Temperature and Solute Molecular Size Effects on the Retention and Enantioselectivity of a Series of D, L Dansyl Amino Acids on a Vancomycin-Based Chiral Stationary Phase

Summary The influence of column temperature (0–28 °C) and solute molecular size on the retention and enantioselectivity of a series of D, L dansyl amino acids with a non-polar side chain (valine, leucine, phenylalanine and tryptophan) were investigated using a vancomycin-based chiral stationary phase (CSP). The enthalpic and entropic terms for the solute-CSP association were determined from the linear vant Hoff plots. Two solute groups were distinguished in relation to these thermodynamic quantities: the solute group I (dansyl valine, dansyl leucine, dansyl phenylalanine) for which large negative values of enthalpic terms were obtained and the solute group II (dansyl tryptophan) for which H value was much less negative. The enthalpy-entropy compensation study revealed that the interaction mechanism was identical for the group I solute enantiomers but changed for D, L dansyl tryptophan. This was further exemplified as the group I compound enantiomers were resolved over the temperature range while the enantiomers of dansyl tryptophan were not separated in the operating conditions. Relationships between both the solute retention factors and apparent enantioselectivity, and the accessible surface area of the amino acid side chain indicated that when the solute molecular size increased (i) the retention was enhanced by the hydrophobic effect and (ii) the chiral discrimination decreased dependent, at least in part, on a steric hindrance phenomenon at the vancomycin aglycone pocket.     

Preparation of chiral amino acid materials and the study of their interactions with 1,1‐Bi‐2‐naphthol

The amino acid tryptophan has been converted into acrylamide monomers using L /D ‐tryptophan methyl ester forming the enantiopure chiral monomers. Attempts were made to polymerize these monomers via reversible addition fragmentation chain transfer (RAFT) polymerization to form poly(tryptophan). Unfortunately, this proved difficult, and instead, a postpolymerization modification route was used by first synthesizing poly(pentafluorophenyl acrylate) via RAFT, which was then substituted with L ‐tryptophan methyl ester to give poly(L ‐tryptophan). The interactions of the newly synthesized tryptophan monomers, as well as previously reported phenylalanine monomers, were studied in the presence of rac‐BINOL. It has been shown that the enantiomers of tryptophan have a stronger interaction with BINOL than phenylalanine and this has been attributed to the larger π system on the side chain. By monitoring the shifts and splitting of the phenolic protons of BINOL, it has been observed that S‐BINOL interacts more favorably with L ‐monomer enantiomers and R‐BINOL with D ‐monomer enantiomers. Similar interactions have also been seen with poly(phenylalanine) and the newly synthesized poly(tryptophan) materials. © 2012 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem, 2012     

支持向量回归同时测定苯丙氨酸、酪氨酸和色氨酸的研究

在波长200~400 nm范围内,测定酪氨酸、色氨酸和苯丙氨酸混合体系的吸光度,用连续小波变换(CWT)对光谱数据进行预处理,再用支持向量回归(SVR)方法进行建模,建立了支持向量回归紫外分光光度法同时测定酪氨酸、色氨酸和苯丙氨酸的方法,用所建方法对模拟样品进行了测定。结果表明,酪氨酸、色氨酸和苯丙氨酸预测结果的回收率在98%~102%之间,测定结果准确。     

Selective determination of amino acids using flow injection analysis coupled with chemiluminescence detection

The determination of the amino acids proline, histidine, tyrosine, arginine, phenylalanine and tryptophan using flow injection analysis (FIA) with chemiluminescence detection is described. Proline was the only amino acid to exhibit chemiluminescence with the tris(2,2-bipyridyl)ruthenium(III) reaction at pH 10. While, histidine was found to selectively enhance the reaction of luminol with Mn(II) salts in a basic medium. Acidic potassium permanganate chemiluminescence was able to selectively determine tyrosine at pH 6.75. Low pressure separations using a C18 guard column allowed the simultaneous determination of tyrosine and tryptophan or phenylalanine and tryptophan with acidic potassium permanganate and copper(II)-amino acid-hydrogen peroxide chemiluminescence, respectively. Precision for each method was less than 3.9% (R.S.D.) for five replicates of a standard (1×10⁻⁵ M) and the detection limits ranged between 4×10⁻⁹ and 7×10⁻⁶ M. Preliminary investigations revealed that the methodology developed was able to selectively determine the individual amino acids in an equimolar mixture of the 20 naturally occurring amino acids.     

Selective transport of D,L‐tryptophan through poly(L‐glutamic acid) membranes

Permeability coefficients of D ‐ and L ‐tryptophan (D ‐, L ‐Trp) were estimated for poly(L ‐glutamic acid) (PLG) membranes immersed in aqueous ethanol. D ‐tryptophan was selectively transported (the maximum permeability ratio was 2.6) depending on the amount and the species of crosslinking agent, and on the composition of immersing solvent. It is suggested that hydrogen bonding between uncharged permeates and carboxyl and/or amide groups of PLG is an essential factor for the selective transport. © 1999 John Wiley & Sons, Inc. J Polym Sci B: Polym Phys 37: 1035–1041, 1999     

Thermal vaporization of biological nanoparticles: fragment-free vacuum ultraviolet photoionization mass spectra of tryptophan, phenylalanine-glycine-glycine, and beta-carotene

A simple, new way to introduce fragile biomolecules into the gas phase via thermal vaporization of nanoparticles is described. The general utility of this technique for the study of biomolecules is demonstrated by coupling this source to tunable synchrotron vacuum ultraviolet radiation. Fragment-free photoionization mass spectra of tryptophan, phenylalanine-glycine-glycine, and beta-carotene are detected with signal-to-noise ratios exceeding 100. The 8.0 eV photoionization mass spectrum of tryptophan nanoparticles vaporized at 373 K is dominated by a single parent ion peak that exhibits a 20-fold enhancement over the methylene indole fragment ion. The degree of dissociative photoionization of tryptophan can be precisely controlled either by the thermal energy imparted into the neutral tryptophan molecule or by the energy of the ionizing photon. The results reveal how approximately 0.5 eV changes in internal energy affect both the photoionization mass spectrum of tryptophan and the appearance energy of the daughter ion fragments. This method allows the ionization energies of glycine (9.3 +/- 0.1 eV), tryptophan (7.3 +/- 0.2 eV), phenylalanine (8.6 +/- 0.1 eV), phenylalanine-glycine-glycine (9.1 +/- 0.1 eV), and beta-carotene (<7.0 eV) molecules to be determined directly from the photoionization efficiency spectra.     

Expedient synthesis of threo-beta-hydroxy-alpha-amino acid derivatives: phenylalanine, tyrosine, histidine, and tryptophan

An expedient synthesis of enantiomerically pure threo-beta-hydroxy-alpha-amino acid derivatives of phenylalanine, tyrosine, histidine, and tryptophan is described. The NBS-mediated radical bromination of the N,N-di-tert-butoxycarbonyl protected alpha-amino acids and subsequent treatment with silver nitrate in acetone provided the trans-oxazolidinones predominantly. Cesium carbonate catalyzed hydrolysis then generated the beta-hydroxy amino acid derivatives in excellent overall yield.     

Racemic D,L-asparagine causes enantiomeric excess of other coexisting racemic D,L-amino acids during recrystallization: a hypothesis accounting for the origin of L-amino acids in the biosphere

When recrystallizations were performed using a mixture of 12 D,L-amino acids (alanine, aspartic acid, arginine, glutamic acid, glutamine, histidine, leucine, methionine, serine, valine, phenylalanine, and tyrosine) with excess D,L-asparagine, all amino acids with the same configuration as asparagine were preferentially co-crystallized, indicating that it is the nature of a mixture of racemic amino acids to produce a spontaneous high enantiomeric excess.     

石英微米晶粒电泳微柱的制备及其电泳分离

研究了内径2 mm石英管填充均匀石英微米晶粒的电泳微柱制备及其电泳分离的可行性.石英微米晶粒用水热法合成.含30%甲醇的1.5 mmol/L Na2HPO4为电泳缓冲液(pH 11.5),对无需衍生的色氨酸、苯丙氨酸和酪氨酸进行了微柱电泳分离和紫外吸收检测.检出限分别为0.038、 0.21和0.20 μmol/L; 色氨酸的分离效率为4.4 × 104塔板数/m,电泳微柱的样品容量达到35 μL, 且有较好的分离重现性.对未填充石英微粒, 填充平均粒度360 μm石英砂和长9 μm石英微米晶粒的电泳微柱的热效应进行了讨论.实验结果表明,在电泳微柱中填充石英微米晶粒可抑制大柱径电泳的热效应,增大样品容量,提高检测灵敏度.此微柱电泳技术可作为现场、实时和便携式电动流动全分析系统的高效分离手段,适合大体积和低浓度样品分析.     

Kinetics of gas-phase hydrogen/deuterium exchange and gas-phase structure of protonated phenylalanine, proline, tyrosine and tryptophan

Site-specific rate constants for the gas-phase hydrogen/deuterium (H/D) exchange of four, three, five and five hydrogen atoms in protonated phenylalanine (Phe), proline (Pro), tyrosine (Tyr) and tryptophan (Trp), respectively, were determined from matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry (MALDI-FTICRMS) experiments with D(2)O, D(2)S, and CH(3)OD as deuterating agents. No H/D exchange was observed with D(2)S. For exchange with both CD(3)OD and D(2)O, which is about ten times slower in the latter, results indicate for all compounds protonation of the alpha-amino group in agreement with theoretical results. Also, with both reagents, all compounds exchange at the COOH site more than ten times faster than at the protonation site, with OH and NH sites of Tyr and Trp, respectively, exchanging slowest. The observation of H/D exchange despite the high differences in proton affinities between the amino acids and deuterating agent exceeding 200 kJ mol(-1) is in agreement with lowering of the barrier for proton transfer through hydrogen bonding proposed by Lebrilla and coworkers.     

Coordination properties of 3-functionalized beta-cyclodextrins. Thermodynamic stereoselectivity of copper(II) complexes of the A,B-diamino derivative and its exploitation in LECE

The bis-amino AB derivative of beta-cyclodextrin on the secondary rim was synthesised and spectroscopically characterised by different techniques. Its binary systems both with protons and copper(II) were thermodynamically characterised by pH-metric potentiometry. In addition the ternary systems with each of the enantiomers of tryptophan and alanine were investigated. A thermodynamic stereoselectivity was observed for the tryptophan enantiomers and this was exploited to separate them by capillary electrophoresis through a ligand exchange mechanism (LECE). LECE separation of racemates of phenylalanine and tyrosine was also obtained.     

Complex formation of tetrakis(4-sulfonatophenyl)porphyrin with γ-cyclodextrin, phenylalanine, and tryptophan in aqueous solution

In pH 10.3 buffer, in which phenylalanine and tryptophan are in an anionic form, the interactions of tetrakis(4-sulfonatophenyl)porphyrin (TSPP) with γ-cyclodextrin (γ-CD) and phenylalanine (or tryptophan) have been examined by means of absorption and fluorescence spectroscopy. A 1:1 inclusion complex is formed between TSPP and γ-CD. TSPP forms 1:2 complexes with l- and d-phenylalanine (LPhe and DPhe), while TSPP forms 1:1 complexes with l- and d-tryptophan (LTrp and DTrp). In TSPP solution containing γ-CD and LPhe (or DPhe), the 1:1:2 γ-CD–TSPP–LPhe (or –DPhe) inclusion complex is formed, while the 1:1:1 γ-CD–TSPP–LTrp (or –DTrp) inclusion complex is formed for LTrp (or DTrp). The equilibrium constants for the formation of the complexes have been evaluated from the fluorescence intensity change of TSPP. The equilibrium constants are nearly the same for the optical isomers of phenylalanine and tryptophan, respectively, indicating that the optical isomers are not discriminated through the complexation. For the LPhe complexes, the equilibrium constants have also been evaluated at a fixed ionic strength of 0.2 mol dm^?3 using NaCl. 3-Phenyl-1-propanol and l-phenylalaninol, which are analogous to phenylalanine in molecular structure, have been examined for the complexation with TSPP and γ-CD. In contrast to phenylalanine, the stoichiometries of their binary complexes with TSPP and their ternary inclusion complexes with γ-CD and TSPP are 1:1 and 1:1:1, respectively.     

平行因子算法用于酪氨酸、色氨酸和苯丙氨酸的同时定性与定量测定

酪氨酸、色氨酸和苯丙氨酸是天然氨基酸中仅有的会发光的组分。本文将三维荧光分析法与平行因子（PARAFAC）算法相结合，在激发波长为205-290nm(5nm为间隔），发射波长为270-380nm（5nm为间隔）对该体系进行了分辨研究，分辨结果与实际浓度一致。该方法分辨速度快，易于编程实现，且分辨效率高，解决了三者同时分辨难的问题，进一步说明了化学计量学在生物化学中具有广阔的应用前景。     

Polarized Raman spectra and intensities of aromatic amino acids phenylalanine,tyrosine and tryptophan

The prominent marker bands in the Raman spectra of the aromatic proteinogenic amino acids phenylalanine, tryptophan and tyrosine have been reinvestigated. Previous studies have been extended by measuring intensities against NaClO₄ as an external standard. Raman spectra were divided into isotropic (trace scattering of symmetric vibrations) and anisotropic (quadrupole scattering of antisymmetric or symmetric vibrations). These intensity and polarization properties of the marker bands were followed through pH changes from about 11–13 to 1–2.     

Characterization of transient species in laser photolysis of aromatic amino acids using acetone as photosensitizer

The photochemical processes of aromatic amino acids were investigated in aqueous solution using acetone as photosensitizer by KrF (248 nm) laser flash photolysis. Laser-induced transient species were characterized according to kinetic analysis and quenching experiments. The intermediates recorded were assigned to the excited triplet state of tryptophan, the radicals of tryptophan and tyrosine. The excited triplet state of tryptophan produced via a triplet-triplet excitation transfer and the radicals arising from electron transfer reaction has been identified. Neither electron transfer nor energy transfer between triplet acetone and phenylalanine can occur in photolysis of phenylalanine aqueous solution which contains acetone. Furthermore, triplet acetone-induced radical transformation: Trp/N-Tyr→Trp-Tyr/O was observed directly in photolysis of dipeptide (Trp-Tyr) aqueous solution containing acetone, and the transformation resulting from intramolecular electron transfer was suggested.     

Sorption of Amino Acids from Aqueous Solutions by a Polarized Carbon Adsorbent

The adsorption and electrosorption of tyrosine, tryptophan, and phenylalanine on an A-4 model carbon adsorbent from 0.1 N K₂SO₄solutions were investigated at various acidity values of the medium. It was found that the sorption of tyrosine and tryptophan is significantly affected by both the polarization potential of the adsorbent and pH of the solution. For phenylalanine, electrosorption dependences were not measured because of its electrochemical instability under experimental conditions. The electrosorption effect was demonstrated for the low-molecular-weight protein, pepsin.     

Determination of the enantiomeric purity of levodopa, methyldopa, carbidopa and tryptophan by use of chiral mobile phase high-performance liquid chromatography

Chiral mobile phase high-performance liquid chromatography was used successfully for the determination of the enantiomeric purity of levodopa, methyldopa, carbidopa and tryptophan. The method investigated uses phenylalanine and copper sulfate in the mobile phase and a C18 column. Linearity, precision, accuracy, detection limit and interference from expected impurities were assessed. The method is also applicable to the measurement of enantiomeric purity in levodopa tablets and capsules.     

Study on the interaction mechanism between aromatic amino acids and quercetin

In this paper, we selected quercetin and aromatic amino acids (tryptophan, tyrosine, phenylalanine) as the research objects to investigate the change rules in the reaction process. The thermodynamic functions (Ka, ΔG, and ΔS) of the interactions between quercetin and aromatic amino acids (tryptophan, tyrosine, phenylalanine) were measured by isothermal titration calorimetry. The values of binding constant (Ka) reached maximum at 25°C; the entropies and Gibbs free energies were both negative at different temperatures. The kinetic parameters of quercetin and amino acids in the interaction process was determined by microcalorimetry. The results inferred that the driving force of the reaction was hydrogen bond or van der Waals force.     

Local structure of beta-hairpin isotopomers by FTIR, 2D IR, and ab initio theory

The 12-residue tryptophan zipper beta-hairpin (SWTWENGKWTWK) and two (13)C-isotopomers were examined in the amide-I region using FTIR and femtosecond two-dimensional infrared (2D IR) spectroscopies. Spectroscopic features of the labeled transitions with (13)C-substituted amide unit present in the terminal or turn region of the hairpin, including their frequency shifts and distributions, line broadenings, orientations, and anharmonicities of diagonal peaks, allow the peptide local structure and local environment to be examined. The results suggest a larger structure fluctuation in the terminal region than in the turn region as a result of the side chain effect and solvent-peptide interaction. The results also suggest that the uncoupled amide-I modes are not degenerate and that this is likely to be a common situation for solvated polypeptides. In addition, the amide-I states in the terminal and turn regions were found to be delocalized over several neighboring amide units. Cross-peaks between the various labeled and unlabeled structural regions were clearly observed in the 2D IR correlation spectra, allowing them to be characterized for monitoring structural changes. These results illustrate the sensitivity of 2D IR to the local environment of solvated peptides. The simulated 1D and 2D IR spectra of the hairpin, obtained by using the vibrational exciton model incorporating coupling constants from quantum chemical computations and semiempirical calculations, were found to reproduce the essential features of the experimental results.     

Efficient Oligomerization of Aromatic Amino Acids Induced by Gaps in Four-Helix Bundles of DNA or RNA

The formation of peptides from amino acids is one of the processes associated with life. Because of the dominant role of translation in extant biology, peptide-forming processes that are RNA induced are of particular interest. We have previously reported the formation of phosphoramidate-linked peptido RNAs as the products of spontaneous condensation reactions between ribonucleotides and free amino acids in aqueous solution. We now asked whether four-helix bundle (4HB) DNA or RNA folding motifs with a single- or double-nucleotide gap next to a 5’-phosphate can act as reaction sites for phosphoramidate formation. For glycine, this was found to be the case, whereas phenylalanine and tryptophan showed accelerated formation of peptides without a covalent link to the nucleic acid. Free peptides with up to 11 tryptophan or phenylalanine residues were found in precipitates forming in the presence of gap-containing DNA or RNA 4HBs. Control experiments using motifs with just a nick or primer alone did not have the same effect. Because folded structures with a gap in a double helix are likely products of hybridization of strands formed in statistically controlled oligomerization reactions, our results are interesting in the context of prebiotic scenarios. Independent of a putative role in evolution, our findings suggest that for some aromatic amino acids an RNA-induced pathway for oligomerization exists that does not have a discernable link to translation.