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1.
Soluble polyacrylate (PA), a polydisperse mixture of polyacrylate polymers, is strongly adsorbed and biodegradable. Biotic fate studies were carried out with once-through columns containing sand colonized with anaerobic biomass previously grown in a methanogenic fluidized bed. A fraction of soluble PA having a weight-average molecular weight of 16,700 and a range of molecular weight from 103 to 105 was biologically removed and mineralized to CO2. Due to its polydisperse nature, the breakthrough curve had a gradual increase to an apparent steady-state removal of approximately 60% near one day when the liquid detention time was 21 minutes. Modeling successfully explained the observed breakthrough result when the fraction was divided into components having a wide range of retardation factors (R): about 25% was strongly adsorbed (R=200 and 500), 45% was moderately adsorbed (R=50 and 100), and 30% was weakly adsorbed (R=1–10). In this study, in which active biomass already was present from utilization of a primary substrate (glucose here), equilibrium adsorption increased the time to breakthrough, which also reduced the exiting concentration by increasing the substrate contact time.  相似文献   

2.
Phosphatidic acid (PA) from swine and beef RBCs was isolated by chromatography on silicic acid columns. It comprised about 1 per cent of the total lipid phosphate in RBCs, but was eluted nearly pure from columns. An uncharacterized inositide accounted for 5 to 10 per cent of the phosphate in the PA-containing fraction. When cells were incubated with HP32O4=, the fraction containing PA became more radioactive than any of the other fractions obtained. However, analysis of the labeled material by paper chromatography showed that most of the P32 was in the inositide, not in PA. With the assumption of kinetic homogeneity for cellular PA, compartmental analysis of the kinetics of tracer incorporation showed that PA turnover is 3 to 4 orders of magnitude too slow to account for sodium extrusion by these cells.  相似文献   

3.
Abstract: The carotid injection technique, used previously to quantitate the kinetics of blood-brain barrier transport of metabolic substrates, may be modified to analyze the rate of cerebral glucose utilization. A 0.2-ml solution of [14C]glucose (GF) and [3H]methylglucose (M), an internal reference, is rapidly injected into the carotid artery, followed by microwave fixation of brain at various times up to 4 min after injection. The brain radioactivity is separated into a fraction containing neutral hexoses (GF and M) and a fraction containing metabolites of glucose. The GF/M ratio is related to the rate constant (k3) of brain glucose utilization by the simple, linear equation: In(GF/M) = In(GF°/M°) –k3t, where GF°/M°= the brain uptake index of glucose, relative to methylglucose, at 5-15 s after injection, and t= the time after carotid injection, e.g., 1–4 min. It is assumed that (a) the rate of influx due to recirculation of label is minimal during the 4-min circulation period; and (b) the rate constants of glucose efflux (k2) and methylglucose efflux (k2*) are identical. Independent estimates of k2 and k2* showed these parameters to be identical: k2= 0.14 + 0.08 min-I; k2*= 0.14 ± 0.02 min-I. A logarithmic plot of GF/M ratios versus time was linear (r = 0.99), and was described by the slope k2= 0.21 ± 0.02 min?1. Assuming glucose is uniformly distributed in brain, then the glycolytic rate = k3× brain glucose = (0.21 min?1) (2.6 μmol g?1) = 0.55 μmol min?1 g?1 for the cortex of the barbiturate-anesthetized rat. These studies provide the basis for a simple method of measurement of regional brain glycolysis that does not require either the use of correction factors, e.g., the lumped constant, or the use of differentially labeled glucose.  相似文献   

4.
Summary 1. Primary production by Chara vulgaris and by epipelic and epilithic algal assemblages was measured in a semiarid, Mediterranean stream (Chicamo stream, Murcia, Spain) during one annual cycle. 2. The rates of gross primary production (GPP) and community respiration (CR) were determined for each algal assemblage using oxygen change in chambers. The net daily metabolism (NDM) and the GPPd?1 : CR24 ratio were estimated by patch‐weighting the assemblage‐level metabolism values. 3. Gross primary production and CR showed significant differences between assemblages and dates. The highest rates were measured in summer and spring, while December was the only month when there were no significant differences in either parameters between assemblages. GPP was strongly correlated with respiration, but not with algal biomass. 4. Chara vulgaris showed the highest mean annual metabolic rates (GPP = 2.80 ± 0.83 gC m?2 h?1, CR = 0.76 ± 0.29 gC m?2 h?1), followed by the epilithic assemblage (GPP = 1.97 ± 0.73 gC m?2 h?1, CR = 0.41 ± 0.12 gC m?2 h?1) and epipelic algae (GPP = 1.36 ± 0.22 gC m?2 h?1, CR = 0.39 ± 0.06 gC m?2 h?1). 5. The epipelic assemblage dominated in terms of biomass (82%) and areal cover (88%), compared with the other primary producers. Epipelic algae contributed 84% of gross primary production and 86% of community respiration in the stream. 6. Mean monthly air temperature was the best single predictor of macrophyte respiration and of epipelic GPP and CR. However, ammonium concentration was the best single predictor of C. vulgaris GPP, and suspended solid concentration of epilithon GPP and CR. 7. Around 70% of the variation in both mean GPP and mean CR was explained by the mean monthly air temperature alone. A multiple regression model that included conductivity, PAR and nitrates in addition to mean monthly air temperature, explained 99.99% of the variation in mean CR. 8. Throughout the year, NDM was positive (mean value 7.03 gC m?2 day?1), while the GPP : CR24 ratio was higher than 1, confirming the net autotrophy of the system.  相似文献   

5.
Labeled glucose solution was introduced into cut fiber flax plants (45–50 cm high) under a pressure of 0.1 bar for 30 min, 1, and 2 h using a special device. The highest quantities of labeled carbon were revealed in the woody tissue. Sucrose made up a considerable proportion in low molecular weight products of [2-14C]-glucose transformation (23.5%). Metabolism of labeled glucose in the leaves exposed to sunlight yielded a set of metabolites similar to products of 14CO2 photoassimilation. In the shade, the pattern of 14C distribution in labeled compounds of the alcohol/water soluble fraction was similar to that in the light in mature leaves; while in juvenile leaves, 14C content decreased in sucrose and increased in organic and amino acids. In the shade, the incorporation of 14C into starch and hot water soluble polysaccharides increased at the expense of the acetone fraction (lipids and pigments), water/salt soluble proteins, and cellulose. Low light conditions increased the radioactivity ratio of sparingly soluble (KOH and Triton X-100 soluble) proteins to albumins and globulins. We propose that the synthesis of components of the photosynthetic apparatus in juvenile leaves is directly powered by photosynthesis and the photosynthesis of sucrose and the polymers compete for photosynthetic ATP. Appearance of sucrose in the xylem is due to its release from the phloem to the stem apoplast and the radial transfer to the xylem, where it is transported to the upper part of the shoot with the transpiration stream.__________Translated from Izvestiya Akademii Nauk, Seriya Biologicheskaya, No. 3, 2005, pp. 294–299.Original Russian Text Copyright © 2005 by Chikov, Avvakumova, Bakirova, Khamidullina.  相似文献   

6.
Rhizodeposition, i.e. the release of carbon into the soil by growing roots, is an important part of the terrestrial carbon cycle. However thein situ nature and dynamics of root-derived carbon in the soil are still poorly understood. Here we made an investigation of the latter in laboratory experiments using13CO2 pulse chase labelling of wheat (Triticum aestivum L.). We analyzed the kinetics of13C-labelled carbon and more specially13C carbohydrates in the rhizosphere. Wheat seedlings-soil mesocosms were exposed to13CO2 for 5 hours in controlled chambers and sampled repeatedly during two weeks for13C/C analysis of organic carbon. After a two-step separation of the soil from the roots, the amount of total organic13C was determined by isotope ratio mass spectrometry as well as the amounts of13C in arabinose, fructose, fucose, glucose, galactose, mannose, rhamnose and xylose. The amount and isotopic ratio of monosaccharides were obtained by capillary gas chromatography coupled with isotope ratio mass spectrometry (GC/C/IRMS) after trimethyl-silyl derivatization. Two fractions were analyzed : total (hydrolysable) and soluble monomeric (water extractable) soil sugars. The amount of organic13C found in the soil, expressed as a percentage of the total photosynthetically fixed13C at the end of the labelling period, reached 16% in the day following labelling and stabilised at 9% after one week. We concluded that glucose under the form of polymers was the dominant moietie of rhizodeposits. Soluble glucose and fructose were also present. But after 2 days, these soluble sugars had disappeared. Forty percent of the root-derived carbon was in the form of neutral sugars, and exhibited a time-increasing signature of microbial sugars. The composition of rhizospheric sugars rapidly tended towards that of bulk soil organic matter.  相似文献   

7.
The number of metabolically active bacteria was measured with nalidixic acid over two annual cycles at three depths in the epilimnion of hypertrophic Hartbeespoort Dam, South Africa. Concurrent measurements were made of water temperature, DOC, phytoplankton production of dissolved (EDOC) and particulate organic carbon, chlorophyll a and the uptake of glucose (Vmax). The objective was to determine the dominant factors correlated to the number of metabolically active bacteria and the relationship between active bacterial numbers and heterotrophic activity.The number of active bacteria was usually highest at the surface and ranged between 0.70 and 6.82 x 106 cells ml–1. The dominant factors correlated to the number of bacteria at the surface were water temperature (r = 0.65, n = 54, p<0.001), primary production (r = 0.53, n = 51, p<0.001) and EDOC (r = 0.37, n = 45, p = 0.005). Surface Vmax for glucose ranged between 0.11 and 4.0 µgC 1–1 h–1 and was positively correlated to the number of active bacteria (r = 0.61, n = 53, p<0.001). The specific activity index (10–12 µgC cell–1 h–1) varied between 80 and 2290 at the surface and was most strongly correlated to EDOC (r = 0.70, n = 48, p<0.001). Relationships between active bacterial numbers, water temperature, phytoplankton activity and glucose uptake were also found at two additional depths within the epilimnion. These data suggest that bacterial populations in nutrient enriched lakes contain a large number of metabolically active cells with high individual activity as a result of enhanced phytoplankton growth.  相似文献   

8.
Human placental microsomes exhibit uptake of d-[3H]glucose which is sensitive to inhibition by cytochalasin B (apparent Ki = 0.78 /gm M). Characterization of [3H]cytochalasin B binding to these membranes reveals a glucose-sensitive site, inhibited by d-glucose with an ED50 = 40 mM. The glucose-sensitive cytochalasin B binding site is found to have a Kd = 0.15μM by analysis according to Scatchard. Solubilization with octylglucoside extracts 60–70% of the glucose-sensitive binding component. Equilibrium dialysis binding of [3H]cytochalasin B to the soluble protein displays a pattern of inhibition by d-glucose similar to that observed for intact membranes, and the measurement of an ED50 = 37.5 mM d-glucose confirms the presence of the cytochalasin B binding component, putatively assigned as the glucose transporter. Further evidence is attained by photoaffinity labelling; ultraviolet-sensitive [3H]cytochalasin B incorporation into soluble protein (Mr range 42 000-68 000) is prevented by the presence of d-glucose. An identical photolabelling pattern is observed for incorporation of [3H]cytochalasin B into intact membrane protein, confirming the usefulness of this approach as a means of identifying the presence of the glucose transport protein under several conditions.  相似文献   

9.
The utility of recombinant herpes simplex virus type 1 (HSV-1) vectors may be expanded by manipulation of the virus envelope to achieve cell-specific gene delivery. To this end, an HSV-1 mutant virus deleted for glycoprotein C (gC) and the heparan sulfate binding domain of gB (KgBpKgC) was engineered to encode different chimeric proteins composed of N-terminally truncated forms of gC and the full-length erythropoietin hormone (EPO). Biochemical analyses demonstrated that one gC-EPO chimeric molecule (gCEPO2) was posttranslationally processed, incorporated into recombinant HSV-1 virus (KgBpKgCEPO2), and neutralized with antibodies directed against gC or EPO in a complement-dependent manner. Moreover, KgBpKgCEPO2 recombinant virus was specifically retained on a soluble EPO receptor column, was neutralized by soluble EPO receptor, and stimulated proliferation of FD-EPO cells, an EPO growth-dependent cell line. FD-EPO cells were nevertheless refractory to productive infection by both wild-type HSV-1 and recombinant KgBpKgCEPO2 virus. Transmission electron microscopy of FD-EPO cells infected with KgBpKgCEPO2 showed virus endocytosis leading to aborted infection. Despite the lack of productive infection, these data provide the first evidence of targeted HSV-1 binding to a non-HSV-1 cell surface receptor.  相似文献   

10.
We report some properties of Protein PA which has been isolated from the soluble fraction of a chlB mutant after anaerobic growth in the presence of KNO3. This protein has been identified by its capacity to reactivate nitrate reductase present in the soluble fraction of a chlA mutant by the complementation process. The presence of active Protein PA in the chlB mutant is independent of the presence of oxygen or of nitrate during growth. In contrast, the addition of sodium tungstate to the growth medium leads to the formation of inactive Protein PA which is not able to activate nitrate reductase in the chlA-soluble extract by complementation. Inactive Protein PA has been quantitated immunologically. The partial purification of Protein PA has been achieved from various chlorate-resistant mutants (chlA?chlG). The establishment of particular complementation systems comprising the soluble extracts of chlA or chlB mutants and partially purified Protein PA from soluble fractions of different chlorate-resistant mutants, has allowed the quantitative estimation of this protein. The analysis by ‘rocket immunoelectrophoresis’ using an antiserum specific for Protein PA has shown that inactive Protein PA is present in approximately equivalent amounts in the chlA, chlE, chlG and chlD mutants  相似文献   

11.
By the example of glucose uptake by the soil bacteria Pseudomonas aureofaciens BS1393(pBS216) and Rhodococcus sp. 3–30 immobilized on a solid-phase surface (quartz sand), their growth parameters were determined: growth rate (doubling time), total CO2 production, CO2 production per cell, lag period with respect to substrate uptake, respiratory quotient. The growth of P. aureofaciens and Rhodococcus sp. on glucose revealed (1) differences of the lag period with respect to substrate (lag time of ~4 h for P. aureofaciens and ~26 h for Rhodococcus sp.); (2) differences between the maximal rates of CO2 production (~50 μg C-CO2 g?1 sand h?1 for P. aureofaciens and ~8.5 μg C-CO2 g?1 sand h?1 for Rhodococcus sp.); (3) differences in CO2 production per cell (~1.94 × 10?9 μM CO2/CFU for P. aureofaciens and more than ~3.4 × 10?9 μM CO2/CFU for Rhodococcus sp.). The kinetics of the metabolic CO2 isotopic composition was shown to be determined by the difference in the carbon isotopic characteristics of products in the cell. Upon introduction of glucose into the medium (the preparatory stage of the metabolism), the uptake of intracellular 13C-depleted products (lipids) is noted; at the stage of the maximal cell growth rate, introduced glucose is mainly metabolized; and at the final stage, upon exhaustion of substrate, the “stored” products—the lipid fraction—get involved in the metabolism. At the maximal rate of glucose uptake, the CO2 carbon isotopic fractionation coefficient relative to organic products of microbial biosynthesis was determined to be α = 1.009 ± 0.002.  相似文献   

12.
The metabolism of exogenously added d-myo-[1-3H]inositol 1,4,5-trisphosphate (IP3) has been examined in microsomal membrane and soluble fractions of carrot (Daucus carota L.) cells grown in suspension culture. When [3H]IP3 was added to a microsomal membrane fraction, [3H]IP2 was the primary metabolite consisting of approximately 83% of the total recovered [3H] by paper electrophoresis. [3H]IP was only 6% of the [3H] recovered, and 10% of the [3H]IP3 was not further metabolized. In contrast, when [3H]IP3 was added to the soluble fraction, approximately equal amounts of [3H]IP2 and [3H]IP were recovered. Ca2+ (100 micromolar) tended to enhance IP3 dephosphorylation but inhibited the IP2 dephosphorylation in the soluble fraction by about 20%. MoO42− (1 millimolar) inhibited the dephosphorylation of IP3 by the microsomal fraction and the dephosphorylation of IP2 by the soluble fraction. MoO42−, however, did not inhibit the dephosphorylation of IP3 by the soluble fraction. Li+ (10 and 50 millimolar) had no effect on IP3 metabolism in either the soluble or membrane fraction; however, Li+ (50 millimolar) inhibited IP2 dephosphorylation in the soluble fraction about 25%.  相似文献   

13.
Mechanical treatments of cell suspensions of Escherichia coli K 12 strain PA 601, and its two mutants chl A and chl B, in a buffer without Mg2+ lead to partial solubilization of membrane-bound ATPase. After ultracentrifugation of cell-free extracts, ATPase can be recovered in the soluble fraction. Contrary to membrane ATPase, the soluble enzyme has the following properties: (1) it is insensitive to N,N′-dicyclohexylcarbodiimide; (2) heat-inactivation kinetics show a reactivation in the first 3 min and the half-time is 15 min; (3) ADP is a substrate. In the course of complementation between soluble fractions of mutants chl A and chl B, a part of soluble ATPase is incorporated into the newly formed particles. The specific activity of these particles is nearly the same as that of native particles; the ATPase bound to native membrane and the ATPase bound to the newly-formed particles both have the same biochemical properties.  相似文献   

14.
Following application of 3H-Gibberellin A20 (GA20) to roots of G2 pea seedlings and homogenization of the roots, about 3% of the radioactivity in the tissue could be precipitated from a 30,000 × g supernatant with trichloroacetic acid (TCA) (soluble fraction) while about 5% of the radioactivity pelleted at 30,000 × g (particulate fraction). The radioactivity in the particulate fraction was soluble in sodium dodecyl sulfate (SDS), but was not dialyzable and was insoluble in ethanol. Electrophoresis of the soluble fraction gave only one band of radioactivity, while that of the particulate fraction gave multiple bands. Acid hydrolysis of the soluble fraction released radioactivity that ran coincident with acid-treated GA20 on silicic-acid column chromatography. The particulate fraction gave numerous radioactive peaks following acid hydrolysis, two of which were coincident with GA20 and GA29 (hydroxylation product of GA20) on silicic acid chromatography. Treatment of the particulate and soluble fractions with RNase, DNase, and proteases showed a significant solubilization of radioactivity only with the proteases, suggesting that the GA is bound to a proteinaceous macromolecule. Complete proteolytic hydrolyis followed by thin layer chromatography showed 65% of the radioactivity from the soluble fraction running separately from free GAs or the individual amino acids; the particulate fraction gave mainly (60%) free GAs on enzymatic hydrolysis and much smaller amounts (17%) in a position separate from that of the GAs or amino acids. Binding of 3H-GA to protease-sensitive material was obtained with biologically active 3H-GA20 and 3H-GA1.  相似文献   

15.
The rate of ADP-glucose formation from [14C]glucose 6-phosphate and ATP by the soluble fraction of lysed chloroplasts is studied as a function of the levels of metabolites (3-phosphoglycerate, orthophosphate, hexose monophosphate, and ATP) as determined in whole chloroplasts of Spinacia oleracea in light and dark.  相似文献   

16.
Two soluble hexokinases and a particulate hexokinase have been separated and partially purified from spinach leaves. One of the soluble hexokinases showed a high affinity for glucose (Km = 63 μM) which was far greater than that for fructose (Km = 9.1 mM). However, with saturating fructose the activity was twice that with saturating glucose. The particulate hexokinase showed kinetic properties similar to those of this soluble hexokinase. The second soluble hexokinase was distinct in that it was much more active with fructose than with glucose at all concentrations tested, although the Km values for these hexoses (210 μM and 71 μM respectively) were similar. The activity of this hexokinase was stimulated by the monovalent cations K+ and NH4+.  相似文献   

17.
It was to be shown whether during the biogenesis of microbodies some of their components were already present in the cell prior to the organelle's assembly. To this end, the occurrence and properties of catalase in soluble and particular fractions of ripening cucumber seeds were examined. Homogenates of seeds from ripening fruits were fractionated by isopycnic density gradient centrifugation, and thus catalase was found in three different fractions: as a soluble enzyme in the gradient supernatant, as a membrane fraction at density d=1.18 kg l-1, and in association with microbodies. In the early steps of seed formation, catalase was detected at density d=1.18 kg l-1 and in the gradient supernatant. At a later stage of seed maturation, however, catalase was primarily associated with microbodies which exhibited an equilibrium density of d=1.23 kg l-1. M r as well as subunit M r of catalase were determined, and their close immunological relationship to leaf peroxisomal catalase and glyoxysomal catalase was demonstrated. Biosynthesis of catalase at different stages of seed maturation was investigated by in vivo labeling with l-[35S]methionine, l-[14C]leucine and -[3H]aminolaevulinic acid. Electrophoretic analysis of de novo synthesized catalase subunits revealed the occurrence of a heavy form (M r 57,500) in the soluble fraction; this form was preferentially labeled. A light form, M r 53,500, was detected in microbodies and also in the soluble fraction. The findings lend support to the hypothesis that the rate of catalase synthesis is highest in an early stage of seed formation, when globulins have already been formed, but before de novo synthesis of malate synthase has commenced. Prior to microbody assembling, a cytoplasmic pool of catalase was labeled.Abbreviations EDTA Na2-ethylenediaminotetraacetate - Hepes 4-(2-Hydroxyethyl)-1-piperazineethanesulfonic acid - M r molecular weight  相似文献   

18.
A glycolipid fraction was isolated from Corynebacterium ovis (C. pseudotuberculosis). It had [α]D25 = + 63.2° (C = 0.5, CHCl3) and m.p. 43–46°C; the sugar content was 26%, determined as trehalose. Alkaline hydrolysis of the isolated fraction found trehalose as the sole water-soluble component, while glucose was found only after acid hydrolysis of the aqueous phase. Saturated and unsaturated short-chain mycolic acids with carbon atoms ranging from C30 to C36 were the constituents of the fatty acid moiety. The glycolipid fraction of C. ovis is therefore assumed to be a mixture of trehalose esters in which the trehalose molecule is esterified by saturated and unsaturated short-chain (C30–C36) mycolic acids.  相似文献   

19.
The subcellular localization of enterokinase is controversial. In this study, enterokinase was extracted from a soluble fraction and a brush border fraction of rat small intestine by differential centrifugation. The soluble fraction contained 41% of the initial enterokinase activity while the brush border fraction contained only 4.6% of the initial activity. In contrast, alkaline phosphatase monitored as a brush border marker, yielded 26.3 in the brush border fraction and only 6% in the soluble fraction. Further separation of the soluble fraction on a Sepharose 4B column revealed three peaks of enterokinase activity. One small peak (3%) of a bound enzyme (Mr, 2·10?6) and two larger peaks of free enzyme (Mr, 3·105 and 9·105). In contrast, alkaline phosphatase major fraction was in a high molecular weight peak of bound enzyme. When the brush border fraction was chromatographed only a single peak of bound enterokinase and alkaline phosphatase were found. In the lower part of the small intestine, no brush border-bound enterokinase was found, while the peak of alkaline phosphatase was the same as in the upper intestine. These data suggest that enterokinase activity in the rat intestine is mainly in a free form localized in the mucin and soluble fraction and to a negligible extent in the brush border.  相似文献   

20.
Labeling pattern of translocated stachyose in squash   总被引:4,自引:4,他引:0       下载免费PDF全文
Hendrix JE 《Plant physiology》1968,43(10):1631-1636
One mature blade of each squash plant was continuously labeled with 14CO2 for 15, 30, or 70 minutes in light. The ethanol soluble materials from serial sections of petioles were extracted and separated by paper chromatography. The ratios of label in the various components of this fraction were determined. Stachyose, which contained the major portion of the label of this fraction, was hydrolyzed and the resultant hydrolysate was separated by paper chromatography. Specific activities of the hexoses derived from stachyose were determined. It was found that the glucose and fructose moieties of stachyose became labeled at the same rates; however, the galactose moiety became labeled more rapidly.  相似文献   

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