Effects of Various Amino Acids on Methionine-Induced Hyperhomocysteinemia in Rats

Rats were fed diets supplemented with 1% L-methionine with and without 2.5% various amino acids for 7 d to determine what amino acids other than glycine, serine, and cystine can suppress methionine-induced hyperhomocysteinemia. L-Glutamic acid, L-histidine, and L-arginine significantly suppressed methionine-induced enhancement of plasma homocysteine concentrations, but the mechanisms underlying the effect of these amino acids are thought not to be identical.     

Kinetics of luxury uptake of phosphate by algae-dominated benthic communities

The uptake of phosphate by benthic communities, dominated by living algae, previously exposed to different levels of external nutrient loading, exhibited first-order kinetics with respect to the intracellular P-deficit. This deficit is the difference between the maximum and the actual intracellular P-concentration.The maximum storage capacity of P per unit of dry weight was positively correlated to the level of external nutrient loading, whereas the phosphate uptake rate constant was negatively correlated.The observed internal P concentrations in the benthic layer of test ditches over a period of two and a half years, indicated a slight decrease towards a minimum value in a ditch with a low external P-input. In a medium loaded ditch the internal P-concentration did not change significantly. In a high loaded ditch increasing internal P-concentrations over time were observed, towards P-saturation of the benthic community.     

AMINO ACID AND UREA UPTAKE IN TEN SPECIES OF CHLOROPHYTA1

The Uptake of radioartively-labelled mixed amino acids, arginine, lysine, leucine, glutamic acid and urea was examined in six species of Volvocales and four species of Chlarococcales grown in nitrate-containing medium. Nonradioactive amino acids in excess were used to estimate specificity of amino and carriers in selected cases. All ten species possess salurable (hence, carrier-mediated) systems for uptake of both arginine and urea. In all Volvacales and one Chlorococcales, the arginine-speciftc carrier (which also transported lysine with lower efficiency) was the only amino acid carrier detected. Three species of Chlororoccales appear to possess a separate carrier for lysine and two of these appear to possess at least one additional carrier that is involved in uptake of non-basic amino acids.     

Mechanisms and specificity of amino acid transport in Taenia crassiceps larvae (Cestoda)

The absorption of lysine, arginine, phenylalanine and methionine by Taenia crassiceps larvae is linear with respect to time for at least 2 min. Arginine uptake occurs by a mediated system and diffusion, and arginine, lysine and ornithine (in order of decreasing affinity) are completely competitive inhibitors of arginine uptake. The basic amino acid transport system has a higher affinity for l-amino acids than d-amino acids, and blocking the α-amino group of an amino acid destroys its inhibitory action. Phenylalanine uptake by T. crassiceps larvae is inhibited in a completely competitive fashion by serine, leucine, alanine, methionine, histidine, phenylalanine, tyrosine and tryptophan (in order of increasing affinity). Methionine apparently binds non-productively to the phenylalanine (aromatic amino acid-preferring) transport system. l-methionine uptake by larvae is inhibited more by d-alanine and d-valine than by their respective l-isomers, while d- and l-methionine inhibit l-methionine uptake equally well. The presence of an unsubstituted α-amino group is essential for an inhibitor to have a high affinity for the methionine transport system. Uptake of arginine, phenylalanine and methionine is Na⁺-insensitive, and both phenylalanine and methionine are accumulated by larvae against a concentration difference in the presence or absence of Na⁺. Arginine accumulation is precluded by its rapid metabolism to proline, ornithine and an unidentified compound.     

Chemical Modification of Aspartic Acid and Arginine Residues in Horseradish Peroxidase Role of ASP 43 and ARG 38 in the Catalytic Cycle

In an attempt to alter the catalytic properties of horseradish peroxidase (HRP, EC 1.11.1.7), aspartic, glutamic and arginine residues were modified using ethanedithiol and diacetyl. Modification of Asp and Glu led to a marked increase in V_max along with denaturation of the protein. The thiol groups introduced were thought to be responsible, despite being situated on the periphery of the molecule as shown by the modification of the apoenzyme. The role of Arg 38 in the activation of hydrogen peroxide was indicated by the modifications of both enzyme and apoenzyme. An amino acid residue close to Arg 38 was thought to take over its function after blocking the group.     

苯丙酮尿症(PKU)患者血清的氨基酸定量分析(英文)

目的 PKU患者血清中游离氨基酸组成及定量分析。方法 应用日立835-50型氨基酸自动分析仪进行测定。结果 七例PKU患者血清中Phe患者血清中Phe (1.5480.080) 和Glu (0.4680.098) 的含量分别为正常组Phe (0.1070.014) 和Glu (0.1330.046) 的15倍和3.5倍,差异非常显著 (p<0.001)。Arg (0.0330.046) 为正常组 (0.1130.025) 的1/4 (p<0.001)。但患者血清中Tyr (0.0500.016) 与正常值 (0.0470.008) 相比则无显著性差异 (p>0.05)。结论 用氨基酸分析方法可准确测定PKU血清中各种氨基酸含量,为PKU诊断提供了可靠的数据: 并发现除Phe外,还有Arg的异常降低及Glu的异常升高,其发生机理及临床意义有待进一步研究。     

Glutamine synthetase of Dunaliella primolecta. Partial characterization and possible adenylation control in relation to nitrogen nutrient levels

Glutamine synthetase (GS, E.C. 6.3.1.2.) of the unicellular alga Dunaliella primolecta has been partially purified by gel filtration and affinity chromatography. The molecular weight of the enzyme has been estimated at 480,000, comprising eight subunits of 60,000 each. The kinetic behaviour of the enzyme exhibits a biphasic profile of substrate saturation, corresponding to a negative cooperativity process. Alanine, carbamoyl phosphate and glucosamine exert a strong inhibitory effect. The feedback control is cumulative. The effect of Mn²⁺ and Mg²⁺ has been studied. The results suggest the existence of an adenylation process and the possibility of a role of Dunaliella GS in the overall control of nitrogen assimilation.     

Capacities and constraints of amino acid utilization in Arabidopsis

Various amino acids, including both L- and D-enantiomers, may be present in soils, and recent studies have indicated that plants may access such nitrogen (N) forms. Here, the capacity of Arabidopsis to utilize different L- and D-amino acids is investigated and the constraints on this process are explored. Mutants defective in the lysine histidine transporter 1 (LHT1) and transgenic plants overexpressing LHT1 as well as plants expressing D-amino acid-metabolizing enzymes, were used in studies of uptake and growth on various N forms. Arabidopsis absorbed all tested N-forms, but D-enantiomers at lower rates than L-forms. Several L- but no D-forms were effective as N sources. Plants deficient in LHT1 displayed strong growth reductions and plants overexpressing LHT1 showed strong growth enhancement when N was supplied as amino acids, in particular when these were supplied at low concentrations. Several D- amino acids inhibited growth of wild-type plants, while transgenic Arabidopsis-expressing genes encoding D-amino acid-metabolizing enzymes could efficiently utilize such compounds for growth. These results suggest that several amino acids, and in particular L-Gln and L-Asn, promote growth of Arabidopsis, and increased expression of specific amino acid transporters enhances growth on amino acids. The efficiency by which transgenic plants exploit D-amino acids illustrates how plants can be engineered to utilize specific N sources otherwise inaccessible to them.     

土壤氮磷对四季竹叶片氮磷化学计量特征和叶绿素含量的影响

植物器官化学计量特征可以把环境和器官功能性状联系起来, 从而为探索环境作用于植物器官功能的内在机制及器官功能的调控提供可能。通过土壤氮(N)、磷(P)添加设置土壤不同全N和全P浓度的盆栽实验, 分析了土壤和四季竹(Oligostachyum lubricum)叶片N、P化学计量特征及叶片叶绿素含量间的关系。实验设置的土壤不同全N和全P浓度包括对照(全N: 421.76 mg·kg^-1, 全P: 37.35 mg·kg^-1, 1N1P)、全N和全P浓度分别是对照相应浓度的2倍和2倍(2N2P)、2倍和3倍(2N3P)、2倍和4倍(2N4P)、3倍和2倍(3N2P)、3倍和3倍(3N3P)、3倍和4倍(3N4P)、4倍和2倍(4N2P)、4倍和3倍(4N3P)、4倍和4倍(4N4P)共10个处理。结果表明: 土壤N含量分别与叶片N含量和叶片N : P呈极显著正相关, 而土壤P含量与叶片P含量及叶片N : P均无显著性相关。叶片N : P随土壤N : P的增大而增大, 但其增加速率小于土壤N : P的增加速率。相同土壤N : P (11.29)条件下, 生长在2N2P处理和3N3P处理土壤中的立竹叶片N : P无显著差异, 但均显著高于对照(1N1P)并显著低于4N4P处理。土壤不同全N浓度对叶片N : P的影响与相应浓度N和P处理对叶片N : P的影响具有相同的规律。叶片N : P是影响叶片叶绿素含量的主要因素。分析发现: 土壤全N较土壤全P对四季竹叶片N、P化学计量特征具有更大的影响, 并且在土壤全N供应充足时四季竹叶片存在对N的奢侈吸收。N、P添加前土壤N是影响四季竹生长的主要限制元素。     

On-line gas analysis in animal cell cultivation: II. Methods for oxygen uptake rate estimation and its application to controlled feeding of glutamine

Different methods for oxygen uptake rate (OUR) determinations in animal cell cultivation were investigated using a high quality mass spectrometer. Dynamic measurements have considerable disadvantages because of disturbances of the growing cells by the necessary variations of dissolved oxygen concentration. Only infrequent discrete measurements are possible using this method. Stationary liquid phase balance yielded better results with much higher frequency. Gas phase balancing has the advantage of not requiring dissolved oxygen measurement and knowledge of K(L)a, both of them are easily biased. It was found that simple gas phase balancing is either very inaccurate (error larger than expected signal) or very slow, with gas phase residence times of several hours. Therefore, a new method of aeration was designed. Oxygen and CO(2) transfer are mainly achieved via sparging. The gas released to the headspace is diluted with a roughly 100-fold stream of an inert gas (helium). Through this dilution, gas ratios are not changed for O(2), CO(2), Ar, and N(2). The measurement of lower concentrations (parts per million and below) is easy using mass spectrometry with a secondary electron multiplier. With this new method an excellent accuracy and sufficient speed of analysis were obtained. All these on-line methods for OUR measurement were tested during the cultivation of animal cells. The new method allowed better study of the kinetics of animal cell cultures as was shown with a hybridoma cell line (HFN 7.1, ATCC CRL 1606) producing monoclonal antibodies against human fibronectin. With the aid of these methods it was possible to find a correlation between a rapid decrease in oxygen uptake rate (OUR) and glutamine concentration. The sudden decrease in OUR can be attributed to glutamine depletion. This provided a basis for the controlled addition of glutamine to reduce the formation of ammonia produced by hydrolysis. This control method based on OUR measurement resulted in increased cell concentration and threefold higher product concentration. (c) 1995 John Wiley & Sons, Inc.