Amino Acid Molar Ratio Research Articles

Synthesis and performance of plant-based ionic liquids in water-based drilling fluids

In this paper, the ionic liquids Met-1, Met-2, Asp-1 and Asp-2 were prepared by acid-base neutralization reactions with different molar ratios of amino acids (methionine and aspartic acid) and concentrated sulfuric acid as raw materials. Four ionic liquids were formulated into different concentrations of inhibitors, and their hydration inhibition and inhibition mechanism were investigated by anti-expansion experiment, linear swelling experiment and flocculation experiment. The results showed that when the molar ratio of aspartic acid to concentrated sulfuric acid was 1:1.5, which was named Asp-2, and the concentration of Asp-2 was 1.0%, the inhibitor achieved the best hydration inhibition performance on clay with 91.95% anti-swelling rate and 21.37% linear swelling rate. Infrared spectra (FT-IR) analysis, thermogravimetric analysis (TGA), and laser particle size analysis were used to investigate the inhibition mechanism of Asp-2. The results showed that the average particle size of the hydrated soil samples increased by 0.282 μm and the median particle size increased by 2.101 μm after the addition of Asp-2 inhibitor, indicating that Asp-2 inhibitor played a good inhibitory role in the drilling fluid.

Non-salt based co-amorphous formulation produced by freeze-drying

Amino acids-based co-amorphous system (CAM) has shown to be a promising approach to overcome the dissolution challenge of biopharmaceutics classification system class II drugs. To date, most CAM formulations are based on salt formation at a 1:1 M ratio and are prepared by mechanical activation. However, its use in medicinal products is still limited due to the lack of in-depth understanding of non-ionic based molecular interactions. There are also limited studies on the effect of drug-to-co-former ratio, the development of more scalable, less aggressive, manufacturing processes such as freeze drying and its dissolution benefits. This work aims to investigate the effect of the ratio of tryptophan (a model non-ionic amino acid) to indomethacin (a model drug) on a non-salt-based CAM prepared via freeze-drying with the tert-butyl alcohol-water cosolvent system. The CAM material was systemically characterized at various stages of the freeze-drying process using DSC, UV–Vis, FT-IR, NMR, TGA and XRPD. Dissolution performance and physical stability upon storage were also investigated. Freeze-drying using the cosolvent system has been successfully shown to produce CAMs. The molecular interactions involving H-bonding, H/π and π–π between compounds have been confirmed by FT-IR and NMR. The drug release rate for formulations with a 1.5:1 drug: amino acid molar ratio (or 1:0.42 wt ratio) or below is found to be significantly improved compared to the pure crystalline drug. Furthermore, formulation with a 2.3:1 drug:amino acid molar ratio (or 1:0.25 wt ratio) or below have shown to be physically stable for at least 9 months when stored at dry condition (5% relative humidity, 25 °C) compared to the pure amorphous indomethacin. We have demonstrated the potential of freeze-drying using tert-butyl alcohol-water cosolvent system to produce an optimal non-salt-based class II drug-amino acid CAM.

Aliphatic amino acids as possible hydrogen bond donors for preparing eutectic solvents

Since their first introduction in the literature about two decades ago, deep eutectic solvents have competed with ionic liquids due to their lower production cost, simple synthesis, and favorable biodegradability. Many studies showed their suitability in wide-ranging applications involving mass and heat transfer, catalysis as well as in the pharmaceutical industries. Very few amino acids-based eutectic solvents have been reported including neutral and acidic-basic amino acids. In this work, a group of four aliphatic-type amino acids were successfully coupled as hydrogen bond donors with tetrabutylammonium chloride forming new eutectic solvents at different molar compositions. Three different salts to amino acid molar ratios were prepared and characterized in terms of their phase-change behavior and basic physicochemical properties within the temperature range of 313.15–353.15 K. Besides, their thermal phase change nature, as well as the ionic strength and infra-red spectra, have also been studied. Results showed an inverse linear relation for density, surface tension, refractive index, and pH. While an inverse exponential relation was noticed for the viscosity and proportional exponential relation for the conductivity with the increase of temperature. However, the molar ratio trends did not follow a specific pattern.

Acetone-assisted co-processing of meloxicam with amino acids for enhanced dissolution rate

Meloxicam is a widely used non-steroidal anti-inflammatory agent. However, its erratic and poor dissolution delays its onset of action. Dissolution enhancement of such medicine is essential to obtain rapid pain relief. Amino acids showed high potential to enhance the dissolution rate of drugs after co-processing. Accordingly, the objective of this work was to investigate the effect of co-processing of meloxicam with arginine, cysteine, and glycine on its crystalline structure and dissolution rate. Meloxicam was mixed with increasing molar ratios of amino acids before acetone-assisted kneading. The resulting products were examined using Fourier transform infrared spectroscopy, differential scanning calorimetry, and X-ray diffraction in addition to monitoring the dissolution behavior. Combined instrumental analysis indicated salt formation with a possibility of further crystalline changes at high concentration of amino acids. Salt formation and crystalline structure modification were associated with a significant increase in the dissolution rate of meloxicam. The study introduced amino acids as potential excipients for enhanced dissolution of meloxicam after wet co-processing.

Influence of pH and aurate/amino acid ratios on the tuneable optical features of gold nanoparticles and nanoclusters

Blue- and green-emitting gold nanostructures have been synthesized by spontaneous interactions of the tetrachloroaurate anions (AuCl4−) with L-histidine (His) and L-tryptophan (Trp) at 37°C in aqueous medium. The dominant role of AuCl4−/amino acid molar ratios as well as the pH in the formation of gold nanoparticles (Au NPs) or nanoclusters (Au NCs) was confirmed by numerous spectroscopic techniques. We demonstrate that without additional reducing agents the His and Trp show different reduction behaviour with AuCl4− which supports the fact that the unique optical features originate from diverse structures. Because of the pH-dependent complexation and reduction capability of the studied amino acids the dominant presence of polynuclear fluorescent gold(I) complexes (quantum yield (QY%) ∼4%) with a well-ordered structure is presumable for His-directed Au system confirmed by mass spectrometry, X-ray diffraction and infrared experiments. The existence of d=8 nm-sized Au NPs or subnanometer-sized (d<2nm) Au NCs (Au3-Au5) that have tuneable blue or green photoluminescence (λemission=470–500nm) was identified in the samples containing Trp depending on the applied AuCl4−/Trp ratios. The blue-emitting Au/His nanohybrid system as potential bioimaging agent was used to visualize fluorescently albumin- and chitosan-based carrier composite particles.

Release of selected amino acids from zinc carriers.

The paper deals with the results of an investigation of the release of selected amino acids (histidine, tryptophan, tyrosine) from model suspensions prepared by co-precipitation with zinc chloride. It has been proven that the influence of the Zn(II)/amino acid molar ratio on dissolution profiles of the tested amino acids and dissolution half-life (t1/2) of histidine or tryptophan is significant. The amount of amino acid in the dispersed phase (supporting dose) is a determinant of the amino acid release profile. There is a minimal supporting dose (30.0 μmol of histidine or 17.4 μmol of tryptophan) that provides release of similar amounts of amino acid (4.1-4.6 μmol of histidine or 8.7-9.9 μmol of tryptophan) after the same time intervals. The tyrosine release profiles follow first order kinetics since the supporting dose (0.9-11.2 μmol) is limited by the tyrosine low solubility in water.

Monitoring potential molecular interactions of adenine with other amino acids using Raman spectroscopy and DFT modeling

We report on the modes of inter-molecular interaction between adenine (Ade) and the amino acids: glycine (Gly), lysine (Lys) and arginine (Arg) using Raman spectroscopy of binary mixtures of adenine and each of the three amino acids at varying molar ratios in the spectral region 1550–550cm−1. We focused our attention on certain specific changes in the Raman bands of adenine arising due to its interaction with the amino acids. While the changes are less apparent in the Ade/Gly system, in the Ade/Lys or Ade/Arg systems, significant changes are observed, particularly in the Ade Raman bands that involve the amino group moiety and the N7 and N1 atoms of the purine ring. The ν(N1–C6), ν(N1–C2), δ(C8–H) and δ(N7–C8–N9) vibrations at 1486, 1332, 1253 and 948cm−1 show spectral changes on varying the Ade to amino acid molar ratio, the extent of variation being different for the three amino acids. This observation suggests a specific interaction mode between Ade and Lys or Arg, which is due to the hydrogen bonding. The measured spectral changes provide a clear indication that the interaction of Ade depends strongly on the structures of the amino acids, especially their side chains. Density functional theory (DFT) calculations were carried out to elucidate the most probable interaction modes of Ade with the different amino acids.

Renal transplantation promptly restores excretory function but disturbed L-arginine metabolism persists in patients during the early period after surgery

The synthesis and whole body metabolism of L-arginine (Arg) are disturbed in renal diseases. Renal transplantation represents the best therapy in the end-stage of these diseases. In the present we compared alterations of plasma Arg and related compounds with renal excretory function in patients with end-stage renal disease, before and after kidney transplantation. Arg, asymmetric dimethylarginine (ADMA), citrulline (Cit), glutamine (Gln), ornithine (Orn), phenylalanine (Phe), tyrosine (Tyr), urea, creatinine, albumin, and nitrate were analyzed in patients before, immediately after (0-time) and 1, 2, 3, 7 and 14 days following living donors kidney transplantation. Healthy subjects were controls. Glomerular filtration rate (GFR) and amino acid molar ratios were calculated. Before transplantation creatinine, urea, Cit, Gln, ADMA, and nitrate were above, while GFR and Arg were below controls, confirming disturbed excretory and metabolic renal functions in patients with renal disease. Renal transplantation promptly normalized creatinine, urea, GFR, Cit, and nitrate. However, regardless of increased molar Phe/Tyr ratios, indicating increased net protein catabolism in peripheral tissues, low Arg and elevated ADMA concentrations persisted throughout the examined period. Alterations of other amino acids also suggest similarly disturbed Arg metabolism in patients after kidney transplantation. In conclusion, renal transplant promptly restored its excretory function, but increased net protein catabolism, disturbed Arg metabolism and endothelial dysfunction in entire body of these patients were not improved throughout the early period after the operation. That has to be considered in their therapy.

Profiles of serum amino acids to screen for catabolic and inflammation status in calves with Mycoplasma bronchopneumonia.

The aim of the present study was to investigate the relationships between serum amino acid profiles in normal and calves with Mycoplasma bronchopneumonia. Serum free amino acid concentrations in serum obtained from 34 calves with or without Mycoplasma bronchopneumonia were determined by high-performance liquid chromatography. The calves with Mycoplasma were characterized by significantly lower total amino acid and total essential amino acid concentrations and molar ratios of branched-chain amino acid (BCAA) to aromatic amino acid (BCAA/AAA) and BCAA to tyrosine (BTR), and by a significantly higher molar ratio of serine phosphorylation (SPR). The proposed diagnostic cutoffs for BCAA/AAA, BTR and SPR in serum based on ROC analysis for detection of catabolic states associated with Mycoplasma bronchopneumonia were set at <1.75, <2.86 and >0.85, respectively. Our results suggest that determining the profiles of amino acids, especially BTR and SPR, could provide useful diagnostic information in terms of predicting protein catabolism in Mycoplasma bronchopneumonia.

Detection of TPN contamination of dried blood spots used in newborn and metabolic screening and its impact on quantitative measurement of amino acids

BackgroundMarkers derived from dextrose (d-glucose) are observed in the MS/MS-based acylcarnitine profiles from dried-blood spots of some premature infants receiving intravenous nutrition. The presence of these markers at m/z 325, 399 and 473 are thought to arise from contamination of blood by total parenteral nutrition (TPN) solutions during specimen collection from premature infants. These solutions contain high concentrations of amino acids and as a result, false-positive screening results for amino acid disorders may occur. This study investigates quantitative parameters of dextrose and amino acids in blood samples enriched with different TPN solutions. MethodsWhole blood collected in heparin was enriched with three different TPN solutions containing 5, 10 or 12.5% dextrose and amino acids that were originally prepared for delivery of 2.5, 3 or 4g/kg/day of Premasol® then spotted onto filter paper cards. Acylcarnitine and amino acid profiles using MS/MS were obtained. Ion ratios of dextrose relative to specific acylcarnitine stable isotope internal standards and amino acid concentrations were obtained. ResultsThe ion ratios for each of the dextrose markers at m/z 325, 399 and 473 exhibit linearity with the concentration of the dextrose component of TPN added to blood. The lowest detectable dextrose concentration added to blood was 7.6mmol/l at 1:80v/v TPN in blood. Furthermore, the concentrations of amino acids were linear with the concentration of the amino acid component of TPN added to blood. At the lowest detectable concentrations of dextrose marker, the amino acid concentrations were at or above the values considered abnormal in newborn screening laboratories. The molar ratios of amino acids approached the relative quantity of amino acid in the TPN solution with increasing enrichments in blood. ConclusionsDetection of the combinations of dextrose markers, very high elevations of amino acids and unusual molar ratios can be used to reject a specimen as improperly collected rather than declaring it a false positive and hence reduce false positive rates. This process enhances efficiency, reduces parental anxiety, and improves positive predictive values.

Heat and moisture promoted deterioration of raw silk estimated by amino acid analysis

Abstract Silks are amongst the most precious artifacts of our textile heritage. While most silk fabrics are made from degummed silk, some in collections, such as ancient Chinese Juan, are composed of raw or just partially degummed silk. Such silks with remnant sericin gum may require special consideration for their optimum preservation. Conservators and curators then need to know which of the silks in their collections are in this category, and what effect the sericin has on the ageing of silk fibres. In the study reported here, the artificial heat and moisture ageing of raw silk was investigated by means of amino acid analysis. Amongst various amino acid molar ratios, Asp/Gly proved a valuable indicator of residual sericin gum on aged silk. Measurement of the tyrosine content was also useful for gauging the conservation state of silk in some circumstances. The results of the amino acid analyses when combined with those for tensile tests suggested that sericin did not inhibit fibroin deterioration under the ageing conditions employed.

Speciation of the Ternary Complexes of Vanadium(III)–Dipicolinic Acid and the Amino Acids Cysteine, Histidine, Aspartic and Glutamic Acids in 3.0 mol⋅dm−3 KCl at 25 °C

In this work we present results for the speciation of the ternary complexes formed in the aqueous vanadium(III)–dipicolinic acid and the amino acids cysteine (H2cys), histidine (Hhis), aspartic acid (H2asp) and glutamic acid (H2glu) systems (25 °C; 3.0 mol⋅dm−3 KCl as ionic medium), determined by means of potentiometric measurements. The potentiometric data were analyzed with the least-squares program LETAGROP, taking into account the hydrolysis of vanadium(III), the acid-base reactions of the ligands, and the binary complexes formed. Under the experimental conditions (vanadium(III) concentration = 2–3 mmol⋅dm−3 and vanadium(III): dipicolinic acid: amino acid molar ratio 1:1:1, 1:1:2 and 1:2:1), the following species [V(dipic)(H2asp)]+, [V(dipic)(Hasp)], [V(dipic)(asp)]−, [V(dipic)(asp)(OH)]2−, and [V(dipic)(asp)(OH)2]3− were found in the vanadium(III)–dipicolinic acid–aspartic acid system. In the vanadium(III)–dipicolinic acid–glutamic acid system [V(Hdipic)(H2glu)]2+, [V(dipic)(H2glu)]+, [V(dipic)(Hglu)], [V(dipic)(Hglu)(OH)]−, and [V(dipic)(Hglu)(OH)2]2− were observed. In the vanadium(III)–dipicolinic acid–cysteine system the complexes [V(dipic)(H2cys)]+, [V(dipic)(Hcys)], [V(dipic)(cys)]−, and [V(dipic)(cys)(OH)]2− were present. And finally, in the vanadium(III)–dipicolinic acid–histidine system the complexes [V(Hdipic)(Hhis)]2+, \([\mathrm{V}(\mathrm{dipic}) (\mathrm{Hhis})]^{+}\), [V(dipic)(his)], [V(dipic)(his)(OH)]−, and [V(dipic)(his)(OH)2]2− were observed. The stability constants of these complexes were determined. The species distribution diagrams as a function of pH are briefly discussed.

Diversity of Innate Immune Recognition Mechanism for Bacterial Polymeric meso-Diaminopimelic Acid-type Peptidoglycan in Insects

In Drosophila, the synthesis of antimicrobial peptides in response to microbial infections is under the control of the Toll and immune deficiency (Imd) signaling pathway. The Toll signaling pathway responds mainly to the lysine-type peptidoglycan of Gram-positive bacteria and fungal β-1,3-glucan, whereas the Imd pathway responds to the meso-diaminopimelic acid (DAP)-type peptidoglycan of Gram-negative bacteria and certain Gram-positive bacilli. Recently we determined the activation mechanism of a Toll signaling pathway biochemically using a large beetle, Tenebrio molitor. However, DAP-type peptidoglycan recognition mechanism and its signaling pathway are still unclear in the fly and beetle. Here, we show that polymeric DAP-type peptidoglycan, but not its monomeric form, formed a complex with Tenebrio peptidoglycan recognition protein-SA, and this complex activated the three-step proteolytic cascade to produce processed Spätzle, a Toll receptor ligand, and induced Drosophila defensin-like antimicrobial peptide in Tenebrio larvae similarly to polymeric lysine-type peptidoglycan. Monomeric DAP-type peptidoglycan induced Drosophila diptericin-like antimicrobial peptide in Tenebrio hemocytes. In addition, both polymeric and monomeric DAP-type peptidoglycans induced expression of Tenebrio peptidoglycan recognition protein-SC2, which is DAP-type peptidoglycan-selective N-acetylmuramyl-l-alanine amidase that functions as a DAP-type peptidoglycan scavenger, appearing to function as a negative regulator of the DAP-type peptidoglycan signaling by cleaving DAP-type peptidoglycan in Tenebrio larvae. Taken together, these results demonstrate that molecular recognition mechanism for polymeric DAP-type peptidoglycan is different between Tenebrio larvae and Drosophila adults, providing biochemical evidences of biological diversity of innate immune responses in insects.

High-performance liquid chromatographic method for determination of amino acids by precolumn derivatization with 4-chloro-3,5-dinitrobenzotrifluoride

This work presents an high-performance liquid chromatography method for the determination of amino acids after precolumn derivatization with 4-chloro-3,5-dinitrobenzotrifluoride (CNBF) which can readily react with both primary and secondary amines. The precolumn derivatization conditions, including the CNBF concentration, reaction pH, temperature and reaction time were investigated for method optimization. In pH 9.0 borate buffer, the reaction of amino acids with CNBF was carried out at 60 °C for 30 min, the optimized concentration of CNBF was 70 mmol L −1 and the molar ratio of amino acids to CNBF was 1:5.25. The chromatographic separation of 19 amino acids derivatives was performed on a Kromasil ODS C 18 column (250 mm × 4.6 mm, 5 μm) with good reproducibility, and ultraviolet detection was applied at 260 nm. The mobile phase was a mixture of phase A (acetonitrile) and phase B (acetate buffer, acetonitrile, triethylamine; 82.8:17:0.2, pH 4.9), and the flow rate was 0.4 mL min −1. The separation of all the labeled amino acids was achieved within 45 min at room temperature by gradient elution mode. The method linearity, calculated for each amino acid, had a correlation coefficient higher than 0.9979, in concentrations ranging from 9.60 to 3330.00 μmol L −1. The detection limits of amino acids were 2.40–6.50 μmol L −1, at a signal-to-noise ratio of 3. The proposed method was applied for the determination of amino acids in beer with recoveries of 97.0–103.9% and relative standard deviations of 2.62–4.22%, respectively. This method showed good accuracy and repeatability that can be used for the quantification of amino acids in real samples.

2,7-Dimethyl-3,8-dinitrodipyrazolo[1,5-a:1′,5′-d]pyrazine-4,9-dione: A new labelling reagent for liquid chromatographic analysis of amino acids

The use of 2,7-dimethyl-3,8-dinitrodipyrazolo[1,5-a:1′,5′-d]pyrazine-4,9-dione as pre-column reagent for LC analysis of amino acids is proposed. The compound reacts (30 min at 68 °C in presence of 0.04 M triethylamine in a dimethylsulfoxide–water mixture) with primary and secondary amino function and the stable resulting adducts can be chromatographed under reversed-phase conditions and detected at λ = 280 nm. The derivatization conditions were optimized by a series of experiments. The effect of temperature, triethylamine concentration and reagent on the reaction was investigated. The yield of the glycine derivative was found to be quantitative at a reagent amino acid molar ratio of about 6 by comparison with an authentic specimen of synthesized glycine adduct. Application of the method to quality control of commercially available oral polyaminoacid formulations is described.

Identification of Mn2+-binding Aspartates from α, β, and γ Subunits of Human NAD-dependent Isocitrate Dehydrogenase

The human NAD-dependent isocitrate dehydrogenase (IDH), with three types of subunits present in the ratio of 2alpha:1beta:1gamma, requires a divalent metal ion to catalyze the oxidative decarboxylation of isocitrate. With the aim of identifying ligands of the enzyme-bound Mn(2+), we mutated aspartates on the alpha, beta, or gamma subunits. Mutagenesis target sites were based on crystal structures of metal-isocitrate complexes of Escherichia coli and pig mitochondrial NADP-IDH and sequence alignments. Aspartates replaced by asparagine or cysteine were 206, 230, and 234 of the alpha subunit and those corresponding to alpha-Asp-206: 217 of the beta subunit and 215 of the gamma subunit. Each expressed, purified mutant enzyme has two wild-type subunits and one subunit with a single mutation. Specific activities of WT, alpha-D206N, alpha-D230C, alpha-D234C, beta-D217N, and gamma-D215N enzymes are 22, 29, 1.4, 0.2, 7.3 and 3.7 micromol of NADH/min/mg, respectively, whereas alpha-D230N and alpha-D234N enzymes showed no activity. The K(m,Mn(2+)) for alpha-D230C and gamma-D215N are increased 32- and 100-fold, respectively, along with elevations in K(m,isocitrate). The K(m,NAD) of alpha-D230C is increased 16-fold, whereas that of beta-D217N is elevated 10-fold. For all the mutants K(m,isocitrate) is decreased by ADP, indicating that these aspartates are not needed for normal ADP activation. This study demonstrates that alpha-Asp-230 and alpha-Asp-234 are critical for catalytic activity, but alpha-Asp-206 is not needed; alpha-Asp-230 and gamma-Asp-215 may interact directly with the Mn(2+); and alpha-Asp-230 and beta-Asp-217 contribute to the affinity of the enzyme for NAD. These results suggest that the active sites of the human NAD-IDH are shared between alpha and gamma subunits and between alpha and beta subunits.

Network analysis of plasma and tissue amino acids and the generation of an amino index for potential diagnostic use

Few studies exist on the use of metabolic profiling of amino acids to examine underlying physiologic and disease states. We aimed to introduce a new method for studying relations among amino acids and to generate a diagnostic index, or amino index, based on amino acid concentrations. For network analysis, 35 Fischer-344 rats were randomly divided into 7 groups and fed diets containing 5%, 10%, 15%, 20%, 30%, 50%, or 70% protein. Amino acid concentrations in plasma and various organs were used to derive correlation coefficients that were then used to construct correlation networks. To build a diagnostic index for diabetic rats, the plasma amino acid concentrations of diabetic and normal rats were analyzed by using a novel algorithm developed to generate amino acid-based indexes. Plasma amino acid concentrations from human growth hormone transgenic rats and insulin-treated diabetic rats were used to evaluate the index obtained for diabetes. Dimethylnitrosamine-treated Sprague-Dawley rats were used to generate an index for hepatic fibrosis. The scatter plots of plasma amino acid concentrations showed distinct patterns in different organs that were due to the different protein contents of the diets. Network analysis showed that data-driven networks for blood and tissue could be obtained. We derived a diagnostic index for the discrimination of diabetic rats with both sensitivity and specificity >97% and another surrogate index for liver hydroxyproline with a correlation of r2= 0.85. Correlation-based network analysis may help to uncover specific physiologic conditions or states. A novel approach using amino acid molar ratios was shown to generate indexes that can be used to separate animal disease models and monitor the progression of a disease parameter. Some of the methods described here may be applicable to the clinical setting.

Towards an understanding of (bio)silicification: the role of amino acids and lysine oligomers in silicification

In order to understand the role that proteins play in the generation of well regulated biosilica structures we need to understand the contribution of the components, singly and in combination. To this end we have performed a systematic study of the effect of amino acids and small peptide oligomers on silica formation from aqueous solution. Silicas produced from a potassium silicon catecholate salt at ca. pH 7 in the presence of the amino acids (Gly, Arg, Asn, Gln, Glx, Ser, Thr, Tyr, Pro, Ala, Lys) at a 2 Si : 1 amino acid molar ratio have shown that these amino acids affect the kinetics of small oligomer formation, the growth of aggregate structures and the morphology and surface properties of the silicas produced. The effects seen during the early stages of oligomer formation carry through to the properties of the particles and aggregates produced after extended periods of reaction. The behaviour of the amino acids relates to the pI and hydrophobicity of the individual amino acids. The presence of the nitrogen containing amino acids generates larger particles and the presence of amino acids containing hydroxyl and hydrophobic groups generates silicas with smaller particles than are produced for silicas produced in the absence of amino acids. An extensive study of the effect of the number of lysine and glycine units per peptide was also performed (for lysine, 1–5 and ca. 150 and for glycine, 1,4,5). Increasing the number of glycine units per additive molecule had little effect on the kinetics, aggregation, sample morphology, surface area and porosity of the silicas produced. A distinct relationship between the number of lysine units per additive molecule and an increased rate of oligomer formation, aggregate growth and a reduction in silica surface area and broadening of pore sizes was observed. A distinct change over in behaviour, particularly in regard to the porosity characteristics of the silica produced was noted for between (lys) 3 and (lys) 4 as well as this being the smallest size of peptide that was incorporated into the siliceous material formed. Aggregation was observed to accelerate exponentially over the full range of lysine oligomers used. Consecutive sequences of the same amino acid residues were shown to produce effects much larger than the sum of the effects of the individual residues, and at extremes mediate macroscopic morphological changes. The consequences of these findings for biosilicification are discussed. It is clear that all amino acid functional groups in proteins that are accessible to silica during the stages of formation from orthosilicic acid through to the final material have a role to play in determining the physical nature and structure of the material that forms.

Synthesis and characterization of amino acid-functionalized hydroxyapatite nanorods

Synthesis of aqueous colloids of amino acid-functionalized hydroxyapatite (HAP; Ca10(PO4)6(OH)2) nanoparticles with rod-like morphology and uniform size was achieved by hydrothermal crystallization at 80 °C and pH 9 in highly supersaturated solutions with a Ca2+∶amino acid molar ratio of 1∶2. Stable suspensions of positively charged nanorods, less than 80 nm in length and ca. 5 nm in width, were obtained in the presence of glycine, alanine, serine, lysine and arginine, and together with FTIR spectroscopy data this was consistent with a general model in which the α-carboxylate of the amino acid was preferentially bound to the crystal surfaces. Variations in the size and shape of the nanoparticles with amino acid used were consistent with differences in the strength of binding at the HAP surfaces. In comparison, stable colloids could not be prepared with aspartic acid due to a relatively low surface charge; instead, the precipitate consisted of highly elongated nanorods, ca. 150 × 3.5 nm in dimension. XRD textural analysis suggested a model for nanorod formation based on the oriented aggregation and fusion of primary crystallite domains specifically along the c axis direction. Colloids consisting of HAP/alanine or HAP/arginine nanoparticles were cross-linked in situ by thermal, hydrothermal or chemical polymerisation of surface-attached and unbound amino acids to produce aggregated nanostructures with variable degrees of long-range order.

Amino acid acceptor identity switch of Escherichia coli tmRNA from alanine to histidine in Vitro

According to a trans-translation model, tmRNA facilitates the resumption of translation that has been stalled on the ribosome with the 3′ end of a terminator-less mRNA, to produce a chimera polypeptide of the nascent peptide and the tmRNA-encoding 11 amino acid-tag. The first alanine residue of the tag-sequence is encoded neither by mRNA nor by tmRNA. This alanine is a key molecule for this model, in which it is derived from the alanine moiety aminoacylated to tmRNA. This is supported only by the observation that a point mutation at the third base-pair position of the acceptor stem of Escherichia coli tmRNA that deprives it of its aminoacylation ability causes abolishment of tag-peptide synthesis in vitro. Here, we made an E. coli tmRNA mutant with a completely switched amino acid acceptor identity from alanine to histidine by transplanting the upper half of the acceptor stem of tRNAHis. This histidine acceptor tmRNA mutant still retained an ability of tag-specific amino acid incorporation into the polypeptide in an in vitro poly(U)-dependent tag-peptide synthesis system, with an altered amino acid composition. Histidine, which is not a constituent of the original tag-peptide, was incorporated into the mutant-directed tag. The molar ratio of amino acids incorporated is consistent with that in the tag-sequence with the only expected change being the first amino acid from alanine to histidine. These results indicate that the first alanine residue of the tag-peptide is actually derived from that aminoacylated to tmRNA and is substitutable by other amino acids during the trans-translation processes.