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1.
TheN-linked carbohydrate chains of the-subunit of highly purified urinary human chorionic gonadotropin have been re-investigated. The oligosaccharides were released enzymatically by peptide-N 4-(N-acetyl--glucosaminyl)asparagine amidase-F, and fractionated by a combination of FPLC and HPLC. As a result of the application of improved fractionation methods, apart from the earlier reported carbohydrate chains, also small amounts of trisialo tri- and tri-antennary oligosaccharides were found. The primary structures of the latter carbohydrate chains have been determined by 500-MHz1H-NMR spectroscopy to beAbbreviations hCG human chorionic gonadotropin - hCG- -subunit - hCG- -subunit - PNGase-F peptide-N 4-(N-acetyl--glucosaminyl)asparagine amidase-F (E.C. 3.5.1.52) - endo-F endo--N-acetylglucosaminidase-F (E.C. 3.2.1.96) - SDS sodium dodecyl sulphate - PAGE polyacrylamide gel electrophoresis - CBB coomassie brilliant blue R 250 - GlcNAc N-acetylglucosamine - NeuAc N-acetylneuraminic acid - Man mannose - Gal galactose - Fuc fucose  相似文献   

2.
The study deals with a comparative analysis of the relative abundances of the carbon isotopes 12C and 13C in the metabolites and biomass of the Burkholderia sp. BS3702 and Pseudomonas putida BS202-p strains capable of utilizing aliphatic (n-hexadecane) and aromatic (naphthalene) hydrocarbons as sources of carbon and energy. The isotope compositions of the carbon dioxide, biomass, and exometabolites produced during the growth of Burkholderia sp. BS3702 on n-hexadecane (13C = –44.6 ± 0.2) were characterized by the values of 13CCO 2 = –50.2 ± 0.4, 13Cbiom = –46.6 ± 0.4, and 13Cexo = –41.5 ± 0.4, respectively. The isotope compositions of the carbon dioxide, biomass, and exometabolites produced during the growth of the same bacterial strain on naphthalene (13C = –21 ± 0.4) were characterized by the isotope effects 13CCO 2 = –24.1 ± 0.4, 13Cbiom = –19.2 ± 0.4, and 13Cexo = –19.1 ± 0.4, respectively. The possibility of using the isotope composition of metabolic carbon dioxide for the rapid monitoring of the microbial degradation of petroleum hydrocarbons in the environment is discussed.  相似文献   

3.
Araus  J.L.  Casadesús  J.  Asbati  A.  Nachit  M.M. 《Photosynthetica》2001,39(4):591-596
The relationship between ash content and carbon isotope discrimination () was studied in durum wheat (Triticum durum Desf.) grown in a Mediterranean region (Northwest Syria) under three different water regimes (hereafter referred to as environments). In two of these environments, 144 genotypes were cultivated under rain-fed conditions. In the third environment, 125 genotypes were cultivated under irrigation. Ash content was measured in the flag leaf about 3 weeks after anthesis, whereas was analysed in mature kernels. Total transpiration of the photosynthetic tissues of the culm contributing, from heading to maturity, to the filling of kernels was also estimated. Leaf ash content, expressed either on dry matter or leaf area basis or as total ash per blade, correlated positively (p< 0.001) with in the three environments. However, this relationship was not the result of a positive correlation across genotypes between and tissue water content. Moreover, only a small part of the variation in across genotypes was explained by concomitant changes in ash content. When all genotypes across the three environments were plotted, and ash content followed a non-linear relationship (r 2 = 74), with tending to a plateau as the ash content increased. However, for the set of genotypes and environments combined, total ash content per leaf blade was positively and linearly related (r 2 = 0.76) with the accumulated culm transpiration. The non-linear nature of the relationship between ash content and is sustained by the fact that culm transpiration also showed a non-linear relationship with kernel . Therefore, differences in leaf ash content between environments, and to a lesser extent between genotypes, seem to be brought about by variations in accumulated transpiration during grain formation.  相似文献   

4.
Summary To investigate the impact of SOS induction on the distribution of spontaneous mutation, 111 recA441-mediated mutations were characterized at the DNA sequence level in the lacI gene of Escherichia coli. A 2.6-fold enhancement in lacI mutation frequency was observed after induction of the SOS system in the absence of mutagenic treatment, and specific classes of mutational events were induced. G : C C : G, G : C T : A and A : T T : A transversion events were specifically enhanced after SOS induction. A preferential 5-Y-Purine-3 neighbouring base specificity for these transversion events is reported here (normalised for mutation of the purine residue). In addition, a preference for transversion events at 5-C/GTGG-3 sequences is also observed. Fifty events were recovered at the lacI frameshift hotspot site and were equally represented by 4 bp addition and deletion events. This 1:1 ratio deviates significantly from the 4:1 distribution characteristic of spontaneous frameshift mutation in the RecA+ background and is a consequence of the fourfold induction of the (–)4 event. This abberrant distribution was confirmed by oligomeric probing of 474 independent recA441-mediated spontaneous lacI mutations.  相似文献   

5.
Uptake ofl-[35S]cysteic acid (L-CA) in rat synaptic membrane vesicles was investigated. Preincubation with either 10 mMl-glutamic acid (L-Glu), 25 mM L-CA, 10 mMdl-homocysteic acid, or 25 mMdl-2-amino-4-phosphonobutyrate on membrane vesicles enhanced L-[35S]CA and L-[3H]Glu uptake. Na+ (5 mM) and omission of Cl from the assay medium decreased L-[35S]CA uptake into both 10 mM L-Glu-loaded and non-loaded membrane vesicles. The anion transport blockers, 4-acetamide-4-isothiocyano-2,2-disulfonic acid stibene (SITS) and 4,4-diisothiocyano-2,2-disulfonic acid stilbene (DIDS), inhibited L-[35S]CA uptake in a dose-dependent manner. The maximal uptake rate for L-[35S]CA was decreased by 50 M SITS, while the apparent Km value of L-CA was not changed. SITS increased the EC50 value of Cl for L-[35S]CA uptake from 5 mM to 10 mM with reduction of the maximal effect. These results suggested that L-[35S]CA uptake into synaptic membrane vesicles was mediated by a SITS-sensitive hetero-exchange transport with non-labeled substrates.Abbreviations SITS 4-Acetamide-4-isothiocyano-2,2-disulfonic acid stilbene - DIDS 4,4-Diisothiocyano-2,2-disulfonic acid stilbene - CA Cysteic acid - APB 2-Amino-4-phosphonobutyrate - CSA Cysteine sulfinic acid - EGTA Ethyleneglycol bis(aminoethylether) tetraacetate - GABA -Aminobutyric acid  相似文献   

6.
The plant growth substance N6-(2-isopentenyl) adenine (i6Ade) significantly inhibits the rates of ethanol oxidation and acetaldehyde reduction in vitro by cell-free extracts of Zymomonas mobilis and of an Escherichia coli recombinant strain, containing the Z. mobilis adhB gene. The two-substrate kinetics of ethanol oxidation (forward) is affected by increasing values of dissociation constants for coenzyme and coenzyme —enzyme complexes in the presence of i6Ade, whereas the reaction maximum velocity (V m) remains unchanged and reflects the competitive type of inhibition. Changes of the kinetic constants of acetaldehyde reduction (back) are similar, except the increasing value of V m and correspond to the CIS (competitive inhibition and stimulation) type of inhibition. The estimated values of inhibition constants of the forward and back reactions are 0.39 ± 0.05 mM and 0.19 ± 0.06 mM, respectively.  相似文献   

7.
Modification of lignin by Geotrichum klebahnii   总被引:2,自引:0,他引:2  
13C-NMR spectroscopic analysis indicates that the yeast-like species Geotrichum klebahnii is an efficient microorganism for lignin biodegradation. This strain modified beechwood lignin even if it was the only carbon source by C-C side chain cleavage, C-oxidations, aromatic ring cleavage and reductive reactions. The obtained results outline prospective application of G. klebahnii for biotechnological pre-treatment of lignocellulosic materials.  相似文献   

8.
The effect of pH and transmembrane pH on the efficiency of the proton pump of the mitochondrialbc 1 complex bothin situ and in the reconstituted state was studied. In both cases the H+/e ratio for vectorial proton translocation by thebc 1 complex respiring at the steady state, under conditions in which the transmembrane pH difference (pH) represents the only component of the proton motive force (p), was significantly lower than that measured under level flow conditions. The latter amounts, at neutral pH, to 1 (2 including the scalar H+ release). In the reconstituted system steady-state pH was modulated by changing the intravesicular buffer as well as the intra/extra-liposomal pH. Under these conditions the H+/e ratio varied inversely with the pH. The data presented show that pH exerts a critical control on the proton pump of thebc 1 complex. Increasing the external pH above neutrality caused a decrease of the level flowH +/e ratio. This effect is explained in terms of proton/electron linkage inb cytochromes.  相似文献   

9.
4-Methylumbelliferyl 6-O-benzyl--d-lactoside (6Bn-MU-Lac) and some related compounds were synthesizedvia different selective reactions including phase-transfer glycosylation. Their suitability as substrates for a fluorometric assay of ceramide glycanase (CGase) was evaluated. Among others, the 6Bn-MU-Lac, which is resistant to exogalactosidase, was found to be a suitable substrate for routine assay of the CGase activity. For American leech CGase, theK m value is 0.232 mM at pH 5. Abbreviations: CGase, ceramide glycanase; Gal, galactose; Glc, Glucose; Lac, lactose; MU, 4-methylumbelliferone; MU-Lac, 4-methylumbelliferyl -d-lactoside; bBn-Lac, 6-O-benzyl-lactose; 6Bn-MU-Lac, 4-methylumbelliferyl 6-Obenzyl--d-lactoside; 46Bd-MU-Lac, 4-methylumbelliferyl 4,6-O-benzylidene--d-lactoside; MU-Cel, 4-methylumbellifery -d-cellobioside; 46Bd-MU-Cel, 4-methylumbelliferyl 4,6-O-benzylidene--d-cellobioside; TLC, thin layer chromatography;1H-NMR, proton nuclear magnetic resonance; GSL, glycosphingolipids; CSA, 10-camphorsulfonic acid. See Scheme 1 for chemical structures.  相似文献   

10.
Summary A 3D triple resonance experiment has been designed to provide intraresidual and sequential correlations between amide nitrogens and -carbons in uniformly 13C15N-labeled proteins. In-phase 13C magnetization is transferred to the aliphatic side-chain protons via the side-chain carbons using a CC-TOCSY mixing sequence. Thus, the experiment alleviates the resonance assignment process by providing information about the amino acid type as well as establishing sequential connectivities. Leaving the carbonyl spins untouched throughout the transfer from 13C to 1H leads to E.COSY-type cross peaks, from which the 3JH co coupling constants can be evaluated. The pulse sequence is applied to oxidized Desulfovibrio vulgaris flavodoxin.  相似文献   

11.
Three new major, race-specific, resistance genes to powdery mildew (Erysiphe graminis f. sp. hordei) were identified in three barley lines, RS42-6*O, RS137-28*E, and HSY-78*A, derived from crosses with wild barley (Hordeum vulgare ssp. spontaneum). The resistance gene origining from wild barley in line RS42-6*O, showed a recessive mode of inheritance, whereas the other wild barley genes were (semi)-dominant. RFLP mapping of these three genes was performed in segregating F2 populations. The recessive gene in line RS42-6*O, was localized on barley chromosome 1S (7HS), while the (semi)-dominant genes in lines RS137-28*E, and HSY-78*A, were localized on chromosomes 1L (7HL) and 7L (5HL), respectively. Closely linked RFLP clones mapped at distances between 2.6cM and 5.3 cM. Hitherto, specific loci for powdery mildew resistance in barley had not been located on these chromosomes. Furthermore, tests for linkage to the unlocalized resistance gene Mlp revealed free segregation. Therefore, these genes represent new loci and new designations are suggested: mlt (RS42-6*O), Mlf (RS137-28*E), and Mlj (HSY-78*A). Comparisons with mapped QTLs for mildew resistance were made and are discussed in the context of homoeology among the genomes of barley (H-vulgare), wheat (Triticum aestivum), and rye (Secale cereale). Duplications of RFLP bands detected in the neighbourhood of Mlf and mlt might indicate an evolutionary interrelationship to the Mla locus for mildew resistance.  相似文献   

12.
Natural abundance of 15N in tropical plants with emphasis on tree legumes   总被引:6,自引:0,他引:6  
Natural abundance of 15N ( 15N) of leaves harvested from tropical plants in Brazil and Thailand was analyzed. The 15N values of non-N2-fixing trees in Brazil were +4.5±1.9, which is lower than those of soil nitrogen (+8.0±2.2). In contrast, mimosa and kudzu had very low 15N values (–1.4+0.5). The 15N values of Panicum maximum and leguminous trees, except Leucaena leucocephala, were similar to those of non-N2-fixing trees, suggesting that the contribution of fixed N in these plants is negligible. The 15N values of non-N2-fixing trees in Thailand were +4.9±2.0. Leucaena leucocephala, Sesbania grandiflora, Casuarina spp. and Cycas spp. had low 15N values, close to the value of atmospheric N2 (0), pointing to a major contribution of N2 fixation in these plants. Cassia spp. and Tamarindus indica had high 15N values, which confirms that these species are non-nodulating legumes. The 15N values of Acacia spp. and Gliricidia sepium and other potentially nodulating tree legumes were, on average, slightly lower than those of non-N2-fixing trees, indicating a small contribution of N2 fixation in these legumes.  相似文献   

13.
Adventitious shoot regeneration was compared among leaf, stem and petal explants of carnation (Dianthus caryophyllus L.) cv. Scania on MS medium containing different concentrations of 6-benzyladenine (BA) and -naphthaleneacetic acid (NAA). High frequency regeneration was obtained only from petal explants on the media containing 5 to 10 M BA with or without 5 M NAA. Among the cytokinins tested, N-2-chloro-4-pyridyl-N-phenylurea and N-1,2,3-thiadiazol-5-yl-N-N-phenylurea were more effective than BA, kinetin, N6-2-isopentenyl adenine and zeatin on regeneration from petal explants. Although, high frequency shoot regeneration was obtained from all petal explants harvested from various developmental stages of buds, a significant decrease in regeneration capacity was observed in the explants obtained from fully-opened flowers. High frequency shoot regeneration was also obtained from the petal explants of cvs. Coral. Lena, Nora and White Sim, and an interspecific cultivar Eolo using the method developed in this study.Abbreviations NAA -naphthaleneacetic acid - BA 6-benzyladenine - GA3 gibberellic acid - 2iP N6-2-isopentenyl adenine - KT-30 N-2-chloro-4-pyridyl-N-phenylurea (also called 4PU) - TDZ N-1,2,3-thiadiazol-5-yl-N-phenylurea (also called thidiazuron)  相似文献   

14.
Sporangia were accumulated in autotrophically and mixotrophically growing cultures of the Chlamydomonas reinhardtii mutant strain ls entering the stationary phase. Such an accumulation of sporangia was never observed in stationary-phase cultures of wildtype strains. Sporangia harvested from stationary-phase cultures of the mutant strain ls released their zoospores after being resuspended in fresh culture medium. Liberation of zoospores was also observed during fixation of these sporangia with glutaraldehyde and OsO4. Release of zoospores during fixation was prevented by pretreatment with 3 mol·l–1 LiCl. Ultrastructural analyses of these LiCl-pretreated sporangia revealed that they contained abnormal sporangial walls: sporangia containing sporangia and sporangia surrounded by additional multilayered cell walls have been observed. Similar abnormal cell-wall structures were found in sporangia accumulated at the end of the dark period, when the mutant strain ls was grown photoautotrophically under a 12 h light-12 h dark regime with suboptimal aeration. When grown under optimal conditions, this particular mutant did not show any abnormal wall structures.This work has been supported by a grant from the Deutsche Forschungsgemeinschaft. The authors thank Mrs. C. Adami for the photographic work.  相似文献   

15.
The major pentasaccharides Fuc(1-2)[GalNAc(1-3)]Gal(1-4)[Fuc(1-3)]Glc and Fuc(1-2) [Gal(1-3)]Gal(1-4)[Fuc(1-3)]Glc, which are normally present in the urine of bloodgroup A Leb and B Leb healthy subjects, were each found to be contaminated by a minor component when analysed by1H-NMR. The determination of these structures, Fuc(1-2) [GalNAc(1-3)]Gal(1-3)[Fuc(1-4)]Glc and Fuc(1-2) [Gal(1-3)]Gal(1-3)[Fuc(1-4)]Glc, was based on the results of methylation analysis and1H/13C-NMR spectroscopy.Abbreviations HPLC high performance liquid chromatography - GLC gas liquid chromatography - NMR nuclear magnetic resonance - COSY correlation spectroscopy - Gal d-galactopyranose - GalNAc 2-acetamido-2-deoxy-d-galactopyranose - Glc d-glucopyranose - Fuc l-fucopyranose - LNDFH I lacto-N-difucohexaose I (Leb determinant  相似文献   

16.
T. M. Butt  A. Beckett 《Protoplasma》1984,120(1-2):61-71
Summary A detailed account of the ultrastructure and behaviour of the spindle pole body (SPB) of the entomophthoraceous fungusErynia neoaphidis is presented for the first time.The SPB consists of extranuclear (ENC) and intranuclear (INC) components. The ENC is a saucepan-shaped structure which lies in a pocket of the nuclear envelope. It is composed of a forked, fibrillar handle and a shallow, cylindrical pan. The pan has a wall of two layers, both of which are thickened with a regular periodicity so that they appear to be beaded. It is postulated that the pan is formed from rough endoplasmic reticulum and that it synthesizes the amorphous, electron-dense material coating the ENC.The INC is a saucer-shaped, electron-dense plaque in which the ends of the spindle microtubules terminate. During metaphase, a clear zone separates the INC from the nuclear envelope and persists until telophase. The roles of the amorphous, electron-dense material and the clear zone as well as the method of SPB replication are discussed.  相似文献   

17.
Hubert Felle 《Planta》1981,152(6):505-512
In the aquatic liverwort Riccia fluitans, membrane depolarization (m), change in membrane conductance (gm), and current-voltage (I-V) characteristics in the presence of different amino acids as well as the uptake of 14C-labeled amino acids were measured. L-isomers of the tested amino acids generate larger electrical effects (m, gm) than D-isomers, and the I-V characteristics show that the positive electrical inward-current of 20 mA m-2 generated by 0.5 mM D-serine is only about 50% of the current generated by adding 0.5 mM L-serine. Whereas - and -amino acids rapidly depolarize the membrane to the same extend, with -aminobutyric acid (-AB) and dipeptides no significant electrical effects have been measured. The uptake kinetics of 14C-labeled amino acids display three components: (I) A saturable high-affinity component with Ks-values of 48 M D-alanine, 12 M -aminoisobutyric acid (AIB), 9 M L-alanine, 8 M L-proline, and 6 M L-serine, respectively; (2) an apparently linear low-affinity component, and (3) an also linear but unspecific component at concentrations >20 times the given Ks-value. Uptake of 14C-labeled AIB can be inhibited competitively by all tested neutral amino acids, the L-isomers being more effective than the D-isomers, as well as by ammonium or methylamine. Vice versa, AIB competitively inhibits uptake of L-serine and L-alanine. It is concluded that an uncharged stereospecific carrier for the investigated amino acids exists in the plasmalemma of Riccia fluitans. Accumulation ratios of about 50 suggest secondary active transport driven by a transmembrane electro-chemical gradient (mainly m) which is generated by the electrogenic proton pump. It is suggested that this carrier binds to the amino group forming either a charged binary complex with positively charged amines (Felle 1980), or an uncharged complex with -AB or dipeptides, whereas electrogenic transport of - and -amino acids is mediated by a ternary carrier complex, probably charged by a proton.Symbols and Abbreviations m membrane potential (mV) - Eco equilibrium potential (mV) of the transport system - gm membrane (slope) conductance (Sm-2) - gm change in gm - I-V curve current-voltage curve - AIB -aminoisobutytric acid - -AB -aminobutyric acid  相似文献   

18.
2D NMR spectroscopy and J coupling constant analysis are applied to resolve the structure of two photoproducts of thymidylyl-(35)-thymidine. These products are cyclobutane type thymine dimers possessing the cis-syn (the predominant one) and trans-syn geometry. The cis-syn is formed in an ANTI-ANTI conformation about the N-glycosyl linkages and resembles the normal base-stacked configuration. The glycosidic conformation in solution of the 5 terminal fragment differs from the crystal in which the less common SYN conformation is observed. In this isomer only the sugar pucker of the 3 terminal fragment is changed substantially with respect to the dinucleotide. The trans-syn isomer is formed in a SYN-ANTI glycosidic conformation. In this isomer the sugar puckers of both deoxyribose rings are affected and a preference for a pure 2-endo conformation is observed.Abbreviations dTpdT 2-deoxythymidylyl-(35)-2-deoxythymidine - dTp[]dT cyclobutane type photodimers of dTpdT - dTp- and dTp[]- their 5' terminal fragments (fragment A) - -pdT and-[]pdT their 3 terminal fragments (fragment B) - RP-HPLC reversed-phase high-performance liquid chromatography - COSY two-dimensional correlated spectroscopy - 2D NOE two-dimensional nuclear Overhauser spectroscopy  相似文献   

19.
Structural analogues of the NADP+ were studied as potential coenzymes and inhibitors for NADP+ dependent malic enzyme from Zea mays L. leaves. Results showed that 1, N6-etheno-nicotinamide adenine dinucleotide phosphate ( NADP+), 3-acetylpyridine-adenine dinucleotide phosphate (APADP+), nicotinamide-hypoxanthine dinucleotide phosphate (NHDP+) and -nicotinamide adenine dinucleotide 2: 3-cyclic monophosphate (23NADPc+) act as alternate coenzymes for the enzyme and that there is little variation in the values of the Michaelis constants and only a threefold variation in Vmax for the five nucleotides. On the other hand, thionicotinamide-adenine dinucleotide phosphate (SNADP+), 3-aminopyridine-adenine dinucleotide phosphate (AADP+), adenosine 2-monophosphate (2AMP) and adenosine 2: 3-cyclic monophosphate (23AMPc) were competitive inhibitors with respect to NADP+, while -nicotinamide adenine dinucleotide 3-phosphate (3NADP+), NAD+, adenosine 3-monophosphate (3AMP), adenosine 2: 5-cyclic monophosphate (25AMPc), 5AMP, 5ADP, 5ATP and adenosine act as non-competitive inhibitors. These results, together with results of semiempirical self-consistent field-molecular orbitals calculations, suggest that the 2-phosphate group is crucial for the nucleotide binding to the enzyme, whereas the charge density on the C4 atom of the pyridine ring is the major factor that governs the coenzyme activity.Abbreviations NADP+ 1, N6-etheno-nicotinamide adenine dinucleotide phosphate - NHDP+ nicotinamide-hypoxanthine dinucleotide phosphate - APADP+ 3-acetylpyridine-adenine dinucleotide phosphate - SNADP+ thionicotinamide-adenine dinucleotide phosphate - AADP+ 3-aminopyridine-adenine dinucleotide phosphate - 23NADPc+ -nicotinamide adenine dinucleotide 2: 3-cyclic monophosphate - 3NADP+ -nicotinamide adenine dinucleotide 3-phosphate - 2AMP adenosine 2-monophosphate - 3AMP adenosine 3-monophosphate - 23AMPc adenosine 2: 3 monophosphate cyclic - A adenosine - RuBP ribulose 1,5-bisphosphate - SCF-MO Self-Consistent Field-Molecular Orbitals (method)  相似文献   

20.
The lipopolysaccharides (LPSs) extracted from the outer membrane of Azospirillum brasilense Sp245 and its Omegon-Km mutants KM018 and KM252 with a hot aqueous solution of phenol were found to differ in the content of carbohydrates, glucosamine, and total phosphorus and in the proportion of octadecenoic and hexadecanoic acids in the lipid moieties of the LPSs. The carbohydrate moieties of the LPSs were heterogeneous in charge. The analysis of the O-specific polysaccharides (O-PSs) of the mutants KM018 and KM252 by gas–liquid chromatography, IR spectroscopy, and NMR spectroscopy showed that they are composed of the same linear pentasugar repeating units 2)--D-Rhap-(1 3)--D-Rhap-(1 3)--D-Rhap-(1 2)--D-Rhap-(1 2)--D-Rhap-(1 as the O-PSs of the parent strain Sp245. The reported differences in the biological activity of the LPSs of the parent and mutant strains can be due to their different chemical composition.  相似文献   

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