茚三酮与氨基酸显色反应再探

<正> 一、前言迄今为止,茚三酮与氨基酸显色反应的机理仍不十分清楚。尤其已报道的操作中,该反应对各种氨基酸选择性极低,因而使本法的应用受到了一定限制。此外,据文献报道:茚三酮与氨基酸显色反应不仅仅发生于α-氨基酸、对于β氨基酸、伯胺类等亦能进行;金属离子会干扰显色反应等。对茚三酮与氨基酸显色反应进行深     

生物催化氨基酸C—N裂解反应的研究进展

氨基酸脱氨酶能催化系列氨基酸C—N裂解反应生成对应的α 酮酸和氨,是代谢途径及生物催化的重要酶.综述了近年来催化氨基酸C—N裂解反应的5'磷酸吡哆醛介导的苏氨酸脱氨酶、黄素腺嘌呤二核苷酸介导的L氨基酸脱氨酶和L氨基酸氧化酶,以及这些关键酶应用于多酶级联反应中以生产α 羟基酸、α 酮酸、D氨基酸等精细化学品的研究进展.此...     

蛋白质酸水解终点的决定

在蛋白质酸水介制准氨基酸的水介过程中,同时存在着蛋白质水介生成氨基酸,和已生成累积的氨基酸发生分解破坏的两个反应。本文讨论了此二反应的动力学过程,并提出正确决定水解终点以避免水介过头的设想。     

蛋白质酸水解终点的决定

在蛋白质酸水介制准氨基酸的水介过程中,同时存在着蛋白质水介生成氨基酸,和已生成累积的氨基酸发生分解破坏的两个反应。本文讨论了此二反应的动力学过程,并提出正确决定水解终点以避免水介过头的设想。     

氨基酸的电分析进展

氨基酸电分析研究是生命科学中令人关注的课题。本文就各种氨基酸电分析化学研究的最新进展进行了综述。主要叙述了电活性氨基酸的直接伏安检测和非电活性氨基酸通过衍生化反应和电致化学发光进行的间接检测 ,并对其前景进行了展望。     

混合醇体系中测定氨基酸的茚三酮光度法

氨基酸一般利用自动分析仪定量测定,但需高精密仪器,而基层研究单位一般难以采用,而且分析也不需要如此精密。利用茚三酮能够定量测定氨基酸,除脯氨酸和羟脯氨酸与茚三酮形成黄色化合物外,其它所有氨基酸均与茚三酮反应形成蓝紫色物质,溶液颜色深浅与氨基酸含量成正比。据此本文对氨基酸的茚三酮光度法测定进行了研究和改进。在混合醇体系中以醋酸钠-醋酸为缓冲溶液,加入抗坏血酸以增加反应的灵敏度,100℃水浴20分钟完成显色反应。并对酸度、试剂用量、反应时间等因素进行了研究,拟定了用于混合氨基酸总量测定的操作程序。     

氨基酸的鉴别

<正> 常见的20余种氨基酸的结构和性质相似,尚未见到系统地鉴别各种氨基酸的报道,为此我们应用各种化学反应、薄层层析、紫外、红外吸收光谱相结合的方法,系统地鉴别了18种氨基酸。1.化学鉴别法:我们采用了14种化学反应方法,根据反应产生的气体、颜色、沉淀、气味及变旋性等来鉴别不同的氨基酸。其中有的是所有氨基酸共同的反应,有的是某几种氨基酸共同的反应,更重要的是有的是某一种氨基酸特有的反应。     

氨基酸的鉴别

<正> 常见的20余种氨基酸的结构和性质相似,尚未见到系统地鉴别各种氨基酸的报道,为此我们应用各种化学反应、薄层层析、紫外、红外吸收光谱相结合的方法,系统地鉴别了18种氨基酸。1.化学鉴别法:我们采用了14种化学反应方法,根据反应产生的气体、颜色、沉淀、气味及变旋性等来鉴别不同的氨基酸。其中有的是所有氨基酸共同的反应,有的是某几种氨基酸共同的反应,更重要的是有的是某一种氨基酸特有的反应。     

炒焦对山楂中氨基酸含量的影响分析

为探索炒焦对山楂中氨基酸的影响及山楂炒焦过程中氨基酸的变化,对炒焦前后山楂中氨基酸含量的变化进行了分析.测定结果显示,炒焦前后山楂中氨基酸的种类不变,各种氨基酸含量变化不一;方差分析显示炒焦前后氨基酸的含量变化不显著,炒焦对山楂中氨基酸的影响较小.由于山楂在炒焦过程具备发生美拉德反应的物质基础和客观条件,推测山楂中的氨...     

苦瓜干制前后氨基酸组分的变化

经酸水解,利用氨基酸自动分析仪测定干制加工前后的苦瓜中总氨基酸和游离氨基酸含量。结果表明干制前后的苦瓜在氨基酸质量分数上有明显差别,总氨基酸质量分数分别为11.99%和10.87%,游离氨基酸分别为2.36%和0.70%。结论:干制前后的苦瓜中氨基酸质量分数差别明显,其中碱性氨基酸质量分数的变化尤其显著,总氨基酸中精氨酸质量分数下降50%,在游离氨基酸中下降78%;同时还原糖质量分数也由干制前的4.86%下降为干制后的1.86%,说明苦瓜干制过程中发生了美拉德反应,造成了氨基酸和还原糖含量的下降。     

Sporulation of auxotrophic mutants ofBacillus subtilis in suboptimal concentrations of essential amino acids

The effect of the amino acid concentration on sporulation of five mutants ofBacillus subtilis requiring histidine, isoleucine, leucine, lysine and methionine for growth was studied. Low amino acid concentrations, permitting about 10–20% of maximum growth, inhibit the ability of the bacteria to sporulate on completing vegetative growth. In medium concentrations (from 25% of maximum growth), leucine, isoleucine, histidine and methionine induce sporulation, while the concentration of the other nutrients in the medium is still adequate for further vegetative growth. The percentage of sporulation depends on the amino acid concentration and is characteristic for each amino acid. The lysine mutant strain 1399 was not capable of sporulation unless cultivated in the presence of the lysine concentration optimal for growth. The effect of amino acid deficiency depends on the cultivation method.     

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.     

Water Deficit-Induced Changes in Concentrations in Proline and Some Other Amino Acids in the Phloem Sap of Alfalfa

Changes in amino acid composition of alfalfa (Medicago sativa L.) phloem sap were studies in response to a water deficit. Sap was collected by stylectomy. As the leaf water potential ([psi]) decreased from -0.4 to -2.0 MPa, there was significant increase of the total amino acid concentration, due to that of some amino acids: proline, valine, isoleucine, leucine, glutamic acid, aspartic acid, and threonine. Asparagine concentration, which is the main amino acid assayed in the phloem sap of alfalfa (it accounts for 70% of the total content), did not vary with the plant water status. The other amino acid concentrations remained stable as [psi] varied; in particular, [gamma]-amino butyric acid concentration remained unchanged, whereas it varied in response to wounding. The more striking change in the sieve tubes was the accumulation of proline, which was observed below a [psi] threshold value of about -0.9 MPa (concentration x60 for a decrease of [psi] from -0.9 to -2.0 MPa). The role of such changes in phloem sap amino acid concentration in osmotic adjustment of growing tissues is discussed.     

Optimization of the culture medium composition for the antibody response of mouse spleen cells

Summary The Mishell-Dutton culture system for in vitro primary antibody response of mouse spleen cells was used to optimize the amino acid composition of RPMI 1640 media. Each of the 20 amino acids was tested over a broad range of concentrations always leaving the remaining 19 amino acids unaltered (i.e. at the formula recommended concentration). In several instances, higher plaque-forming cell responses were obtained with an amino acid concentration that was either higher or lower than that recommended: (a) the optimum concentration for valine, glutamine, and lysine lies considerably above the recommended one, (b) the optimum concentration for leucine as well as for several other amino acids lies below the recommended concentration, and (c) the optimum concentration for arginine corresponds exactly to the recommended concentration. The second round of optimization, i.e. combining of two conditions that individually yielded an improved response often caused a decrease of response. The possibility is discussed that for an optimal response a ratio of two or several amino acids rather than the absolute concentration of any one amino acid is of importance. The Basel Institute for Immunology was founded and is supported by F. Hoffman-La Roche & Co., Ltd.     

Gustatory responses of eel palatine receptors to amino acids and carboxylic acids

The gustatory receptors of the eel palate were found to be extremely sensitive to amino acids and carboxylic acids. The results obtained are as follows: (a) 11 amino acids which are among naturally occurring amino acids elicited responses in the palatine nerve, but 9 amino acids did not elicit a response even at a high concentration. The effect of D-amino acids was always much less than that of their corresponding L-isomers. There was no appreciable difference in the effectiveness of an alpha-amino acid (alpha-alanine) and beta-amino acid (beta-alanine). (b) The threshold concentrations of the most potent amino acids (arginine, glycine) were between 10(-8) and 10(-9) M. A linear relation between the magnitude of the response and log stimulus concentration held for a wide concentration range for all the amino acids examined. (c) The palatine receptors responded sensitively to various carboxylic acid solutions whose pH was adjusted to neutral. The threshold concentrations varied between 10(-4) and 10(-7) M. The magnitude of the response at 10(-2) M increased with an increase of carbon chain length. (d) The extent of cross-adaptation was examined with various combinations of amino acids. A variety of the response patterns showing complete cross-adaptation, no cross-adaptation, or synergetic interaction was observed. The synergetic interaction was also observed when one amino acid below its threshold concentration was added to the other amino acid below its threshold concentration was added to the other amino acid. No cross-adaptation was observed between amino acids and fatty acids. (e) The treatment of the palate with papain led to loss of the responses to arginine, glycine, and histidine without affecting those to proline and acetic acid. The treatment with pronase E eliminated selectively the response to proline. The possibility that the eel gustatory receptors are responsible for sensing food at a distance was discussed.     

The role of free amino acids in osmoregulation of cricket blood (Acheta domesticus)

The blood volume increased during normal feeding, and did not decrease during fasting at the end of the stadium. The unexpectedly high blood volume of starved crickets might be an adaptation to increase chances for moulting via stretch receptor stimulation.The amount of blood amino acids was not changed by feeding, but increased with fasting or starvation. Thus amino acid levels in the blood were not directly related to amino acid input from the gut.The blood protein concentration did not change during starvation, but the amino acid concentration was 33% higher in starved crickets that drank water as opposed to those given saline to drink. Thus amino acid levels in the blood were not related directly to blood protein concentration.The blood amino acid concentration was 19–22 mM/l in response to salt intake by feeding crickets or starved crickets drinking saline. The concentration was 32–38 mM/l when the crickets were fasting prior to and after ecdysis or when starved with water to drink during the time when they would normally be feeding. The increase of amino acids during fasting was due to a proportional increase in all amino acids augmented by a 3 × increase in tyrosine. The increase during salt depleting starvation was due to a doubling of the two predominant amino acids proline and glycine. Proline and glycine were not increased in starved crickets drinking saline, thus starvation was not the reason for the increase. This is the first instance where specific amino acids have been implicated in osmoregulation.     

Intraspecific Variation in the Amino Acid Content of Floral Nectar

In order to evaluate the constancy of amino acid composition and concentration, several randomly collected samples of genuine floral nectar from different flowers of several plant species were analysed. Although there seems to exist a rough pattern of species-specific nectar amino acid composition, amino acid concentration varied strongly from sample to sample. Apart from the lack of constancy in concentration of floral amino acids, our samples do not show amino acid concentration values as assigned typical for the pertinent pollination mode by other workers.     

Effect of pH, temperature, and potassium sorbate on amino acid uptake in Salmonella typhimurium 7136

The effect of sorbate on L-serine and L-histidine uptake in Salmonella typhimurium was studied at various pH levels, temperatures, and amino acid and sorbate concentrations. Low pH had an apparent synergistic effect on amino acid uptake inhibition caused by sorbate. The relationship between sorbate concentration and the amount of amino acid uptake inhibition was not linear. Compared with L-histidine, L-serine uptake was more sensitive to changes in pH, temperature, and sorbate concentration. Various degrees of amino acid uptake inhibition by sorbate may be related to differences between amino acid transport systems. The results of this study suggest that sorbate acts as a noncompetitive inhibitor of amino acid uptake in S. typhimurium.     

Effect of pH, temperature, and potassium sorbate on amino acid uptake in Salmonella typhimurium 7136.

The effect of sorbate on L-serine and L-histidine uptake in Salmonella typhimurium was studied at various pH levels, temperatures, and amino acid and sorbate concentrations. Low pH had an apparent synergistic effect on amino acid uptake inhibition caused by sorbate. The relationship between sorbate concentration and the amount of amino acid uptake inhibition was not linear. Compared with L-histidine, L-serine uptake was more sensitive to changes in pH, temperature, and sorbate concentration. Various degrees of amino acid uptake inhibition by sorbate may be related to differences between amino acid transport systems. The results of this study suggest that sorbate acts as a noncompetitive inhibitor of amino acid uptake in S. typhimurium.     

Capillary electrophoresis for the determination of major amino acids and sugars in foliage: application to the nitrogen nutrition of Sclerophyllous species

Amino acids and sugars are probably the most commonly measured solutes in plant fluids and tissue extracts. Chromatographic techniques used for the measurement of such solutes require complex derivatization procedures, analysis times are long and separate analyses are required for sugars and amino acids. Two methods were developed for the analysis of underivatized sugars and amino acids by capillary electrophoresis (CE). Separation of a range of sugars and amino acids was achieved in under 30 min, with good reproducibility and linearity. In general, there was close agreement between amino acid analyses by CE and HPLC with post-column derivatization. An alternative, more rapid method was optimized for the common neutral sugars. Separation of a mixture of fructose, glucose, sucrose, and fucose (internal standard) was achieved in less than 5 min. How the source of N applied (nitrate or ammonium) and its concentration (8.0 or 0.5 mM) affects the amino acid and sugar composition of leaves from Banksia grandis Willd. and Hakea prostrata R. Br. was investigated. The amino acid pool of Banksia and Hakea were dominated by seven amino acids (aspartic acid, glutamic acid, asparagine, glutamine, serine, proline, and arginine). Of these, asparagaine and glutamine dominated at low N-supply, whereas at high N-supply the concentration of arginine increased and dominated amino-N. Plants grown with nitrate had a greater concentration of proline relative to plants with ammonium. In Banksia the concentration of amides was greatest and arginine least with a nitrate N-source, whereas in Hakea amides were least and arginine greatest with nitrate N-source. The concentration of sugars was greater in Banksia than Hakea and in both species at greater N-supply.