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1.
Summary Candida wickerhamii growing on cellobiose produced -glucosidase with high activity against -nitrophenyl glucoside (PNPG) but low activity against cellobiose. -glucosidase production was constitutive, and was repressed by -glucosides and glucose. -glucosides containing an aromatic moiety in the aglycon were the best substrates for -glucosidase indicating that the enzyme is an aryl--glucosidase. A -glucosidase from C. wickerhamii cells was purified by (NH4)2SO4 precipitation, dialysis, ion-exchange chromatography and gel filtration. The purified enzyme was homogeneous as shown by sodium-dodecyl-sulphate polyacrylamide gel electrophoresis and discontinuous gel electrophoresis. The purified enzyme hydrolysed PNPG but not cellobiose. The Km of the enzyme was 0.185 mM. Glucose inhibited the enzyme competitively and the Ki was 7.5 mM. The apparent molecular mass was 97,000. The optimum pH and temperature for enzyme activity were between pH 7 and 7.4 and 40°C respectively. At temperatures of 45°C and greater the enzyme was inactivated. The activation energy of the enzyme was 29.4 kJ · mol-1.  相似文献   

2.
The relative contribution of autotrophic carbon sources (aquatic macrophytes, flooded forest, phytoplankton) for heterotrophic bacterioplankton was evaluated in a floodplain lake of the Central Amazon. Stable carbon isotopes (13C) were used as tracers. Values of 13C of different autotrophic sources were compared to those of dissolved organic carbon (DOC) and those of bacterially produced CO2.The percentage of carbon derived from C4 macrophytes for bacterially produced CO2 was the highest, on average 89%. The average 13C value of CO2 from bacterial respiration was –18.5 ± 3.3. Considering a fractionation of CO2 of 3 by bacterial respiration, 13C value was –15.5, near C4 macrophyte 13C value (–13.1).The average value of total DOC 13C was –26.8 ± 2.4. The percentage of C4 macrophytes carbon for total DOC was on average 17%. Considering that bacteria consume mainly carbon from macrophytes, the dominance of C3 plants for total DOC probably reflects a faster consumption of the former source, rather than a major contribution of the latter source.Heterotrophic bacterioplankton in the floodplain may be an important link in the aquatic food web, transferring the carbon from C4 macrophytes to the consumers.  相似文献   

3.
A catalytic fragment, 1-298, derived from limited chymotryptic digestion of phosphorylaseb kinase (Harris, W.R.et al., J. Biol. Chem., 265: 11740–11745, 1990), is reported to have about six-fold greater specific activity than does the subunit-calmodulin complex. To test whether there is an inhibitory domain located outside the catalytic core of the subunit, full-length wild-type and seven truncated forms of were expressed inE. coli. Recombinant proteins accumulate in the inclusion bodies and can be isolated, solubilized, renatured, and purified further by ammonium sulfate precipitation and Q-Sepharose column. Four out of seven truncated mutants show similar ( 1-353 and 1-341) or less ( 1-331 and 1-276) specific activity than does the full-length wild-type , 1-386. Three truncated forms, 1-316, 1-300, and 1-290 have molar specific activities approximately twice as great as those of the full-length wild-type and the nonactivated holoenzyme. All recombinant s exhibit similarK m values for both substrates, i.e., about 18M for phosphorylaseb and about 75 M for MgATP. Three truncated s, 1-316, 1-300, and 1-290, have a 1.9- to 2.5-fold greater catalytic efficiency (V max/K m) than that of the full-length wild-type and a 3.5- to 4.5-fold greater efficiency than that of the truncated 1-331. This evidence suggests that there is at least one inhibitory domain in the C-terminal region of , which is located at 301-331· 1-290, but not 1-276, which contains the highly conserved kinase domain, is the minimum sequence required for the subunit to exhibit phosphotransferase activity. Both 1-290 and 1-300 have several properties similar to full-length wild-type , including metal ion responses (activation by free Mg2+ and inhibition by free Mn2+) pH dependency, and substrate specificities.  相似文献   

4.
Summary Organic pyrophosphates such as UppA and NAD are formed when a solution containing a nucleotide, a nucleoside 5-polyphosphate, Mg2+ and imidazole are allowed to dry out. We suggest that this synthesis may have occured concurrently with oligonucleotide formation.Abbreviations Im Imidazole - CDI 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride - EDTA ethylenediaminetetraacetic acid - A adenosine - U uridine - pnA adenosine 5-poly-phosphate containing n phosphate residues - pU uridine 5-phosphate - AppA P1,P2-diadenosine 5-pyrophosphate - UppA P1-(uridine 5)-P2-(adenosine 5)-pyrophosphate - ImpA adenosine 5-phosphorimidazolide - NMN nicotinamide mononucleotide - NAD nicotinamide-adenine dinucleotide  相似文献   

5.
Expression of a chimeric stilbene synthase gene in transgenic wheat lines   总被引:11,自引:0,他引:11  
A chimeric stilbene synthase (sts)gene was transferred into wheat. Stilbene synthases play a role in the defence against fungal diseases in some plant species (e.g. groundnut or grapevine) by producing stilbenetype phytoalexins like resveratrol. Resveratrol is also claimed to have positive effects to human health. Embryogenic scutellar calli derived from immature embryos of the two commercial German spring wheat cultivars Combi and Hanno were used as target tissue for cotransformation by microprojectile delivery. The selectable marker/reporter gene constructs contained the bargene either driven by the ubiquitinpromoter from maize (pAHC 25, also containing the uidAgene driven by the ubiquitinpromoter), or by the actinpromoter (pDM 302) from rice. The cotransferred plasmid pStil 2 consisted of a grapevine stscoding region driven by the ubiquitin promoter. Eight transgenic Combi and one Hanno TOplant were obtained and, except one Combi TOplant, found to be cotransformants due to the integration of both the stsgene and the selectable marker or reporter genes. Expression of the stsgene was proven by RTPCR, and, for the first time, by detection of the stilbene synthase product resveratrol by HPLC and mass spectrometry. The stsgene was expressed in four of the seven transgenic Combi T_oplants. Two of the respective T1progenies segregated in a Mendelian manner were still expressing the gene. Investigations into methylation of the stsgene showed that in three nonexpressing progenies inactivation was paralleled by methylation.  相似文献   

6.
Subfamilies of voltage-activated K+ channels (Kv1-4) contribute to controlling neuron excitability and the underlying functional parameters. Genes encoding the multiple subunits from each of these protein groups have been cloned, expressed and the resultant distinct K+ currents characterized. The predicted amino acid sequences showed that each subunit contains six putative membrane-spanning -helical segments (S1-6), with one (S4) being deemed responsible for the channels' voltage sensing. Additionally, there is an H5 region, of incompletely defined structure, that traverses the membrane and forms the ion pore; residues therein responsible for K+ selectivity have been identified. Susceptibility of certain K+ currents produced by the Shaker-related subfamily (Kv1) to inhibition by -dendrotoxin has allowed purification of authentic K+ channels from mammalian brain. These are large (Mr 400 kD), octomeric sialoglycoproteins composed of and subunits in a stoichiometry of ()4()4, with subtypes being created by combinations of subunit isoforms. Subsequent cloning of the genes for 1, 2 and 3 subunits revealed novel sequences for these hydrophilic proteins that are postulated to be associated with the subunits on the inner side of the membrane. Coexpression of 1 and Kv1.4 subunits demonstrated that this auxiliary protein accelerates the inactivation of the K+ current, a striking effect mediated by an N-terminal moiety. Models are presented that indicate the functional domains pinpointed in the channel proteins.  相似文献   

7.
Summary In normal, non-expanding toad epidermis more cells are produced than needed to replace cells lost by moulting. By implication, cell deletion additional to moulting must take place. This paper deals with the mechanisms by which the surplus of cells is deleted, taking advantage of the fact that the ratio between cell birth rate (K b) and the rate of desquamation (K d), which in normal toads is 2 to 3, can be manipulated. In toads deprived of the pars distalis of the pituitary gland it is decreased to 0.2 to 0.3, and in toads with hydrocortisone pellets implanted into the subcutaneous lymph space it is increased to 7 to 10. Thus, structures candidates for the morphological manifestation of the deletion process should occur rarely in toads in which the pars distalis has been removed and frequently in toads with hydrocortisone pellets implanted. Categorization and enumeration of such structures by light microscopy in the epidermis from operated, normal, and hormone-treated toads were performed. The incidence of structures referred to as dark cells and omega-figures were found to correlate relatively well with the K b/Kd-ratio. A subsequent ultrastructural analysis — on a cell-by-cell basis — of dark cells showed these to reflect various stages of apoptosis. The duration of the apoptotic process was calculated to be approximately 7 h. Light- and electron microscopy of omega-figures combined with histochemical observations of PSA-lectin binding were interpreted as reflecting a release of cells from the basal epidermis and their final elimination within the dermis. It is concluded (i) that apoptosis is an important mechanism of controlled cell deletion, (ii) that emigration to, and elimination in, the dermis is a possible deletion mechanism, and (iii) that necrosis is unlikely to play a role in controlled cell deletion.Supported in part by the Danish National Science Research Council (grant no. 11-6498) (PB)Part of this work was presented at the XVth Meeting of the European Study Group for Cell Proliferation, Sundvollen, Norway, 16–20 September 1987  相似文献   

8.
The dynamics of coupled biological oscillators can be modeled by averaging the effects of coupling over each oscillatory cycle so that the coupling depends on the phase difference between the two oscillators and not on their specific states. Average phase difference theory claims that mode locking phenomena can be predicted by the average effects of the coupling influences. As a starting point for both empirical and theoretical investigations, Rand et al. (1988) have proposed d/dt= — K sin ), with phase-locked solutions =arcsin( /K), where is the difference between the uncoupled frequencies and K is the coupling strength. Phase-locking was evaluated in three experiments using an interlimb coordination paradigm in which a person oscillates hand-held pendulums. was controlled through length differences in the left and right pendulums. The coupled frequency c was varied by a metronome, and scaled to the eigenfrequency v of the coupled system K was assumed to vary inversely with c. The results indicate that: (1) and K contribute multiplicatively to (2) =0 or = regardless of K when =0; (3) 0 or regardless of when K is large (relative to ); (4) results (1) to (3) hold identically for both in phase and antiphase coordination. The results also indicate that the relevant frequency is c/v rather than c. Discussion high-lighted the significance of confirming =arcsin(/K) for more general treatments of phase-locking, such as circle map dynamics, and for the 11 phase-entrainment which characterizes biological movement systems.  相似文献   

9.
The conformational properties of the oligosaccharide chain of GM1 ganglioside containingN-glycolyl-neuraminic acid, -Gal-(1-3)--GalNAc-(1-4)-[-Neu5Gc-(2-3)]--Gal-(1-4)--Glc-(1-1)-Cer, were studied through NMR nuclear Overhauser effect investigations on the monomeric ganglioside in dimethylsulfoxide, and on mixed micelles of ganglioside and dodecylphosphocholine in water. Several interresidual contacts for the trisaccharide core--GalNAc-(1-4)-[-Neu5Gc-(2-3)]--Gal-were found to fix the relative orientitation of the three saccharides, while the glycosidic linkage of the terminal -Gal-was found to be quite mobile as the -Gal-(1-3)--GalNAc-disaccharide exists in different conformations. These results are similar to those found for two GM1 gangliosides containingN-acetyl-neuraminic acid and neuraminic acid [1].Abbreviations Ganglioside nomenclature is in accordance with Svennerholm [23] and the IUPAC-IUB Recommendations [24] GM3(Neu5Ac) II3Neu5AcLacCer, -Neu5Ac-(2-3)--Gal-(1-4)--Glc-(1-1)-Cer - GM3(Neu5Gc) II3Neu5GcLacCer, -Neu5Gc-(2-3)--Gal-(1-4)--Glc-(1-1)-Cer - GM1(Neu5Ac) II3Neu5AcGgOse4Cer, -Gal-(1-3)--GalNAc-(1-4)-[-Neu5Ac-(2-3)]--Gal-(1-4)--Glc-(1-1)-Cer - GM1(Neu5Gc) II3Neu5GcGgOse4Cer, -Gal-(1-3)--GalNAc-(1-4)-[-Neu5Gc-(2-3)]--Gal-(1-4)--Glc-(1-1)-Cer - GM1(Neu) II3NeuGgOse4Cer, -Gal-(1-3)--GalNAc-(1-4)-[-Neu-(2-3)]--Glc-(1-1)-Cer - GD1a IV3Neu5AcII3Neu5AcGgOse4Cer, -Neu5Ac-(2-3)--Gal-(1-3)--GalNAc-(1-4)-[-Neu5Ac-(2-3)]--Gal-(1-4)--Glc-(1-1)-Cer - GalNAc-GD1a IV4GalNAcIV3Neu5AcII3Neu5AcGgOse4Cer, -GalNAc-(1-4)-[-Neu5Ac-(2-3)]--Gal-(1-3)--GalNAc-(1-4)-[-Neu5Ac-(2-3)]--Gal-(1-4)--Glc-(1-1)-Cer - Neu neuraminic acid - Neu5Ac N-acetyl-neuraminic acid - Neu5Gc N-glycolyl-neuraminic acid - Cer ceramide  相似文献   

10.
Aspartate transaminase (AST) activity in the camel tick Hyalomma dromedarii was followed throughout embryogenesis. During purification of AST to homogeneity, ion exchange chromatography lead to four separate forms (termed I, II, III and IV). AST II with the highest specific activity was pure after chromatography on Sephacryl S-300. The molecular mass of AST II was 52KDa for the native enzyme, composed of one subunit of 50KDa. AST II had a Km value of 0.67mM for -ketoglutarate and 15.1mM for aspartate. AST II had a pH optimum of 7.5 with heat stability up to 50°C for 15min. The enzyme was activated by MnCl2, and inhibited by CaCl2, MgCl2, NiCl2, and ZnCl2.  相似文献   

11.
Ghosts prepared from rat basophilic leukemia cells (RBL cell ghosts) and permeabilized with -toxin fromS. aureus are a simplified system for the study of FcRI-mediated activation of phospholipase C (PLC). This activity is dependent upon ATP and magnesium, and is enhanced by the addition of another compound containing an energetic phosphate group, either phosphoenolpyruvate (PEP) or phosphocreatine (PCr). This effect appears to be specific for PEP and PCr in that other compounds with energetic phosphate bonds including fructose 1,6-bisphosphate and additional ATP are not effective. On the contrary, GTP--S, an activator of G proteins, activates PLC in the presence of ATP alone and this is not further enhanced by the addition of PEP. In addition to FcRI and GTP--S, two other stimuli lead to enhanced activity of PLC in permeabilized RBL cell ghosts: 1) an inhibitor of tyrosine phosphatases (Na3VO4) and 2) an analog of adenosine (NECA). Data presented here extend previous results to show that activation of PLC by GTP--S is not enhanced either by the addition of PCr or by the addition of a more MgATP. Further new findings include the observations that activation of PLC by Na3VO4 is augmented by PEP and PCr in a fashion similar to that observed for FcRI-mediated activation of PLC and that activation of PLC by NECA shows even more marked dependency on PEP than does activation by FcRI or Na3VO4. Together, these experiments demonstrate that antigen, Na3VO4, GTP--S, and NECA can all activate PLC in permeabilized RBL cell ghosts in the presence of ATP and that these activities are differentially affected by the addition of a second compound with an energetic phosphate bond.Abbreviations DNP25BSA bovine serum albumin derivitized with 25 dinitrophenyl groups per mole of BSA - FcRI high affinity receptor for IgE - GTP--S guanosine-5-O-(3-thiotriphosphate) - IPs inositol phosphate(s) - K2 Pipes dipotassium Pipes - Na3VO4 sodium vanadate - NECA 5-(N-Ethyl)carboxamidoadenosine - PCr phosphocreatine - PEP phosphoenolpyruvate - PLC phospholipase C specific for phosphoinositides - RBL rat basophilic leukemia  相似文献   

12.
1. We have investigated the effect of tityus gamma (TiTX ) scorpion toxin on the release of [3H] dopamine in rat brain prefrontal cortical slices. The stimulatory effect of TiTX on the release of [3H]dopamine was dose/time-dependent with an EC50 of 0.01M.2. Tetrodotoxin blocked the TiTX -induced release of [3H]dopamine, indicating the dependency for Na+ channels.3. EGTA had no effect on the TiTX -induced release of [3H]dopamine, indicating the process is independent of extracellular calcium. Release of [3H]dopamine evoked by TiTX was inhibited by 57% by BAPTA, a chelator of intracellular calcium.4. Xestospongin and 2-APB, putative blockers of IP3-sensitive release of intracellular calcium stores, caused an equal and significant inhibition of 24% of the TiTX -induced release of [3H]dopamine, while the slight inhibition evoked by dantrolene, a putative blocker of ryanodine-sensitive calcium store was not significant.5. Nomifensine and ascorbic acid, blockers of dopamine transporter (DAT), caused an inhibition of 27 and 29%, respectively, on the toxin-induced release of [3H]dopamine suggesting that most of the TiTX -induced release of dopamine is not due to the reversal of Na+ gradient.6. In conclusion the majority of the TiTX -induced release of [3H]dopamine is exocytotic and mobilizes calcium from the intracellular IP3-sensitive calcium stores.  相似文献   

13.
Mattres  Dieter 《Zoomorphology》1962,51(4):375-546
Ohne ZusammenfassungAlphabetisches Verzeichnis der im Text gebranchten Abkürzungen AK Anbieten der Kopfgrube () - AR-seitig außenrandseitig (auf Elytre bezogen) - EO Elytralorgan (EO-Arten = Arten mit Elytralorganen im männlichen Geschlecht) - f Flucht () - F Flucht () - FA, fa frontale Auseinandersetzung (, ) - FS, fs Fühlertrillern bzw. Frontalspiel (, ) - gk Grubenknabbern () - IR-seitig innenrandseitig (auf Elytre bezogen) - K Kopulation - KG Kopfgrube (KG-Arten = Arten mit Kopfgrube im männlichen Geschlecht) - KI Abdomenkitzeln () - KV Kopulationsversuch () - LP-Feld den weiblichen Labialpalpen korreliertes Drüsenporenfeld - MP-Feld den weiblichen Maxillarpalpen korreliertes Drüsenporenfeld - 180 Drehung des um 180° - ob Organbeißen () - ok Organknabbern () - OZ Organzuwendung () - P Prüfung der Kopulationsbereitschaft durch - RB rückwärtige Berührung durch - SLV Seitwärtslauf nach vorn () - SLH Seitwärtslauf nach hinten () - U Umrundung () - vl Vorwärtslauf () - 180 Drehung des um 180° Habilitationsschrift.  相似文献   

14.
Summary A comparison of the mRNAs for rabbit and human-hemoglobins shows that synonymous changes in codons have accumulated three times as rapidly as nucleotide replacements that produced changes in amino acids. This agrees with predictions based on the so-called neutral theory. In addition, seven codon changes that appear to be single-base changes (according to maximum parsimony) are actually two-base changes. This indicates that the construction of primordial sequences is of limited significance when based on inferences that assume minimum base changes for amino acid replacements.  相似文献   

15.
J. L. Karihaloo 《Genetica》1987,73(3):217-221
Three cultivated varieties of Narcissus tazetta, Grand Soleil d'Or, Chinese Sacred Lily and Cypri, are triploid (2n = 3x+30) with the basic number 10. Grand Soleil d'Or has three homomorphic sets, each comprising 2 long submetacentrics, 4 long acrocentrics and 4 short acrocentrics. Karyotypes of the other two varieties are heteromorphic. Both possess one telocentric satellited chromosome. In addition, Cypri shows translocation between two chromosomes belonging to the seventh and eighth triplets. The number of secondary constrictions varies between 3 (Chinese Sacred Lily) and 4 (Grand Soleil d'Or and Cypri) which is also the number of nucleoli observed in the respective varieties.  相似文献   

16.
Recently, it has been found that plants, including tomato (Lycopersicon esculentum), express the Lewis-a epitope, Gal1,3(Fuc1,4)GlcNAc, on some N-glycans. By searching the EST database, it was possible to identify a tomato cDNA encoding a protein, designated FucTC, of 413 amino acids with homology to plant and mammalian 1,3/4-fucosyltransferases. The cDNA was expressed in Pichia pastoris and the recombinant enzyme was found to transfer fucose from GDP-Fuc (Km 16 M) to lacto-N-tetraose (Gal1,3GlcNAc1,3Gal1,4Glc; Km 80 M) as well as to 1,3- and 1,4-galactosylated N-glycans. It is concluded that FucTC is responsible for the biosynthesis of Lewis-a on N-glycans in tomato.  相似文献   

17.
Three alcohol dehydrogenase (ADH) isozymes from embryos of the durum wheat cultivar Bijaga Yellow having the variantAdh-Alb allele were purified using (NH4)2SO4 precipitation, gel filtration, and ion-exchange chromatography. ADH is a dimeric enzyme. The variant isozyme ADH-1-1, which is a homodimer composed of b monomers, was compared with ADH-1-5 (homodimer composed of a monomers), the product ofAdh-B1, and the ADH-1-3 isozyme (ba heterodimer) on a number of parameters includingK m, substrate specificities, and molecular weights. No appreciable differences among the three isozymes were found, except for the faster electrophoretic mobility of bb dimers (ADH-1-1). The results indicate that the variant isozyme is the result of a mutation altering only the charge of the isozyme.  相似文献   

18.
Extraneuronal catecholamine uptake was investigated in isolated quiescent rat myocardial cells. By administration of (3H-)(–)noradrenaline concentration of 22 nmol/l up to 1000 mol/l the following data were obtained: (1) The KM of the uptake process amounted to 260 mol/l, the Vmax to 4.24 nmol/(10 min × mg Protein) corresponding to 179 nmol/(min × gWWt)(WWT = Wet Weight). (2) The uptake was largely inhibited by the uptake2-inhibitors corticosterone (100 mol/l), isoprenaline (IC so = 30.6 mol/l), and O-methylisoprenaline (IC50 = 2.1 pmol/l), but not by the uptake1-inhibitors cocaine (100 mol/l) and desipramine (10 mol/l). (3) The affinity-values KM and IC50 closely agreed with those already known, but the Vmax-value was higher than those obtained in whole rat hearts by a factor of at least 1.79. This is caused presumably by the voltage dependence of the uptake mechanism and the resulting inhibition of uptake 2 during the periods of depolarisation in beating hearts of other studies.  相似文献   

19.
Summary Two glasshouse experiments are described in which the effects of applying starter phosphate fertilizer, 1 cm beneath the seeds, on early growth and nutrient concentrations of lettuce (Lactuca sativa L. cv. Avondefiance) in well fertilized soil were determined. In Experiment 1 various rates of starter P in the form of NH4H2PO4 were applied to soil containing a range of rates of incorporated triple superphosphate. Although there was little response of lettuce dry weight to the incorporated triple superphosphate there was a large response (about 65% increase after 36 days) to the starter. N and P concentrations within the plants were increased by the starter treatments whereas K concentration was reduced. The per cent P in the plants at 36 days from sowing could account for 60% of the variation in plant dry weight. In Experiment 2 the starter P was added as either the Ca, Na or K salt, with or without added (NH4)2SO4. Adding the starter P without ammonum increased the P concentration of the plants by an average of 12% and the dry weight by an average of 39% at 30 days from sowing. The addition of ammonium ions increased plant concentrations of P, Mg and N but decreased plant K concentration. The effect of the ammonium ions on growth depended on the form of phosphate supplied as the starter. This variation in effect of ammonium ions was attributed to the effects of other starter ions on the relative concentration of ammonium in the soil solution.  相似文献   

20.
The influence of some ions in pre-growth culture medium on chromate reduction by resting cells of Agrobacterium radiobacter strain EPS-916 was investigated. The reduction was dependent on the Fe2+ content of the culture medium: the higher the iron content, the lower the reduction rate. The cells showed maximum chromate reduction when pre-grown in the presence of 0.243 m Mg2+, 20 m Ca2+ and 3.6 m Mn2+. Chromate reduction was not affected by the addition of MgCl2, CdCl2, ZnCl2, MnCl2, Na2SO4 (1000 m), and Na2MoO4 (100 m) to the activity assays. However, activity was inhibited by the presence of Na2SO4 (10 mm), Na2MoO4 (200 m) and ferric citrate.  相似文献   

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