Biosynthesis of Protein Amino Acids in Plant Tissue Culture II Further Isotope Competition Experiments Using Protein Amino Acids

The behavior of exogenously supplied alanine, proline, glutamine, glutamic acid and aspartic acid in isotope competition experiments using cells of Paul's Scarlet Rose grown in the presence of glucose-U-¹⁴C was examined at higher concentrations than were used in part I of this series.     

The Mass Spectra of Thiohydantoins Derived from Some Amino Acids

One of the bound forms of vitamin B₆ occurring in rice bran was isolated in a faintly yellowish syrup by repeating ion-exchange and paper-partition chromatographic techniques. The behaviors of the isolate on thin-layer and Aminex A–5 column chromatograms were coincident with those of synthetic pyridoxine-β-d-glucoside which was obtained by Königs-Knorr condensation of α⁴,3-O-isopropylidene pyridoxine and 2,3,4,6-tetra-O-acetyl-α-d-glucopyranosyl bromide. On acid hydrolysis, the isolate gave pyridoxine and glucose. Glucose was proved to bind to the 5-hydroxymethyl group of pyridoxine, because the isolate did not react with 2,6-dichloroquinone chlorimide in the presence of boric acid. An equimolar amount of pyridoxine and d-glucose was produced with an equivalent consumption of the isolate by the action of β-glucosidase. No essential difference between the isolated and synthetic preparations could be detected in UV- and NMR-spectral features. Thus, the chemical structure of the isolate was identified as 5′-O-(β-d-glucopyranosyl) pyridoxine.     

Catabolism of Amino Acids by Megasphaera elsdenii LC1

The amino acids in an acid hydrolysate of casein were catabolized more extensively by Megasphaera elsdenii than those in an enzymic hydrolysate. Threonine and serine were most actively degraded, but no resultant increase in growth yield occurred. Branched-chain volatile fatty acid production, which increased as the dilution rate of a glucose-limited chemostat decreased, seemed to be associated with maintenance rather than with growth.     

Stereochemistry and Reactions of Aziridinyl- phosphinothionates Derived from Amino Acids

Aziridinylphosphinothionates were prepared from optically active ethyl hydrogen phenylphos- phonothionates and l-bromo-2-alkanamines derived from leucine or valine. The aziridine ring was opened by the action of some nucleophiles. Refluxing the aziridinylphosphinothionates in acetone with sodium iodide caused hydrolysis accompanied by the rearrangement of the sulfur atom to give β -mercaptoethylphosphonamidates. The reaction mechanism was discussed with stereochemical considerations. The insecticidal activity of the products was also examined.     

Anti‐barnacle Activity of Isocyanides Derived from Amino Acids

Creation of new potent antifouling active compounds is important for the development of environmentally friendly antifouling agents. Fifteen isocyanide congeners derived from proteinogenic amino acids were synthesized, and the antifouling activity and toxicity of these compounds against cypris larvae of the barnacle Balanus amphitrite were investigated. All synthesized amino acid‐isocyanides exhibited potent anti‐barnacle activity with EC₅₀ values of 0.07 – 10.34 μg/ml after 120 h exposure without significant toxicity. In addition, seven compounds showed more than 95% settlement inhibition of the cypris larvae at 10 μg/ml after 120 h exposure without any mortality observed. Considering their structure, these amino acid‐isocyanides would eventually be biodegraded to their original nontoxic amino acids. These should be useful for further research focused on the development of environmentally friendly antifoulants.     

The Hydrophobic Temperature Dependence of Amino Acids Directly Calculated from Protein Structures

The hydrophobic effect is the main driving force in protein folding. One can estimate the relative strength of this hydrophobic effect for each amino acid by mining a large set of experimentally determined protein structures. However, the hydrophobic force is known to be strongly temperature dependent. This temperature dependence is thought to explain the denaturation of proteins at low temperatures. Here we investigate if it is possible to extract this temperature dependence directly from a large set of protein structures determined at different temperatures. Using NMR structures filtered for sequence identity, we were able to extract hydrophobicity propensities for all amino acids at five different temperature ranges (spanning 265-340 K). These propensities show that the hydrophobicity becomes weaker at lower temperatures, in line with current theory. Alternatively, one can conclude that the temperature dependence of the hydrophobic effect has a measurable influence on protein structures. Moreover, this work provides a method for probing the individual temperature dependence of the different amino acid types, which is difficult to obtain by direct experiment.     

大豆蛋白对急性放射性肠炎大鼠血清氨基酸的影响

为确定大豆蛋白对急性放射性肠炎大鼠血清氨基酸的影响 ,用SD大鼠复制急性放射性肠炎动物模型 ,给予含大豆蛋白饲料 ,测量血清氨基酸含量。结果显示 :使用大豆蛋白后细菌移位率显著降低 ,血LPS含量和肠通透性均未见升高 ,回肠绒毛高度、肠全层厚度和粘膜厚度均高于实验对照组 ,血清谷氨酰胺和支链氨基酸的含量均显著高于对照组。说明大豆蛋白使大鼠血清谷氨酰胺和支链氨基酸的含量明显升高 ,可以提高急性放性射肠炎时的肠屏障功能。     

Biosynthesis of Protein Amino Acids in Plant Tissue Culture. III. Studies on the Biosynthesis of Arginine

Evidence from isotope competition studies and enzymic studies indicates that n-acetyl glutamic semialdehyde, alpha-n-acetyl-l-ornithine, l-ornithine and l-citrulline are intermediates between glucose and arginine in cells of Paul's Scarlet Rose. Evidence for the presence of alpha-n-acetyl-ornithine aminotransferase (E. C. 2.6.1.11) in cell-free extracts was obtained.     

由铬革屑制备可溶性蛋白及氨基酸的研究

成品铬革屑中蛋白质含量在 70 %以上 ,处理成小颗粒后与水、Ca(OH) 2 按 1∶4 0∶0 .4的比例混合 ,90℃反应 5h ,可溶性蛋白转化率近 6 0 % ,且在升温后加入Ca(OH) 2 有利于退鞣。以 6mol/L盐酸水解皮革蛋白粉制备复合氨基酸 ,水解 14h ,水解液以HD -I树脂脱色 ,氨基酸损失较少 ,且动态脱色效率明显高于静态脱色 ,采用 717树脂脱酸可获得pH4 .5 - 5的复合氨基酸溶液 ,总得率为 6 0 .1%。     

Protein Hypersaline Adaptation: Insight from Amino Acids with Machine Learning Algorithms

Traditional bioinformatics methods performed systematic comparison between the halophilic proteins and their non-halophilic homologues, to investigate the features related to hypersaline adaptation. Therefore, proposing some quantitative models to explain the sequence-characteristic relationship of halophilic proteins might shed new light on haloadaptation and help to design new biocatalysts adapt to high salt concentration. Five machine learning algorithm, including three linear and two non-linear methods were used to discriminate halophilic and their non-halophilic counterparts and the prediction accuracy was encouraging. The best prediction reliability for halophilic proteins was achieved by artificial neural network and support vector machine and reached 80 %, for non-halophilic proteins, it was achieved by linear regression and reached 100 %. Besides, the linear models have captured some clues for protein halo-stability. Among them, lower frequency of Ser in halophilic protein has not been report before.     

Analysis of Protein Three-Dimension Structure Using Amino Acids Depths

The issue of amino acid depth in proteins gives important insights to our understanding of protein’s three-dimensional structure. There has already been much research done in mathematical and statistical sciences regarding the general definitions, properties and algorithms describing the particle depth of spatially extended systems. We constructed a method of calculating the amino acids depths and applied it to a set of 527 protein structures. We propose the introduction of amino acid depth tendency factors for three-dimensional structures of proteins. The depth tendency factors relate not only to the hydrophobicity indices but also to the electrostatic charge. We found a relationship between the protein size and the number of residues using the distance between the deepest residue and surface residues. We made a prediction regarding the number of residues on the surface of a protein, the deepest amino acid, and the average depth, all of which are fitted well to a linear functional relationship with the length of the protein. Finally, we have predicted the depths of multiple peptides in protein’s three-dimension structure. Electronic supplementary material The online version of this article () contains supplementary material, which is available to authorized users.     

Selective Acceleration of Arachidonic Acid Reincorporation into Brain Membrane Phospholipid Following Transient Ischemia in Awake Gerbil

Abstract: Awake gerbils were subjected to 5 min of forebrain ischemia by clamping the carotid arteries for 5 min and then allowing recirculation. Radiolabeled arachidonic or palmitic acid was infused intravenously for 5 min at the start of recirculation, after which the brains were prepared for quantitative autoradiography or chemical analysis. Dilution of specific activity of the acyl-CoA pool was independently determined for these fatty acids in control gerbils and following 5 min of ischemia and 5 min of reperfusion. Using a quantitative method for measuring regional in vivo fatty acid incorporation into and turnover within brain phospholipids and determining unlabeled concentrations of acyl-CoAs following recirculation, it was shown that reperfusion after 5 min of ischemia was accompanied by a threefold increase compared with the control in the rate of reincorporation of unlabeled arachidonate that had been released during ischemia, whereas reincorporation of released palmitate was not different from the control. Selective and accelerated reincorporation of arachidonate into brain phospholipids shortly after ischemia may ameliorate specific deleterious effects of arachidonate and its metabolites on brain membranes.     

用阴离子交换树脂脱酸从蛋白质水解液制备混合氨基酸的研究

<正> 本文报导了用阴离子交换树脂脱酸法从猪血水解液制备混合氨基酸的新工艺。该工艺与阳离子交换树脂吸附工艺相比有着工艺简单、产品得率高(67%)和消耗化工原料少的优点。产品中氨基酸的总含量约为80%,其中Lys约8%,Thr约3.5%,His约5.5%。动物试验的结果表明该产品无毒。在小麦面粉中加入3%的该产品,可使小麦面粉蛋白质的利用效率提高一倍。据初步估算,投入批量生产后,该产品的出厂价格在10元以下。从经济的角度考虑,该产品用于医药、食品和饲料是有希望的。     

Metabolic Engineering for Microbial Production of Aromatic Amino Acids and Derived Compounds

Metabolic engineering to design and construct microorganisms suitable for the production of aromatic amino acids and derivatives thereof requires control of a complicated network of metabolic reactions that partly act in parallel and frequently are in rapid equilibrium. Engineering the regulatory circuits, the uptake of carbon, the glycolytic pathway, the pentose phosphate pathway, and the common aromatic amino acid pathway as well as amino acid importers and exporters that have all been targeted to effect higher productivities of these compounds are discussed.     

Biosynthesis of Protein Amino Acids in Plant Tissue Culture IV Isotope Competition Experiments using Glucose-U-C and Potential Intermediates

The isotope competition method with glucose-U-¹⁴C as a carbon source has been used to determine whether or not selected compounds contribute carbon to the biosynthesis of protein amino acids in cells derived from Paul's Scarlet Rose. Of 48 compounds tested, 15 contributed carbon to protein amino acids. The results show for the first time that homoserine is an intermediate in threonine biosynthesis; that homoserine, cystathionine and homocysteine are intermediates in methionine biosynthesis and that histidinol is an intermediate in histidine biosynthesis in plant cells.