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1.
An enzyme has been discovered in Escherichia coli that catalyzes the conversion of the triphosphate ester of 2-amino-4-hydroxy-6-(d- erythro-1′,2′,3′-trihydroxypropyl)-7,8-dihydropteridine, (i.e. d- erythro-dihydroneopterin triphosphate) to an epimer of this compound, l- threo-dihydroneopterin triphophate. The enzyme, which is here named “d- erythro-dihydroneopterin triphosphate 2′-epimerase,” needs a divalent cation (Mg 2+ or Mn 2+ is most effective) for maximal activity. Its molecular weight is estimated at 87 000–89 000. Little or no activity can be detected if either the monophosphate or the phosphate-free form of the substrate is incubated with the enzyme. Evidence is presented to establish that all three phosphate residues of the substrate are retained in the product and that the product is of the l- threo configuration. 相似文献
2.
Three-dimensional, single-crystal, X-ray diffraction methods were used to determine the structure of the calcium chloride complex of α-d-allopyranosyl α-d-allopyranoside. The crystal is monoclinic with cell dimensions: a = 16.262(5), b = 8.345(5), c = 8.298(5)Å, β = 98.428(5)Å, and z = 2. The space group is P2 1. The structure was solved by three-dimensional Patterson and Fourier methods, and was refined by least-squares techniques to give a conventional discrepancy factor of R = 0.026; 2435 diffractometer-read reflections were used. The cation was found to be in 9-fold co-ordination to O-1, O-2, O-3, O-2′, O-3′, and four water molecules. 相似文献
3.
NMR titration curves have been recorded for all the 13C resonances of cis and transN-acetyl -dl -proline in 2H 2O. the measured p K2H values are 3.4 ± 0.8 and 4.13 ± 0.08 respectively; the free energy of ionization for the trans isomer being (3.8 kJ/mole) greater than for the cis. The ionization shifts of the two isomers differ significantly only at the acetyl carbonyl and C γ positions. It is suggested that these are related to conformational changes which stabilize the trans form at low p 2H. 相似文献
4.
A capillary gas chromatographic—mass spectrometric method for the simultaneous determination of stable isotopically labelled l-histidine (l-[3,3- 2H 2,1′,3′- 15N 2]histidine, l-His-[M + 4]) and urocanic acid ([3- 2H,1′,3′- 15N 2]urocanic acid, UA-[M + 3]) in human plasma was developed using dl-[2,3,3,5′- 2H 4,2′- 13C,1′,3′- 15N 2]histidine (dl-His-[M + 7]) and [2,3,5′- 2H 3,2′- 13C,1′,3′- 15N 2]urocanic acid (UA-[M + 6]) as internal standards. l-Histidine and urocanic acid were derivatized to αN-(trifluoroacetyl)- imN-(ethoxycarbonyl)-l-histidine n-butyl ester and imN-(ethoxycarbonyl)urocanic acid n-butyl ester. Quantification was carried out by selected ion monitoring of the molecular ions of the respective derivatives of l-His-[M + 4], dl-His-[M + 7], UA-[M + 3] and UA-[M + 6]. The sensitivity, specificity, precision and accuracy of the method were demonstrated to be satisfactory for measuring plasma concentrations of l-His-[M + 4] and UA-[M + 3] following administration of trace amounts of l-His-[M + 4] to humans. 相似文献
5.
The cyclic dodecapeptide PV, cyclo-(d-Val-l-Pro-l-Val-d-Pro )3, a structural analogue of the ion-carier valinomycin, increase the cation permeability of lipid bilayer membranes. This paper reports the results of two types of relaxation experiments, namely relaxation of the membrane current after a voltage jump and decay of the membrane voltage after a charge pulse in lipid bilayer membranes exposed to PV. From the relaxation data, the rate constant for the translocation of the ion carrier complex across the membrane, as well as the partition coefficient of the complex between water and membrane solution interface were computed and found to be about one order of magnitude less than the comparable values for valinomycin (Val). Furthermore, the dependence of the initial membrane conductivity on ion concentration was used to evaluate the equilibrium constant, K, of complexation between PV and some monovalent cations in water. The values of K yield the following selectivity sequence of PV: Na + < NH 4+ < K + < Cs + < Rb +. These and earlier results are consistent with the idea that PV promotes cation movement across membranes by the solution complexation mechanism which involves complexation between ion and carrier in the aqueous phase and transport of the carrier across the membrane. In the particular form of the solution complexation mechanism operating here, the PV present in the PV-cation complex carrying charge across the membrane derives from the side from which the current is flowing (cis-mechanism). As shown previously, valinomycin, in contrast to PV, acts by an interfacial complexation mechanism in which the Val in the Val-cation complex derives from the side toward which current is flowing (trans-mechanims). The comparison of the kinetic properties of these two closely related compounds yields interesting insights into the relationship between chemical structure and function of ion carriers. 相似文献
6.
When aromatic hydrocarbon ( Ah)-responsive and -non-responsive strains of mice were pretreated with 3-methylcholanthrene (MC) or 2,3,7,8-tetrachlorodibenzo- p-dioxin (TCDD), vitamin C reduced the microsomal aryl hydrocarbon hydroxylase (AHH) activity. The AHH inhibitors 7,8-benzoflavone (7,8-BF) and 3-methylsulfonyl-3′,4,4′,5-tetrachlorobiphenyl (3-MSF-3′,4,4′,5-tetraCB) showed various inhibitory effects depending upon the types of microsomes, whereas vitamin C exhibited inhibition irrespective of the types of microsomes. 7,8-BF and 3-MSF-3′,4,4′,5-tetraCB as well as vitamin C suppressed the reverse mutation of the Salmonella typhimurium tester strains TA98 and TA100 induced by benzo[ a]pyrene. 相似文献
7.
A panel of six complementary monodeoxy and mono- O-methyl congeners of methyl β- d-mannopyranosyl-(1→2)-β- d-mannopyranoside ( 1) were synthesized by stereoselective glycosylation of monodeoxy and mono- O-methyl monosaccharide acceptors with a 2- O-acetyl-glucosyl trichloroacetimidate donor, followed by a two-step oxidation–reduction sequence at C-2′. The β- manno configurations of the final deprotected congeners 2–7 were confirmed by measurement of 1JC1,H1 heteronuclear and 3J1′,2′ homonuclear coupling constants. These disaccharide derivatives will be used to map the protective epitope recognized by a protective anti- Candida albicans monoclonal antibody C3.1 (IgG3) and to determine its key polar contacts with the binding site. 相似文献
8.
A general method for the preparation of 2′-azido-2′-deoxy- and 2′-amino-2′-deoxyarabinofuranosyl-adenine and -guanine nucleosides is described. Selective benzoylation of 3-azido-3-deoxy-1,2- O-isopropylidene-α-d-glucofuranose afforded 3-azido-6- O-benzoyl-3-deoxy-1,2- O-isopropylidene-α-d-glucofuranose ( 1). Acid hydrolysis of 1, followed by oxidation with sodium metaperiodate and hydrolysis by sodium hydrogencarbonate gave 2-azido-2-deoxy-5- O-benzoyl-d-arabinofuranose ( 3), which was acetylated to give 1,3-di- O-acetyl-2-azido-5- O-benzoyl-2-deoxy-d-arabinofuranose ( 4). Compound 4 was converted into the 1-chlorides 5 and 6, which were condensed with silylated derivatives of 6-chloropurine and 2-acetamido-hypoxanthine. The condensation reaction gave α and β anomers of both 7- and 9-substituted purine nucleosides. The structures of the nucleosides were determined by n.m.r. and u.v. spectroscopy, and by correlation of the c.d. spectra of the newly prepared nucleosides with those published for known purine nucleosides. 相似文献
9.
Optically pure 2-acetamido-2-deoxy-3- O-α-L-fucopyranosyl-α-D-glucose was synthesized by the Koenigs-Knorr reaction of 2- O-benzyl-3,4-di- O- p-nitrobenzoyl-α-L-fucopyranosyl bromide with benzyl 2-acetamido-4,6- O-benzylidene-2-deoxy-α-D-glucopyrainoside. Reaction of 2,3,4-tri- O-acetyl-α-L-fucopyranosyl bromide gave the β-L-fucopyranosyl anomer. In contrast to the stereospecificity shown in this reaction by these two bromides, 2,3,4-tri- O-benzyl-α-L-fucopyranosyl bromide afforded a mixture of α-L and β-L anomers in almost equimolar proportions. The disaccharides synthesized were crystallized and characterized, and their optical purity demonstrated by g.l.c. of the per(trimethylsilyl) ethers of the corresponding alditols. 相似文献
10.
The X-ray diffraction patterns, 13C CP MAS NMR spectra, and powder X-ray diffraction analyses were obtained for selected p-nitrophenyl glycosides: α- and β- d-galactopyranosides ( 1 and 2), α- and β- d-glucopyranosides ( 3 and 4), and α- and β- d-mannopyranosides ( 5 and 6). In X-ray diffraction analysis of 1 and 2, characteristic shortening and lengthening of selected bonds were observed in the molecules of 1 due to anomeric effect, and in the crystal lattice of 1 and 2, hydrogen bonds of complex network were detected. In the crystal asymmetric unit of 1 there were two independent molecules, whereas in 2 there was one molecule. For 1 and 3– 6 the number of resonances in solid-state 13C NMR spectra exceeded the number of the carbon atoms in the molecules, while for 2 there were distinct singlet resonances in its solid-state NMR spectrum. Furthermore, the powder X-ray diffraction (PXRD) performed for 1– 3 and 5 revealed that 1, 3, and 5 existed as single polymorphs proving that the doublets observed in appropriate solid-state NMR spectra were connected with two non-equivalent molecules in the crystal asymmetric unit. On the other hand 2 existed as a mixture of two polymorphs, one of them was almost in agreement with the calculated pattern obtained from XRD (the difference in volumes of the unit cells), and the subsequent unknown polymorph existed in small amounts and therefore it was not observed in solid-state NMR measurements. 相似文献
11.
The gene encoding α-methylserine aldolase was isolated from Bosea sp. AJ110407. Sequence analysis revealed that the predicted amino acid sequence encoded by the 1320-bp open reading frame was 65.0% similar to the corresponding sequence of the enzyme isolated from Ralstonia sp. AJ110405. The gene was expressed in Escherichia coli, and the recombinant enzyme was purified. Gel filtration revealed the molecular mass of the purified enzyme to be approximately 78 kDa, suggesting that the enzyme is a homodimer. The enzyme exhibited a specific peak at 429 nm in the spectrum and contained 1 mol pyridoxal 5′-phosphate per mole of the subunit. The Vmax value was 1.40 μmol min −1 mg −1, and the Km value was 1.5 mM for the reaction wherein formaldehyde was released from α-methyl- l-serine. This enzyme could also catalyze the reverse reaction, i.e., the synthesis of α-methyl- l-serine from l-alanine and formaldehyde. This activity was inhibited in the excess of formaldehyde; however, α-methyl- l-serine was efficiently produced from l-alanine in the presence of formaldehyde. This method was also applicable for producing α-ethyl- l-serine from l-2-aminobutyric acid. 相似文献
12.
Benzoylation of benzyl 2-acetamido-2-deoxy-4,6- O-isopropylidene-α-d-glucopyranoside, benzyl 2-deoxy-2-(dl-3-hydroxytetradecanoylamino)-4,6- O-isopropylidene-α-d-glucopyranoside, and benzyl 2-deoxy-4,6- O-isopropylidene-2-octadecanoylamino-β-d-glucopyranoside, with subsequent hydrolysis of the 4,6- O-isopropylidene group, gave the corresponding 3- O-benzoyl derivatives ( 4, 5, and 7). Hydrogenation of benzyl 2-acetamido-4,6-di- O-acetyl-2-deoxy-3- O-[d-1-(methoxycarbonyl)ethyl]-α-d-glucopyranoside, followed by chlorination, gave a product that was treated with mercuric actate to yield 2-acetamido-1,4,6-tri- O-acetyl-2-deoxy-3- O-[d-1-(methoxycarbonyl)ethyl]-β-d-glucopyranose ( 11). Treatment of 11 with ferric chloride afforded the oxazoline derivative, which was condensed with 4, 5, and 7 to give the (1→6)-β-linked disaccharide derivatives 13, 15, and 17. Hydrolysis of the methyl ester group in the compounds derived from 13, 15, and 17 by 4- O-acetylation gave the corresponding free acids, which were coupled with l-alanyl-d-isoglutamine benzyl ester, to yield the dipeptide derivatives 19–21 in excellent yields. Hydrolysis of 19–21, followed by hydrogenation, gave the respective O-( N-acetyl-β-muramoyl-l-alanyl-d-isoglutamine)-(1→6)-2-acylamino-2-deoxy-d-glucoses in good yields. The immunoadjuvant activity of these compounds was examined in guinea-pigs. 相似文献
13.
Injection of 3,5,3′ l-triiodothyronine (15 μg/100 g) induces a biphasic enhancement of rat heart ornithine decarboxylase (EC. 4.1.17) activity after 4 and 21 hours. This induction is observed after each daily injection, but to a lesser extent.The properties of partially purified basal enzyme and induced enzyme, at 21h, after single injections have been compared. 1) Affinity for ornithine is the same for both enzymes, but affinity for pyridoxal-phosphate is 40-fold higher for the induced one. 2) Thermostability studies suggest that basal and induced enzymes have different conformations. 3) The two enzymes have similar immunoreactivity. 4) The comparisons of the time-dependent activity curve after injection and of the antigen/activity ratio suggests that triiodothyronine induces the synthesis of new molecules of enzymes and that an inhibition of the enzyme activity also occurs which explains the biphasic induction.
RésuméL'injection de 3,5,3′ l-triiodothyronine (15 μg/100 g) induit une augmentation biphasique de l'activité ornithine décarboxylase (EC: 4.1.1.17) de cœur de rat à la 4 e et 21 e heure. Ce phénomène se reproduit après chaque injection quotidienne, mais de manière moins intense.Les enzymes, basale et induite à la 21 e heure après une injection de T 3, ont été partiellement purifiées et leurs propriétés comparées: 1) L'affinité des deux enzymes pour l'ornithine est identique, mais l'affinité pour le pyridoxal-phosphate est 40 fois plus élevée pour l'enzyme induite. 2) Les études de thermodénaturation suggèrent que les enzymes de base et induite présentent des conformations différentes. 3) Les deux enzymes présentent des immunoréactivités similaires. 4) Une comparaison de la courbe d'activité et du rapport antigène/activité enzymatique au cours du temps, après induction suggèrent que la triiodothyronine induit la synthèse de novo de l'enzyme, mais qu'intervient également un phénomène d'inhibition qui explique l'aspect biphasique de l'induction.
Mots-clés: régulation; ornithine décarboxylase; triiodothyronine; antizyme Keywords: regulation; ornithine decarboxylase; triiodothyronine; antizyme 相似文献
14.
Starting from 3β-hydroxy-17-oxo-16,17-secoandrost-5-ene-16-nitrile ( 1), the new 16,17-secoandrostane derivatives 4– 9 were synthesized. On the other hand, 3β-hydroxy-17-oxa- d-homoandrost-5-ene-16-one ( 10) yielded the new d-homo derivatives 12, 13 and 15. In vitro antiproliferative activity of selected compounds against three tumor cell lines (human breast adenocarcinoma ER+, MCF-7, human breast adenocarcinoma ER−, MDA-MB-231, prostate cancer AR−, PC-3, and normal fetal lung fibroblasts, MRC-5) was evaluated. Compounds 3 and 12 showed strong antiproliferative activity against PC-3 cells, the IC 50 values being 2 μM and 0.55 μM, respectively. Compounds 6 (10 μM) and 14 (9 μM) showed moderate activity against MDA-MB-231 cells. The synthesized compounds 1– 3, 5– 8, 10 and 12– 15 were not toxic to normal fetal lung fibroblasts cells, MRC-5. 相似文献
16.
The dapE-encoded N-succinyl- l, l-diaminopimelic acid desuccinylase (DapE) is a critical bacterial enzyme for the construction of the bacterial cell wall. A screen biased toward compounds containing zinc-binding groups (ZBG’s) including thiols, carboxylic acids, boronic acids, phosphonates and hydroxamates has delivered a number of micromolar inhibitors of DapE from Haemophilus influenzae, including the low micromolar inhibitor l-captopril (IC 50 = 3.3 μM, Ki = 1.8 μM). In vitro antimicrobial activity was demonstrated for l-captopril against Escherichia coli. 相似文献
17.
An N-acetyl-β-d-hexosaminidase has been purified from primary wheat leaves ( Triticum aestivum L.) by freeze-thawing, (NH 4) 2SO 4 precipitation, methanol precipitation, gel filtration, cation exchange chromatography and affinity chromatography on concanavalin A-Sepharose. The activity of the purified preparations could be stabilised by addition of Triton X-100 and the enzyme was stored at -20°C without significant loss of activity. The enzyme hydrolysed pNP-β-d-GlcNAc (optimum pH 5.2, Km 0.29 mM, Vmax 2.56 μkat mg −1) and pNP-β-d-GalNAc (optimum pH 4.4, Km 0.27 mM, Vmax 2.50 μkat mg −1). Five major isozymes were identified, with isoelectric points in the range 5.13–5.36. All five isozymes possessed both N-acety-β-d-glucosaminidase and N-acetyl-β-d-galactosaminidase activity. Inhibition studies and mixed substrate analysis suggested that both substrates are catalysed by the same active site. Both activities were inhibited by GlcNAc, 2-acetamido-2-deoxygluconolactone, GalNAc and the ions of mercury, silver and copper. The Kis for inhibition of N-acetyl-β-d-glucosaminidase activity were: GlcNAc (15.3 mM) and GalNAc (3.4mM). For inhibition of N-acety-β-d-galactosaminidase activity the corresponding values were: GlcNAc (18.2 mM) and GalNac (2.5 mM). The enzyme was considerably less active at releasing pNP from pNP-β-d-(GlcNAc) 2 and pNP-β-d-(GlcNAc) 3 than from pNP-β-d-GlcNAc. The ability of the N-acetyl-β-d-hexosaminidase to relase GlcNAc from chitin oligomers (GlcNAc) 2 (optimum pH 5.0) and (GlcNAc) 3−6 (optimum pH 4.4) was also low. Analysis of the reaction products revealed that the initial products from the hydrolysis of (GlcNAc) n were predominantly (GlcNAc) n−1 and GlcNAc. 相似文献
18.
Through the screening of microorganisms capable of utilizing α-methylserine, three representative strains belonging to the bacterial genera Paracoccus, Aminobacter, and Ensifer were selected as potent producers of α-methylserine hydroxymethyltransferase, an enzyme that catalyzes the interconversion between α-methyl- l-serine and d-alanine via tetrahydrofolate. Among these strains, Paracoccus sp. AJ110402 was selected as the strain exhibiting the highest α-methylserine hydroxymethyltransferase activity. The enzyme was purified to homogeneity from a cell-free extract of this strain. The native enzyme is a homodimer with apparent molecular mass of 85 kDa and contains 1 mol of pyridoxal-5′-phosphate per mol of the subunit. The Km for α-methyl- l-serine and tetrahydrofolate was 0.54 mM and 73 μM, respectively. The gene from Paracoccus sp. AJ110402 encoding α-methylserine hydroxymethyltransferase was cloned and expressed in Escherichia coli. Sequence analysis revealed an open reading frame of 1278 bp, encoding a polypeptide with a calculated molecular mass of 46.0 kDa. Using E. coli cells as whole-cell catalysts, 9.7 mmol of α-methyl- l-serine was stereoselectively obtained from 15 mmol of d-alanine and 13.2 mmol of formaldehyde. 相似文献
19.
2-Acetamido-3,4,6-tri- O-acetyl-2-deoxy-α,β-D-glucopyranosylammonium phosphate was prepared by the action of crystalline phosphoric acid on 2-acetamido-1,3,4,6-tetra- O-acetyl-β-D-glucopyranose. The α-D anomer ( 3) was the main product, and was isolated pure by preparative thin-layer chromatography or by removal of the β-D anomer ( 6) by selective acid hydrolysis. Ficaprenyl phosphate was prepared from ficaprenol, obtained as an isomeric mixture (mainly C 55) from an extract of Ficus elastica. Compound 3 was converted into the free acid and then into the tributyl-ammonium salt, which was treated with P1-diphenyl P2-ficaprenyl pyrophosphate to give the acetylated pyrophosphate diester 8, characterized by analytical, spectral, and hydrogenolytic studies. Deacetylation of 8 gave the synthetic “lipid intermediate”, P1-(2-acetamido-2-deoxy-D-glucopyranosyl) P2-ficaprenyl pyrophosphate ( 9), the properties of which were compared with those of natural substances considered to be active in the biosynthesis of teichoic acids. 相似文献
20.
d-Amino acid oxidase is a FAD-dependent enzyme that catalyses the conversion of the d-enantiomer of amino acids into the corresponding α-keto acid. Substrate specificity of the enzyme from the yeast Rhodotorula gracilis was investigated towards aromatic amino acids, and particularly synthetic α-amino acids.A significant improvement of the activity ( Vmax,app) and of the specificity constant (the Vmax,app/ Km,app ratio) on a number of the substrates tested was obtained using a single-point mutant enzyme designed by a rational approach. With R. gracilis d-amino acid oxidase the complete resolution of d, l-homo-phenylalanine was obtained with the aim to produce the corresponding pure l-isomer and to use the corresponding α-keto acid as a precursor of the amino acid in the l-form. 相似文献
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