Determination of isokinetic ratios necessary for equimolar incorporation of carboxylic acids in the solid-phase synthesis of mixture-based combinatorial libraries

The methods used to study the relative reaction rates of 45 different aliphatic and aromatic carboxylic acids when coupled to resin-bound amino acid amides is described. Competition experiments involving the coupling of incoming carboxylic acids to resin-bound amino acid amides were performed. The relative composition of each N-acylated amino acid amide in the resulting mixtures was compared to controls prepared by physically mixing equal aliquots of individual compounds in order to study the relative reaction rates of the incoming carboxylic acids. The ratios of the incoming carboxylic acids were then iteratively adjusted to yield as close to equimolar products as possible. As expected, the steric and electronic nature of the incoming carboxylic acids was found to influence their relative reaction rates. The steric hindrance of the resin-bound amino acid appears to have a proportional effect on the reaction rates of the incoming carboxylic acids. N-acylated amino acid amides in the final mixtures, prepared using the final isokinetic ratios, were found to be approximately equimolar.     

Metabolites ofStreptomyces noursei

Branched chain amino acids are degraded byStreptomyces noursei to the corresponding amides: valine being the precursor of isobutyramide, leucine of isovaleramide and isoleucine of anteisovaleramide. The formation of amides is pyridoxalphosphate dependent and is catalyzed by peroxidase. Furthermore it was found that branched chain amino acids were also degraded via the corresponding 2-oxoacids into acylCoA which can be dehydrogenated to unsaturated acylCoA or hydrolyzed to acids and CoASH.     

Differential effect of amino acid residues on the stability of double helices formed from polyribonucleotides and its possible relation to the evolution of the genetic code

The interaction of amino acid residues with polyribonucleotides was characterized by measurements of melting temperatures (tm) for poly(A).poly(U) and poly(I).poly(C) as functions of the concentrations of various amino acid amides. The amides of hydrophilic amino acids lead to a continuous increase of tm with increasing concentration, whereas amides of hydrophobic amino acids induce a decrease of tm at low concentrations (approximately 1 mM) followed by an increase at higher concentrations. Analysis of the data by a simple site model provides the affinity of each ligand for the double helix relative to that for the single strands. This parameter decreases in the order Ala greater than Gly greater than Ser greater than Asn greater than Pro greater than Met, Val greater than Ile, Leu for poly(A).poly(U) and Ala, Gly, Ser greater than Asn greater than Pro greater than Val greater than Ile, Met, Leu for poly(I).poly(C). The special effects of hydrophobic amino acids may be related to the similarity of the codons for these amino acids. A simple model for assignment of codons to amino acids is proposed.     

EFFECTS OF ENVIRONMENTAL FACTORS ON PHOTOSYNTHESIS PATTERNS IN PHAEODACTYLUM TRICORNUTUM (BACILLARIOPHYCEAE). I. EFFECT OF NITROGEN DEFICIENCY AND LIGHT INTENSITY1

Cultures of the marine diatom Phaeodactylum tricornutum Bohlin incorporated, a large proportion of the total fixed carbon (50% or more) into amino acids and amides during short periods of photo-assimilation of ¹⁴C-labelled carbon dioxide. Although increasing nitrogen limitation in a nitrate-limited chemostat had little significant effect on the proportion of C incorporated into amino acids and amides combined, it did affect the distribution of radioactivity within individual compounds of this group. In particular, increasing degrees of N deficiency reduced the proportion incorporated into amides to almost undetectable levels, reduced the proportion in alanine and increased the proportion in glutamic acid. Also, increasing N limitation decreased the relative synthesis of sugar phosphates and increased the proportion of C assimilated into intermediates of the tricarboxylic acid cycle. Reduced light intensity did not have any significant effect on the proportion of C incorporated into the total amino acids and amides, but did cause a decrease in the radioactivity     

Nutrient status effects on loss of amides and amino acids from pine roots

Summary Loss of amides and amino acids from pine roots was two and a half times greater from roots grown in phosphate-deficient nutrient solution than from those grown in complete nutrient solution and ten times greater than from plants grown in nitrogen-deficient solution. The major factor in increased loss from phosphate-deficient plants was an increase in free amides and amino acids in these plants. Increased amide and amino acid loss from phosphate-deficient plants may have important consequences in micro-organism—plant interactions.     

Differential effect of amino acid residues on the stability of double helices formed from polyribonucleotides and its possible relation to the evolution of the genetic code

Summary The interaction of amino acid residues with polyribonucleotides was characterized by measurements of melting temperatures (t_m) for poly(A) poly(U) and poly(I)poly(C) as functions of the concentrations of various amino acid amides. The amides of hydrophilic amino acids lead to a continuous increase of tm with increasing concentration, whereas amides of hydrophobic amino acids induce a decrease of t_m at low concentrations (1 mM) followed by an increase at higher concentrations. Analysis of the data by a simple site model provides the affinity of each ligand for the double helix relative to that for the single strands. This parameter decreases in the order Ala>Gly>Ser>Asn>Pro>Met, Val>Ile, Leu for poly(A) poly(U) and Ala, Gly, Ser>Asn>Pro>Val>Ile, Met, Leu for poly(I)poly(C). The special effects of hydrophobic amino acids may be related to the similarity of the codons for these amino acids. A simple model for assignment of codons to amino acids is proposed.     

Tissue specificity of assimilation and transport of nitrogen in the root ofZea mays L. seedlings. III. Interaction of radial and longitudinal transport

Roots obtained from 4-day maize seedlings were allowed to absorb uniformly labelled¹⁴C-suorose only with their middle part while the apical and basal parts did not contact the medium. The amino acids and their amides produced in the cortex at the expense of nitrates and the¹⁴C-sucrose carbon moved radially to the stele where they were selectively distributed in the polar directions of transport. “Primary” amino acids moved predominantly in the apical direction while amides and basic amino acids were transported mainly in the opposite direction. When the nutrient medium was devoid of nitrogen sources, the amino acids entering the stele from the cortex were transported mostly to the root tip, indicating a significant role of young tissues in consuming organic nitrogen compounds when they are in short supply in the root.     

Variations in the content of free and bound amino acids during dormancy and the first cell cycle in Helianthus tuberosus tuber explants

Abstract

Qualitative and quantitative analysis of free and bound amino acids and amides during dormancy and the most important phases of the first cell cycle was carried out in tubers of Helianthus tuberosus.

In the dormant tuber arginine was confirmed to be the most abundant amino acid. A high amount of asparagine was also present; on the contrary glutamine was found in very low concentrations. During the progression of dormancy, all the free amino acids and amides declined while aspartic and glutamic acid increased.

During the G₁ phase of the first cell cycle induced by 2,4-D, all the free amino acids and amides decreased with the exception of glutamic acid.

At 18, 20, 24 h of activation with 2,4-D, corresponding to the S phase and the beginning of mitosis, bound amino acids were also determined. In these phases of the cell cycle they increased reaching a maximum at 20 h; on the other hand the free amino acid and amide content, especially aspartic acid, asparagine and arginine, decreased with the exception of glutamic acid, alanine and phenylalanine.     

Ontogenetic Changes in the Alcohol-soluble Amino Acid Fraction of the Grain, Leaves and Roots of Triticum aestivum (var. Rideau) Following Vernalization and Seedling Growth

Changes in the free alcohol-soluble amino acid fraction of theseparate organs of Rideau wheat (Triticum aestivum L.) preconditionedby vernalization were determined after 1 and 3 weeks' growthand compared with comparable organs of non-vernalized controlplants. The major components of the fraction were composed of the acidicand neutral amino acids together with asparagine and glutamine.Cystine and methionine were rarely present in more than traceamounts. Changes coincident with ontogency and growth varied considerablyboth between all seedling parts and between comparable organsof the control and vernalized series. Generally, vernalizationwas paralleled by higher levels of the acidic amino acids andthe amides. Coleoptile senescence was characterized by depletionof the total fraction but individual amino acids increased withageing. The pattern of change varied with ontogeny, age andprior grain treatment. However, within each category significantvariations in the level of one or more amino acids could berelated to the physiological status of the tissue. The amides,glutamine and asparagine were consistently absent from the stemapex and the very young fifth and sixth leaves. In stem apextissue, 17 of the commonly occurring amino acids were generallyfound present while, in coleoptiles and young leaves only eightamino acids were present.     

Stability decrease of RNA double helices by phenylalanine-, tyrosine- and tryptophane-amides. Analysis in terms of site binding and relation to melting proteins.

The amides of L-phenylalanine, L-tyrosine and L-tryptophane decrease the melting temperatures tm of poly(A)*poly(U) and poly(I)*poly(C) double helices at low concentrations (1 mM), whereas high concentrations finally lead to an increase of tm. This dependence of the tm-values upon the ligand concentration can be represented quantitatively by a simple site binding model, providing binding parameters for the interaction between the amides and the nucleic acids both in the double- and the single-stranded conformation. According to these data the affinity to the single strands is higher than that to the double strands and increases in the series Phe less than Tyr less than Trp. The binding constants decrease with increasing salt concentration as expected for an interaction driven by electrostatic attraction. However, part of the interaction is also due to stacking between the aromatic amides and the nucleic acid bases. The present results indicate a direct correlation between the presence of aromatic amino acids at the binding site of helix destabilising proteins and the properties of simple derivatives of these amino acids. Furthermore the results suggest that very simple peptides containing aromatic amino acids served as a starting point for the evolution of helix destabilising proteins.     

The mechanism of action of dipeptidyl aminopeptidase. Inhibition by amino acid derivatives and amines; activation by aromatic compounds.

A variety of amino acid and peptide amides have been shown to be inhibitors of dipeptidyl aminopeptidase. Among these compounds derivatives of strongly hydrophobic amino acids are the strongest inhibitors (Phe-NH2, Ki = 1.0 +/- 0.2 mM), while amides of basic amino acids were somewhat less effective (Lys-NH2, Ki = 36 +/- 3 mM). Short chain amino acid amides are notably weaker inhibitors (Gly-NH2, Ki = 293 +/- 50 mM). The interaction of the side chains of compounds with the enzyme appears to be at a site other than that at which the side chain of the amino-penultimate residue of the substrate interacts since the specificity of binding is different. Primary amines have been shown to inhibit, e.g., butylamine, Ki = 340 +/- 40 mM, and aromatic compounds have been shown to stimulate activity toward Gly-Gly-NH2 and Gly-Gly-OEt (phenol, 35% stimulation of activity at a 1:1 molar ratio with the substrate). The data suggest that inhibition involves binding at the site occupied by the free alpha-amino group and the N-terminal amino acid.     

Synthesis of cinnamoyl and hydroxycinnamoyl amino acid conjugates and evaluation of their antioxidant activity.

Fifteen amides of cinnamic, ferulic and sinapic acids with natural and unnatural C-protected amino acids have been synthesized. The amides (E)-N-(feruloyl)-L-tyrosine methyl ester (10), (E)-N-(feruloyl)-L-phenylalanine t-butyl ester (11), (E)-N-(sinapoyl)-L-tyrosine methyl ester (13) and (E)-N-(sinapoyl)-L-phenylalanine t-butyl ester (15) with a free carboxyl group of amino acids have been found in nature. The rest of the compounds are unknown. The hydroxycinnamoyl amino acid conjugates have been studied for their antioxidant activity (AOA) in bulk phase lipid autoxidation. The highest AOA has been found for the compounds 11 and 15, which contain the same phenylalanine moiety.     

SPPS of protected peptidyl aminoalkyl amides.

Monophthaloyl diamines derived from naturally occurring amino acids were attached through their free amino functions to resins of the trityl type. The phthaloyl groups were removed by hydrazinolysis, and peptide chains were assembled using Fmoc/tBu-amino acids on the liberated amino functions. The peptidyl aminoalkyl amides obtained were cleaved from the resins by mild acidolysis, with the tBu-side chain protection remaining intact.     

Formation and distribution of free amino acids in Cunninghamella elegans mycelium during growth on media with glucose and dodecane]

The growth, assimilation of organic nitrogen compounds, and production and distribution of free amino acids were studied during the growth of Cunninghamella elegans (--) 1204 on the defined medium 12 containing glucose and dodecane and without an additional carbon source (control). The culture utilized all leucine, glycine, and asparagine after three days of the growth, irrespective of the source of carbon in the medium. Glutamic acid was assimilated only in the presence of glucose. Some 15 free amino acids and 2 amides were detected in the cells, and also some correlations in the composition of the pool in the presence of different sources of carbon in the medium. The substitution of dodecane for glucose decelerates metabolism of free intracellular amino acids and stimulates their accumulation in the cells, especially if there is a deficiency of carbon sources: the overall content of amino acids and amides on the medium with glucose is from 2.2 to 4.1 mg/g; with dodecane, from 9.8 to 16.5 mg/g; in the control, from 16.4 to 25.8 mg/g.     

Changes in γ-Methyleneglutamic acid, γ-Methyleneglutamine and other amino acids and amides during fruit growth of Tribulus terrestris

Abstract

Distribution and metabolism of γ-methyleneglutamic acid, γ-methyleneglutamine and other amino acids and amides has been studied during fruit growth of Tribulus terrestris. The largest concentration of free amino acids and amides has been observed in fruit stage 1. The marked decline in the amount of γ-methyleneglutamic acid and γ-methyleneglutamine after fruit stage 1 may indicate their rapid utilization along with asparagine and glutamine during fruit growth. In leaf and in different fruit growth stages, γ-methyleneglutamic acid dominated over γ-methyleneglutamine.     

Thermochemistry of N-C bonds hydrolysis in amides,peptides, N-acetyl amino acids and high-energy N-C bonds hydrolysis in N-acetyl imidazole and urea

• 1.1. The reaction enthalpies of hydrolysis of amides, peptides and N-acetyl amino acids were calculated for both ionized and un-ionized forms of reaction components.
• 2.2. The average values of reaction enthalpies of amides, peptides and N-acetyl amino acids hydrolysis were essentially different from each other for ionized forms of reaction components and were equal for un-ionized forms of reaction components in the error interval.
• 3.3. As an example of high-energy N-C bonds N-acetyl imidazol and urea were discussed. It was found that the reaction enthalpies of hydrolysis of above compounds were different from analogous thermodynamic values of hydrolysis of amides, peptides and N-acetyl amino acids for any forms of components.

     

New enzymatic method of chiral amino acid synthesis by dynamic kinetic resolution of amino acid amides: use of stereoselective amino acid amidases in the presence of alpha-amino-epsilon-caprolactam racemase

D- and L-amino acids were produced from L- and D-amino acid amides by D-aminopeptidase from Ochrobactrum anthropi C1-38 and L-amino acid amidase from Pseudomonas azotoformans IAM 1603, respectively, in the presence of alpha-amino-epsilon-caprolactam racemase from Achromobacter obae as the catalyst by dynamic kinetic resolution of amino acid amides.     

Condensation of amino acids to form peptides in aqueous solution induced by the oxidation of sulfur(iv): An oxidative model for prebiotic peptide formation

Condensation of amino acids to peptides is an important step during the origin of life. However, up to now, successful explanations for plausible prebiotic peptide formation pathways have been limited. Here we report that the oxidation of sulfur (IV) can induce the condensation reaction of carboxylic acids and amines to form amides, and the condensation reaction of amino acids to form peptides. This might be a general reaction contributing to prebiotic peptide formation.     

Free Amino Acids in the Leaves of Salvinia natans and Azolla filiculoides Grown in Light and Dark

Free amino acids were determined quantitatively by thin-layer chromatography and identified by various chromatographic methods in the leaves of two floating water ferns. Special interest was paid to the amount of γ-amino butyric acid. Water soluble nitrogen compounds increase in the leaves of both plants during the growth of three to four weeks in darkness. Compounds especially accumulating are ammonia and the amides asparagine and glutamine. The amount of most proper amino acids is reduced in the dark, except γ-amino butyric acid, which, in addition to amides, is the main amino compound increasing in darkness. 0.1 M HCl extracts the greatest amount of nitrogen compounds of low molecular weight from the dried leaves of the experimental plants. If we denote the amount of the dissolved amino compounds as 100 per cent, we can say that water extracts 96 per cent of the amino compounds, and that an 80 per cent ethanol solution extracts 79 per cent of these compounds.