Effect Of Asparagine Research Articles

Effect of asparagine, corncob biochar and Fe(II) on anaerobic biological treatment under low temperature: Enhanced performance and microbial community dynamic.

To ensure the efficiency of anaerobic biological treatment technology at lower temperature will expand the application of anaerobic reactor in practical industrial wastewater treatment. Through a batch experiment, asparagine, corncob biochar and Fe2+ were selected as strengthening measures to analyze the effects on the anaerobic sludge characteristics, microbial community and functional genes in the low temperature (15°C). Results showed that after 21 days, asparagine began to promote chemical oxygen demand (COD) removal by the anaerobic treatment, with highest COD removal rate (81.65%) observed when the asparagine concentration was 1mmol/L. When adding 3g biochar, 25mg/L Fe2+, and the combination of biochar and Fe2+, the COD removal rates reached to 82%, 92% and 97%, respectively. In the presence of asparagine, both biochar and Fe2+ alone or in combination increased the activity of protease (16.35%-120.71%) and coenzyme F420 (5.63%-130.2%). The relative abundance of Proteobacteria and Methanobacterium increased in the presence of biochar and Fe2+. In addition, the KEGG results showed that the combined addition of biochar and Fe2+ enhanced bacterial replication and repair and promoted amino acid metabolism of archaea.

Fumarate hydratase-deficient renal cell carcinoma cells respond to asparagine by activation of the unfolded protein response and stimulation of the hexosamine biosynthetic pathway

BackgroundThe loss-of-function mutation of fumarate hydratase (FH) is a driver of hereditary leiomyomatosis and renal cell carcinoma (HLRCC). Fumarate accumulation results in activation of stress-related mechanisms leading to upregulation of cell survival-related genes. To better understand how cells compensate for the loss of FH in HLRCC, we determined the amino acid nutrient requirements of the FH-deficient UOK262 cell line (UOK262) and its FH-repleted control (UOK262WT).MethodsWe determined growth rates and survival of cell lines in response to amino acid depletion and supplementation. RNAseq was used to determine the transcription changes contingent on Asn and Gln supplementation, which was further followed with stable isotope resolved metabolomics (SIRM) using both [U- 13C,15N] Gln and Asn.ResultsWe found that Asn increased the growth rate of both cell lines in vitro. Gln, but not Asn, increased oxygen consumption rates and glycolytic reserve of both cell lines. Although Asn was taken up by the cells, there was little evidence of Asn-derived label in cellular metabolites, indicating that Asn was not catabolized. However, Asn strongly stimulated Gln labeling of uracil and precursors, uridine phosphates and hexosamine metabolites in the UOK262 cells and to a much lesser extent in the UOK262WT cells, indicating an activation of the hexosamine biosynthetic pathway (HBP) by Asn. Asn in combination with Gln, but not Asn or Gln alone, stimulated expression of genes associated with the endoplasmic reticulum (ER) stress and the unfolded protein response (UPR) in UOK262 to a greater extent than in FH-restored cells. The changes in expression of these genes were confirmed by RT-PCR, and the stimulation of the UPR was confirmed orthogonally by demonstration of an increase in spliced XBP1 (sXBP1) in UOK262 cells under these conditions. Asn exposure also increased both the RNA and protein expression of the HBP regulator GFPT2, which is a transcriptional target of sXBP1.ConclusionsAsn in the presence of Gln induces an ER stress response in FH-deficient UOK262 cells and stimulates increased synthesis of UDP-acetyl glycans indicative of HBP activity. These data demonstrate a novel effect of asparagine on cellular metabolism in FH-deficient cells that could be exploited therapeutically.

OsASN1 Plays a Critical Role in Asparagine-Dependent Rice Development.

Asparagine is one of the important amino acids for long-distance transport of nitrogen (N) in plants. However, little is known about the effect of asparagine on plant development, especially in crops. Here, a new T-DNA insertion mutant, asparagine synthetase 1 (asn1), was isolated and showed a different plant height, root length, and tiller number compared with wild type (WT). In asn1, the amount of asparagine decreased sharply while the total nitrogen (N) absorption was not influenced. In later stages, asn1 showed reduced tiller number, which resulted in suppressed tiller bud outgrowth. The relative expression of many genes involved in the asparagine metabolic pathways declined in accordance with the decreased amino acid concentration. The CRISPR/Cas9 mutant lines of OsASN1 showed similar phenotype with asn1. These results suggest that OsASN1 is involved in the regulation of rice development and is specific for tiller outgrowth.

Effect of asparagine, cysteine, citrulline, and glutamine on in vitro rooting and biochemical constituents in cherry rootstocks

Effects of four amino acids, L-asparagine, L-cysteine, L-citrulline, and L-glutamine in different concentrations (0, 0.5, 1, and 2 mg dm-3) combined with 2 mg dm-3 indole-3-butyric acid, on in vitro rooting and biochemical constituents of cherry rootstocks CAB-6P (Prunus cerasus L.) and Gisela 6 (P. canescens × P. cerasus) were investigated. In CAB-6P, root number and root fresh mass (FM) were maximum at 0.5 mg dm-3 cysteine. All amino acids reduced root length in CAB-6P and root number as well as root FM in Gisela 6. In Gisela 6, 0.5 mg dm-3 asparagine or 2 mg dm-3 glutamine reduced root length. In CAB-6P, 100 % rooting was achieved in the control and with 1 and 2 mg dm-3 cysteine or 1 mg dm−3 citrulline. In Gisela 6, the rooting percentage was maximum (76.92 %) with 0.5 mg dm−3 asparagine. Callus FM in CAB-6P was the greatest at 1 mg dm−3 and in Gisela 6 at 2 mg dm−3 citrulline. Callusing was 100 % in the majority of treatments for CAB-6P and 92.31 % for Gisela 6 with 0.5 or 2 mg dm−3 citrulline. Cysteine, citrulline, and glutamine diminished chlorophyll content in Gisela 6 whereas in CAB-6P all four amino acids hardly affected it. Carotenoid and porphyrin content in CAB-6P was decreased due to asparagine (0.5 or 1 mg dm−3). Porphyrin content in CAB-6P was also reduced by adding 0.5 or 1 mg dm−3 cysteine or 2 mg dm−3 citrulline. In Gisela 6, all amino acids decreased carotenoid and porphyrin content. In CAB-6P, all treatments except 0.5 mg dm−3 glutamine or 2 mg dm−3 asparagine increased leaf sucrose content. In roots, both sucrose and proline content were increased only at 1 mg dm−3 cysteine whereas in leaves only 0.5 mg dm−3 asparagine caused a 3-fold increase in proline content. A decrease in root proline in CAB-6P was observed due to asparagine, citrulline, or glutamine. In Gisela 6, decreased leaf sucrose and proline content was recorded at 2 mg dm−3 cysteine. All amino acids did not alter root sugar content remarkably whereas root proline content was raised by adding 0.5 mg dm−3 glutamine or 1 mg dm−3 cysteine.

Micellization Behavior of a Cationic Gemini Surfactant, Pentanediyl-1,5-Bis(Dimethylcetylammonium Bromide): Effect of Asparagine and Temperature

Herein, we report the effect of amino acid and temperature on the micellization of a cationic gemini surfactant, pentanediyl-1,5-bis(dimethylcetylammonium bromide), 16-5-16. In aqueous solution, the critical micelle concentration (CMC) of 16-5-16 was measured by the conductivity method at different fixed temperatures and a fixed concentration of asparagine. An increasing trend of CMC values was found with both higher (asparagine) concentrations (i.e., increases with increasing asparagine concentration) and higher temperatures (i.e., increases with increasing temperature). The thermodynamics (viz. standard Gibbs energy (#x00029;, standard enthalpy (), and standard entropy ()) of micellization of the gemini surfactant (16-5-16) were evaluated, which indicated less stability of the 16-5-16 solution in the presence of amino acid (asparagine).

Specific nutrient supplementation of defined serum-free medium for the improvement of CHO cells growth and t-PA production

Recombinant CHO TF70R cells are able to grow and produce t-PA on serum-free medium BIOPRO1 (BioWhitaker Europe, Belgium). The purpose of the present study was to determine the effect of medium supplementation with vitamins, lipids, and specific amino acids on cell growth, t-PA production and biological functionality. Among vitamins, only biotin, folic acid, cobalamine and benzoic acid were required for improving growth and t-PA production. Lipid supplement allowed a significant increase cell concentration and t-PA specific activity and concentration, though its specific production rate decreased slightly. Medium supplementation with proline, serine and asparagine had also positive effects on cell growth. Besides, the addition of asparagine (even in the presence of glutamine) was essential for the production and biological quality of the t-PA. This systematic approach for media supplementation produced an increase in cell concentration around 100% and in t-PA production around 80%, with no detrimental effect on its biological activity. The effect of asparagine on t-PA production was unexpected and needs to be further studied. The above modifications of the production medium did not produce a significant effect on the metabolism of the main carbon and energy sources (glucose and glutamine) and the level of by-product formation (lactate and ammonia).

Glutamine transport in isolated epithelial intestinal cells. Identification of a Na+-dependent transport mechanism, highly specific for glutamine.

L-glutamine transport was evaluated in isolated cells from the guinea-pig small intestine by measuring [(3)H]- L-glutamine uptake. Villous and crypt cells expressed Na(+)-dependent and Na(+)-independent transport mechanisms. Glutamine transport systems were identified using various amino acids and analogues as inhibitors. In both villous and crypt cells, 2-(methylamino)-isobutyrate (MeAIB), a system A inhibitor, did not inhibit Na(+)-dependent glutamine influx. 2-Aminobicyclo(2,2,1)heptane-2-carboxylate (BCH), a system B(0) and B(0,+) substrate, had no effect on Na(+)-dependent influx. Serine, cysteine and threonine, system ASC inhibitors, reduced Na(+)-dependent influx by 50%. Asparagine, but not histidine, system N inhibitors, reduced Na(+)-dependent glutamine influx by 50%, however the effect of asparagine was not additive to that of threonine. The remaining Na(+)-dependent glutamine influx (50%) was only inhibited by glutamine itself, by Na(+) substitution ( N-methyl-glucamine, K(+), Li(+)) or by external pH reduction. Phenyl-acetyl-glutamine (PAG), a synthetic amino acid analogue, also inhibited this Na(+)-dependent, threonine-insensitive glutamine influx (IC(50) 2.45 mM). The Na(+)-independent uptake was partially inhibited by BCH, a system L inhibitor, and other neutral amino acids, but was not affect by PAG. Our results suggest that glutamine is transported in both villous and crypt cells by the Na(+)-independent system L, by the Na(+)-dependent system ASC and by an as yet undescribed Na(+)-dependent transport mechanism, highly specific for glutamine.

Interaction of asparagine and EGF in the regulation of ornithine decarboxylase in IEC-6 cells.

Our laboratory has shown that asparagine (ASN) stimulates both ornithine decarboxylase (ODC) activity and gene expression in an intestinal epithelial cell line (IEC-6). The effect of ASN is specific, and other A- and N-system amino acids are almost as effective as ASN when added alone. In the present study, epidermal growth factor (EGF) was unable to increase ODC activity in cells maintained in a salt-glucose solution (Earle's balanced salt solution). However, the addition of ASN (10 mM) in the presence of EGF (30 ng/ml) increased the activity of ODC 0.5- to 4-fold over that stimulated by ASN alone. EGF also showed induction of ODC with glutamine and alpha-aminoisobutyric acid, but ODC induction was maximum with ASN and EGF. Thus the mechanism of the interaction between ASN and EGF is important for understanding the regulation of ODC under physiological conditions. Therefore, we examined the expression of the ODC gene and those for several protooncogenes under the same conditions. Increased expression of the genes for c-Jun and c-Fos but not for ODC occurred with EGF alone. The addition of ASN did not further increase the expression of the protooncogenes, but the combination of EGF and ASN further increased the expression of ODC over that of ASN alone. Western analysis showed no significant difference in the level of ODC protein in Earle's balanced salt solution, ASN, EGF, or EGF plus ASN. Addition of cycloheximide during ASN and ASN plus EGF treatment completely inhibited ODC activity without affecting the level of ODC protein. These results indicated that 1) the increased expression of protooncogenes in response to EGF is independent of increases in ODC activity and 2) potentiation between EGF and ASN on ODC activity may not be due to increased gene transcription but to posttranslational regulation and the requirement of ongoing protein synthesis involving a specific factor dependent on ASN.

Translational control mechanism of ornithine decarboxylase by asparagine and putrescine in primary cultured hepatocytes

Asparagine stimulated the translation of ornithine decarboxylase (ODC) mRNA more than 10-fold in cultured hepatocytes which had been pretreated with glucagon in simple salt/glucose medium. Putrescine suppressed the increase in the rate of ODC synthesis caused by asparagine without significant change in the amount of ODC mRNA, suggesting that putrescine inhibited the effect of asparagine at least in part at the level of translation. Polysomal distribution of ODC mRNA was analyzed to examine the site of translational regulation by these effectors. In uninduced hepatocytes, most of the ODC mRNA was sedimented slightly after the 40 S ribosomal subunit. This ODC mRNA was sequestered from translational machinery since it was not shifted to the polysome fraction when peptide elongation was specifically inhibited by a low concentration of cycloheximide. In asparagine-treated cells, 40% of total ODC mRNA was in the polysomal fraction and formed heavier polysomes, indicating that asparagine stimulated both recruitment of ODC mRNA from the untranslatable pool and the initiation steps of translation. Putrescine did not change the distribution pattern of ODC mRNA on polysomes significantly. Thus, 30% of ODC mRNA remained on polysomes even when ODC synthesis was completely inhibited by putrescine. Paradoxically more than 70% of ODC mRNA was shifted into polysomes by putrescine in the presence of low concentrations of cycloheximide. These results, together with changes in the polysome profile, suggested that putrescine nonspecifically stimulated the recruitment of ODC mRNA from the untranslatable pool, whereas it specifically inhibited its translation at both the initiation and the elongation steps.

The effect of asparagine and adenine on the glutamine requirement for growth of human peripheral lymphocytes

Quantitative growth responses of lymphocytes directly isolated from individual subjects in a newly developed chemically-defined, protein-free medium are used to demonstrate that supplements of both L-asparagine and a purine source, but neither alone, significantly reduce the quantitative requirement for L-glutamine for growth. This system is useful for exploring individual differences in quantitative glutamine requirements and adequacy of asparagine and purine biosynthesis.

Cultured hepatoma cells for the study of enzyme regulation: induction of ornithine decarboxylase by insulin and asparagine.

The induction and decay of ornithine decarboxylase (ODC) by insulin and asparagine in cultures of H4-II-EC3 (H35) hepatoma cells was studied in a modified Waymouth medium in the presence of fetal bovine serum (FBS) and in serum-free media. The insulin response was enhanced by the presence of asparagine although the effect of asparagine was not so much on the initial increase as it was on a slowing of the decline after the maximum was reached at 6 to 8 h after the supplements were added together with fresh medium. In all cases the initial ODC activity was zero at zero time for addition of media and supplements, and, after reaching the maximum, activity declined to near zero by 24 h. Fetal bovine serum gave induction that followed a similar time course but was inferior to the combination of insulin plus asparagine and, in fact, FBS inhibited the latter response. Putrescine (the product formed from ornithine by ODC), at 10(-5) M, markedly inhibited the induction of ODC by insulin or FBS, but the inhibition was less when asparagine was present.

Metabolism of polyamines by cultured glioma cells. Effect of asparagine on γ-aminobutyric acid concentrations

The activity of ornithine decarboxylase (EC 4.1.1.17) increased in confluent cultures of glioma C6BU-1 cells 3 h after adding a complete serum-containing medium, and was maximal 5 h later. The activity of S-adenoxyl-L-methionine decarboxylase (EC 4.1.1.50) increased soon after addition of the complete medium to the cells, and reached its peak after 11 h. The activity of diamine oxidase (EC 1.4.3.6) also increased soon after adding complete medium and was maximal 8h later, when the activity of ornithine decarboxylase reached its peak. The increase in the activity of S-adenosyl-L-methionine decarboxylase was accompanied by changes in cellular spermidine and spermine concentrations, whereas the increase in the activity of diamine oxidase was followed by the accumulation of gamma-aminobutyric acid, which was detected both in the cells and in the medium. Asparagine enhanced the utilization of radioactive putrescine by glioma cells suspended in buffered-salt/glucose solution and increased intracellular and extracellular gamma-aminobutyric acid concentrations. Radioactive putrescine was converted into spermidine and spermine by glioma cells after addition of a serum-containing medium, but not after adding buffered--salt/glucose solutions, in the presence or absence of asparagine. The kinetics of ornithine decarboxylase 'induction' and the half-life of the enzyme differed in cells incubated with buffered asparagine solutions and serum-containing media.

Effect of asparagine and glutamine deficiency on normal and leukemic cells.

Normal and leukemic human blood cells and cells from mouse lymphoma 6C3HED were incubated in media that lacked L-asparagine or L-glutamine or both. Viable cell counts and the incorporation of radioactive L-leucine into cell proteins were measured before and after incubation. The absence of either L-asparagine or L-glutamine from the media produced a delayed cytocidal effect on 6C3HED cells and caused a decreased uptake of L-leuclne, Cells from patients with acute leukemia also required L-asparagine and Lglutamine for survival at 37° C for 2–5 days, but seemed less sensitive than the 6C3HED cells. L-Glutamine but not L-asparagine deficiency killed lymphocytes from 5 of 6 patients with chronic leukemia and inhibited the uptake of LL-leucine. L-Asparagine deficiency had no effect, and L-glutamine deficiency only minimal effect, on normal lymphocytes as measured by percent survival or L-leucine uptake. Transformation of normal lymphocytes by pokeweed mitogen was, however, completely inhibited by a deficiency of L-glutamine but not by a deficiency of L-asparagine.

Effect of asparagine on growth and protein synthesis in weanling rats.

Effect of asparagine on growth and protein synthesis in weanling rats.

トビイロウンカの吸汁におよぼすアスパラギンの効果

トビイロウンカの吸汁におよぼすアスパラギンの効果

The Effect of Asparagine on the Growth of a Gram-Positive Coccus

SUMMARY A Pediococcus sp., unusual in that it needed reducing conditions for growth, would not grow in a medium based on acid-hydrolysed casein but grew when asparagine was added. Neither glutamine nor aspartic acid + an ammonium salt satisfied this requirement. When the acid-hydrolysed casein was replaced by a mixture of amino acids, the omission of asparagine caused a decrease in growth in the presence of aspartic acid + an ammonium salt. No growth occurred in the absence of both aspartic acid and asparagine.