不同来源仙草的氨基酸比较分析

采用盐酸水解法分析不同来源仙草的氨基酸组成及含量差异,结果表明供试仙草均含有17种氨基酸和7种必需氨基酸,氨基酸总量为4.75％～13.65％,17种氨基酸总量和7种必需氨基酸总量的高低顺序都是9＞10＞5＞8＞1 ＞6＞4＞3＞2＞7,均以9号仙草含量最高,7号仙草最低.各种氨基酸含量高低顺序相似,以谷氨酸、天门冬氨...     

Amino acid transport in a polyaromatic amino acid auxotroph of Saccharomyces cerevisiae

The initiation of growth of a polyaromatic auxotrophic mutant of Saccharomyces cerevisiae was inhibited by several amino acids, whereas growth of the parent prototroph was unaffected. A comparative investigation of amino acid transport in the two strains employing (14)C-labeled amino acids revealed that the transport of amino acids in S. cerevisiae was mediated by a general transport system responsible for the uptake of all neutral as well as basic amino acids. Both auxotrophic and prototrophic strains exhibited stereospecificity for l-amino acids and a K(m) ranging from 1.5 x 10(-5) to 5.0 x 10(-5) M. Optimal transport activity occurred at pH 5.7. Cycloheximide had no effect on amino acid uptake, indicating that protein synthesis was not a direct requirement for amino acid transport. Regulation of amino acid transport was subject to the concentration of amino acids in the free amino acid pool. Amino acid inhibition of the uptake of the aromatic amino acids by the aromatic auxotroph did not correlate directly with the effect of amino acids on the initiation of growth of the auxotroph but provides a partial explanation of this effect.     

商品鹿鞭中氨基酸含量比较

采用氨基酸自动分析仪测定不同品种与产地的9份鹿鞭药材中氨基酸成分的种类及含量。这9份样品中均含17种氨基酸,其中人体必需氨基酸6种。17种氨基酸中以甘氨酸含量最高,必需氨基酸中亮氨酸含量最高;比较不同样品的含量,以辽宁西丰的梅花鹿鞭的总氨基酸含量最高,黑龙江梅花鹿鞭的必需氨基酸含量最高。结论为不同产地鹿鞭中氨基酸含量有一定差异,该方法可用于鹿鞭药材的质量评价。     

ACCELERATED EFFLUX OF [14C] AND [3H] AMINO ACIDS FROM SUPERFUSED SLICES OF RAT BRAIN1

Abstract— The rate of spontaneous efflux of several isotopically-labelled amino acids was measured in superfused slices of rat brain. The group specificity of amino acid efflux was studied by examining the pattern of accelerated efflux in the presence of extracellular unlabeled amino acid. The mediated efflux of γ-aminobutyric acid and l -glutamic acid was highly specific. Less specificity was found for amino acids in the small neutral, large neutral and basic groups. The efflux of γ-aminobutyric acid and l -glutamic acid was accelerated by structurally similar amino acids which are known to depress or excite motor neurons. This finding raises the possibility that the physiological effects of non-specific amino acids during local iontophoresis may be secondary to an accelerated release of more specific transmitter amino acids from neighbouring cells.     

Neutral amino acids in the brain: changes in response to food ingestion

Abstract— The brain levels of each of the aromatic and branched-chain amino acids change 2 h after fasting rats begin to consume either a carbohydrate-fat diet or a similar diet containing 18% or 40% protein. Carbohydrate-fat ingestion elevates the concentrations of each of the aromatic amino acids in brain, while substantially depressing those of the branched-chain amino acids. The inclusion of protein in this diet suppresses the increases in brain aromatic amino acids and attenuates the decreases in the branched-chain amino acids. The changes in the brain level of each neutral amino acid following the ingestion of any of these diets correlate extremely well with the effects of the diet on the serum neutral amino acid pattern, specifically on the serum concentration ratio of each neutral amino acid to the sum of the other neutral amino acids. The diet-induced changes in the brain level of each of the amino acids also correlate surprisingly well with the calculated rate of brain influx for each amino acid.     

Mechanism of amino Acid uptake by sugarcane suspension cells

The amino acid carriers in sugarcane suspension cells were characterized for amino acid specificity and the stoichiometry of proton and potassium flux during amino acid transport.

Amino acid transport by sugarcane cells is dependent upon three distinct transport systems. One system is specific for neutral amino acids and transports all neutral amino acids including glutamine, asparagine, and histidine. The uptake of neutral amino acids is coupled to the uptake of one proton per amino acid; one potassium ion leaves the cells for charge compensation. Histidine is only taken up in the neutral form so that deprotonation of the charged imidazole nitrogen has to occur prior to uptake. The basic amino acids are transported by another system as uniport with charge-compensating efflux of protons and potassium. The acidic amino acids are transported by a third system. Acidic amino acids bind to the transport site only if the distal carboxyl group is in the dissociated form (i.e. if the acidic amino acid is anionic). Two protons are withdrawn from the medium and one potassium leaves the cell for charge compensation during the uptake of acid amino acids. Common to all three uptake systems is a monovalent positively charged amino acidproton carrier complex at the transport site.

     

Amino acid transport and metabolism in nitrogen-starved cells of Saccharomyces cerevisiae.

Nitrogen-starved yeast derepress a general amino acid permease which transports basic and hydrophobic amino acids. Although both groups of amino acids are metabolized, the derivatives of the basic amino acids are retained by the cells, whereas those of the hydrophobic amino acids are released as acidic and neutral deaminated derivatives. The release of the deaminated derivatives of the hydrophobic amino acids only occurs in the presence of glucose, which presumably produces amino acceptors. The accumulation of intracellular amino acids results in trans-inhibition of the uptake of exogenous amino acids whether the intracellular amino acid is a basic amino acid or the product of intracellular transamination from a hydrophobic amino acid. Variation of permease and transaminase activity was measured during growth under repressed (ammonia-grown) and derepressed (proline-grown) conditions. Maximum levels for both activities occurs at the mid-exponential phase.     

Comparison of Protein Precipitants for the Determination of Free Amino Acids in Plasma

The present study was undertaken to select the most preferred deproteinizing agent for microbiological or chromatographic determination of free amino acids in plasma. The highest quantities of amino acids were obtained from samples treated with picric acid, ethanol or sulfosalicylic acid. It was also found from examining the recoveries of ¹⁴C-labeled amino acids added to plasma following treatment with picric acid, ethanol and sulfosalicylic acid that one kind of deproteinizing agent is not equally effective with all amino acids. Picric acid treatment gave the most reliable results for determination of amino acids except the basic amino acids, alanine and tryptophan. Sulfosalicylic acid was the most recommended deproteinizing agent for basic amino acids and ethanol was good for the assay of alanine and tryptophan. The response of d-amino acids to the deproteinizing agents tested was not always similar to that of their l-isomers.     

基于游离氨基酸的组分及特征比较四种食用菌与四种果蔬的营养与风味特征

为科学评价食用菌与果蔬两类食材的营养特征与口感风味,本研究选取4种常见食用菌（金针菇、斑玉蕈、香菇、双孢蘑菇）和4种常见果蔬（苹果、香蕉、胡萝卜、番茄）,采用氨基酸自动分析仪测定其游离氨基酸,系统比较了两类食材的游离氨基酸组分比例及呈味特征。主成分和聚类分析表明 4种食用菌和4种果蔬显著分为两类,食用菌类游离氨基酸总量平均是果蔬类的3倍以上,4种食用菌中各游离氨基酸的比例均衡。4种食用菌的必需氨基酸/(必需氨基酸+非必需氨基酸)在40%左右,必需氨基酸/非必需氨基酸接近或大于60%,均接近理想值。4种食用菌所含的呈味氨基酸（鲜味、甜味、芳香族、苦味）均是4种果蔬的2倍以上,TAV值最显著的是：谷氨酸、苯丙氨酸、甘氨酸、苏氨酸、组氨酸、赖氨酸、缬氨酸、精氨酸和丙氨酸。这些氨基酸分属在五大氨基酸家族,在食用菌与果蔬中代谢途径相同;只有赖氨酸在二者中的合成途径不同。结果表明供试的食用菌比果蔬有优质的氨基酸比例和丰富的呈味组分,此差异体现在众多氨基酸的代谢层面,而非个别氨基酸导致。     

Studies on Amino Acid Sequence Determination from N-Terminal of Peptide by Methylthiohydantoin

Methylisothiocyanate reacts with the amino group of amino acid in alkaline solution to give methylthiocarbamyl amino acid. This is converted to the methylthiohydantoin derivative (MTH-amino acid) by subsequent acidification.

Gas chromatographical identification of 20 amino acids were examined by making MTH-amino acids. Among them, ten amino acids (glycine, alanine, valine, leucine, isoleucine, proline, methionine, phenylalanine, serine and threonine) were successfully identified. Aspartic acid and glutamic acid could be identified as the methyl esters of the MTH-amino acids.     

Amino acid transport systems required for diazotrophic growth in the cyanobacterium Anabaena sp. strain PCC 7120.

Uptake of 16 amino acids by the filamentous, heterocyst-forming cyanobacterium Anabaena sp. strain PCC 7120 was characterized with regard to kinetic parameters of transport, intracellular accumulation of the transported amino acids, and sensitivity of the transport process to energy metabolism inhibitors. Mutants resistant to certain toxic analogs of some amino acids were isolated that were impaired in amino acid transport. Results obtained in this study, together with those reported previously (A. Herrero and E. Flores, J. Biol. Chem. 265:3931-3935, 1990), suggest that there are at least five amino acid transport systems in strain PCC 7120: one high-affinity, active system for basic amino acids; one low-affinity, passive system for basic amino acids; two high-affinity, active systems with overlapping, but not identical, specificities for neutral amino acids; and one putative system for acidic amino acids. Some of the amino acid transport mutants were impaired in diazotrophic growth. These mutants were unable to develop a normal percentage of heterocysts and normal nitrogenase activity in response to nitrogen stepdown. Putative roles for the amino acid transport systems in uptake of extracellular amino acids, recapture of amino acids that have leaked from the cells, and intercellular transfer of amino acids in the filaments of Anabaena sp. strain PCC 7120 are discussed.     

Utilization of amino acids during development of the African migratory locust embryo

Summary The metabolic fate of amino acids introduced into the locust egg by the addition of amino acids after oviposition is compared to that of amino acids derived from the maternal haemocoel. It was found that amino acids from the two sources did not always follow the same pattern of utilization. Topical application resulted in some of the applied amino acids being converted into other amino acids and ammonia. When the labelled amino acid was administered to the egg via the maternal haemocoel, radioactivity was restricted to the amino acid orginally provided.     

Amino Acid Transport in Pseudomonas aeruginosa

Properties of the transport systems for amino acids in Pseudomonas aeruginosa were investigated. Exogenous (14)C-labeled amino acids were shown to equilibrate with the internal native amino acid pool prior to incorporation into protein. When added at low external concentrations, the majority of the amino acids examined entered the protein of the cell unaltered. The rates of amino acid transport, established at low concentrations with 18 commonly occurring amino acids, varied as much as 40-fold. The transport process became saturated at high external amino acid concentrations, was temperature-sensitive, and was inhibited by sodium azide and iodoacetamide. Intracellular to extracellular amino acid ratios of 100- to 300-fold were maintained during exponential growth of the population in a glucose minimal medium. When the medium became depleted of glucose, neither extracellular nor intracellular amino acids could be detected.     

Structure of thermal polymers of amino acids

When glutamic acid is a predominant amino acid in a thermally polymerized mixture of amino acids, pyro Glu is exclusively found at the N-terminal end of the poly-amino acid polymer. It probably initiates the polymerization process. Lysine-containing polymers will probably contain epsilon N-(glutamyl)L-lysine cross links which may account for the higher molecular weights observed in these polymers (100-200 000). Incorporation of some amino acids facilitates the incorporation of others. When utilizing mixtures of three to eight amino acids with glutamic acid as one of the amino acids, some fractions are obtained which include all the amino acids in the polymerization mixture. The biosynthesis of glutathione, gramicidin, tyrocidine and cell-wall polypeptides has demonstrated that non-random amino acid sequence peptides can be biologically synthesized without the direct participation of nucleic acids. That is, the enzymes appear to provide adequate chemical specificity to form non-random amino acid sequence peptides. The properties and replication of the scrapie agent may provide us with more profound insight as to the evolution of purely physical-chemical systems into biological systems.     

Synthesis of N-carboxyalkyl and N-aminoalkyl functionalized dipeptide building units for the assembly of backbone cyclic peptides.

To improve the assembly of backbone cyclic peptides, N-functionalized dipeptide building units were synthesized. The corresponding N-aminoalkyl or N-carboxyalkyl amino acids were formed by alkylation or reductive alkylation of amino acid benzyl or tert-butyl esters. In the case of N-aminoalkyl amino acid derivatives the aldehydes for reductive alkylation were obtained from N,O-dimethyl hydroxamates of N-protected amino acids by reduction with LiAlH4. N-carboxymethyl amino acids were synthesized by alkylation using bromoacetic acid ester and the N-carboxyethyl amino acids via reductive alkylation using aldehydes derived from formyl Meldrums acid. Removal of the carboxy protecting group leads to free N-alkyl amino acids of very low solubility in organic solvents, allowing efficient purification by extraction of the crude product. These N-alkyl amino acids were converted to their tetramethylsilane-esters by silylation with N,O-bis-(trimethylsilyl)acetamide and could thus be used for the coupling with Fmoc-protected amino acid chlorides or fluorides. To avoid racemization the tert-butyl esters of N-alkyl amino acids were coupled with the Fmoc-amino acid halides in the presence of the weak base collidine. Both the N-aminoalkyl and N-carboxyalkyl functionalized dipeptide building units could be obtained in good yield and purity. For peptide assembly on the solid support, the allyl type protection of the branching moiety turned out to be most suitable. The Fmoc-protected N-functionalized dipeptide units can be used like any amino acid derivative under the standard conditions for Fmoc-solid phase synthesis.     

Effects of Bestatin and Leupeptin on Protein Degradation in Perfused Rat Hindquarters: Accumulation of Acid Soluble Peptides in Muscle during Bestatin Perfusion

Microbial protease inhibitors, bestatin and leupeptin, were perfused through hindquarters, and the effects of these inhibitors on the amino acid release and the accumulation of acid soluble peptides were studied using normal and Streptozotocin-induced diabetic rats. Both inhibitors depressed the amino acid release from the hindquarters of normal rats. However, leupeptin, unlike bestatin, failed to suppress the release of amino acids in diabetic rats. Bestatin caused an accumulation of acid soluble peptides in perfused skeletal muscle. However, leupeptin did not show this effect. The amino acid composition and the N-terminal amino acids were analyzed on the acid soluble peptides accumulated after bestatin perfusion. Branched-chain amino acids were preferentially accumulated as the acid soluble peptides, and more than half of the total amounts of these amino acids were located in the N-terminus. From these results, it was concluded that bestatin-sensitive protease(s), probably leucine aminopeptidase and/or arylamidase, play an important role in the degradation process of skeletal muscle proteins, especially in the steps to degrade acid soluble peptides into free amino acids.     

Substrate-dependent regulation of intracellular amino acid concentrations in cultured bovine aortic endothelial cells

Amino acid deprivation induces adaptive changes in amino acid transport and the intracellular amino acid pool in cultured cells. In this study intracellular amino acid levels were determined in cultured bovine aortic endothelial cells (EC) deprived of L-arginine or total amino acids for 1, 3, 6 and 24 h. Amino acid concentrations were analyzed by reverse phase HPLC after precolumn derivatisation. Under normal culture conditions levels of L-arginine L-citrulline, total essential and non-essential amino acids were 840 +/- 90 microM, 150 +/- 40 microM, 11.4 +/- 0.9 mM and 53.3 +/- 3.4 mM (n = 9), respectively. In EC deprived of L-arginine or all amino acids for 24 h L-arginine and L-citrulline levels were 200 microM and 50 microM, and 670 microM and 100 microM Deprivation of L-arginine or total amino acids induced rapid (1 h) decreases (30 - 50%) in the levels of other cationic (lysine, ornithine) and essential branched-chain (valine, isoleucine, leucine) and aromatic (phenylalanine, tryptophan) amino acids. L-glutamine was reduced markedly in EC deprived of total amino acids for 1 h - 6 h but actually increased 3-fold in EC deprived of L-arginine for 6 h or 24 h. Arginine deprivation resulted in a rapid decrease in the total intracellular amino acid pool, however concentrations were restored after 24 h. Increased amino acid transport and/or reduced protein synthesis may account for the restoration of amino acid levels in EC deprived of L-arginine. The sustained reduction in the free amino acid pool of EC deprived of all amino acids may reflect utilization of intracellular amino acids for protein synthesis.     

Influence of amino acid supply on ribosomes and protein synthesis of perfused rat liver

1. The livers of rats were perfused in situ with medium containing mixtures of amino acids in multiples of their concentration in normal rat plasma. The incorporation of labelled amino acid into protein of the liver and of the perfusing medium increased with increasing amino acid concentration. During 60min. perfusions, labelling of liver protein reached a plateau, and labelling of medium protein was inhibited when the initial concentration of the amino acid mixture was more than ten times the normal plasma value. 2. Examination of polysome profiles derived from livers perfused without amino acids in the medium showed that the number of large aggregates was decreased and the number of small aggregates, particularly monomers and dimers, was increased with time of perfusion. The addition of amino acids to the perfusion medium reversed this polysome shift to an extent that was dependent on the initial concentration of amino acids. Polysome profiles derived from livers perfused for 60min. with ten times the normal plasma concentration of amino acids were essentially the same as the polysome profiles of normal non-perfused livers. 3. The ability of ribosome preparations from perfused livers to incorporate amino acids into protein in vitro decreased with increasing time of perfusion when no amino acids were added to the medium, but increased as the concentration of amino acids in the perfusion medium was increased. 4. The ability of cell sap from perfused livers to support protein synthesis in vitro was not influenced by the amino acid concentration of the perfusion medium. 5. Livers were perfused for 60min. with medium containing amino acid mixtures at ten times the normal plasma concentration but deficient in one amino acid. Maximal incorporation of labelled amino acid into liver protein, the stability of the polysome profile and the ability of ribosome preparations to incorporate amino acids into protein were found to depend on the presence of 11 amino acids: arginine, asparagine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, threonine, tryptophan and valine. A mixture of these 11 amino acids, at ten times their normal plasma concentration, stimulated the incorporation of labelled amino acid into liver protein, stabilized the polysome profile and increased the ability of ribosome preparations to incorporate amino acids into protein to the same extent as the complete mixture. 6. It is concluded that the availability of certain amino acids plays an important role in the control of protein synthesis, possibly by stimulating the ability of ribosomes to become, and to remain, attached to messenger RNA.     

Influence of triiodothyronine and dexamethasone on renal amino acid handling in rats loaded with various amino acid mixtures

Summary In adult female rats, the influence of dexamethasone or triiodothyronine on renal amino acid handling was investigated in amino acid loaded animals. Amino acids were administered intravenously as two mixtures, each containing four amino acids to overload amino acid reabsorption capacity. Bolus injections of both mixtures were followed by temporary increase in fractional excretion of the administered amino acids as well of the amino acids which were not covered in the mixtures. The administration of the two mixtures was followed by different interactions between various amino acid carriers.After dexamethasone pretreatment (60µg/100g b.wt. for 3 days, once daily) a stimulation of the renal amino acid handling could be shown. Triiodothyronine (20µg/100g b.wt. for 3 days, once daily) did not increase tubular reabsorption capacity for amino acids. It even increased fractional amino acid excretion in amino acid loaded rats as a sign of enhanced amino acid metabolism in the kidney and/or increased amino acid uptake into the tubular cells from the luminal site.     

Analysis of amino acids in nectar from pitchers of Sarracenia purpurea (Sarraceniaceae)

Sarracenia purpurea L. (northern pitcher plant) is an insectivorous plant with extrafloral nectar that attracts insects to a water-filled pitfall trap. We identified and quantified the amino acids in extrafloral nectar produced by pitchers of S. purpurea. Nectar samples were collected from 32 pitchers using a wick-sampling technique. Samples were analyzed for amino acids with reverse-phase high-performance liquid chromatography with phenylisothiocyanate derivatization. Detectable amounts of amino acids were found in each of the 32 nectar samples tested. Mean number of amino acids in a nectar sample was 9 (SD = 2.2). No amino acid was detected in all 32 samples. Mean amount of amino acids in a nectar sample (i.e., amount per wick) was 351.4 ng (SD = 113.2). Nine amino acids occurred in 20 of the 32 samples (aspartic acid, cysteine, glutamic acid, glycine, histidine, hydroxyproline, methionine, serine, valine) averaging 263.4 ng (SD = 94.9), and accounting for ~75% of the total amino acid content. Nectar production may constitute a significant cost of carnivory since the nectar contains amino acids. However, some insects prefer nectar with amino acids and presence of amino acids may increase visitation and capture of insect prey.