C6‑HSL enhances BPA tolerance in Pseudomonas asiatica: an integrative Raman‑DIP and transcriptomic study

Bisphenol A (BPA) and p-nitrophenol (PNP) are emerging contaminants of soils due to their wide presence in agricultural and industrial products. Thus, the present study aimed to integrate morpho-physiological, ionic homeostasis, and defense- and antioxidant-related genes in the response of tomato plants to BPA or PNP stress, an area of research that has been scarcely studied. In this work, increasing the levels of BPA and PNP in the soil intensified their drastic effects on the biomass and photosynthetic pigments of tomato plants. Moreover, BPA and PNP induced osmotic stress on tomato plants by reducing soluble sugars and soluble proteins relative to control. The soil contamination with BPA and PNP treatments caused a decline in the levels of macro- and micro-elements in the foliar tissues of tomatoes while simultaneously increasing the contents of non-essential micronutrients. The Fourier transform infrared analysis of the active components in tomato leaves revealed that BPA influenced the presence of certain functional groups, resulting in the absence of some functional groups, while on PNP treatment, there was a shift observed in certain functional groups compared to the control. At the molecular level, BPA and PNP induced an increase in the gene expression of polyphenol oxidase and peroxidase, with the exception of POD gene expression under BPA stress. The expression of the thaumatin-like protein gene increased at the highest level of PNP and a moderate level of BPA without any significant effect of both pollutants on the expression of the tubulin (TUB) gene. The comprehensive analysis of biochemical responses in tomato plants subjected to BPA and PNP stress illustrates valuable insights into the mechanisms underlying tolerance to these pollutants.

Catalyst: Is the Amino Acid a New Frontier for Biorefineries?

Hypoxia impairs brain function by incompletely defined mechanisms. Mild hypoxia, which impairs memory and judgment, decreases acetylcholine (ACh) synthesis, but not the levels of ATP or the adenylate energy charge. However, the effects of mild hypoxia on the synthesis of the glucose-derived amino acids [alanine, aspartate, gamma-amino butyric acid (GABA), glutamate, glutamine, and serine] have not been characterized. Thus, we examined the incorporation of [U-14C]glucose into these amino acids and ACh during anemic hypoxia (injection of NaNO2), hypoxic hypoxia (15 or 10% O2), and hypoxic hypoxia plus hypercarbia (15 or 10% O2 with 5% CO2). In general, the synthesis of the amino acids and of ACh declined in parallel with each type of hypoxia we studied. For example, anemic hypoxia (75 mg/kg of NaNO2) decreased the incorporation of [U-14C]glucose into the amino acids and into ACh similarly. [Percent inhibition: ACh (57.4), alanine (34.4), aspartate (49.2), GABA (61.9), glutamine (59.2), glutamate (51.0), and serine (36.7)]. A comparison of several levels (37.5, 75, 150, 225 mg/kg of NaNO2) of anemic hypoxia showed a parallel decreased in the flux of glucose into ACh and into the amino acids whose synthesis depends on mitochondrial oxidation: GABA (r = 0.98), glutamate (r = 0.99), aspartate (r = 0.96), and glutamine (r = 0.97). The synthesis of the amino acids not dependent on mitochondrial oxidation did not correlate as well with changes in ACh metabolism: serine (r = 0.68) and alanine (r = 0.76). The decreases in glucose incorporation into ACh and into the amino acids with hypoxic hypoxia (15% or 10% O2) or hypoxic hypoxia with 5% CO2 were very similar to those with the two lowest levels of anemic hypoxic. Thus, and explanation of the brain's sensitivity to a decrease in oxygen availability must include the alterations in the metabolism of the amino acid neurotransmitters as well as ACh.

Synthesis of non-canonical branched-chain amino acids in Escherichia coli and approaches to avoid their incorporation into recombinant proteins

Effects of Bisphenol A on expression of genes related to amino acid transporters, insulin- like growth factor, aquaporin and amino acid release by porcine trophectoderm cells

SummaryA study has been made of the assimilation of ammonium salts by young barley seedlings grown under controlled conditions in water culture. The synthesis of amino acids, amides and proteins in the roots was traced by means of isotopic labelling with 15N. Chromatographic analysis on ion‐exchange resins was used to separate the free and combined amino acids of the root tissues.There was evidence of the synthesis of proteins in the roots under the experimental conditions. After short periods of assimilation, the soluble amides and amino acids, especially glutamine and glutamic acid, were highly labelled with 15N, indicating a rapid synthesis of these constituents. Subsequently, the abundance of 15N in the amino acids combined in the root proteins showed a marked increase. The data are consistent with the occurrence of an extensive synthesis and interconversion of amino acids and their utilization as precursors in the synthesis of proteins.The importance of glutamic acid and glutamine in the assimilation of nitrogen and synthesis of amino acids is shown by the rapid incorporation of 15N into both the amide and amino groups. Other amino acids are more slowly labelled and may be formed secondarily by transamination or interconversion. γ‐Aminobutyric acid appears to play a significant part in the metabolism of the roots, since it is formed at an early stage of assimilation. The labelling of γ‐aminobutyric acid with 15N indicates that, in barley seedlings, it is a secondary product, rather than a precursor of glutamic acid or glutamine.

The influence of different N sources on fermentation rate and de novo amino acid synthesis by rumen micro-organisms was investigated in vitro using rumen fluid taken from four sheep receiving a mixed diet comprising (g/kg DM): grass hay 500, barley 299.5, molasses 100, fish meal 91, minerals and vitamins 9.5. Pancreatic casein hydrolysate (P; comprising mainly peptides with some free amino acids; 10 g/l), free amino acids (AA; casein acid hydrolysate + added cysteine and tryptophan; 10 g/l), or a mixture of L-proline, glycine, L-valine and L-threonine (M; 0.83 g/l each) were added to diluted (1:3, v/v), strained rumen fluid along with 15NH4Cl (A; 1.33 g/l) and 6.7 g/l of a mixture of starch, cellobiose and xylose (1:1:1, by weight). P and AA, but not M, stimulated net gas production after 4 and 8 h incubation (P < 0.05) in comparison with A alone. P increased microbial-protein synthesis (P < 0.05) compared with the other treatments. All of the microbial-N formed after 10 h was synthesized de novo from 15NH3 in treatment A, and the addition of pre-formed amino acids decreased the proportion to 0.37, 0.55, and 0.86 for P, AA, and M respectively. De novo synthesis of amino acids (0.29, 0.42 and 0.69 respectively) was lower than cell-N. Enrichment of alanine, glutamate and aspartate was slightly higher than that of other amino acids, while enrichment in proline was much lower, such that 0.83-0.95 of all proline incorporated into particulate matter was derived from pre-formed proline. Glycine, methionine, lysine, valine and threonine tended to be less enriched than other amino acids. The form in which the amino acids were supplied, as P or AA, had little influence on the pattern of de novo synthesis. When the concentration of peptides was decreased, the proportion of microbial-N formed from NH3 increased, so that at an initial concentration of 1 g peptides/l, similar to the highest reported ruminal peptide concentrations, 0.68 of cell-N was formed from NH3. Decreasing the NH3 concentration at 1.0 g peptides/l caused proportionate decreases in the fraction of cell-N derived from NH3, from 0.81 at 0.53 g NH3-N/l to 0.40 at 0.19 g NH3-N/l. It was concluded that different individual amino acids are synthesized de novo to different extents by mixed rumen micro-organisms when pre-formed amino acids are present, and that the source of N used for synthesis of cell-N and amino acids depends on the respective concentrations of the different N sources available; however, supplementing only with amino acids whose synthesis is lowest when pre-formed amino acids are present does not stimulate fermentation or microbial growth.

Metabolite profiles and activities of key enzymes in the metabolism of organic acids, nitrogen and amino acids were compared between chlorotic leaves and normal leaves of 'Honeycrisp' apple to understand how accumulation of non-structural carbohydrates affects the metabolism of organic acids, nitrogen and amino acids. Excessive accumulation of non-structural carbohydrates and much lower CO(2) assimilation were found in chlorotic leaves than in normal leaves, confirming feedback inhibition of photosynthesis in chlorotic leaves. Dark respiration and activities of several key enzymes in glycolysis and tricarboxylic acid (TCA) cycle, ATP-phosphofructokinase, pyruvate kinase, citrate synthase, aconitase and isocitrate dehydrogenase were significantly higher in chlorotic leaves than in normal leaves. However, concentrations of most organic acids including phosphoenolpyruvate (PEP), pyruvate, oxaloacetate, 2-oxoglutarate, malate and fumarate, and activities of key enzymes involved in the anapleurotic pathway including PEP carboxylase, NAD-malate dehydrogenase and NAD-malic enzyme were significantly lower in chlorotic leaves than in normal leaves. Concentrations of soluble proteins and most free amino acids were significantly lower in chlorotic leaves than in normal leaves. Activities of key enzymes in nitrogen assimilation and amino acid synthesis, including nitrate reductase, glutamine synthetase, ferredoxin and NADH-dependent glutamate synthase, and glutamate pyruvate transaminase were significantly lower in chlorotic leaves than in normal leaves. It was concluded that, in response to excessive accumulation of non-structural carbohydrates, glycolysis and TCA cycle were up-regulated to "consume" the excess carbon available, whereas the anapleurotic pathway, nitrogen assimilation and amino acid synthesis were down-regulated to reduce the overall rate of amino acid and protein synthesis.

Effect of induced rumination on cortisol habituation to repeated acute stress

Dynamics of the photosynthesis of carbon compounds II. Amino acid synthesis

The metabolism of acetate was investigated in the nerve-electroplaque system of Torpedo marmorata. In intact fragments of electric organ, radiolabeled acetate was incorporated into acetylcholine (ACh), acetylcarnitine (ACar), and three amino acids: aspartate, glutamate, and glutamine. These compounds were identified by TLC, high-voltage electrophoresis, column chromatography, and enzymic tests. The system responsible for acetate transport and incorporation into ACh displayed a higher affinity but a lower Vmax than that involved in the synthesis of ACar and amino acids. Choline, when added to the medium, increased the rate of acetate incorporation into ACh but decreased (at concentrations greater than 10(-5) M) that into ACar and amino acids. Monofluoroacetate slightly depressed ACh and ACar synthesis from external acetate but inhibited much more the synthesis of amino acids. During repetitive nerve stimulation, the level of the newly synthetized [14C]ACh was found to oscillate together with that of endogenous ACh, but the level of neither [14C]ACar nor the 14C-labeled amino acids exhibited any significant change as a function of time. This means that there is probably no periodic transfer of acetyl groups between ACh and the investigated metabolites in the course of activity. Acetate metabolism was also tested in the electric lobe (which contains the cell bodies of the neurons innervating the electric organ) and in Torpedo synaptosomes (which are nerve terminals isolated from the same neurons). Radioactive pyruvate and glutamine were also assayed in some experiments for comparison with acetate. These observations are discussed in connection with ACh metabolism under resting and active conditions in tissues where acetate is the preferred precursor of the neurotransmitter.

Dehydroalanine modification sees the light: a photochemical conjugate addition strategy

Metabolic and proteomic mechanism of bisphenol A degradation by Bacillus thuringiensis

Synthetic amino acids have particular appeal to extend our understanding of protein structure and stabilization into the realm of folded, non biological polymers. Chemists are now beginning to design biomimetic amino acids that can form both secondary and tertiary structures. An overview of the synthesis of non-protein amino acids recently used in the design of conformationally well-defined foldamers—artificial oligomers that also adopt well-defined folded conformations— are discussed.

Peptides exhibit properties based on the amino acids present in the peptide and structure the peptide assumes in its environment. For example, peptides have been shown to be antimicrobial when they are amphipathic and in an $\alpha$-helical shape. The mechanism for this antimicrobial activity is subject to dispute. One theory states that the peptides work by forming a "pore" through the cell membrane which causes a depolarization of the membrane and the cell lysis because of osmosis. Another theory states that the peptides merely compromise the integrity of the membrane by solubilizing the membrane. In order for the "pore" theory of lytic activity to occur, the peptides must arrange themselves in a discrete aggregated fashion. The goal of this research is to explore the nature of the aggregated state of designed peptides through the use of CD spectroscopy and fluorescent labeling. Several peptides were designed using a minimalist approach and were measured using CD spectroscopy to explore how the placement of leucines in the peptides cause a change in their propensity for aggregation. Using this information, the site for an incorporation of a fluorescently labeled peptide was chosen. The synthesis of the fluorescently labeled amino acid presented an opportunity to explore different types of glycine templates and to find a quicker method for making the amino acid in high yields. After the synthesis of the amino acid, it was incorporated into the peptides and CD and fluorescence studies were accomplished. The result shows the peptides, when aggregated, align themselves in an antiparallel fashion.