Surface active properties of isobornylphenols in systems with different degrees of complexity

Interrelations between the structure of the semi-synthetic phenolic antioxidants — isobornylphenols and their surface active properties were studied in the chemical (the lecithin aggregation in hexane) and biological (the incubation with the blood erythrocytes) model systems. It has been shown that all studied compounds are able to affect the lecithin aggregation in hexane: the share of the main fraction of the L micelles decreases with increasing the share of particles of greater size. The effect substantially depends on hindered OH group and the presence of the intramolecular hydrogen bond in molecule. The cytotoxic properties of isobornylphenols (the concentration is 100 μM) are predominantly due to the molecule structure. The interrelation between the aggregate size of the main fraction of L in the presence of the studied compounds and the discocyte share during mice blood erythrocyte incubation in their presence for 4 h is revealed. Thus, this provides the possibility to assume that the ability of the different biological active substances to affect the lecithin aggregation in non-polar solvent could be used as a model system for the initial assessment of their surface active properties.     

Surface and aggregate properties of an amphiphilic derivative of carboxymethylchitosan

A new kind of amphiphilic derivative of carboxymethylchitosan, a group of (2-hydroxyl-3-butoxyl)propylcarboxymethylchitosans (HBP-CMCHS), has been synthesized, and the surface and aggregate properties have been studied by means of surface tension, surface pressure and fluorescence measurements. HBP-CMCHS can adsorb on the surface to decrease the surface tension of the solution. The adsorption film was quite stable, which can make the relative compressed pressure increase dramatically with the decrease of the surface area. In solution, hydrophobic aggregations were identified by the decrease in the ratio of the fluorescence emission intensity of the first and third pyrene vibronic peaks ( I(1)/ I(3)). Results showed that the aggregation began to form at a concentration similar to that of the polymer transfer to the air-water interface. Aggregate formation of the polymers is a gradually compact process with hydrophobic associations. Increase of DS and addition of NaCl to the HBP-CMCHS solution can make the surface tension decrease, make the aggregation occur at lower concentration, and make the aggregation more hydrophobic.     

Surface properties of crosslinked erythrocytes as studied by counter-current distribution in aqueous polymer two-phase systems

The bifunctional imidoester dimethyl suberimidate hydrochloride can stabilize rat red blood cells (RBCs) by membrane protein crosslinking, and in that way they can be used as carrier systems for exogenous substances. Counter-current distribution fractionation in charge-sensitive dextran-polyethyleneglycol two-phase systems has been used to detect slight changes in surface charge in stabilized cells. A decrease in the surface charge of crosslinked RBCs and an apparent masking of the age-related cell surface properties have been found to result from the protein crosslinking. Digitonin treatment used to permeabilize crosslinked RBCs produces a significant decrease of the cell surface charge while the age-related surface properties do not seem to be modified by the treatment.     

Surface properties of aqueous solutions of L-leucine

The surface tension, sigma, of solutions of L-leucine (CH3)2CHCH2CH(NH2)COOH in water, as well as in aqueous solutions of NaOH and HCl were measured in the temperature range between 278 and 308 K using the Wilhelmy plate method. L-Leucine was found to be a very weak surfactant, which can be understood if assuming strong interactions of this solute with the water structure. Striking differences were observed in the surface entropy of L-leucine solutions in water, 0.5 M HCl and 0.5 M NaOH. Moreover, surface activity of the solute is much lower than that supposed taking into account the hydrophobicity of this amino acid. It was concluded that the observed phenomena are caused by the water structure changes close to the side chain of leucine, caused by enforced hydrophobic hydration, i.e. formation of clathrate-like hydrates.     

Surface properties of membrane systems. Transport of staphylococcal delta-toxin from aqueous to membrane phase

Hemolytic delta-toxin from Staphylococcus aureus was soluble in either water, methanol or chloroform/methanol (2 : 1, v/v). The toxin spread readily from distilled water into films with pressures (pi) of 10 dynes/cm on water and 30 dynes/cm on 6 M urea; from chloroform/methanol it produced 40 dynes/cm pressure on distilled water. The toxin adsorbed barely from water (pi = 1 dyne/ cm) but it did rapidly from 6 M urea (pi = 35 dynes/cm). The protein films had unusually high surface potentials, which increased with the film pressure and decreased with increasing both pH and urea concentration in the aqueous phase. The fluorescence of 1-aniline 8-naphthalene sulfonate with delta-toxin was much greater than that with RNAase and dipalmitoyl phosphatidylcholine itself, indicating probably a marked lipid-binding character of the toxin. By circular dichroism the alpha-helix content of delta-toxin was 42% in water, 45% in methanol, 24% in 6 M urea. Infrared spectroscopy showed predominant alpha-helix in both 2H2O and deuterated chloroform/methanol as well as in films spread from either solvent on 2H2O. In spreading from 6 M [2H]urea, in which the major infrared absorption was that of [2H]urea with peaks at 1600 and 1480 cm(-1), the delta-toxin film showed prevalently non-alpha-helix structures with major peak intensities at 1633 cm(-1) > 1680 cm(-1), indicating the appearance of new beta-aggregated and beta-antiparallel pleated sheet structures in the film. The data prove that (1) high pressure protein films can consist of alpha-helix as well as non-alpha-helix structures and, differently from another cytolytic protein, melittin, delta-toxin does not resume the alpha-helix conformation in going into the film phase from the extended chain in 6 M urea; (2) conformational changes are important in the transport of proteins from aqueous to lipid or membrane phase; (3) delta-toxin is by far more versatile in structural dynamics and more surface active than alpha-toxin.     

Physical properties of detergent-based aqueous two-phase systems

The physical behavior of the binary phase systems of the non-ionic polyoxyethylene detergent Agrimul NRE 1205 and water was investigated. This technical detergent can be used for the large-scale recovery of biomolecules in detergent based aqueous two-phase systems. The phase diagram was determined. It shows significant and unexpected differences to highly purified detergents. Very similar to neat detergents the phase diagram can be influenced by auxiliary chemicals thus shifting the entire phase diagram in general to lower temperatures. This was demonstrated by lowering the cloud-point by various additions. The concentration factor, as an important parameter of a first capture step in purification was investigated and modeled. Auxiliary chemicals, temperature change and change in detergent concentration also influence the viscosity and density of the phases. These experimental data are shown. They can help to explain the separation behavior of proteins. In large-scale separations aqueous two-phase systems are separated using disc-stack centrifuges. It is demonstrated that this is not a feasible method for detergent-based aqueous two-phase extraction and the physical reason is presented.     

The structure and thermotropic properties of pure 1,2-diacylgalactosylglycerols in aqueous systems

Pure 3-sn-monogalactosyldilinolenoylglycerol and 3-sn-digalactosyldilinol-enoylglycerol have been isolated from bean leaves. Distearoyl derivatives have been prepared by catalytic hydrogenation of the unsaturated galactolipids. The unsaturated lipids form stable monomolecular films at the air/water interface which are similar to liquid-expanded phospholipid monolayers. The limiting areas were about 0.57 nm² and 0.62 nm² for the mono- and digalactosyldiacyl-glycerols, respectively. The saturated galactolipids formed condensed monolayers that were relatively unstable. The surface pressure-area isotherm of the digalactosyl derivative was more expanded than that of the monogalactosyldiacylglycerol especially at low surface pressures. Low-angle X-ray diffraction and freeze-fracture electron microscopy studies of the monogalactosyldiacylgly-cerols showed that an hexagonal-type structure was formed by the unsaturated lipid in aqueous systems, whilst the saturated lipid was arranged in a lamellar configuration. Both digalactosyldiacylglycerols form lamellar structures in water. A gel-to-liquid-crystalline phase transition of distearoyldigalactosylgly-cerol was observed at about 51°C by fluorescence depolarization measurements, using 1,6-diphenylhexatriene, and by differential scanning calorimetry. The saturated monogalactosyldiacylglycerol did not form dispersions suitable for fluorescence probe studies of a phase transition. A complex pattern of endotherms was observed for this lipid by differential scanning calorimetry.     

Surface properties of irreversibly sickled cells differ from those of the bulk of sickle cells Studies by partitioning in aqueous phase systems

Counter-current distribution (CCD) of red blood cells (RBC) from individuaks with homozygous sickle cell (HbSS) disease in a charge-sensitive aqueous dextran-poly(ethylene glycol) phase system, which fractionates cells on the basis of surface properties, indicates that the percentage of irreversibly sickled cells (ISC) increases and the percentage of reticulocytes decreases with increasing cell partition ratios. The high partition ratios of ISC correspond to those of older RBC when RBC from normal individuals are subjected to CCD. Our results thus indicate that ISC differ in surface properties from those of the bulk of sickle RBC (including reticulocytes) in the population and that the difference is, most likely, charge-related. While the question as to whether ISC are indeed old cells has not yet been unequivocally answered, this view finds support in the fact that the independent parameters of ISC surface properties, as reflected by partition ratios, and densities correlate as they do in older RBC from normal individuals.     

Production and surface-active properties of microbial surfactants

A new source of surface-active compounds produced by microbes was investigated. These biologically derived surfactants (biosurfactants) were found to be extracellular products of two newly isolated Corynebacterium species, which were gorwn on kerosene (Imperial Oil No.9). Batch-growth kinetic studies were carried out in a 14 liter fermentor and the production of biosurfactants was found to be cell growth associated. The surface tension of the whole microbial broth was reduced to a minimum of about 30 dyn/cm, as compared with the surface tension for distilled water of 72 dyn/cm. The crude biosurfactants were recovered from the cell-free broth, freeze-dried, redissolved in water, and their surface-active properties were studied. The biosurfactants reported here were found to be nontoxic and their ability to lower the surface tension of water was found to be comparable to that of sodium dodecylbenzene sulfonate, common commercial synthetic surfactant.     

Role of surfactants in optimizing fluorene assimilation and intermediate formation by Rhodococcus rhodochrous VKM B-2469

Biodegradation of fluorene by Rhodococcus rhodochrous VKM B-2469 was investigated and optimized by adding non-ionic surfactants to the liquid media. The utilization of 1-1.5% Tween 60 or 1% Triton X100 allowed to solubilize 1 mM fluorene over 150 times more than in water medium (from 9-11 microM to above 1.5 mM at 28 degrees C). We observed that Tween 60 was useful to enhance the fluorene biodegradation rates further supporting R. rhodochrous VKM B-2469 growth as an additional carbon source and to decrease fluorene toxicity for bacterial cells whereas Triton X100 resulted to be toxic for this strain. An additional enzyme induction step before starting the bioconversion process and the increase of incubation temperature during fluorene bioconversion led to further improvements in rates of fluorene utilization and formation of its intermediates. In the optimized conditions 1 mM fluorene was degraded completely within 24h of incubation. Some intermediates in fluorene degradation built up during the process reaching maxima of 31% for 9-hydroxyfluorene, 2.1% for 9-fluorenone and 1.9% for 2-hydroxy-9-fluorenone (starting from 1 mM substrate). In the presence of Tween 60 the appearance and following conversion of 2-hydroxy-9-fluorenone was observed for R. rhodochrous VKM B-2469 revealing the existence of a new pathway of 9-fluorenone bioconversion.     

耕作方式和秸秆还田对华北地区农田土壤水稳性团聚体分布及稳定性的影响

本研究基于10年耕作措施的定位试验,利用湿筛法研究了不同耕作措施和秸秆还田条件下农田土壤水稳性团聚体的分布规律,并利用平均重量直径（MWD）和几何平均直径（GMD）评价了不同处理水稳性团聚体的稳定性。研究结果表明,不同耕作措施处理的水稳性团聚体在0~10cm,10~20cm和20~30cm土层表现出不同的分布趋势,随着土层的加深,各处理水稳定团聚体的分布呈粒径逐渐减小、分布范围逐渐扩大趋势,0~10cm土层的水稳性团聚体多集中于2~5mm粒径范围,10~20cm土层水稳性团聚体多集中在0.5~5mm粒径范围,以0.5~1mm最多,而20~30mm,则广泛分布在0.25~5mm粒径范围。通过MWD和GMD值可以看出,免耕、耙耕和旋耕措施更能有效地保护表层（0~10cm）土壤水稳定团聚体的稳定性,常规耕作和深松处理则显著降低了20~30cm水稳性团聚体稳定性。秸秆还田显著增加了土壤有机碳（SOC）含量,且显著影响了土壤表层水稳性团聚体的稳定性。作用力分析结果表明,在0~10cm表层,秸秆作用在土壤表层显著影响了水稳定大团聚体的数量及其稳定性（P<0.001）,多元回归分析说明其与土壤有机碳含量达到了极显著水平（P<0.01）,且同时也受到来自耕作和秸秆交互效应的影响（P<0.01）;但在10~20cm土层,影响其数量分布的主要因素是不同的耕作措施及耕作和秸秆的交互效应（P<0.001）;而在20~30cm,耕作措施、秸秆和两者的交互效应共同对水稳性大团聚体数量产生重要的影响,但主要作用力来自耕作措施（P<0.001）和两者的交互效应（P<0.001）。免耕秸秆还田措施能显著提高土壤的水稳性团聚体的比例和稳定性。     

Detection of surface charge-related properties in model membrane systems by aqueous two-phase partition

Unilamellar vesicles composed of phosphatidylcholine (PC) and either phosphatidic acid (PA) or phosphatidylglycerol (PG) partition to the upper poly(ethylene glycol) (PEG)-rich phase of a charge-sensitive 5%:5% (w/w) PEG 8000/Dextran T-500 phase system containing 10 mM sodium phosphate at pH 7, consistent with the vesicles bearing a net negative charge. When prepared in the presence of a pH gradient (interior acidic), PC/PA vesicles exhibit an increased partition to the top PEG-rich phase, consistent with a redistribution of the PA from the inner to the outer monolayer of the vesicle bilayer. Conversely, when prepared in the presence of a pH gradient (interior basic), PC/PG vesicles exhibit a decreased top-phase partition, consistent with a redistribution of the PG from the outer to the inner monolayer of the vesicle bilayer. Unilamellar vesicles composed of PC and stearylamine partition to the lower dextran-rich phase of a 5%:5% (w/w) PEG 8000/Dextran T-500 phase system containing 10 mM sodium phosphate at pH 8.5, consistent with the vesicles bearing a net positive charge. When prepared in the presence of a pH gradient (interior acidic), conditions under which the stearylamine is trapped on the inner monolayer of the bilayer, the vesicles now partition predominantly to the interface in a manner similar to vesicles composed of PC alone. These results demonstrate that partitioning in aqueous two-phase polymer systems is a sensitive method for monitoring the asymmetry of charged lipids in model membrane systems and also suggests that partitioning in charge-sensitive systems depends only on the physical nature of the exterior surface of the membrane.     

Hydrophobic surface properties of erythrocytes studied by affinity partition in aqueous two-phase systems

Summary Erythrocytes from various species have been partitioned in aqueous two-phase systems consisting of water, dextran, poly-(ethylene glycol), salt and buffer. The terminal hydroxyl groups of the latter polymer were esterified with palmitic, oleic, linoleic and linolenic acids, as well as with deoxycholic acid. In a two-phase system containing unesterified poly(ethylene glycol) the erythrocytes are exclusively in the dextran-rich lower phase. When the poly(ethylene glycol) is esterified the red blood cells collect at the interface and/or in the poly(ethylene glycol)-rich upper phase depending on the type and concentration of esterified acid. Palmitate ester is most effective in increasing the affinity of the cells for the upper phase, followed by oleate, linolate, linolenate, and deoxycholate esters. The partition behaviour of erythrocytes from various species differs considerably. Two groups can be distinguished: one consisting of erythrocytes from dog, guinea pig and rat, the other from human, sheep and rabbit. This division can be correlated to the content of sphingomyelin and phosphatidyl choline in the erythrocyte membranes.     

Simultaneous existence of different environments in aqueous clay systems and its possible role in prebiotic synthesis

Summary The formation of packets of parallel oriented platelets and separating distances of several angstrom units in montmorillonite-water systems produces an intrinsic inhomogeneity with respect to the proton donating power of internal and external zones. Stable packets can be induced by both inorganic and organic molecules or ions, in suspensions or in drying-out systems. The coexistence of zones with different proton donating power was demonstrated by the pH-sensitive color reaction of benzidine, where stable packets of montmorillonite platelets were formed by the use of either paraquat or diquat.The close proximity of the two types of zones, which can be of the order of several angstroms, produces the conditions which were defined by Katchalsky as essential for the polymerization of amino acids. Since these environmental conditions are quite common in nature, both at present and in prebiotic times, it is proposed that the inhomogeneity of clay-water systems with respect to proton donating power should be taken into account in both theoretical and experimental efforts to demonstrate the catalytic activity of clays in prebiotic synthesis.     

Pseudomonas aeruginosa PAO1 as a model for rhamnolipid production in bioreactor systems

Rhamnolipids are biosurfactants with interesting physico-chemical properties. However, the main obstacles towards an economic production are low productivity, high raw-material costs, relatively expensive downstream processing, and a lack of understanding the rhamnolipid production regulation in bioreactor systems. This study shows that the sequenced Pseudomonas aeruginosa strain PAO1 is able to produce high quantities of rhamnolipid during 30 L batch bioreactor cultivations with sunflower oil as sole carbon source and nitrogen limiting conditions. Thus PAO1 could be an appropriate model for rhamnolipid production in pilot plant bioreactor systems. In contrast to well-established production strains, PAO1 allows knowledge-based systems biotechnological process development combined with the frequently used heuristic bioengineering approach. The maximum rhamnolipid concentration obtained was 39 g/L after 90 h of cultivation. The volumetric productivity of 0.43 g/Lh was comparable with previous described production strains. The specific rhamnolipid productivity showed a maximum between 40 and 70 h of process time of 0.088 g_RL/g_BDMh. At the same time interval, a shift of the molar di- to mono-rhamnolipid ratio from 1:1 to about 2:1 was observed. PAO1 not only seems to be an appropriate model, but surprisingly has the potential as a strain of choice for actual biotechnological rhamnolipid production.     

Membrane surface properties of normal and malignant cells: partition in aqueous two-polymer phase systems

The partition of normal and malignantly transformed fibroblast lines and cell lines initiated from malignant human astrocytomas and a benign ganglioneuroma has been examined in aqueous dextran-polyethylene glycol phase system containing phosphate buffer with a low phosphate/sodium chloride ratio. The malignant astrocytomas showed a significantly lower partition coefficient as compared with the benign ganglioneuroma. Treatment of astrocytoma cells with dexamethasone caused an increase in the partitioning of the cell population. No differences were found in the partition behaviour of normal BHK-21 cells and their malignant transformants, the TRES fibrosarcoma cells. Polyoma and simian virus-transformed 3T3 fibroblasts showed partition ratios similar to the untransformed cells. Dexamethasone pre-treatment had no effect on the partition behaviour of these cells. The significance of these observations has been discussed in relation to the surface hydrophobicity and the neoplastic state.     

Effects of surfactants on emulsification and secondary structure of lysozyme in aqueous solutions

This paper reports our investigations on the secondary structure of lysozyme in aqueous solution with D₂O and comparing systems emulsified with ionic and non-ionic amphiphiles, respectively. The opposite effect of concentration of surfactants on the lysozyme aqueous system was studied and the activity of lysozyme by the turbidimetric assay with different kinds of surfactant was compared. The effect of different surfactants on emulsification volume formed was observed and discussed. The different interactive patterns between lysozyme and ionic and non-ionic surfactants, such as sodium dodecyl sulfate (SDS), cetyl trimethyl ammonium bromide (CTAB) and Triton-X100 (Isooctoylphenoxypolyethoxy ethanol), respectively, was studied with FTIR. A simple relationship between the secondary structure and denaturation of protein is given for the emulsification of lysozyme in aqueous solutions. It is important for understanding the stabilization and avoiding the emulsions in the extraction process that the interactions between the proteins and surfactants which leads to the formation of such complexes are characterized. The utility of surfactants as a de-emulsifying agent for the commercial process to extract penicillin G from the filtrate of fermentation broth was consistent with these results.