Radiation-induced inactivation of dihydroorotate dehydrogenase in dilute aqueous solution.

The inactivation of dihydroorotate dehydrogenase by gamma irradiation in dilute aqueous solution has been investigated. The activity of the enzyme decreased exponentially as a function of the absorbed dose under aerated and nitrous oxide-saturated conditions. The contributions of the individual radical species derived from water radiolysis were estimated from the inactivation results observed under aerated, argon-saturated, and nitrous oxide-saturated conditions. The hydrogen atom and hydroxyl radical were found to be important in enzyme inactivation. The effect of selected inorganic radical anions such as Br.2-, I.2-, and (SCN).2- on the enzyme activity was also studied, and the results implicate the possible involvement of cysteine and tyrosine residues in the catalytic activity of dihydroorotate dehydrogenase. Changes in the kinetic parameters (Michaelis-Menten constant, Km, and maximal velocity, Vmax) due to irradiation under the conditions investigated suggest that radiation-induced inactivation is due to modification of the substrate binding sites and that of the active site residues in the enzyme. Evidence for the reduction of iron-sulfur centers in the enzyme during the inactivation process has been put forward from the difference spectrum of the irradiated dihydroorotate dehydrogenase. It has also been shown by electrophoretic studies that radiation-induced inactivation was not due to any fragmentation of the protein structure or the formation of any intermolecular crosslinking.     

Oligonucleotides as probes for studying polymerization reactions in dilute aqueous solution

We have prepared a [³²P]-labeled oligonucleotide probe carrying a free primary amine at its 3′terminus. This probe is used to initiate polymerization of aziridine (ethyleneimine) in aqueous solution. The nature of the oligomeric products and the kinetics of their formation are then monitored by gel electrophoresis. Our results are generally consistent with those obtained using conventional techniques. We have also investigated the effect of polyanionic templates on the rate of oligomerization of aziridine. We find that water-soluble polyanions generally accelerate the polymerization. The sodium salt of polymethacrylic acid is the most effective of the templates that we studied. The methods introduced in this paper should be applicable to a variety of polymerization reactions in aqueous solution. They should greatly simplify the screening of potentially prebiotic polymerization reactions. Correspondence to: L.E. Orgel on sabbatical leave     

The role of water in the thermodynamics of dilute aqueous solutions

Water plays a role in the thermodynamics of dilute aqueous solutions that is unusual in two ways. First, knowledge of hydration equilibrium constants of species is not required in calculations of thermodynamic properties of biochemical reactants and reactions at specified pH. Second, since solvent provides an essentially infinite source of oxygen atoms in a reaction system where water is a reactant, oxygen atoms are not conserved in the reaction system in dilute aqueous solutions. This is related to the fact that H₂O is omitted in equilibrium expressions for dilute aqueous solutions. Calculations of the standard transformed Gibbs energies of formation of total carbon dioxide and total ammonia at specified pH are discussed, and the average bindings of hydrogen ions by these reactants are calculated by differentiation. Since both of these reactants are involved in the urease reaction, the apparent equilibrium constants and changes in the numbers of hydrogen ions bound are calculated for this reaction as functions of pH.     

Investigation of uranium recovery from dilute aqueous solutions using silk fibroin

The uranium uptake ability of silk fibroin was investigated. High ability to uptake uranium from nonsaline water containing 2.500 mg of uranium was observed with the silk fibroin tested. The uranium uptake was very rapid and was dependent on pH, uranium concentration, temperature, and retention time. Almost all uranium taken up is easily eluted with 1 mol/L CH₃COONH₄. This biomatrix, therefore appears to have potential for use in a commercial process for uranium recovery from uranium-containing waste water.     

On the temperature-induced coil to globule transition of poly-N-isopropylacrylamide in dilute aqueous solutions

Poly-N-isopropylacrylamide (PNIPAM) is a chemical isomer of poly-leucine, having the polar peptide group in the side-chain rather than in the backbone. It has been demonstrated experimentally that PNIPAM dissolved in aqueous solution undergoes a collapse transition from coil to globule on increasing temperature above the θ-point. By a careful reviewing of existing experimental data, we emphasize that such coil to globule collapse has to be considered an intramolecular first-order transition, analogous to the cold renaturation of small globular proteins. The main theoretical approaches to the coil to globule collapse in homopolymers are discussed briefly, and a critical comparison between the existing models is performed. We point out that, as a general result, the coil to globule collapse is expected to be a first-order transition for rigid and semi-rigid macromolecules. Finally, taking advantage of the analogy between the coil to globule collapse of PNIPAM and the cold renaturation of small globular proteins, we try to clarify some important and intriguing aspects of protein thermodynamics. This leads to the conclusion that the amphiphilic nature of polypeptide chain plays the fundamental role for the existence of two temperature-induced conformational transitions.     

Hydration of nucleosides in dilute aqueous solutions. Ultrasonic velocity and density measurements

The values of the concentration increments of the ultrasound velocity and their temperature slopes, apparent molar volumes, apparent molar expansibilities, apparent molar adiabatic compressibilities and their temperature gradients for 12 nucleosides and their analogs, as well as for ribose and deoxyribose, have been obtained using precision measurements of ultrasound velocity and density. The following hydration parameters for the atomic groups of the nucleosides, reflecting the state of water in the hydration shells of these groups, have been analyzed: (1) the contribution of ribose to the values of the concentration increment of ultrasound velocity A, the apparent molar volumes phi v and apparent molar adiabatic compressibilities phi ks of nucleosides; (2) contributions of the CH3, NH2 and O = ... -H groups of nucleic bases to the A, phi v and phi ks values of nucleosides and free nucleic bases; (3) contributions of the 2'-OH group of ribose to the values of A, phi v and phi ks nucleosides; (4) changes in the A values of nucleosides and free nucleic bases upon their protonation and deprotonation. Data have been obtained on the mutual influence of the atomic groups of nucleosides on their hydration. It is shown that the GC pairs of free deoxynucleosides undergo hydration more vigorously than the AT pairs, which contrasts with the relation of the degree of hydration of the GC and AT pairs of the double helix.     

A study of dilute aqueous solutions of humic acids by scanning microcalorimetry

It has been found by reversed-phase chromatography that humic acids obtained from vermicomposts of different duration of vermicomposting consist of a hydrophilic and a hydrophobic fractions, the hydrophobic fraction having a substantially lower content of charged, probably carboxylic, groups. A change in the sign of the temperature dependence of the heat capacity of dilute aqueous solutions of humic acids at ∼58°C has been found by differential scanning microcalorimetry, which indicates an increase in the hydration of hydrophobic groups. A jump-wise increase in heat capacity in the temperature range from 86 to 90°C was also found, which is perhaps due to hydration of hydrophobic groups in the interior of “micelles“, because of “devitrification” of the hydrophobic nucleus of micelle-like structures. It was shown that increasing the duration of vermicomposting leads to an increase in the relative content of the hydrophobic fraction of humic acids and in the cooperativity of the thermodynamic transition, which manifests itself as the jump of heat capacity, which probably results from the increase in the “micelle” size.     

Radiation-induced conversion of 5,6-dihydropyrimidines to the corresponding pyrimidines in N2O-saturated aqueous solutions

The effect of transition metal salts on the radiation-induced conversion of 5,6-dihydropyrimidines to the corresponding parent pyrimidines was studied in N2O-saturated aqueous solution at pH 7.0. The yield of the pyrimidines increased in sigmoidal forms with the increased one-electron reduction potential of the transition metal salts. The radiolysis of 5,6-dihydroorotic acid suggested that the 6-yl radical of the acid undergoes oxidation by transition metal salts to give orotic acid, whereas the corresponding 5-yl radical readily liberates CO2 to give uracil radical anion.     

Separation of dilute aqueous butanol and acetone solutions by pervaporation through liquid membranes

A simultaneous extraction-stripping process is proposed for separating volatile products from fermentation broths, it is based on pervaporation through a liquid membrane supported with a hydrophobic porous membrane. The liquid membrane prepared with oleyl alcohol was selected as the most suitable for separating volatile products resulting from acetone-butanol fermentation. The separation performance and stability of the oleyl alcohol liquid membrane were investigated by using dilute aqueous butanol and acetone solutions. The oleyl alcohol liquid membrane was found to be superior by far in both selectivity and permeability of butanol to the better known silicone rubber membrane in its high selectivity for alcohols. Using the oleyl alcohol liquid membrane, the dilute aqueous butanol solutions of around 4 g/L obtained in acetone-butanol fermentation could be concentrated up to 100 times. The stability of this liquid membrane was also quite good as long as the surface tension of the feed solution was less than the critical surface tension of the support membrane. No change in the separation performance was found after the continuous usage in a long period of 100 h.     

Radiation-induced glyoxal formation in sugar solutions

Aqueous solutions (4 %) of glucose, fructose, and sucrose were exposed to gamma irradiation in the dose range of 2.2 to 24.0 megarads. The Gglyoxa1 values at 2.2 megarads were 0.35, 0.18, and 0.06 for glucose, fructose, and sucrose, respectively. These values decreased at higher dose levels. The glyoxal concentration of the irradiated solutions did not appreciably change during a 2-week postirradiation storage at room temperature.     

Selective recovery of uranium and thorium ions from dilute aqueous solutions by animal biopolymers

Selective actinide ion recovery from dilute, aqueous, multication waste streams is an important problem. The recovery of uranium (U) and thorium (Th) by various animal biopolymers was examined. Of four species of biopolymers tested, a high uptake of uranium and thorium was found in hen eggshell membrane (ESM) and silk proteins, with the maximum uranium and thorium recovery exceeding 98% and 79%, respectively. The uptake of U and Th was significantly affected by the pH of the solution. The optimum pH values were 6 and 3 for the uptake of U and Th, respectively. The effect of temperature differed with the metal. The uptake of U decreased with increasing temperature (30–50°C), whereas the Th uptake increased with increasing temperature. Selective recovery of U and Th from dilute aqueous binary and multimetal solutions was also examined. ESM and silk proteins tested were effective and selective for removing each metal by controlling the pH and temperature of the solution. In multimetal systems, the order of sorption of ESM proteins was preferential: U > Cu > Cd > Mn > Pb > Th > Ni > Co > Zn at pH 6 and Th > U > Cu > Pb > Cd > Mn > Co > Ni = Zn at pH 3. These biopolymers appear to have potential for use in a commercial process for actinide recovery from actinide-containing wastewater.     

Removal of metal cyanides from aqueous solutions by suspended and immobilized cells of Rhizopus oryzae (MTCC 2541)

This paper presents a study on biodegradation and simultaneous adsorption and biodegradation (SAB) of zinc and iron cyanides by Rhizopus oryzae (MTCC 2541), with a brief process review. Granular activated carbon was used for the immobilization of Rhizopus oryzae (MTCC 2541) for the SAB study. pH and temperature were optimized at an initial cyanide concentration of 100 mg/L for biodegradation and SAB. The microbes adapted to grow at maximum cyanide concentration were harvested and their ability to degrade cyanide was measured in both biodegradation and SAB. The removal efficiency of the SAB process was found to be better as compared to the biodegradation process. In the case of biodegradation, removal was found up to a maximum cyanide concentration of 250 mg CN^?/L for zinc cyanide and 200 mg CN^?/L for iron cyanide, whereas in the case of SAB, about 50% removal of cyanide at 400 mg CN^?/L zinc cyanide and 300 mg CN^–/L iron cyanide was possible. It was found that the SAB process is more effective than biodegradation.     

The effect of autoclaving on the dispersibility and stability of three neutral polysaccharides in dilute aqueous solutions

The dispersibility of three neutral polysaccharides, oat β-glucan, detarium xyloglucan and dextran in a dilute water–cadoxen mixture was studied by viscosity measurement. It was found that intrinsic viscosity measurement, with water–cadoxen mixtures as solvents, is a useful tool to distinguish polymer degradation from disruption of supramolecular aggregates. This approach, in conjunction with size exclusion chromatography, was used to study the effects of heat and pressure treatment on the dispersibility and stability of three polysaccharides in aqueous solutions. Autoclaving treatment at 121°C for 15 min may reduce the degree of aggregation. Following autoclaving in aqueous solution, the Huggins constants decreased from 0.66 to 0.42 for oat β-glucan and from 0.63 to 0.56 for detarium xyloglucan. It remains the same for dextran, indicating good solubility of this polymer in water. The current treatment did not cause evident changes in molecular weight and structures to detarium xyloglucan and dextran. However, degradation occurred with oat β-glucan. The Burchard–Stockmayer–Fixman approach was applied to estimate the unperturbed dimension of oat β-glucan and detarium xyloglucan on samples after autoclaving. The characteristic ratio C_∞ was found to be 7.3 for detarium xyloglucan and 4.7 for oat β-glucan, corresponding to the Kratky–Porod persistence lengths of 2.0 and 1.2 nm, respectively.     

Chemical and biological removal of cyanides from aqueous and soil-containing systems

A study was conducted to determine the effect of various factors on the rate and extent of potassium cyanide and potassium hexacyanoferrate (II, complex form) removal from aqueous and soil-containing systems. In a sterile aqueous system at neutral pH, the concentration of free cyanide was reduced by 42% in 334 h as a result of the protonation of CN^– and the volatilization of the HCN formed. In the presence of aerobic mixed consortium of the Institute of Gas Technology and a methylotrophic culture, Isolate 3, the concentration of free cyanide was reduced by 59% and 66% in 357 h, respectively, as a result of combined chemical conversion and microbial degradation. In the sterile aqueous system amended initially with the complex form of cyanide, a less-than-20% reduction in cyanide occured. The sorption equilibria for free and complex cyanides in slurries of the topsoil and manufactured gas plant (MGP) soil was reached in less than 22 and 4 days, respectively. The extent of desorption of cyanides from topsoil and MGP soil into water decreased with time. In sterile systems containing topsoil and MGP soil that were previously equilibrated to cyanides, only a 2% reduction in cyanide concentration occurred in 336 h due to chemical conversion. In the presence of microbial cultures, the concentration of cyanide was reduced by less than 15% and 7% in the slurries of topsoil and MGP soil, respectively. The comparison of the rate and extent of cyanide removal from the aqueous and soil-containing systems in the presence of micro-organisms suggests that cyanides were retained by the solid phase of the soil-containing systems and therefore were less available for biodegradation.