimental tests (electrochemical analysis and XPS test) and density functional theory calculation suggest that the electron-rich imidazole ring of polybenzimidazle can enhance Li+ mobility electrostatic attraction interaction while the block the PF6− mobility via electrostatic repulsion interaction. Therefore, high Li+ transference number of 0.76 was obtained for the neat polybenzimidazole-based polymer electrolyte. As a proof of concept, the Li/LiFePO4 cell with the polybenzimidazole-based polymer electrolyte/1.0 M LiPF6− ethylene carbonate/dimethyl carbonate (v:v = 1:1) electrolyte exhibits excellent rate capability of >100 mAh g−1 at 6 C (1 C = 170 mA g−1) and superior cycle stability of 1000 cycles.

Structural, optical and rheological behavior investigations of graphene oxide/glycerol based lyotropic liquid crystalline phases

Herein, we report the dispersion and stabilization of graphene oxide (GO) in the polar organic solvent glycerol (C3H8O3). The GO/glycerol (0.5 mg/mL and 1 mg/mL) dispersions are found stable for about 4 months. Strong hydrogen bonding and electrostatic repulsive interactions amid solvent and solute may be responsible for the stable dispersion. Moreover, such interactions and the non-covalent bonding may also lead to the organization of GO sheets in the self-sustained anisotropic supramolecular lyotropic structures in glycerol. Two dimensional lamellar smectic lyotropic phases with surface chirality have been observed for 0.5 mg/mL and 1 mg/mL, GO/glycerol dispersions. Partial reduction of GO into rGO has been observed and confirmed by UV–Visible measurement. Graphene-based lyotropic structures exhibit ionic conductivity of micron order (S/cm) and also possesses conduction activation energy of range 13–14 kJ/mol. GO/glycerol-based Lyotropic dispersions show non-Newtonian behaviour along with the decrease in the viscosity as the GO concentration is increased. The maximum viscosity noted for these systems is ~630 mPa.s. Viscoelastic behaviour of lyotropic dispersion has been confirmed by dynamic rheology and G'/G'' [G'(storage) and G'' (loss) modulus] crossover. Rheological studies infer that such lyotropic dispersion with high viscosity can be explored for ink-jet printing and printing of flexible membrane.

Application of a Cellular Automaton Method to Model the Structure Formation in Soils Under Saturated Conditions: A Mechanistic Approach

Soil functions are closely related to the structure of soil microaggregates. Yet, the mechanisms controlling the establishment of soil structure are diverse and partly unknown. Hence, the understanding of soil processes and functions requires the connection of the concepts on the formation and consolidation of soil structural elements across scales that are hard to observe experimentally. At the bottom level, the dynamics of microaggregate development and restructuring build the basis for transport phenomena at the continuum scale. By modeling the interactions of specific minerals and/or organic matter, we aim to identify the mechanisms that control the evolution of structure and establishment of stationary aggregate properties. We present a mechanistic framework based on a cellular automaton model to simulate the interplay between the prototypic building units of soil microaggregates quartz, goethite, and illite subject to attractive and repulsive electrostatic interaction forces. The resulting structures are quantified by morphological measures. We investigated shielding effects due to charge neutralisation and the aggregate growth rate in response to the net system charge. We found that the fraction as well as the size of the interacting oppositely charged constituents control the size, shape, and amount of occurring aggregates. Furthermore, the concentration in terms of the liquid solid ratio has been shown to increase the aggregation rate. We further adopt the model for an assessment of the temporal evolution of aggregate formation due to successive formation of particle dimers at early stages in comparison to higher order aggregates at later stages. With that we show the effect of composition, charge, size ratio, time and concentration on microaggregate formation by the application of a mechanistic model which also provides predictions for soil aggregation behaviour in case an observation is inhibited by experimental limitations.

Thermodynamic evaluation of roles of hydrophobic and electrostatic interactions in separation of benzocaine and procaine using polymer (PEO, L35) + MSO4 (M = Na; Li) aqueous biphasic system

The driving force involved in the partition of benzocaine (BZ) and procaine (PR) in macromolecule + electrolyte + water aqueous biphasic systems (ABSs) was investigated. Enthalpy changes using isothermal titration calorimetry and the partition coefficients (K) of both solutes were studied as function of the pH, macromolecule hydrophobicity, and tie-line length of the ABS. The BZ and PR molecules presented distinct partition behavior, and the KBZ and KPR values in poly(ethylene oxide)11–poly(propylene oxide)17–poly(ethylene oxide)11 ABS were as high as 5.14 × 103 and 5.12, respectively. The hydrophobic interactions between BZ and the components of the macromolecule-rich phase (MRP) contributed to the more negative standard enthalpy change of transfer, ΔtrHθ, values of this solute, which ranged from −38.8 to −24.6 kJ mol−1 while the ΔtrHθ values of PR ranged from −10.5 to −8.33 kJ mol−1. At lower pH values, KBZ and KPR decreased from 2.37 × 103 and 354, respectively, at pH = 12.0, to 1.01 × 103 and 2.98, respectively, at pH = 1.0, owing to the increase in electrostatic repulsive interactions between solutes in ionized form and the positive charge species that formed in the MRP. Hydrophobic forces were the main contributors to the partition of the BZ and PR molecules.

Reversible multi-induced transformation between wormlike micelles and gels

A novel CO2 responsive self-assembled surfactants system was prepared using N - [3 - (dimethylamino) propyl] docosanamide (NDPD) and sodium salicylate (NaSal). Rheological measurements were performed to study the phase behaviors of NDPD/NaSal system. It was found that gels and wormlike micelles can be formed in the NDPD/NaSal system. The phase behaviors were found to be influenced by changing temperature or by involving different anions. Through increasing the temperature, gels can transform to wormlike micelles. When HCl and CO2 were used to adjust pH, NDPD/NaSal system performed decidedly different rheological behaviors at the same pH. By theoretical calculation, it was found that the binding energy between NDPD and Cl− is larger than that of NDPD and HCO3−, indicating a stronger shielding ability of Cl− ions. Due to the weakened electrostatic repulsion interactions, the internal structure of NDPD/NaSal switchable system wrapped more tightly through adding HCl compared with adding CO2.

Effect of β-cyclodextrin, alcohol and temperature on the aggregation transitions of cationic/cationic surfactant systems

The mutual transitions between micelles and vesicles induced by β-cyclodextrin (β-CD), alcohol and temperatures, which had rarely been reported in mixed cationic/cationic surfactant systems, was studied in a dodecyltrimethylammonium chloride (DTAC)/didodecyldimethylammonium bromide (DDAB) aqueous solution. The effect of β-CD, temperature and alcohol on this mixed system was investigated by the methods of turbidity, transmission electron microscopy (TEM), dynamic light scattering (DLS), and 1H Nuclear magnetic resonance (1H NMR) measurements. We found that increasing the concentration of β-CD would induce the micelle to vesicle (M-V) transition. While the vesicle to micelle (V-M) transition further occurred with increasing temperature or adding alcohol in this β-CD/cationic/cationic mixed system. The M-V transition with adding β-CD has the mechanism that the inclusion complexation of β-CD to surfactants alters the composition ratio of the DDAB/DTAC systems, which changes the packing parameter. The V-M transition induced by temperature or alcohols is explained as the change of electrostatic repulsion interaction among the similar cationic polar heads in vesicles. The heat-induced V-M transition also verifies the M-V transition mechanism. Moreover, we firstly found that this heat-induced V-M transition was not only related to the percentage of DDAB concentration, but also to the total concentration of surfactant in cationic/cationic mixed surfactant systems.

Decorating surface charge of graphite nanoplate using an electrostatic coupling agent for 3‐dimensional polymer nanocomposite

ABSTRACTHere, we report a facile approach to electrostatically couple the surface charges of graphite nanoplate (GNP) fillers and poly(methyl methacrylate) (PMMA) polymer particles using ethylene maleic anhydride (EMA) copolymer as an electrostatic coupling agent. Our strategy involved switching the intrinsic repulsive electrostatic interactions between the directly exfoliated GNPs fillers and the PMMA particles to attractive electrostatic surface interactions for preparing core(PMMA)‐shell (GNP) precursor in order to optimizing 3‐dimensionally dispersed polymer nanocomposite. As a result, the electrical conductivity of the composites dramatically increased by a factor of 16.7 in the EMA‐coupled GNP/PMMA composites compared with that of the EMA‐free GNP/PMMA composites. In addition, the percolation threshold was also notably reduced from 0.32 to 0.159 vol% after electrostatic coupling of the GNPs fillers and PMMA particles. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2020, 137, 48390.

Disordered RNA chaperones can enhance nucleic acid folding via local charge screening

RNA chaperones are proteins that aid in the folding of nucleic acids, but remarkably, many of these proteins are intrinsically disordered. How can these proteins function without a well-defined three-dimensional structure? Here, we address this question by studying the hepatitis C virus core protein, a chaperone that promotes viral genome dimerization. Using single-molecule fluorescence spectroscopy, we find that this positively charged disordered protein facilitates the formation of compact nucleic acid conformations by acting as a flexible macromolecular counterion that locally screens repulsive electrostatic interactions with an efficiency equivalent to molar salt concentrations. The resulting compaction can bias unfolded nucleic acids towards folding, resulting in faster folding kinetics. This potentially widespread mechanism is supported by molecular simulations that rationalize the experimental findings by describing the chaperone as an unstructured polyelectrolyte.

Adsorption behavior and mechanism of sulfonamides on phosphonic chelating cellulose under different pH effects

Phosphonic chelating fiber (PCCSF) as a novel adsorbent was produced through alkalization, etherification, amination and phosphonation, and then it was applied to adsorb sulfonamides (SAs), such as sulfadiazine (SD), sulfamonomethoxine (SMM) and sulfamethoxazole (SMZ). Specially, their adsorption behavior at different pH values was studied. As a result, PCCSF was provided with amino (NH2 or NH) and PO(OH)2 (PO) groups, and its equilibrium data were generally represented by both Langmuir and Freundlich models. Combining adsorbent-to-solution distribution coefficients (Kd) values and the effect of pH, the primary mechanism suggested that adsorption capacity of PCCSF was lower in strong acid and alkali solution, due to the electrostatic repulsion and hydrophobic interactions. By contrast, its adsorption affinity became more excellent at 3 < pH < 9 owing to the π-π electron-donor-acceptor (EDA) charge-assisted H-bond, Lewis acid-base interaction and charge-assisted H-bond (CAHB).

Designing and construction of novel variants of Chondroitinase ABC I to reduce aggregation rate

Chondroitinase ABC I (cABC I) can degrade inhibitory molecules for axon regrowth at the site of damage after spinal cord injury (SCI). One of the main problems in the practical application is the possibility of structural changes that lead to the inactivation of the enzyme. In current work, three variants of cABC I was designed and constructed by manipulation of a short helix conformation (Gln678-Leu679-Ser680-Gln681); where Gln residues were converted to Glu. According to the enzyme kinetics studies, the catalytic efficiency of the Q681E and double mutant (Q678E/Q681E) increases in comparison with WT enzyme; while that of Q678E decreases. It was also shown that the rate of the inactivation of the enzyme variants over time is greater in WT and Q678E variants than that of the Q681E and double mutant. Negative values of entropy change of thermal inactivation measurements; demonstrate that inactivation of the WT and Q678E variants are mainly originated from aggregation. These observations can be explained by considering the repulsive electrostatic interaction between enzyme molecules that prevents protein aggregation over time. It is concluded that increasing the solubility of the Q681E and double mutant via favorable interactions of surface-exposed charged residues with dipole momentum of water molecules accompanied by the presence of intermolecular repulsive electrostatic interaction leads to decreasing the rate of aggregation in both long-term storage and heat-induced structural changes.