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1.
The activities and kinetics of the enzymes G6PDH (glucose-6-phosphate dehydrogenase) and 6PGDH (6-phosphogluconate dehydrogenase) from the mesophilic cyanobacterium Synechococcus 6307 and the thermophilic cyanobacterium Synechococcus 6716 are studied in relation to temperature. In Synechococcus 6307 the apparent K m's are for G6PDH: 80M (substrate) and 20M (NADP+); for 6PGDH: 90M (substrate) and 25M (NADP+). In Synechococcus 6716 the apparent K m's are for G6PDH: 550M (substrate) and 30M (NADP+); for 6PGDH: 40M (substrate) and 10M (NADP+). None of the K m's is influenced by the growth temperature and only the K m's of G6PDH for G6P are influenced by the assay temperature in both organisms. The idea that, in general, thermophilic enzymes possess a lower affinity for their substrates and co-enzymes than mesophilic enzymes is challenged.Although ATP, ribulose-1,5-bisphosphate, NADPH and pH can all influence the activities of G6PDH and 6PGDH to a certain extent (without any difference between the mesophilic and the thermophilic strain), they cannot be responsible for the total deactivation of the enzyme activities observed in the light, thus blocking the pentose phosphate pathway.Abbreviations G6PDH glucose-6-phosphate, dehydrogenase - 6PGDH 6-phosphogluconate dehydrogenase - G6P glucose-6-phosphate - 6PG 6-phosphogluconate - RUDP ribulose-1,5-bisphosphate - Tricine N-Tris (hydroxymethyl)-methylglycine  相似文献   

2.
Role of mitochondria in ethanol tolerance of Saccharomyces cerevisiae   总被引:7,自引:0,他引:7  
The presence of active mitochondria and oxidative metabolism is shown to be essential to maintain low inhibition levels by ethanol of the growth rate (), fermentation rate (v) or respiration rate () of Saccharomyces cerevisiae wild type strain S288C. Cells which have respiratory metabolism show K i (ethanol inhibition constant) values for , v and , higher (K i>1 M) than those of petite mutants or grande strains grown in anaerobiosis (K i=0.7 M). In addition, the relationship between or v and ethanol concentration is linear in cells with respiratory metabolism and exponential in cells lacking respiration. When functional mitochondria are transferred to petite mutants, the resulting strain shows K i values similar to those of the grande strain and the inhibition of and v by increasing ethanol concentrations becomes linear.  相似文献   

3.
Summary Copper-deficient cells ofPseudomonas stutzeri strain ZoBell synthesize catalytically inactive nitrous oxide (N2O) reductase which is activated by added Cu(II) in the absence of de novo protein synthesis. The apparentK m for the activation process is 0.13 M. Activation is temperature-dependent and is inhibited by Cd(II)(K i 1.27 M) and less strongly by Zn(II), Ni(II), and Co(II). The same metal ions at 20 M have little or no effect on N2O reduction of intact cells. Apo-N2O reductase of transposon Tn5-inducednos mutants with defective Cu-chromophore biosynthesis is not reactivated by Cu(II). N2O reductase of Cu-sufficient and Cu-deficient wild type, and ofnos mutants is localized in the periplasm, the latter providing the likely site of metal incorporation into the apoenzyme.  相似文献   

4.
Endogenous and xenobiotic sulphur-containing convulsant and non-convulsant compounds containing structural moieties of, or bearing a structural resemblance to, GABA and homocysteine were tested in binding studies for their potency in displacing the GABA-mimetic [3H]muscimol from specific, high-affinity sites (K d=3.6 nM;B max=3.94 pmol/mg protein) on freeze-thawed, Triton-treated calf-brain synaptic membranes. The xenobiotic convulsants, 4-mercaptobutyric acid (MBA), 3-mercaptopropionic acid (3-MPA) and 2-mercaptopropionic acid (2-MPA) were found to be two-site competitive inhibitors exhibiting apparent inhibition affinity constants (K i app ) of 5000 M, 3750 M, and 4800 M, respectively; while homocysteic acid (K i app =4800 M) was shown to be a one-site partial competitive inhibitor. Intermediary metabolites of methionine: S-adenosyl-l-homocysteine,l-cysteine, the convulsantl-homocysteine, and its non-convulsant disulphide oxidation product, homocystine, were found to be one-site partial competitive inhibitors exhibitingK i app values of 5750 M, 8350 M, 5000 M, and 510 M, respectively. The endogenous anticonvulsant neuroeffector, taurine, and the tripeptide, reduced glutathione (GSH) were shown to be, respectively, one-site (K i=20 M) and two-site (K i app =4300 M) competitive inhibitors of [3H]muscimol binding. These findings are discussed with regard to a previously proposed mechanism for the convulsant action of homocysteine.  相似文献   

5.
We have compared the biochemical properties of two different Arabidopsis ammonium transporters, AtAMT1;1 and AtAMT1;2, expressed in yeast, with the biophysical properties of ammonium transport in planta. Expression of the AtAMT1;1 gene in Arabidopsis roots increased approximately four-fold in response to nitrogen deprivation. This coincided with a similar increase in high-affinity ammonium uptake by these plants. The biophysical characteristics of this high-affinity system (Km for ammonium and methylammonium of 8 M and 31 M, respectively) matched those of AtAMT1;1 expressed in yeast (Km for methylammonium of 32 M and Ki for ammonium of 1–10 M). The same transport system was present, although less active, in nitrate-fed roots. Ammonium-fed plants exhibited the lowest rates of ammonium uptake and appeared to deploy a different transporter (Km for ammonium of 46 M). Expression of AtAMT1;2 in roots was insensitive to changes in nitrogen nutrition. In contrast to AtAMT1;1, AtAMT1;2 expressed in yeast exhibited biphasic kinetics for methylammonium uptake: in addition to a high-affinity phase with a Km of 36 M, a low-affinity phase with a Km for methylammonium of 3.0 mM was measured. Despite the presence of a putative chloroplast transit peptide in AtAMT1;2, the protein was not imported into chloroplasts in vitro. The electrophysiological data for roots, together with the biochemical properties of AtAMT1;1 and Northern blot analysis indicate a pre-eminent role for AtAMT1;1 in ammonium uptake across the plasma membrane of nitrate-fed and nitrogen-deprived root cells.  相似文献   

6.
The Rubus species R. parviflorus, R. spectabilis and R. strigosus interfere with conifer seedling establishment on forest regeneration sites in Canada and the United States. As a first step towards microbial metabolite-based control, callus and cell suspension cultures of the Rubus species were developed as a bioassay system to detect phytotoxic compounds that may have relevance in a vegetation control context. Rapidly growing friable callus and suspension cultures were obtained from leaf disks of the three weedy Rubus species using similar culture media conditions (modified Murashige and Skoog) but required different plant growth regulators (R. parviflorus, 4.5 M 2,4-D; R. spectabilis, 26.9 M NAA/0.5 M zeatin; and R. strigosus, 12.4 M picloram). Cell growth and health attributes including callus circumference, degree of browning and suspension culture cell viability as measured by the TTC vital stain assay were developed and were rapid and convenient to use. We have established Rubus tissue culture systems that will make it possible for large scale screening of phytotoxic metabolites.  相似文献   

7.
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   

8.
J. Sybenga 《Genetica》1965,36(1):243-252
Whilst reliable estimates of chiasma frequencies can usually not be obtained, the probability (b) of a chromosome arm to be bound by at least one chiasma can often be determined. In the absence of interference this probability equals (1–e –2), where 2 is the average chiasma frequency of the chromosome arm and the average crossover frequency or map length. In the presence of interference is shown to retain its genetic meaning as an additive metric that may describe the chromosome arm or other distinctive chromosome segment in terms of genetic recombination. It is a form of potential map length, comparable to, but numerically different from the regular map length. It is termed provisionally crossing-over potential.A chromosome with armsm andn with crossing-over potentials and will form ring bivalents with a frequency (1–e –2).(1–e –2); open bivalents with a frequency (1–e –2).e –2+(1–e –2).e –2; univalent pairs with a frequencye –2.e –2. Estimates of these frequencies yield equations from which and may be solved. In rye (Secale cereale) their ratio (q) is approximately two and differs from the mitotic arm length ratio of 1.4, indicating localization of chiasmata in the long arms.Graphs are given to show how, with constantq, the relation between the probabilitiesb m andb n of the two arms being bound changes with changing averageb.Data are presented on chiasma frequencies in M I, and compared with the frequencies expected in the absence of interference to give an impression of the degree of interference. Apparent fusion of chiasmata simulates interference.  相似文献   

9.
Jia  Yinsuo  Gray  V.M. 《Photosynthetica》2003,41(4):605-610
We determined for Vicia faba L the influence of nitrogen uptake and accumulation on the values of photon saturated net photosynthetic rate (P Nmax), quantum yield efficiency (), intercellular CO2 concentration (C i), and carboxylation efficiency (C e). As leaf nitrogen content (NL) increased, the converged onto a maximum asymptotic value of 0.0664±0.0049 mol(CO2) mol(quantum)–1. Also, as NL increased the C i value fell to an asymptotic minimum of 115.80±1.59 mol mol–1, and C e converged onto a maximum asymptotic value of 1.645±0.054 mol(CO2) m–2 s–1 Pa–1 and declined to zero at a NL-intercept equal to 0.596±0.096 g(N) m–2. fell to zero for an NL-intercept of 0.660±0.052 g(N) m–2. As NL increased, the value of P Nmax converged onto a maximum asymptotic value of 33.400±2.563 mol(CO2) m–2 s–1. P N fell to zero for an NL-intercept of 0.710±0.035 g(N) m–2. Under variable daily meteorological conditions the values for NL, specific leaf area (L), root mass fraction (Rf), P Nmax, and remained constant for a given N supply. A monotonic decline in the steady-state value of Rf occurred with increasing N supply. L increased with increasing N supply or with increasing NL.  相似文献   

10.
This study reports the analysis of K+ channel activity in bovine periaxolemmal-myelin and white matter-derived clathrin-coated vesicles. Channel activity was evaluated by the fusion of membrane vesicles with phospholipid bilayers formed across a patch-clamp pipette. In periaxolemmal myelin spontaneous K+ channels were observed with amplitudes of 25–30, 45–55, and 80–100 pS, all of which exhibited mean open-times of 1–2 msec. The open state probability of the 50 pS channel in periaxolemmal-myelin was increased by 6-methyldihydro-pyran-2-one. Periaxolemmal-myelin K+ channel activity was regulated by Ca2+. Little or no change in activity was observed when Ca2+ was added to thecis side of the bilayer. Addition of 10 M total Ca2+ also resulted in little change in K+ channel activity. However, at 80 M total Ca2+ all K+ channel activity was suppressed along with the activation of a 100 pS Cl channel. The K+ channel activity in periaxolemmal myelin was also regulated through a G-protein. Addition of GTPS to thetrans side of the bilayer resulted in a restriction of activity to the 45–50 pS channel which was present at all holding potentials. Endocytic coated vesicles, form in part through G-protein mediated events; white matter coated vesicles were analyzed for G proteins and for K+ channel activity. These vesicles, which previous studies had shown are derived from periaxolemmal domains, were found to be enriched in the subunits of G0, Gs, and Gi and the low molecular weight G protein,ras. As with periaxolemmal-myelin treated with GTPS, the vesicle membrane exhibited only the 50 pS channel. The channel was active at all holding potentials and had open times of 1–6 msec. Addition of GTPS to the bilayer fused with vesicle membrane appeared to suppress this channel activity at low voltages yet induced a hyperactive state at holding potentials of 45 mV or greater. The vesicle 50 pS K+ channel was also activated by the 6-methyl-dihydropyron-2-one (20 M).Abbreviations CNPase 2–3 cyclic nucleotide phosphohydrolase - EDTA ethylenediamine N,N,N,N-tetraacetic acid - G-protein GTP(guanosine triphosphate) binding protein - GTPS guanosine 5-O-(3-thiotriphosphate) - MAG myelin associated glycoprotein - Na+ K+ ATPase, Na+ and K+ stimulated adenosine triphosphatase - PLP myelin proteolipid protein Special issue dedicated to Dr. Majorie B. Lees.  相似文献   

11.
Andreas Renz  Mark Stitt 《Planta》1993,190(2):166-175
The substrate dependence and product inhibition of three different fructokinases and three different hexokinases from growing potato (Solanum tuberosum L.) tubers was investigated. The tubers contained three specific fructokinases (FK1, FK2, FK3) which had a high affinity for fructose K m=64, 90 and 100 (M) and effectively no activity with glucose or other hexose sugars. The affinity for ATP (K m=26, 25 and 240 M) was at least tenfold higher than for other nucleoside triphosphates. All three fructokinases showed product inhibition by high fructose (K i=5.7, 6.0 and 21 mM) and were also inhibited by ADP competitively to ATP. Sensitivity to ADP was increased in the presence of high fructose, or fructose-6-phosphate. In certain conditions, the K i (ADP) was about threefold below the K m (ATP). All three fructokinase were also inhibited by fructose-6-phosphate acting non-competitively to fructose (K i=1.3 mM for FK2). FK1 and FK2 showed very similar kinetic properties whereas FK3, which is only present at low activities in the tuber but high activities in the leaf, had a generally lower affinity for ATP, and lower sensitivity to inhibition by ADP and fructose. The tuber also contained three hexokinases (HK1, HK2, HK3) which had a high affinity for glucose (K m=41, 130 and 35 M) and mannose but a poor affinity for fructose (K m=11, 22 and 9 mM). All three hexokinases had a tenfold higher affinity for ATP (K m=90, 280 and 560 M) than for other nucleoside triphosphates. HK1 and HK2 were both inhibited by ADP (K i=40 and 108 M) acting competitively to ATP. HK1, but not HK2, was inhibited by glucose-6-phosphate, which acted non-competitively to glucose (K i=4.1 mM). HK1 and HK2 differed, in that HK1 had a narrower pH optimum, a higher affinity for its substrate, and showed inhibition by glucose-6-phosphate. The relevance of these properties for the regulation of hexose metabolism in vivo is discussed.Abbreviations FK fructokinase - Fru6P fructose-6-phosphate - Glc6P glucose-6-phosphate - HK hexokinase - NTP nucleoside triphosphate - Pi inorganic phosphate - UDPGlc uridine-5-diphosphoglucose This work was supported by the Deutsche Froschungsgemeinschaft (SFB 137). We are grateful to Professor E. Beck (Lehrstuhl für Pflanzenphysiologie, Universität Bayreuth, FRG) for providing laboratory facilities.  相似文献   

12.
DOPA synthesis from phenylalanine was studied in PC12 cells incubated with m-hydroxybenzylhydrazine, to inhibit aromatic L-amino acid decarboxylase. DOPA synthesis rose with increasing concentrations of either phenylalanine or tyrosine; maximal rates (~55 pmol/min/mg protein for tyrosine; ~40 pmol/min/mg protein for phenylalanine) occurred at a medium concentration of ~10 M for either amino acid. The Km for either amino acid was about 1 M (medium concentration). At tyrosine concentrations above 30 M, DOPA synthesis declined; inhibition was observed at higher concentrations for phenylalanine (300 M). These effects were most notable in the presence of 56 mM potassium. Measurements of intracellular phenylalanine and tyrosine suggested the Km for either amino acid is 20–30 M; maximal synthesis occurred at 120–140 M. In the presence of both phenylalanine and tyrosine, DOPA synthesis was inhibited by phenylalanine only at a high medium concentration (1000 M), regardless of medium tyrosine concentration. The inhibition of DOPA synthesis by high medium tyrosine concentrations was antagonized by high medium phenylalanine concentrations (100, 1000 M). Together, the findings indicate that for PC12 cells, phenylalanine can be a significant substrate for tyrosine hydroxylase, is a relatively weak inhibitor of the enzyme, and at high concentrations can antagonize substrate inhibition by tyrosine.  相似文献   

13.
To assess the long-term effect of increased CO2 and temperature on plants possessing the C3 photosynthetic pathway, Chenopodium album plants were grown at one of three treatment conditions: (1) 23 °C mean day temperature and a mean ambient partial pressure of CO2 equal to 350 bar; (2) 34 °C and 350 bar CO2; and (3) 34 °C and 750 bar CO2. No effect of the growth treatments was observed on the CO2 reponse of photosynthesis, the temperature response of photosynthesis, the content of Ribulose-1,5-bisphosphate carboxylase (Rubisco), or the activity of whole chain electron transport when measurements were made under identical conditions. This indicated a lack of photosynthetic acclimation in C. album to the range of temperature and CO2 used in the growth treatments. Plants from every treatment exhibited similar interactions between temperature and CO2 on photosynthetic activity. At low CO2 (< 300 bar), an increase in temperature from 25 to 35 °C was inhibitory for photosynthesis, while at elevated CO2 (> 400 bar), the same increase in temperature enhanced photosynthesis by up to 40%. In turn, the stimulation of photosynthesis by CO2 enrichment increased as temperature increased. Rubisco capacity was the primary limitation on photosynthetic activity at low CO2 (195 bar). As a consequence, the temperature response of A was relatively flat, reflecting a low temperature response of Rubisco at CO2 levels below its km for CO2. At elevated CO2 (750 bar), the temperature response of electron transport appeared to control the temperature dependency of photosynthesis above 18 °C. These results indicate that increasing CO2 and temperature could substantially enhance the carbon gain potential in tropical and subtropical habitats, unless feedbacks at the whole plant or ecosystem level limit the long-term response of photosynthesis to an increase in CO2 and temperature.Abbreviations A net CO2 assimilation rate - C a ambient partial pressure of CO2 - C i intercellular partial pressure of CO2 - Rubisco Ribulose-1,5-bisphosphate carboxylase - VPD vapor pressure difference between leaf and air  相似文献   

14.
Summary Suspensions of log phase cells ofRhodospirillum rubrum at pH 5.5 show a light-induced decrease in the pH of the medium which is reversed during the subsequent dark period. The velocity and magnitude of the pH change were the same whether the cells were bubbled with air, CO2-free air or N2 during experimentation. The pH response is temperature dependent. Phenazine methyl sulfate (PMS) at concentrations above 0.05mm stimulates the light-induced pH change. PMS at 1mm gives a 2-fold increase in the initial rate upon illumination and a 1.5-fold increase in the total change in pH after 2 min of illumination. The inhibition of the proton transport by 10 g/ml antimycin A or 20 m 2-n-heptyl-4-hydroxyquinoline-N-oxide can be partially relieved by PMS. However, inhibition of the light-induced proton transport with 0.5mm 2,4-dinitrophenol or 3 m carbonylcyanide-m-chlorophenylhydrazone (CCCP) cannot be overcome by addition of PMS. Valinomycin, at a concentration of 3 m, caused a slight stimulation of the light-induced proton transport in the presence of 200mm KCl. The inhibition of proton transport by 3 m CCCP was partially relieved with 3 m valinomycin in the presence of 200mm KCl, but the antibiotic was without effect when the cells were suspended in 200mm NaCl. The results are discussed in terms of current theories of the action of PMS, antimycin A, valinomycin, and uncouplers on the light-induced electron flow and photophosphorylation inR. rubrum.  相似文献   

15.
The effects of eight microelements (I, BO3 3–, MoO4 2–, Co2+, Cu2+, Mn2+, Fe2+, Zn2+) on the biosynthesis of camptothecin and the growth of suspension cultures of Camptotheca acuminata were studied. The increase of I to 25 M l–1, Cu2+ to 1 M l–1, Co2+ to 2 M l–1 and MoO4 2– to 10 M l–1 in Murashige and Skoog (MS) medium resulted in 1.66, 2.84, 2.53 and 2.04 times higher of camptothecin yield than that in standard MS medium respectively. Combined treatment of I (25 M l–1), Cu2+ (1 M l–1), Co2+ (2 M l–1) and MoO4 2– (10 M l–1) lead to improve cell dry weight, camptothecin content, and camptothecin yield to 30.56 g l–1, 0.0299%, and 9.15 mg l–1, respectively, which were 20.2, 208.9 and 273.8% increment respectively when compared with those of control.  相似文献   

16.
Elephant apple (Feronia limonia L.). was micropropagated on MS medium containing 4.4 M benzyladenine and 4.6 M kinetin using cotyledon explants taken from in vitro-grown seedlings. Adventitious buds formed on the cotyledon developed into shoots that were rooted in half-strength MS medium containing 0.57 M indoleacetic acid and 0.49 M indolebutyric acid. Plants were successfully established in soil.Abbreviations BA 6-benzyladenine - IAA 3-indoleacetic acid - IBA 3-indolebutyric acid - MS Murashige & Skoog  相似文献   

17.
A pyruvate, orthophosphate dikinase (EC 2.7.9.1) has been isolated from Acetobacter aceti grown on pyruvate as the only source of carbon and energy. The enzyme was purified 65-fold, and its molecular weight was determined to be about 330,000 by gel filtration.The optimum pH was 8.0 in the forward direction [phosphoenolpyruvate (PEP) formation] and 7.1 for the backward reaction (pyruvate production). In both directions Mg2+ was required (forward K m 1.70 mM; reverse K m 0.87 mM) and no other divalent cation was able to replace it. The K m values for pyruvate, ATP, and Pi were 27 M, 0.20 mM, and 0.83 mM, respectively, in the forward direction. The K m values for PEP, AMP, and PPi were 0.13 mM, 6 M, and 62 M, respectively, for the reverse reaction. The substrate-product pairs pyruvate-PEP, ATP-AMP, Pi-PPi were competitive inhibitors to each other in both directions. These product inhibition studies suggest for the enzyme from A. aceti nonclassical three-site Tri (Uni Uni) Ping-Pong kinetics.Abbreviations PEP phosphoenolpyruvate - OAA oxaloacetate - MW molecular weight - SDS sodium dodecyl sulphate - TEMG buffer 50 mM Tris-HCl, pH 7.5, 1 mM EDTA, 5 mM MgCl2, 1 mM glutathione  相似文献   

18.
A photoinduced-H2 production system, coupling cellulose degradation by cellulase and glucose dehydrogenase (GDH) and H2 production with colloidal Pt as a catalyst using the visible light-induced photosensitization of Mg chlorophyll a, has been developed. When the sample solution containing methylcellulose, cellulase, GDH, NAD+, Mg chlorophyll a, Methyl viologen and colloidal Pt was irradiated, continuous H2 production was observed. The amount of H2 production was about 12 mol after 4 h irradiation.  相似文献   

19.
A specific system for taurine transport is present at the early stages of development in both chick and rat retinas. The results obtained with taurine analogs indicate a high degree of specificity of taurine uptake. Two transport systems were detected for the adult rat retina: a high-affinity (K m 21 M) and a low-affinity transport system (K m 312 M). On the other hand, in the adult chick retina, only a low-affinity transport system (K m 580 M) could be detected. Nevertheless, embryo chick retina accumulated [3H]taurine by two different kinetic mechanisms withK m s of 242 M and 21 M for the low- and high-affinity processes, respectively. Taurine uptake systems were absolutely Na+ dependent. The sodium-dependence curve for taurine uptake was sigmoid. These mechanisms appear not to be mediated by a Na+ cotransport system. In spite of the differences observed in taurine uptake in both species, in each of them it closely parallels the changes brought about by the morphological and functional maturation of the retina.  相似文献   

20.
Summary A study has been made of the mineral requirements ofBacillus thuringiensis subsp.israelensis for production of the mosquitocide delta endotoxin. The optimum concentrations of K2HOP4, MgSO4.7H2O and CaCO3 for toxin production are 1g/l, 0.3g/l and 1g/l respectively while the elements Fe, Mn, Cu are required at levels of 2 g/ml, 5 g/ml and 0.25 g/ml respectively.  相似文献   

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