Excitable artificial cells of proteinoid

The proteinoid cells are assembled of thermal polymers of amino acids. Typically, an appropriate mixture of amino acids containing aspartic or glutamic acid is heated at 190°C for 6 h, stirred with water for 2 h, dialyzed during 2 d, and lyophilized. Spheroidal cells are made from such polymer by dissolving it in the water by boiling, and then cooling. Many of them can be made by sonication at room temperature. These artificial cells, ranging from microns to tens of microns in diameter (depending on composition and preparation), have double membranes and various internal compositions. The spherules can microencapsulate dyes, oxidant-reductant compounds or acceptor-donor substances, and can be packed together. Such spherules display electrical polarization and electrical discharges and respond to intra- and extracellular ionic and electric influence upon membrane and action potential. These properties arise from the double membrane structure, asymmetric membrane permeability, and channeling phenomena. Such features as exponential dependence of the steady-state conductance and capacitance as well as negative resistance of the membrane seem to be responsible for the flip-flop alternations of the membrane polarization, rhythmic electric oscillations, and all-or-none action potentials. The presence of such chromophores as pteridine and flavin in polymers constituting these cells is responsible for their photosensitivity.     

DP-sensors based on MF-4SC membrane and silica with hydrophobic surface for determination of phenylalanine,valine, and methionine cations

The influence of modification of MF-4SC membranes with silica (or surface-hydrophobized silica) nanoparticles on their ionic conductivity and diffusion permeability as well as the response stability and sensitivity of the membrane-based DP-sensors for cations of amino acids containing hydrophobic fragment has been studied. The membranes modified with surface-hydrophobized silica exhibited the 2–5 times reduced relative error of determination of the hydrophobic amino acids as compared to the initial membranes.     

Identification of beta-barrel membrane proteins based on amino acid composition properties and predicted secondary structure

Unlike all-helices membrane proteins, beta-barrel membrane proteins can not be successfully discriminated from other proteins, especially from all-beta soluble proteins. This paper performs an analysis on the amino acid composition in membrane parts of 12 beta-barrel membrane proteins versus beta-strands of 79 all-beta soluble proteins. The average and variance of the amino acid composition in these two classes are calculated. Amino acids such as Gly, Asn, Val that are most likely associated with classification are selected based on Fishers discriminant ratio. A linear classifier built with these selected amino acids composition in observed beta-strands achieves 100% classification accuracy for 12 membrane proteins and 79 soluble proteins in a four-fold cross-validation experiment. Since at present the accuracy of secondary structure prediction is quite high, a promising method to identify beta-barrel membrane proteins is presented based on the linear classifier coupled with predicted secondary structure. Applied to 241 beta-barrel membrane proteins and 3855 soluble proteins with various structures, the method achieves 85.48% (206/241) sensitivity and 92.53% specificity (3567/3855).     

Competitive anion transport in desalting of mixtures of organic acids by batch electrodialysis

Desalting and separation of binary and quaternary acid mixtures via batch electrodialysis are investigated in this article. A monoselective cation exchange membrane and either a non-selective or a monoselective anion exchange membrane are employed in the electrodialysis stack. The effects of current density and composition of the initial feed of the electrodialysis stack (employing a non-selective anion exchange membrane) on its performance are studied in experiments involving mixtures of acetic and succinic acids. The effect of the type of the anion exchange membrane on the process performance is examined in desalting experiments involving a mixture of acetic, formic, lactic, and succinic acids. The trends observed in the experiments are interpreted in terms of species-specific parameters (such as molar concentration, charge on ionic species, molecular weight, degree of ionization, and ionic equivalent conductivity) and characteristics of anion exchange membrane used.     

Fractionation of casein hydrolysates using polysulfone ultrafiltration hollow fiber membranes

The objective of this study was to characterize the fractionation profile of casein hydrolysates obtained with polysulfone hollow fiber ultrafiltration membranes. The two-step ultrafiltration process developed by Turgeon and Gauthier [J. Food Sci., 55 (1990) 106] was used: a caseinate solution was submitted to proteolysis with chymotrypsin or trypsin, and the reaction mixture (RM) was subsequently ultrafiltered using a 30 kDa (MWCO) hollow-fiber polysulfone membrane. The total hydrolysate permeating from this first step was further fractionated using a 1 kDa (MWCO) membrane, producing the mixture of polypeptides (retentate) and the amino acid fraction (permeate). The effect of enzyme specificity and of membrane retentivitiy on the total composition (total nitrogen, fat, lactose, minerals) and amino acid profile of the fractions was studied. The overall composition of the fractions was not significantly affected by the nature of the enzyme but the degree of hydrolysis and the molecular weight distribution profile analyses showed a marked effect of the enzyme specificity, with trypsin giving a larger proportion of small peptides (< 200 Da) in the mixture of polypeptides. Amino acid profile analyses provided useful information on the phenomena governing the fractionation of amino acids with a polysulfone membrane: (1) the target amino acids of the enzyme are concentrated in the permeate as a result of their presence in all peptides produced by hydrolysis, (2) polar amino acids are retained by the membrane, (3) non-polar amino acids are not selectively rejected by the membrane. Our results suggest that the charge/hydrophobicity balance of the peptides produced is the predominant factor determining the fractionation of casein hydrolysates.     

Study on glucose oxidase fermentation coupled with membrane dialysis

The repression of glucose oxidase (GOD) biosynthesis by catabolites during fermentation can be alleviated through membrane dialysis fermentation (MDF). The results show that the volumetric enzyme productivity of MDF was two times higher than that of the control (fermentation without dialysis), and its total enzyme activity was increased by 30–50%. The operation conditions of MDF, such as pore size of the membrane, initiating time for membrane dialysis, and volume of dialysate used, were optimized. The content of amino acids and organic acids in the fermentation broth and the dialysate in the reservoir were assessed by amino acid analyzer and ionic chromatography, respectively. The relationship among the contents of pyruvic acid, gluconic acid, and enzyme activity during fermentation was analyzed quantitatively. Furthermore, the effect of membrane dialysis technology applied to the low-yield strain was found to be more effective than that applied to the high-yield strain.     

Identification of β-barrel membrane proteins based on amino acid composition properties and predicted secondary structure

Unlike all-helices membrane proteins, β-barrel membrane proteins can not be successfully discriminated from other proteins, especially from all-β soluble proteins. This paper performs an analysis on the amino acid composition in membrane parts of 12 β-barrel membrane proteins versus β-strands of 79 all-β soluble proteins. The average and variance of the amino acid composition in these two classes are calculated. Amino acids such as Gly, Asn, Val that are most likely associated with classification are selected based on Fishers discriminant ratio. A linear classifier built with these selected amino acids composition in observed β-strands achieves 100% classification accuracy for 12 membrane proteins and 79 soluble proteins in a four-fold cross-validation experiment. Since at present the accuracy of secondary structure prediction is quite high, a promising method to identify β-barrel membrane proteins is presented based on the linear classifier coupled with predicted secondary structure. Applied to 241 β-barrel membrane proteins and 3855 soluble proteins with various structures, the method achieves 85.48% (206/241) sensitivity and 92.53% specificity (3567/3855).     

Electrooxidation of Silver in the Presence of Some Amino Acids

Data on the influence the solution's ionic composition and amino acids (histidine, methionine) have on the anodic dissolution of silver are presented. The dissolution is most effective in intensively agitated nitrate solutions containing methionine in excess of 0.05 M (current efficiency reaches 75%). By analogy with the gold dissolution, acceleration of the silver dissolution is attributed to oxidized methionine complexing with metals.     

Room temperature ionic liquids from 20 natural amino acids

We first succeeded in synthesizing ionic liquids from 20 natural amino acids. Amino acid ionic liquids dissolved native amino acids, despite water-free conditions. Furthermore, these ionic liquids are soluble in various organic solvents, such as chloroform. Effects of acidity, hydrogen bonding ability, and steric factors on the properties of these ionic liquids were analyzed as the function of side groups.     

Retention study of some cation-type compounds using bile acid sodium salts as ion pairing agents in liquid chromatography

The possibility of forming ion-pairs between bile acids (sodium taurocholate, sodium taurodeoxycholate and sodium taurochenodeoxycholate) and different compounds (pralidoxime, obidoxime and pyridostigmine) having a cationic character has been studied in reversed-phase liquid chromatography (RP-LC). This study can be useful in understanding the role of bile acids in the transport of ionic species through hydrophobic membrane. The present study focused on the influence of mobile phase composition on the retention parameters of chosen compounds (percentage of acetonitrile, pH of aqueous component or ionic strength). For constant concentration of bile acids in aqueous component of mobile phase the functional dependencies between the logarithm of the retention factor (k) and the methanol content in the mobile phase followed a binomial pattern (U-shaped), with a minimum positioned within the interval 70-85% methanol.     

Efficient Procedure for the Preparation of Oligomer-Free N α -Fmoc Amino Acids

A two-step method is presented for the peptide-free, high-purity, and high-yield synthesis of N^α-Fmoc amino acids. The first step involves the preparation of stable dicyclohexylammonium–amino acid ionic adduct in acetone. Subsequently, the ionic adducts, on reaction with Fmoc-Nosu under mild alkaline conditions, give dipeptide-free N^α-Fmoc amino acids. The positive charge of the dicyclohexylammonium counterion in the ionic salt has a longer radius, moderating the nucleophilicity of the carboxylate ion of the amino acid and preventing by-products by arresting the formation of mixed anhydrides, the precursors of oligopeptide impurities.     

Adsorption Behavior of Amino Acids on a Stainless Steel Surface

The adsorption behavior of various amino acids on a stainless steel surface was investigated at 30 degrees C and over a pH range of 3-10. Acidic and basic amino acids except histidine adsorbed remarkably at pH 3-4 and 7-10, respectively, and showed Langmuir-type adsorption isotherms. The effects of pH and ionic strength on the adsorption isotherms were investigated to analyze the interactions between amino acids and adsorption sites on the stainless steel. Hydrophobic amino acids and glycine showed only small adsorbed amounts at all pHs tested. For the acidic and basic amino acids, reversibility of the absorption and the influence of the ionic strength on the adsorption behavior were examined. The adsorption isotherms of the derivatives of aspartic acid were also measured in order to examine the contribution of the carboxylic groups of acidic amino acids to the adsorption. Furthermore, a Fourier-transform infrared spectroscopic analysis and semiempirical molecular orbital calculation were carried out to analyze the ionization states and the configuration of the amino acids adsorbed on a stainless steel surface. These investigations suggest that the acidic and basic amino acids adsorb through two electrostatic interactions of two ionized groups in the amino acid with a stainless steel surface. Copyright 2000 Academic Press.     

反萃分散组合液膜分离提取氨基酸

建立了分离提取蛋氨酸、亮氨酸、苯丙氨酸和色氨酸的磷酸二（2-乙基己基）酯(D2EHPA) 煤油-HCl反萃分散组合液膜体系,考察了料液相pH值、载体D2EHPA浓度、液膜相与反萃相体积比、反萃相组成、料液相与反萃分散相流速、传输时间以及支撑膜重复使用次数对氨基酸渗透系数和传输效率的影响。 在优化的条件下,建立的反萃分散组合液膜体系对4种氨基酸均可以获得大于35%的传输效率,其中色氨酸和亮氨酸的传输效率超过了79%,且传输效率呈Et,Trp＞Et,Leu＞Et,Phe＞Et,Met的趋势。 支撑膜重复使用25次,对氨基酸的传输效率没有明显改变。建立的液膜体系对考察的氨基酸展示了较高传输效率和优越的传输选择性,是一种简单和环境友好的分离技术。     

Functional amino acid ionic liquids as solvent and selector in chiral extraction

Amino acid ionic liquids (AAILs) have received great attention due to their potentials in catalysis and separations. In this work, functional AAILs were used as solvent and selector in chiral liquid–liquid extraction for the first time. The AAILs have shown distinct enantioselectivity in amino acid extraction. Using these functional AAILs as acceptor phase and ethylacetate as donor phase, more L-enantiomer of amino acid was extracted into the ionic liquid phase than that of D-enantiomer. The influencing factors, including AAILs structure, copper ion concentration, organic phase and amino acid concentration, were investigated. We found that the enantioselective enrichment of racemic amino acids was achieved through a chiral ligand-exchange mechanism. The enantioselectivity of single-step extraction was up to enantiomeric excess value of 50.6%. Moreover, the functional AAILs were found to be efficient extraction solvents for amino acids. The logarithm of distribution coefficient for L-Phe was in the range of 3.4–3.6 in the ionic liquid–ethylacetate two-phase system. This liquid–liquid extraction approach may extend the application of ionic liquids in chiral separations.     

二(2-乙基己基)磷酸对氨基酸的萃取平衡

二（２－乙基己基）磷酸对氨基酸的萃取平衡曹汉瑾,王德宝,刘沛妍,吴子生,严忠（东北师范大学化学系,长春,１３００２４）关键词氨基酸,二（２－乙基己基）磷酸,萃取平衡,分配比迄今为止,有关氨基酸溶剂萃取的文献报道还不多［１～３］．本实验以二（２－乙基己...     

Separation and Determination of Amino Acids by CE Using 1-Butyl-3-methylimidazolium-Based Ionic Liquid as Background Electrolyte

Capillary electrophoresis using 1-butyl-3-methylimidazolium tetrafluoroborate as background electrolyte for the separation and determination of 11 amino acids was studied. Several parameters, such as ionic liquid concentration, pH of background electrolyte and applied voltage, were optimized. Amino acids were derived with Sanger’s reagent (2,4-dinitrofluorobenzene) and the detection was performed by an ultraviolet absorption detector at 360 nm. Under selected conditions, 11 amino acids were completely separated within 16 min. The RSD values of integrated areas and migration times are <3.03 and <1.64%, respectively. The electroosmotic flow can be influenced by imidazolium cation, and the association between the imidazolium cation and the derived amino acids plays an important role in the separation mechanism. This method not only provides an approach for the separation and determination of amino acids, but is also an extension of the ionic liquid application to capillary electrophoresis.     

Effects of ionic dissociation of membrane polymer on pervaporation performance

Pervaporation of a water/alcohol mixture through a membrane which has ion-exchange capacity has been investigated. Theoretical equations are introduced which relate the degree of ionic dissociation of the polymer to quantities of water and alcohol dissolved in the polymer. From these equations, an equation for selective dissolution R is derived which does not contain an explicit term for ionic dissociation. Dissociation affects selective dissolution only by changing the degree of swelling of the polymer. Reformulating R asymptotically obtains a reciprocal relationship between permselectivity and permeability for a water-selective membrane. Experiments to check the validity of the relationship have been carried out using chitosan membranes neutralized by several acids. The effect of degree of neutralization also has been investigated. Results can be well understood on the supposition that ionic dissociation depends upon the water/alcohol composition, the kind of acid, and the degree of neutralization. Experimental results indicate that the reciprocal relationship is maintained over an appropriate range of feed compositions which confirm the validity of the theoretical equations for the swelling equilibrium of an ionic membrane. © 1994 John Wiley & Sons, Inc.     

Electromembrane extraction of amino acids from body fluids followed by capillary electrophoresis with capacitively coupled contactless conductivity detection

Electromembrane extraction (EME) proved to be a simple and rapid pretreatment method for analysis of amino acids and related compounds in body fluid samples. Body fluids were acidified to the final concentration of 2.5 M acetic acid and served as donor solutions. Amino acids, present as cations in the donor solutions, migrated through a supported liquid membrane (SLM) composed of 1-ethyl-2-nitrobenzene/bis-(2-ethylhexyl)phosphonic acid (85:15 (v/v)) into the lumen of a porous polypropylene hollow fiber (HF) on application of electric field. The HF was filled with 2.5 M acetic acid serving as the acceptor solution. Matrix components in body fluids were efficiently retained on the SLM and did not interfere with subsequent analysis. Capillary electrophoresis with capacitively coupled contactless conductivity detection was used for determination of 17 underivatized amino acids in background electrolyte solution consisting of 2.5 M acetic acid. Parameters of EME, such as composition of SLM, pH and composition of donor and acceptor solution, agitation speed, extraction voltage, and extraction time were studied in detail. At optimized conditions, repeatability of migration times and peak areas of 17 amino acids was better than 0.3% and 13%, respectively, calibration curves were linear in a range of two orders of magnitude (r(2)=0.9968-0.9993) and limits of detection ranged from 0.15 to 10 μM. Endogenous concentrations of 12 amino acids were determined in EME treated human serum, plasma, and whole blood. The method was also suitable for simple and rapid pretreatment and determination of elevated concentrations of selected amino acids, which are markers of severe inborn metabolic disorders.     

Open-tubular capillary electrochromatography-mass spectrometry with sheathless nanoflow electrospray ionization for analysis of amino acids and peptides

A novel, rugged sheathless capillary electrochromatography-electrospray ionization (CEC-ESI) device, in which an open-tubular separation capillary and an electrospray tip are integrated with a Nafion tubing junction, is coupled to mass spectrometry (MS) for the analysis of amino acids and peptides. A stable electrospray was generated at nanoflow rates by applying a positive electrical potential at the Nafion membrane junction. To sustain the stable spray, an electroosmotic flow (EOF) to the spray was supported by coating the fused silica capillary with Lupamin, a high-molecular-weight linear positively charged polyvinylamine (PVAm) polymer, which also minimizes analyte adsorption. Electrochromatographic separation of amino acids and peptides was further enhanced by the chromatographic selectivity of Lupamin stationary phase for these molecules. The device was very reliable and reproducible for CEC-ESI-MS analyses of amino acids and peptides for over a hundred injections. The separation and detection behaviors of amino acids and peptides under different conditions including pH, concentration, and composition of mobile phases on Lupamin-coated and uncoated capillaries have been investigated. The relationship between nano electrospray stability and EOF is discussed.