Mole Of Glycine Research Articles

Bacterial growth yields on EDTA, NTA, and their biodegradation intermediates

Ethylenediaminetetraacetic acid (EDTA) and nitrilotriacetic acid (NTA) are widely used anthropogenic chelating agents for control of metal speciation and are ubiquitous in natural waters and wastewaters. This is the first report of systematic measurement of the growth yields of a mixed culture (BNC1-BNC2) on EDTA and its biodegradation intermediates, and of Aminobacter aminovorans (aka Chelatobacter heintzii) ATCC 29600 on NTA and its biodegradation intermediates. The yields measured for BNC1-BNC2 co-culture were 75.0 g of cell dry weight (CDW) (mole of EDTA)(-1), 68.6 g of CDW (mole of ED3 A)(-1), 51.2 g of CDW (mole of N,N'-EDDA)(-1), 34.5 g of CDW (mole of ED)(-1), 26.3 g of CDW (mole of IDA)(-1), 12.2 g of CDW (mole of glycine)(-1), and 9.7 g of CDW (mole of glyoxylate)(-1). The yields measured for A. aminovorans were 44.3 g of CDW (mole of NTA)(-1), 37.9 g of CDW (mole of IDA)(-1), 15.2 g of CDW (mole of glycine)(-1), and 10.4 g of CDW (mole of glyoxylate)(-1). The biodegradation pathways of EDTA, NTA, and several of their metabolic intermediates include reactions catalyzed by oxygenase enzymes, which may reduce energy available for cell synthesis. Comparison of measured yields with predicted yields indicates that the effect of oxygenase reaction on cell yield can be quantified experimentally as well as modeled based on thermodynamics.

Synthesis of YAG powder by aluminum nitrate–yttrium nitrate–glycine reaction

Yttrium aluminum garnet (YAG) powders of controlled size distribution find application in the synthesis of optically transparent solid state laser components, advanced engineering materials and composites, etc. [1, 2]. Solution combustion is one of the simple methods for synthesizing the powder. In this technique, from a very concentrated solution of the metal nitrates a very porous sponge-like oxide compound is formed due to the evolution of a large amount of gaseous products formed in combustion reaction between an oxidizer (nitrate) and a fuel (urea, glycine etc.) present in the reactant system. The salient feature of combustion reaction is that it is exothermic and self sustaining. In this study the fuel, glycine, is chosen for formation of YAG since the amount of heat evolved and kinetics of the reaction are such that the combustion proceeds smoothly without bursting or spilling out of the product, a desirable aspect from the point of view of scaling up. The oxide formed is weakly agglomerated and needs to be ground into a fine powder before forming into shape. Formation of amorphous precursors for yttria–alumina compound phases by the nitrate–glycine reaction (with lean amount of fuel) and evolution of crystalline phases upon heat treatment has been studied by XRD [3]. However a study of the thermal evolution behavior by TG–DTA of the precursors formed by combustion to obtain temperature regimes of weight loss and heat effects (evolution/absorption) before and during crystallization of YAG is essential in obtaining conditions for formation of chemically pure YAG (Y3Al5O12 oxide compound). A study of the grinding behavior of the agglomerated mass formed is important from the point of view of its dispersion behavior, as the presence of coarse agglomerates reduces sinterability and homogeneity of microstructure of the product formed. As no study on these aspects has been reported yet for the precursors formed by the nitrates–glycine reaction (with stoichiometric amount of fuel), a detailed investigation of the same has been carried out. The stock solutions of aluminum nitrate (1.54 M) and yttrium nitrate (1.17 M) in the required molar ratio (5 : 3) for a batch size of 20 g. of YAG were placed in a pyrex beaker and the required amount of glycine was added. The ideal molar ratio of fuel (glycine) to oxidizer (nitrate ion) to be used was calculated using the following reaction and is 5/9 or 0.556 (i.e., 5 moles of glycine requires 9 moles of nitrate ions) [4]

Novel antibiotics, amythiamicins. II. Structure elucidation of amythiamicin D.

The structure of a unique polythiazole-containing cyclic peptide antibiotic, amythiamicin D, was elucidated by chemical degradations and NMR spectral analyses. Acid hydrolysis of amythiamicin D gave one mole of glycine and three new amino acids. Structures of N-acetyl-O-methyl derivatives of these new amino acids were determined by NMR and UV spectral analyses. Connectivities of these amino acids were determined by HMBC experiments.

Studies of the oxidation of iminodiacetic acid (IDA) and nitrilotriacetic acid (NTA) with lead dioxide suspension in nitric acid

The stoichiometry of the oxidation of IDA or NTA with lead dioxide suspension was studied by polarographic measurement and by derivative polarographic titration. One mole and two moles of Pb(IV) are reduced per mole of IDA and NTA respectively, with moderate speed at room temperature in nitric acid solutions. One mole each of carbon dioxide, formaldehyde and glycine are produced from the oxidation of 1 mole of IDA, and two moles of carbon dioxide, two moles of formaldehyde and one mole of glycine from 1 mole of NTA. The overall reaction in each case may be written as follows: Pb(IV) + IDA + H 2O → Pb(II) + CO 2 + HCHO + H 2NCH 2COOH + 2H + 2Pb(IV) + NTA + 2H 2O → 2Pb(II) + 2CO 2 + 2HCHO + H 2NCH 2COOH + 4H +

Characterization of the two major species of slow reacting substance from rat basophilic leukemia cells as glutathionyl thioethers of eicosatetraenoic acids oxygenated at the 5 position. Evidence that peroxy groups are present and important for spasmogenic activity

The most prominent slow reacting substance from rat basophilic leukemia cells (type I) was characterized by radiochemical, chemical and physical methods and shown to contain a C 20 unsaturated fatty acid oxygenated at the 5 position and a sulfur containing side chain in thioether linkage at the 6 position. Its spasmogenic action on guinea pig ileal muscle was largely inactivated under reducing conditions which suggested that a peroxy group was present and important for contractile activity. This was supported by ferrous thiocyanate analysis. The peroxy group is almost certainly at the 5 position, probably in the form of a peroxy ester or hydroperoxide. Based on amino acid hydrolysis (0.85 moles of glycine and 0.30 moles of glutamic acid per mole SRS), the sulfur containing side chain is apparently a mixture of glutathione and cysteinyl-glycine, but by chromatography the side chain is predominantly glutathione and the low yield of glutamic acid may be due to complexing of its α COOH group in a peroxy ester linkage. The fatty acid moiety has 3 conjugated double bonds, probably at the 7,8, 9,10 and 11,12 positions. Type II SRS, the second major species, differs in that the sulfur containing side chain is linked at the 12 or 13 position and is almost certainly glutathione and in the failure of alkaline borohydride to produce inactivation. These observations strongly implicate the lipoxygenase pathway in slow reacting substance biosynthesis.

Isolation and some properties of arachin subunits.

Six kinds of arachin subunits were isolated by isoelectric focusing in sucrose density gradient in the presence of 6M urea and 0.2M, I-mercaptoethanol. These subunits (S1 to S6) had different isoelectric points (pI 5.8, 6.0, 6.3, 7.1, 7.4 and 8.3) and they were found in parent arachin in the weight ratio of 2.5:2.2:2.6:1.6:1.1:1.0. The molecular weights and N-terminal amino acids of the isolated subunits were 35, 500 and valine (S1), 37, 500 and isol-eucine (S2), 40, 500 and isoleucine (S3) and 19, 500 and glycine (S4, S5 and S6). While, the parent arachin contained approximately one mole of valine, two moles of isoleucine and three moles of glycine as the N-terminal amino acids per 180, 000g. It is, therefore, concluded that arachin consists of six different subunits to form the molecular weight 180, 000. No disulfide bond takes part in the subunit association of arachin, because the dissociation in a urea solu-tion occurred in the absence of β-mercaptoethanol.

Fortimicins A and B, new aminoglycoside antibiotics. II. Isolation, physico-chemical and chromatographic properties.

The aminoglycoside antibiotics fortimicins A and B produced by a naturally occurring strain Micromonospora sp. MK-70 were isolated from its fermentation beer. Fortimicins A and B were isolated as water-soluble, basic, white amorphous powders having molecular formula C17H35N5O6 and C15H32N4O5, respectively. Acid hydrolysis of fortimicin A indicated that it has one mole of glycine in its molecule while fortimicin B has not. Paper chromatography, silica-gel and carbon thin-layer chromatography revealed that fortimicins A and B are novel aminoglycoside antibiotics.

The physiological response of the estuarine clam, Rangia cuneata (Gray), to salinity. II. Uptake of glycine.

1. The effect of salinity on the uptake of dissolved C14-labeled glycine by Rangia cuneata and isolated gill tissue was studied.2. The percentage water in Rangia tissue ranged from 81% at 1‰ S to 73% at 32‰ S, while the ash-free dry weights at these same salinities were 2.5% and 7.6%, respectively. The per cent water and per cent ash values were linear throughout this salinity range.3. The uptake of glycine was relatively uniform at salinities between 6 and 32‰ for whole animals and between 10 and 25‰ for gill tissue.4. At salinities below 6‰ uptake decreased sharply and in a linear fashion to a low at 1‰ S. Suppression of uptake was also exhibited when gill tissue was exposed for 90 minutes at 32‰ S.5. With increasing salinity the percentage utilization of accumulated glycine increased, but the rate of glycine incorporation into the alcohol insoluble fraction decreased.6. Removal of glycine by gill tissue from salinities of 10 to 25‰ occurred at a relatively linear rate up to 90 minutes and the levels obtained were considerably higher than those for whole animals when compared on a basis of cpm per mg ashfree dry weight.7. In a constantly flowing system Rangia at 15‰ S removed 0.16 µ moles of glycine/g/hour for a period of 11 hours, when supplied with 5.2 µ moles of glycine per liter.8. The significance of the relationship between salinity and glycine uptake, and possible explanations for this relationship are discussed.

The covalently-bound flavin of hepatic monoamine oxidase. 2. Identification and properties of cysteinyl riboflavin.

In the previous paper in this series it was shown that a pure flavin pentapeptide isolated from hepatic monoamine oxidase contains 1 mole each of serine and tyrosine and 2 moles of glycine, as well as an additional amino acid which is covalently linked to the 8α carbon of riboflavin. This amino acid has been identified as cysteine and the linkage to the flavin as a thioether on the basis of positive chloroplatinic and negative iodine‐azide tests, performic acid oxidation or reduction with zinc, followed by acid hydrolysis, which yield cysteic acid or cysteine, respectively. The fluorescence properties of the flavin from monoamine oxidase agree with those expected for a thioether and its oxidation products. In regard to optical and electron spin resonance spectra, chemical stability, and fluorescence characteristics the flavin peptide isolated from the enzyme agrees excellently with the properties of synthetic cysteinyl 8α‐riboflavin.

Structural Properties of Guinea Pig Liver Transglutaminase

Evidence is presented that transglutaminase is composed of a single polypeptide chain of molecular weight 80,000 to 90,000. (a) Polyacrylamide gel electrophoresis in the presence of dodecyl sulfate and mercaptoethanol gave a single band with a mobility corresponding to a molecular weight of approximately 85,000. (b) Gel filtration in guanidine HCl of the 14C-carbamidomethyl carboxymethylated enzyme protein showed a single peak of absorbance and radioactivity from which a molecular weight of approximately 85,000 was estimated. (c) Amino-terminal analysis by conventional methods showed no evidence of free α-amino groups. A peptide, believed to contain the amino-terminal residue, was obtained by Pronase digestion and was isolated at levels of 0.75 and 0.8 mole/90,000 g of enzyme. The sequence of this peptide was determined as pyroglutamylalanylaspartylleucine. (d) Digestion by carboxypeptidase A of the carboxymethylated enzyme protein in denaturing solvents released glycine and serine at equal rates to the level of 1 mole/90,000 g of protein. Hydrazinolysis gave approximately 1 mole of glycine. These findings, together with earlier evidence that the molecular weight of the native enzyme is 80,000 to 90,000 and that the enzyme protein contains 17 or 18 —SH groups, but no disulfide bonds, form the basis for the view of an unbridged monomeric structure of transglutaminase. Indication that transglutaminase performs its catalytic functions in the monomeric form was obtained from a comparison of the gel filtration patterns for the enzyme protein in the presence and absence of calcium ion. The identical nature of these patterns is in accord with the suggestion that this metal, which is essential for activation of transglutaminase, does not affect a change in enzyme molecular weight. A revised enzyme purification procedure is presented. Rabbit antiserum against transglutaminase has been prepared and used to characterize the enzyme purified by this procedure as immunologically homogeneous.

Tryptophanyl Transfer Ribonucleic Acid Synthetase from Bovine Pancreas: II. The chemically different subunits

Abstract Bovine pancreas tryptophanyl transfer RNA synthetase, a crystalline enzyme, is shown by filtration on a carefully calibrated Sephadex G-150 column and by equilibrium centrifugation to have a molecular weight of 110,000 (s20,w = 5.6) in the presence of 10-4 M tryptophan. When the enzyme is treated with 4 M guanidine-hydrochloride for several days or with 8 M urea, values for the s20,w become 1.6 and 2.0, respectively. The enzyme dissociates slowly (to s20,w = 3.0) on dialysis against Tris-glycine buffer, pH 8.5, or 0.1 M ammonium bicarbonate, pH 8.8. A very slow dissociation is caused by 0.2 M β-mercaptoethanol which is rapidly speeded up on the addition of 1% sodium dodecyl sulfate. Titration of native enzyme with 5,5'-dithiobis(2-nitrobenzoate) indicates the presence of four free —SH groups; titration in the presence of 8 M urea indicates six —SH groups; titration of sodium borohydride-reduced enzyme indicates 14 —SH groups per molecule of 110,000 molecular weight. The enzyme has been reduced with 0.2 M β-mercaptoethanol in 8 M urea and carboxymethylated with 14C-iodoacetic acid. The carboxymethylated protein is eluted as a single peak in a position corresponding to a particle with a molecular weight of 27,000 to 30,000 from a Sephadex G-150 column. Sedimentation equilibrium experiments show a single molecular weight species of 27,000 (s20,w = 1.9). However, chromatography on carboxymethyl-Sephadex separates the carboxymethylated protein into two distinct radioactive protein peaks. The carboxymethylated proteins were acetylated, digested with trypsin, and subjected to peptide mapping. A total of seven 14C-cysteine peptides and 22 arginine-containing peptides was identified in both protein species, while the amino acid composition of the total unfractionated carboxymethylated protein shows 14 carboxymethylcysteine residues and 44 arginine residues per molecule of 110,000 molecular weight. The amino acid compositions of the two separated carboxymethylated proteins show significant differences, one carrying 6 half-cysteine residues, the other only 1. Catalytic hydrazinolysis of the individual carboxymethylated proteins gives 1 mole of glutamic acid and 1 mole of glycine, respectively, per 27,000 molecular weight. These experiments lead to the conclusion that a molecule of tryptophanyl-tRNA synthetase is made up of four subunits per molecular weight of 110,000. These subunits consist of two kinds of monomers with different primary structures. Tryptophanyl-tRNA synthetase activity is not restricted to the tetrameric protein but is also found in a smaller molecular species (s20,w = 3.0 to 3.3). This molecule is found naturally in preparations of the enzyme and is also formed on dissociation of the tetrameric enzyme. These findings and those reported in the accompanying paper suggest that the catalytic form of the enzyme, the protomer, may be a dimer, and that the tetrameric form has no special functional significance and most likely is an artifact produced by the presence of tryptophan during the purification procedure.

Die aminosäuresequenz des mureins von Microbacterium lacticum

Cell walls of Microbacterium lacticum were purified by digestion with trypsin and extraction with trichloroacetic acid. The amino acid analysis showed that in addition to the usual amino sugars the murein (peptidoglycan) contained glutamic acid, lysine, glycine and d-alanine in the molar ratio of about 1:2:2:1, but no ammonia. The dinitrophenylation of the cell wall yielded ε-DNP-lysine (66% of the total amount of lysine). The hydrazinolysis showed that at least 40% of the lysine, 6% of d-alanine and 2.6% of glycine are C-terminal. From partial acid hydrolysate oligopeptides were isolated by paper chromatography and identified. From these data and those mentioned above, the following scheme of the amino acid sequence and mode of crosslinking of the main part of the murein could be proposed. The tetrapeptide bound to muramic acid showed the sequence Gly-Glu-Lys- d-Ala. The second mole of glycine and of lysine, respectively, is involved in the crosslinking. They form the peptide N 2-glycyllysine which is bound by its glycine end to glutamic acid and by the ε-amino group of lysine to d-alanine of an adjacent tetrapeptide. In contrast to other mureins the ε-amino group of the lysine within the tetrapeptide is free. Only about 60% of the tetrapeptides are actually crosslinked. In the other cases, the ε-amino group of the lysine is free or the crosslinking peptide is incomplete. Various data indicate that some tetrapeptides are also incomplete. In this case the C-terminal d-alanine, and sometimes also the lysine, are missing. The murein of M. lacticum resembles the unusual mureins of Butyribac- terium rettgeri and Corynebacterium poinsettiae.

Studies on the amino acid sequence of tobacco mosaic virus (TMV) protein. I. Fractionation of products of tryptic hydrolysis by countercurrent distribution

Tobacco mosaic virus (TMV) protein (mol. wt. of subunit 18,270) was hydrolyzed with trypsin. The material soluble at pH 4.0 was fractionated by countercurrent distribution with the solvent system 2-butanol-0.1 M dichloroacetic acid. Preliminary investigations on these fractions have shown that nine apparently major tryptic hydrolysis products containing arginine and one containing lysine have been obtained from this fractionation. One arginine-containing peptide of large molecular weight was found in the portion of the digest insoluble at pH 4.0. One of these peptides has been found to contain arginine in both the N- and C-terminal positions. The nature and number of the N- and C-terminal residues produced as a result of the action of trypsin on TMV protein have been investigated. These results have revealed the presence of about one mole each of N-terminal glutamic acid, aspartic acid, serine, threonine, phenylalanine, tyrosine, and arginine and approximately two moles of glycine. Fractional amounts of N-terminal leucine and valine were also found. One mole each of C-terminal threonine and lysine and slightly more than nine moles of C-terminal arginine were determined.

Alkali-catalyzed ring closure of pyruvoylglycine

The product of the treatment of pyruvoylglycine by alkali at 25 ° has been isolated in crystalline form. The compound may exist either as the monohydrate or in the anhydrous form. The empirical composition of the anhydrous product is identical with that of pyruvoylglycine, it possesses a pK′ of 3.8 and forms a monosodium salt. On destructive distillation with lime, products giving characteristic pyrrole reactions are formed. The absorption spectra in the ultraviolet and in the infrared resemble those of pyrrolidone carboxylic acid, and on hydrolysis in 5 N HCl close to 1 mole of dl-alanine is formed. (Pyruvoylglycine under the same conditions yields 1 mole of glycine.) On the basis of these and previous findings, it is concluded that the treatment of pyruvoylglycine by alkali under mild conditions leads to an irreversible ring closure and the formation of the isomeric γ-hydroxypyrrolidone carboxylic acid. This ring closure, leading to the establishment of two centers of asymmetry, would explain in part the origin of the racemic alanine formed on treatment with boiling 5 N HCl.