Influence of alkaline concentration on molecular association structure and viscoelastic properties of curdlan aqueous systems

Curdlan is an extracellular polysaccharide produced from soil microorganism Alcaligens faecalis var. 10C3K, and the linear structure consists of β-1,3-glycoside linkages. Curdlan is not soluble in water but it is soluble in alkaline aqueous solution, and we can obtain the gel when curdlan alkaline solution is heated above 60°C or neutralized by acids. In the present study, the gelation mechanism and dispersing structure of curdlan in the alkaline solutions are studied in terms of correlation between the molecular association structure and viscoelastic properties, using static light scattering and rheological measurements. The degree of association for the curdlan molecules in dilute solution increases with decreasing alkaline concentration. The viscoelastic properties also depend strongly on the alkaline concentration. The concentrated curdlan solution shows almost Newtonian flow at high alkaline concentrations and shows a gel-like behavior at low alkaline concentrations. It was elucidated that the molecular association in the dilute solution reflects on the viscoelastic properties of the concentrated solution and that the gelation mechanism is related to the association structure of curdlan molecules. In the case of lower NaOH concentration systems, the molecular association is likely to consist of a hydrophobic core and hydrophilic surface. The gelation mechanism above 60°C is considered to include the dissociation process of the molecular association and reformation of the network structure. © 1997 John Wiley & Sons, Inc. Biopoly 42: 479–487, 1997     

Dynamic viscoelastic properties of cellulose carbamate dissolved in NaOH aqueous solution

Dynamic viscoelastic properties of cellulose carbamate (CC) dissolved in NaOH aqueous solution were systematically studied for the first time. CC was microwave-assisted synthesized from the mixture of cellulose and urea and then dissolved in 7 wt % NaOH aqueous solution precooled to -7 °C. The obtained CC solution is transparent and has good liquidity. To clarify the rheological behavior of the solution, the CC solutions were investigated by dynamic viscoelastic measurements. The shear storage modulus (G') and loss modulus (G') as a function of the angular frequency (ω), concentration (c), nitrogen content (N %), viscosity-average molecular weight (M(η)), temperature (T), and time (t) were analyzed and discussed in detail. The sol-gel transition temperature of CC (M(η) = 7.78 × 10(4)) solution decreased from 36.5 to 31.3 °C with an increase of the concentration from 3.0 to 4.3 wt % and decreased from 35.7 to 27.5 °C with an increase of the nitrogen content from 1.718 to 5.878%. The gelation temperature of a 3.8 wt % CC solution dropped from 38.2 to 34.4 °C with the M(η) of CC increased from 6.35 × 10(4) to 9.56 × 10(4). The gelation time of the CC solution was relatively short at 30 °C, but the solution was stable for a long time at about 15 °C. Moreover, the gels already formed at elevated temperature were irreversible; that is, after cooling to a lower temperature including the dissolution temperature (-7 °C), they could not be dissolved to become liquid.     

Molecular study of the diffusional process of DMSO in double lipid bilayers

As a way to quantify the diffusion process of molecular compounds through biological membranes, we investigated in this study the dynamics of DMSO through an 1,2-Dipalmitoyl-sn-Glycero-3-Phosphocholine (DPPC) bilayer system. To properly account for the diffusion of DMSO due to a concentration gradient, a double DPPC bilayer was setup for our simulations. In such configuration, the aqueous phases can be explicitly associated with the extra and intracellular domains of the membrane, which is seldom the case in studies of single lipid bilayer due to the periodicity imposed by the simulations. DMSO molecules were initially contained in one of the aqueous phases (extracellular region) at a concentration of 5 wt.%. Molecular dynamics simulation was performed in this system for 95 ns at 350 K and 1 bar. The simulations showed that although many DMSO molecules penetrated the lipid bilayer, only about 10% of them crossed the bilayer to reach the other aqueous phase corresponding to the intracellular region of the membrane. The simulation time considered was insufficient to reach equilibrium of the DMSO concentration between the aqueous phases. However, the simulations provided sufficient information to estimate parameters to apply Fick's Law to model the diffusion process of the system. Using this model, we predicted that for the time considered in our simulation, the concentration of DMSO in the intracellular domain should have been about half of the actual value obtained. The model also predicted that equilibrium of the DMSO concentration in the system would be reached after about 2000 ns, approximately 20 times longer than the performed simulation.     

Molecular study of the diffusional process of DMSO in double lipid bilayers

As a way to quantify the diffusion process of molecular compounds through biological membranes, we investigated in this study the dynamics of DMSO through an 1,2-Dipalmitoyl-sn-Glycero-3-Phosphocholine (DPPC) bilayer system. To properly account for the diffusion of DMSO due to a concentration gradient, a double DPPC bilayer was setup for our simulations. In such configuration, the aqueous phases can be explicitly associated with the extra and intracellular domains of the membrane, which is seldom the case in studies of single lipid bilayer due to the periodicity imposed by the simulations. DMSO molecules were initially contained in one of the aqueous phases (extracellular region) at a concentration of 5 wt.%. Molecular dynamics simulation was performed in this system for 95 ns at 350 K and 1 bar. The simulations showed that although many DMSO molecules penetrated the lipid bilayer, only about 10% of them crossed the bilayer to reach the other aqueous phase corresponding to the intracellular region of the membrane. The simulation time considered was insufficient to reach equilibrium of the DMSO concentration between the aqueous phases. However, the simulations provided sufficient information to estimate parameters to apply Fick's Law to model the diffusion process of the system. Using this model, we predicted that for the time considered in our simulation, the concentration of DMSO in the intracellular domain should have been about half of the actual value obtained. The model also predicted that equilibrium of the DMSO concentration in the system would be reached after about 2000 ns, approximately 20 times longer than the performed simulation.     

Rheological characterization of schizophyllan aqueous solutions after denaturation-renaturation treatment

Schizophyllan (SPG) with a molecular weight of 2.6x10(6), designated SPG-1, is denatured and then renatured at a concentration of 1.8 wt % by alkalization-neutralization. The prepared denatured-renatured samples (DRSPG-1) are diluted to various concentrations and equilibrated for 10 days before rheological and intrinsic viscosity measurements. When concentration (C(p)) is above 0.75 wt %, DRSPG-1 aqueous systems have weak gel-type rheological properties. However, for 0.28 wt % 相似文献   

氨水流加用于粪产碱杆菌热凝胶发酵

热凝胶是粪产碱杆菌(Alcaligenes faecalis)在氮源限制条件下生成的水不溶性胞外多糖,分泌到胞外后就附着在菌体外壁,因此在细胞生长期提高生物量对促进热凝胶合成有重要意义。热凝胶分批发酵时, 起始NH4Cl浓度提高到3.6 g/L时能促进菌体生长和热凝胶合成,但是过量NH4Cl会抑制热凝胶合成,且生物量提高不是很明显。为了进一步提高菌体浓度, 在菌体生长期, 氨水取代NaOH溶液进行流加控制pH为7.0, 随后又用2 mol/L NaOH控制pH 5.6。实验表明, 氨水流加使菌体浓度大大提高,流加24 h使菌体浓度达到18.8 g/L。但是菌体浓度过高也会抑制热凝胶的合成,在氨水流加14 h时,菌体浓度在11.9 g/L左右, 热凝胶产量最高（72 g/L）。     

氨水流加用于粪产碱杆菌热凝胶发酵

热凝胶是粪产碱杆菌(Alcaligenes faecalis)在氮源限制条件下生成的水不溶性胞外多糖,分泌到胞外后就附着在菌体外壁,因此在细胞生长期提高生物量对促进热凝胶合成有重要意义。热凝胶分批发酵时, 起始NH4Cl浓度提高到3.6 g/L时能促进菌体生长和热凝胶合成,但是过量NH4Cl会抑制热凝胶合成,且生物量提高不是很明显。为了进一步提高菌体浓度, 在菌体生长期, 氨水取代NaOH溶液进行流加控制pH为7.0, 随后又用2 mol/L NaOH控制pH 5.6。实验表明, 氨水流加使菌体浓度大大提高,流加24 h使菌体浓度达到18.8 g/L。但是菌体浓度过高也会抑制热凝胶的合成,在氨水流加14 h时,菌体浓度在11.9 g/L左右, 热凝胶产量最高（72 g/L）。     

Molecular dynamics simulations of Leu-enkephalin in water and DMSO.

The structure of Leu-enkephalin (L-Enk) and Met-enkephalin (M-Enk) have frequently been studied, in particular by nuclear magnetic resonance spectroscopy. After more than 20 years of research, it was concluded that enkephalins have no preferred structure in aqueous solution, but that they may have in other solvents. We have performed molecular dynamics simulations of zwitterionic L-Enk in water, and zwitterionic as well as neutral L-Enk dimethyl sulfoxide (DMSO). In water the peptide is very flexible, although there seems to be a preference for compact conformations. In DMSO, the peptide forms a clear salt bridge in the zwitterionic form, but has no preferred conformation in the neutral form. This difference in conformation may provide an explanation for measurements in DMSO in which multiple conformations were found to exist. In this paper we introduce a new formulation for a dihedral angle autocorrelation function, and apply it to study side-chain dynamics in L-Enk. We find that the side-chain dynamics of the large Tyr and Phe residues cannot be adequately sampled in 2.0-ns simulations, while this does seem to be possible for the smaller Leu side chain.     

Tapping mode atomic force microscopy of scleroglucan networks

Tapping mode Atomic Force Microscopy (TmAFM) has been used to study the fungal polysaccharide scleroglucan deposited from aqueous solution and dimethyl sulfoxide (DMSO) onto a mica surface. The solutions from which the microscope samples were produced were prepared by first dissolving the solid scleroglucan in 0.1M NaOH, then neutralizing the solution with HCl, followed by dilution to the required concentration in either water or DMSO. It was found that from the aqueous solution described above, scleroglucan forms networks. Based on a comparison of the denatured-renatured and aqueous solution samples, network formation is due to the imperfect registration between the chains forming the triple helices. The relatively large stiffness of the scleroglucan triple helix is also assumed to contribute to the formation of the extended networks. The triple helix diameter was measured to be 0.92 ± 0.27 nm, which is in the same range as that obtained by other researchers using similar techniques. Denatured scleroglucan, deposited from DMSO onto mica, forms a web-like layer on top of which there are sphere-like structures. These morphologies are believed to be due to triple helix denaturation yielding highly flexible single chains in DMSO, which results in coiling and web-like dense packing of scleroglucan upon deposition onto mica. Most interestingly after addition of water to the samples deposited from DMSO, some of the chains can be renatured into short, stiff rod-like structures which are similar to the structures observed by other researchers. The imaging data for aqueous solution deposition can be analyzed by plotting maximum end-to-end distance versus the perimeter of the networks deposited onto mica. This yields a Flory-like exponent of 0.67, which is almost similar in value to that obtained by other researchers for linear structures of scleroglucan but less than that expected for a polymer chain following a self-avoiding walk (v = 0.75) model on a two-dimensional surface. The fractal dimension that can be used to characterize the networks was determined graphically to be 1.22 ± 0.06. © 1997 John Wiley & Sons, Inc. Biopoly 42: 89–100, 1997     

Solubility and property of chitin in NaOH/urea aqueous solution

NaOH/urea aqueous solution has been used as a solvent for chitin for the first time. Effects of this solvent composition and temperature on the solubility and stability of chitin solution were studied with an optical microscope, from which 8 wt% NaOH/4 wt% urea concentrations were deduced as suitable and −20 °C as the appropriate temperature. The original and regenerated chitin were characterized by viscosity, elemental analysis, FI-IR and X-RD analysis, and the effect of solvent composition and temperature on chitin structure was investigated. It was inferred that 8 wt% NaOH/4 wt% urea solvent under low temperature adventitiously has little effect on chitin structure and the urea is of benefit to the stability of chitin solution. In addition, the rheological properties suggested that chitin aqueous solution in high concentration is a pseudoplastic fluid and that chitin aqueous solution in low concentrations is a Newtonian fluid. This chitin aqueous solution is sensitive to temperature and will transform it to a gel when temperature increases.     

Solvent effects on the cooperative order-disorder transition of aqueous solutions of schizophyllan, a triple-helical polysaccharide

A triple helical polysaccharide schizophyllan in aqueous solution exhibited a highly cooperative transition between ordered and disordered states associated with the conformation of its side chains and nearby water molecules. The transition was followed by optical rotation and calorimetry using water containing additives such as NaOH and DMSO as solvents. The ordered state was stabilized or destabilized depending on the kind and amount of the additive employed; in particular, the addition of DMSO had a remarkable stabilizing effect. This effect was analyzed by means of a statistical mechanical theory of linear cooperative transitions, where DMSO was assumed to interact favorably with the ordered side chains. A small amount of NaOH in a solvent mixture stabilized the ordered state and made the transition curve very gradual. No molecular mechanism was elucidated to account for the role of NaOH.     

DMSO Induces Dehydration near Lipid Membrane Surfaces

Dimethyl sulfoxide (DMSO) has been broadly used in biology as a cosolvent, a cryoprotectant, and an enhancer of membrane permeability, leading to the general assumption that DMSO-induced structural changes in cell membranes and their hydration water play important functional roles. Although the effects of DMSO on the membrane structure and the headgroup dehydration have been extensively studied, the mechanism by which DMSO invokes its effect on lipid membranes and the direct role of water in this process are unresolved. By directly probing the translational water diffusivity near unconfined lipid vesicle surfaces, the lipid headgroup mobility, and the repeat distances in multilamellar vesicles, we found that DMSO exclusively weakens the surface water network near the lipid membrane at a bulk DMSO mole fraction (X_DMSO) of <0.1, regardless of the lipid composition and the lipid phase. Specifically, DMSO was found to effectively destabilize the hydration water structure at the lipid membrane surface at X_DMSO <0.1, lower the energetic barrier to dehydrate this surface water, whose displacement otherwise requires a higher activation energy, consequently yielding compressed interbilayer distances in multilamellar vesicles at equilibrium with unaltered bilayer thicknesses. At X_DMSO >0.1, DMSO enters the lipid interface and restricts the lipid headgroup motion. We postulate that DMSO acts as an efficient cryoprotectant even at low concentrations by exclusively disrupting the water network near the lipid membrane surface, weakening the cohesion between water and adhesion of water to the lipid headgroups, and so mitigating the stress induced by the volume change of water during freeze-thaw.     

Comparison of curdlan and its carboxymethylated derivative by means of Rheology, DSC, and AFM

Curdlan was carboxymethylated in an aqueous alkaline medium using monochloroacetic acid as the etherifying agent. The structure of carboxymethylated curdlan (CMc) was analyzed by FT-IR and NMR spectroscopy, which revealed that the carboxymethyl group was introduced mainly at the C-6 position as well as at the C-2 and C-4 positions. Furthermore, CMc was compared with the native curdlan by using rheology and DSC methods. It was found that in water, both polysaccharides behaved as pseudoplastic fluids and fit the power law and Herschel-Bulkley rheological models well. Both the storage shear modulus G' and the loss shear modulus G' of CMc aqueous solutions decreased and became more frequency dependent with decreasing concentration in comparison with the curdlan aqueous suspensions. The modulus-temperature curve also suggested that the gel characteristic of curdlan has been lost after chemical modification, which is consistent with the DSC results. AFM images revealed differences in the conformation of native and carboxymethylated curdlan, which changed from the aggregation of macromolecules to triple helices. All the experimental results suggest that the hydrogen bonds that bind curdlan with interstitial water to form the micelles have been destroyed completely and that the hydrophobic interactions related to the methylene groups at C-6 formed above 55 degrees C disappeared due to the introduction of the hydrophilic carboxymethyl group.     

Dynamic and extensional properties of starch in aqueous dimethylsulfoxide

The effect of starch composition and concentration on the rheological properties of starch in a mixed solvent, water–DMSO, was investigated in dynamic shear and extensional mode. High amylose corn starch containing 70% amylose and 30% amylopectin, common corn starch containing 25% amylose and 75% amylopectin, and waxy corn starch containing about 99% amylopectin were used in this study. Concentrations of 2, 4, 6, and 8% (w/v) in 10% water-90% DMSO (v/v) were used for each starch type. An increase in the amylopectin content of starch from 30 to 99% caused a change in behavior from semidilute solution to viscoelastic solid at a concentration of 8% (w/v). At a concentration of 2%, an increase in the amylopectin content of starch from 30 to 99% caused a change from Newtonian to incipient gel-like behavior. Behavior at intermediate concentrations of 4 and 6% (w/v) varied from semidilute to critical gel-like with increasing amylopectin content. A power-law relaxation was observed for all concentrations of common and waxy corn starches with the slope decreasing with increase in concentrations. A 2% waxy corn starch solution displayed extension thinning behavior, while a 2% high amylose corn starch solution displayed Newtonian behavior.     

DMSO enhanced conformational switch of an interfacial enzyme

Interfacial proteins function in unique heterogeneous solvent environments, such as water–oil interfaces. One important example is microbial lipase, which is activated in an oil‐water emulsion phase and has many important enzymatic functions. A unique aprotic dipolar organic solvent, dimethyl sulfoxide (DMSO), has been shown to increase the activity of lipases, but the mechanism behind this enhancement is still unknown. Here, all‐atom molecular dynamics simulations of lipase in a binary solution were performed to examine the effects of DMSO on the dynamics of the gating mechanism. The amphiphilic α5 region of the lipase was a focal point for the analysis, since the structural ordering of α5 has been shown to be important for gating under other perturbations. Compared to the closed‐gorge ensemble in an aqueous environment, the conformational ensemble shifts towards open‐gorge structures in the presence of DMSO solvents. Increased width of the access channel is particularly prevalent in 45% and 60% DMSO concentrations (w/w). As the amount of DMSO increases, the α5 region of the lipase becomes more α‐helical, as we previously observed in studies that address water–oil interfacial and high pressure activation. We believe that the structural ordering of α5 plays an essential role on gating and lipase activity.     

The effect of dimethyl sulfoxide on cholesterol and bile acid metabolism in rats

The effect of DMSO on cholesterol and bile acid metabolism was studied in rats. Male Sprague-Dawley rats were randomly assigned to one of two groups and given either tap water or 2% DMSO (v/v) in tap water to drink for 9 days. Both food (stock rat diet) and water were available ad libitum. Animals in both groups gained weight equally throughout the study. They also had similar liver weights (g/100 g body wt) at the end of the study (control: 5.0 +/- 0.1 (N = 6) vs DMSO: 4.9 +/- 0.1 (N = 6]. The activity of hepatic cholesterol 7 alpha-hydroxylase (pmole/mg/min), the rate-limiting enzyme of bile acid biosynthesis, was significantly (P less than 0.005) reduced in the treated animals (control: 9.7 +/- 1.0 (N = 6) vs DMSO: 4.3 +/- 0.7 (N = 6)). Plasma cholesterol (mg/dl) was significantly (P less than 0.005) elevated in the treated animals (control: 90 +/- 3 (N = 6) vs DMSO: 107 +/- 4 (N = 6)), a finding consistent with the reduced CH-7 alpha hydroxylase activity in this group. DMSO treatment did not affect either microsomal cholesterol content or hepatic glutathione content. Thus, this study has shown that DMSO treatment per se can affect cholesterol and bile acid metabolism. However, the precise mechanisms whereby DMSO exerts the observed effects are not known.     

Rheological properties of acid converted waxy maize starches in water and 90% DMSO/10% water

Rheological properties of acid-converted Amioca starches in 90% dimethyl sulfoxide (DMSO)/10% water and 100% water were examined. Rheological flow data was described using Cross and Carreau models while zero-shear viscosities were found for starches that had been acid modified at least 25 min, apparent yield stresses were exhibited by unconverted Amioca dispersions. Dynamic rheological tests showed that the acid modified starch dispersions behaved like Newtonian liquid-like solutions, while unconverted Amioca dispersions behaved like weak gels. The Cox–Merz rule was followed by starches that had been acid modified for at least 45 min. The reduction in molecule size of acid converted starches (+45 min) allowed the Cox–Merz rule to hold as opposed to the highly branched Amioca and 25 min acid converted starches which showed apparent viscosity higher than complex viscosity.     

Solution properties of the acrylamide-modified cellulose polyelectrolytes in aqueous solutions

A novel cellulose-based polyelectrolyte (AM-C) containing acylamino (DS = 0.625) and carboxyl (DS = 0.148) groups was homogeneously synthesized from cellulose with acrylamide in NaOH/urea aqueous solutions. Solution properties of AM-C in aqueous solutions were investigated by laser light scattering, rheometry, and viscometry. The results indicated that AM-C could form large aggregates spontaneously in water with or without the addition of salts by the strong hydrogen bonds and electrostatic interaction between acylamino and carboxyl groups. Steady-shear flow study showed a Newtonian behavior of the solutions in the dilute regime while a shear-thinning behavior as the concentration increases. The critical concentration (c_e) for transition from dilute to concentrated solution was determined to be 0.7 wt %. Aqueous solutions of AM-C displayed good thermo-stability, reversible liquid-like characters attributing to the chemical modification. The derivation from Cox-Merz rule at relatively low concentration was related to the co-existence of single chain and large aggregates of AM-C in dilute regime. As the polymer concentration increased, the AM-C system was transformed into a homogeneous entanglement structure, resulting in the disappearance of deviations from the Cox-Merz rule.     

多糖的甲基化方法及图谱解析

本文对微量多糖的甲基化方法进行了研究。0.1 mg多糖样品用0.3 mL二甲亚砜溶解后,加入10 mg氢氧化钠粉末,室温下超声20 min。冰浴冷却后加入0.1 mL碘甲烷1,8～20℃下超声20 min,再加入0.1 mL碘甲烷,超声20 min。加入1 mL含4 mmol/L Na2S2O3的水,终止甲基化反应。反应液用0.5 mL氯仿提取4次,氯仿提取液用0.5 mL水处理5次。氯仿相用无水硫酸钠脱水后,氮气吹干。该法简便易行,甲基化程度高,适用于易溶(或难溶)于水和二甲亚砜的多糖。此外,本文对部分甲基化的糖醇乙酰酯衍生物的质谱图解析进行了阐述,并对特殊糖类样品的甲基化方法进行了说明。     

The influence of the non-Newtonian properties of blood on the flow in large arteries: unsteady flow in a 90 degrees curved tube.

A numerical and experimental investigation of unsteady entry flow in a 90 degrees curved tube is presented to study the impact of the non-Newtonian properties of blood on the velocity distribution. The time-dependent flow rate for the Newtonian and the non-Newtonian blood analog fluid were identical. For the numerical computation, a Carreau-Yasuda model was employed to accommodate the shear thinning behavior of the Xanthan gum solution. The viscoelastic properties were not taken into account. The experimental results indicate that significant differences between the Newtonian and non-Newtonian fluid are present. The numerical results for both the Newtonian and the non-Newtonian fluid agree well with the experimental results. Since viscoelasticity was not included in the numerical code, shear thinning behavior of the blood analog fluid seems to be the dominant non-Newtonian property, even under unsteady flow conditions. Finally, a comparison between the non-Newtonian fluid model and a Newtonian fluid at a rescaled Reynolds number is presented. The rescaled Reynolds number, based on a characteristic rather than the high-shear rate viscosity of the Xanthan gum solution, was about three times as low as the original Reynolds number. Comparison reveals that the character of flow of the non-Newtonian fluid is simulated quite well by using the appropriate Reynolds number.