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1.
Biofuel cell system employing thermostable glucose dehydrogenase 总被引:1,自引:0,他引:1
Okuda-Shimazaki J Kakehi N Yamazaki T Tomiyama M Sode K 《Biotechnology letters》2008,30(10):1753-1758
Enzyme biofuel cells utilizing glucose dehydrogenase as an anode enzyme were constructed. The glucose dehydrogenase is composed of a catalytic subunit, an electron transfer subunit, and a chaperon-like subunit. Cells, constructed using either a glucose dehydrogenase catalytic subunit or a glucose dehydrogenase complex, displayed power outputs that were dependent on the glucose concentration. The catalytic subunit in the anode maintained its catalytic activity for 24 h of operation. The biofuel cell which composed of glucose dehydrogenase complex functioned successfully even in the absence of an electron mediator at the anode cell. These results indicate the potential application of this thermostable glucose dehydrogenase for the construction of a compartment-less biofuel cell. 相似文献
2.
枯草芽孢杆菌葡萄糖脱氢酶基因的克隆及其序列分析 总被引:4,自引:1,他引:3
根据Lampel报道的葡萄糖脱氢酶基因序列设计合成两条引物,以野生型枯草芽孢杆菌染色体DNA为模板,PCR扩增得到含有葡萄糖脱氢酶基因的大约780bp的DNA片段,将其克隆到pUC-T载体中。序列分析表明,克隆得到的葡萄糖脱氢酶基因含有783bp,编码261个氨基酸的蛋白质。得到的基因序列与文献报道的进行比较,其核苷酸同源率为75.5%,编码氨基酸序列的同源率为83.9%。 相似文献
3.
The ability of a microsomal enzyme, glucose dehydrogenase (hexose 6-phosphate dehydrogenease) to supply NADPH to the microsomal electron transport system, was investigated. Microsomes could perform oxidative demethylation of aminopyrine using microsomal glucose dehydrogenase in situ as an NADPH generator. This demethylation reaction had apparent Km values of 2.61 X 10(-5) M for NADP+, 4.93 X 10(-5) m for glucose 6-phosphate, and 2.14 X 10(-4) m for 2-deoxyglucose 6-phosphate, a synthetic substrate for glucose dehydrogenase. Phenobarbital treatment enhanced this demethylation activity more markedly than glucose dehydrogenase activity itself. Latent activity of glucose dehydrogenase in intact microsomes could be detected by using inhibitors of microsomal electron transport, i.e. carbon monoxide and p-chloromercuribenzoate (PCMB), and under anaerobic conditions. These observations indicate that in microsomes the NADPH generated by glucose dehydrogenase is immediately oxidized by NADPH-cytochrome c reductase, and that glucose dehydrogenase may be functioning to supply NADPH. 相似文献
4.
Enzyme histochemical observations on the segmentation of the proximal tubules in the kidney of the female rat. 总被引:1,自引:0,他引:1
N O Jacobsen 《Histochemistry》1975,43(1):11-32
The segmentation of the proximal tubules in the kidney of the female rat was studied by means of enzyme histochemical reactions and the results compared with those observed in male and recently described by Jacobsen and J0rgensen (1973 a). Reactions were performed for the following soluble, coezyme-dependent oxido-reductases: glucose 6-phosphate dehydrogenase, alpha-glycerophosphate dehydrogenase, 3 alpha-hydroxysteroid dehydrogenase, NAD-as well as NADP-dependent isocitrate dehydrogenases, NAD-dependent malate dehydrogenase, NADP-dependent, decarboxylating malate dehydrogenase, uridine diphosphate glucose dehydrogenase. Measures were taken to reduce enzyme diffusion and eliminate interference from tissue tetrazolium reductases. Furthermore, reactions were performed for a number of less soluble or insoluble enzymes: glucose 6-phosphatase, mitochondrial alpha-glycerophosphate dehydrogenase, beta-hydroxybutyrate dehydrogenase, succinate dehydrogenase and tetrazolium reductases. In the proximal tubules of the female rat all enzymes studied--except beta-hydroxybutyrate dehydrogenase--showed segmental differences, most of them clearly revealing three segments. Sex differences were found concerning all enzymes except uridine diphosphate glucose dehydrogenase and NADP-dependent isocitrate dehydrogenase. The most pronounced sex-related differences were seen in the third segment in which part the male rat showed highest activity in respect to tetrazolium reductases, NAD-dependent isocitrate dehydrogenase, succinate dehydrogenase, beta-hydroxybutyrate dehydrogenase, 3 alpha-hydroxysteroid dehydrogenase and glucose 6-phosphate dehydrogenase and the female in respect to glucose 6-phosphatase, alpha-glycerophosphate dehydrogenases, and NADP-dependent, decarboxylating malate dehydrogenase. A few of the enzymes exhibited minor sex differences in the first two segments. 相似文献
5.
Ralph N. Martins Gilbert B. Stokes Colin L. Masters 《Molecular and cellular biochemistry》1986,70(2):169-175
Summary Liver glucose 6-phosphate dehydrogenase and phosphogluconate dehydrogenase activities were significantly decreased in both diabetic and fasted rats. Treatment of diabetic rats with insulin resulted in liver glucose 6-phosphate dehydrogenase and phosphogluconate dehydrogenase activities that were significantly greater than controls. Insulin promoted an increase in food consumption that was blocked by adrenaline. Insulin, when administered together with adrenaline, restored hepatic glucose 6-phosphate dehydrogenase and phosphogluconate dehydrogenas activities of diabetic animals to control values, without altering food consumption. Brain glucose 6-phosphate dehydrogenase and phosphogluconate dehydrogenase activities were not significantly altered by either dietary restriction, diabetes or insulin treatment. These results demonstrate a dissociation between the action of insulin on hepatic glucose 6-phosphate dehydrogenase activity and its action to increase food intake.Abbreviations NADP+ oxidoreductase, EC 1.1.1.49
Glucose 6-P dehydrogenase, GPD, D-glucose-6-phosphate
- NADP+ 2-oxidoreductase (decarboxylating), EC 1.1.1.44
phosphogluconate dehydrogenase, PGD, 6-phospho-D-gluconate 相似文献
6.
7.
Clark Bublitz 《Biochemical and biophysical research communications》1981,98(2):455-462
The relative substrate specificities of glucose dehydrogenases (E.C. 1.1.1.47) from beef liver and rat liver are very different. The beef enzyme oxidizes glucose more rapidly than either glucose-6-phosphate or galactose-6-phosphate. On the other hand, the dehydrogenase from rat liver prefers the hexose phosphates to glucose.A procedure for estimating the level of glucose dehydrogenase in rat and beef liver is described. The glucose-6-phosphate dehydrogenase activity attributed to glucose dehydrogenases is estimated to be about one-fifth and one-third that of cytoplasmic glucose-6-phosphate dehydrogenase (E.C. 1.1.1.49) in female and male rat liver respectively.A fluorometric adaptation of the less sensitive spectrophotometric assay for glucose dehydrogenase is described. 相似文献
8.
Ivanova EV Ershov AY Laurinavicius V Meskus R Ryabov AD 《Biochemistry. Biokhimii?a》2003,68(4):407-415
The comparative kinetic study of two glucose oxidizing enzymes, FAD-dependent glucose oxidase and PQQ-dependent glucose dehydrogenase, is presented in the artificial electron transfer mediator system based on ruthenium(III) compounds. It is demonstrated that FAD-dependent glucose oxidase and PQQ-dependent glucose dehydrogenase follow Michaelis kinetics in the D-glucose/ruthenium(III) system. PQQ-dependent glucose dehydrogenase is more active than FAD-dependent glucose oxidase in the process of D-glucose oxidation by ruthenium(III) compounds, this being due to the different catalytic mechanisms of these enzymes. 相似文献
9.
When strain C3 of Klebsiella pneumoniae is grown on a minimal medium with excess glucose, isocitrate dehydrogenase, malate dehydrogenase, and succinate dehydrogenase specific activities increase in the last period of the exponential growth phase and in the beginning of the stationary phase. Glucose exhaustion does not alter the development of malate dehydrogenase and succinate dehydrogenase, but specific activities are higher than those obtained with excess glucose. In contrast, glucose exhaustion can be correlated with a decrease of isocitrate dehydrogenase specific activity in the stationary phase. Induction of strain C3 isocitrate dehydrogenase by glucose in complex medium and repression by cAMP in mineral medium were observed. Glucose induction and the NADP/NADPH ratio are suggested as regulatory mechanisms controlling isocitrate dehydrogenase synthesis in the Enterobacteriaceae, but the former appears to be restricted to some Klebsiella strains. 相似文献
10.
Differential Sensitivities of the Two Malate Dehydrogenases and the Maltose Permease to the Effect of Glucose in Saccharomyces carlsbergensis 总被引:6,自引:5,他引:1
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In Saccharomyces carlsbergensis the two malate dehydrogenase activities, which are localized in different compartments of the cell, were found to differ in their response to glucose. The cytoplasmic malate dehydrogenase activity appears to be sensitive to inactivation by very low concentrations of glucose. The mitochondrial malate dehydrogenase activity is only repressed at a higher glucose concentration. Maltose permease is also sensitive to inactivation by glucose. Conditions were found such that the maltose permease was present while the cytoplasmic malate dehydrogenase was inactivated. The different sensitivities of the two malate dehydrogenases and maltose permease to the effect of glucose may explain the preferential use of glucose, maltose, and products of glucose metabolism (2- and 3-carbon skeletons) as carbon sources for growth in the order as mentioned. 相似文献
11.
F. Monot Y. Benoit J. Lemal A. Honorat D. Ballerini 《Biocatalysis and Biotransformation》1989,2(3):161-174
An improved method for the simultaneous production of valine dehydrogenase and glucose dehydrogenase by Bacillus megaterium (ATCC 39118) is described. The highest yields in volumetric activities (8200 U.S-1' of glucose dehydrogenase and 7200 U.S1 of valine dehydrogenase) were obtained using a fed batch cultivation technique with glucose, yeast extract and corn steep liquor in the feed medium. The main characteristics (stability, optimal pH, Michaelis constants, substrate and product inhibitions) of valine dehydrogenase and glucose dehydrogenase from crude extracts were determined. B. megaterium crude extract was suitable for synthesis of L-valine from $aL-keto isovalerate with glucose dehydrogenase as the NADH-regenerating enzyme and the conditions of the conversion have been optimized. $aL-Keto acid was supplied in fed batch mode in order to avoid substrate inhibition and was not involved in side reactions. With the optimized system, a concentration of 95 mM L-valine was obtained in 45 hours with a molar conversion yield close to 100%. 相似文献
12.
Reactivative Action of Ethylenediaminetetraacetic Acid or Dipicolinic Acid on Inactive Glucose Dehydrogenase Obtained from Heated Spores of Bacillus subtilis 总被引:2,自引:2,他引:0
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Partially purified inactive glucose dehydrogenase obtained from spores which were heated at 87 or 90 C for 30 min is converted to an active from by the addition of ethylenediaminetetraacetic acid, dipicolinic acid, or some salts. The molecular weight of the inactive glucose dehydrogenase in the heated spores is about one-half of that of the active glucose dehydrogenase in the intact resting spores. The possibility is discussed that the active glucose dehydrogenase in the intact resting spores divides into subunits and is converted to stable and inactive form during heating of spores at a particular range of temperature (87 to 90 C). 相似文献
13.
R. W. J. Hommes P. T. D. Herman P. W. Postma D. W. Tempest O. M. Neijssel 《Archives of microbiology》1989,151(3):257-260
No holoenzyme pyrroloquinoline quinone (PQQ)-dependent glucose dehydrogenase and only very low apoenzyme levels could be detected in cells of Klebsiella pneumoniae, growing anaerobically, or carrying out a fumarate or nitrate respiration. Low glucose dehydrogenase activity in some aerobic glucose-excess cultures of K. pneumoniae (ammonia or sulphate limitation) was increased significantly by addition of PQQ, whereas in cells already possessing a high glucose dehydrogenase activity (phosphate or potassium limitation) extra PQQ had almost no effect. These observations indicate that the glucose dehydrogenase activity in K. pneumoniae is modulated by both PQQ synthesis and synthesis of the glucose dehydrogenase apo-enzyme.Abbreviations PQQ
2, 7, 9-tricarboxy-1H-pyrrolo-(2,3-f)quinoline-4,5-dione
- WB
Wurster's Blue (1,4-bis-(dimethylamino)-benzene perchlorate) 相似文献
14.
Glucose dehydrogenase from rat liver microsomes was found to react not only with glucose as a substrate but also with glucose 6-phosphate, 2-deoxyglucose 6-phosphate and galactose 6-phosphate. The relative maximum activity of this enzyme was 29% for glucose 6-phosphate, 99% for 2-deoxyglucose 6-phosphate, and 25% for galactose 6-phosphate, compared with 100% for glucose with NADP. The enzyme could utilize either NAD or NADP as a coenzyme. Using polyacrylamide gradient gel electrophoresis, we were able to detect several enzymatically active bands by incubation of the gels in a tetrazolium assay mixture. Each band had different Km values for the substrates (3.0 x 10(-5)M glucose 6-phosphate with NADP to 2.4M glucose with NAD) and for coenzymes (1.3 x 10(-6)M NAD with galactose 6-phosphate to 5.9 x 10(-5)M NAD with glucose). Though glucose 6-phosphate and galactose 6-phosphate reacted with glucose dehydrogenase, they inhibited the reaction of this enzyme only when either glucose or 2-deoxyglucose 6-phosphate was used as a substrate. The Ki values for glucose 6-phosphate with glucose as substrate were 4.0 x 10(-6)M with NAD, and 8.4 x 10(-6)M with NADP; for galactose 6-phosphate they were 6.7 x10(-6)M with NAD and 6.0 x 10(-6)M with NADP. The Ki values for glucose 6-phosphate with 2-deoxyglucose 6-phosphate as substrate were 6.3 x 10(-6)M with NAD and 8.9 x 10(-6)M with NADP; and for galactose 6-phosphate, 8.0 x 10(-6)M with NAD and 3.5 x 10(-6)M with NADP. Both NADH and NADPH inhibited glucose dehydrogenase when the corresponding oxidized coenzymes were used (Ki values: 8.0 x 10(-5)M by NADH and 9.1 x 10(-5)M by NADPH), while only NADPH inhibited cytoplasmic glucose 6-phosphate dehydrogenase (Ki: 2.4 x 10(-5)M). The results indicate that glucose dehydrogenase cannot directly oxidize glucose in vivo, but it might play a similar role to glucose 6-phosphate dehydrogenase. The differences in the kinetics of glucose dehydrogenase and glucose 6-phosphate dehydrogenase show that glucose 6-phosphate and galactose 6-phosphate could be metabolized in quite different ways in the microsomes and cytoplasm of rat liver. 相似文献
15.
Various concentrations of Tergitol NP-10 stimulate mannose-6-phosphatase and glucose dehydrogenase to the same extent in untreated rat liver microsomes. Thus, the latency of glucose dehydrogenase may be used as an alternative to mannose phosphatase as a measure of the integrity of the microsomal membrane. The advantage of using glucose dehydrogenase rather than mannose phosphatase to monitor microsomal integrity is that NADH is more easily measured than Pi. 相似文献
16.
The preparation of nylon-tube-supported hexokinase and glucose 6-phosphate dehydrogenase and the use of the co-immobilized enzymes in the automated determination of glucose.
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Triethyloxonium tetrafluoroborate was used to O-alkylate nylon-tube thus producing the imidate salt of the nylon which was further made to react with 1,6-diaminohexane. 2. Hexokinase (EC 2.7.1.1) and glucose 6-phosphate dehydrogenase (EC 1.1.1.49) were immobilized on the amino-substituted nylon tube through glutaraldeyde and bisimidates. 3. The effect of varying the conditions of O-alkylation and the amount of enzyme immobilized on the activity of nylon tube-hexokinase derivatives was determined. 4. The effect of varying the amount of enzyme immobilized on the activity of nylon-tube-glucose 6-phosphate dehydrogenase derivatives was determined. 5. The thermal stability of nylon-tube-hexokinase and nylon-tube-glucose 6-phosphate dehydrogenase derivatives was studied. 6. Different ratios of hexokinase and glucose 6-phosphate dehydrogenase were co-immobilized on nylon tube, and the rate of conversion of glucose into 6-phosphogluconolactone was compared with the individual activities of the immobilized enzymes. 7. Hexokinase and glucose 6-phosphate dehydrogenase co-immobilized on nylon tube were used in the automated analysis of glucose. 相似文献
17.
This review discusses recent achievements in the field of cofactor regeneration for the nicotinamide cofactors NADH and NADPH. The examples discussed include alcohol dehydrogenases, formate dehydrogenase, glucose dehydrogenase and a hydrogenase. For the reaction either one-phase systems or two-phase systems in combination with an organic solvent are discussed. For the enantioselective reduction of 2-octanone to (R)-2-octanol it could be shown that enzyme coupled NADPH regeneration with glucose dehydrogenase and glucose results in shorter reaction times and higher yields when compared to the substrate coupled regeneration with 2-propanol.ADH: alcohol dehydrogenase; LDH: Lactose dehydrogenase; GDH: Glucose dehydrogenase; FDH: Formate dehydrogenase; LB-ADH: alcohol dehydrogenase from Lactobacillus brevis; HL-ADH: alcohol dehydrogenase from horse liver; TB-ADH: alcohol dehydrogenase from Thermoanaerobicum brockii; PS-GDH: Glucose dehydrogenase from Pseudomonas species; [BMIM][PF6]: Butyl-methyl-imidazoliumhexafluorophosphate 相似文献
18.
The role of nicotinamide–adenine dinucleotide phosphate-dependent malate dehydrogenase and isocitrate dehydrogenase in the supply of reduced nicotinamide–adenine dinucleotide phosphate for steroidogenesis in the superovulated rat ovary
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1. Superovulated rat ovary was found to contain high activities of NADP-malate dehydrogenase and NADP-isocitrate dehydrogenase. The activity of each enzyme was approximately four times that of glucose 6-phosphate dehydrogenase and equalled or exceeded the activities reported to be present in other mammalian tissues. Fractionation of a whole tissue homogenate of superovulated rat ovary indicated that both enzymes were exclusively cytoplasmic. The tissue was also found to contain pyruvate carboxylase (exclusively mitochondrial), NAD-malate dehydrogenase and aspartate aminotransferase (both mitochondrial and cytoplasmic) and ATP-citrate lyase (exclusively cytoplasmic). 2. The kinetic properties of glucose 6-phosphate dehydrogenase, NADP-malate dehydrogenase and NADP-isocitrate dehydrogenase were determined and compared with the whole-tissue concentrations of their substrates and NADPH; NADPH is a competitive inhibitor of all three enzymes. The concentrations of glucose 6-phosphate, malate and isocitrate in incubated tissue slices were raised at least tenfold by the addition of glucose to the incubation medium, from the values below to values above the respective K(m) values of the dehydrogenases. Glucose doubled the tissue concentration of NADPH. 3. Steroidogenesis from acetate is stimulated by glucose in slices of superovulated rat ovary incubated in vitro. It was found that this stimulatory effect of glucose can be mimicked by malate, isocitrate, lactate and pyruvate. 4. It is concluded that NADP-malate dehydrogenase or NADP-isocitrate dehydrogenase or both may play an important role in the formation of NADPH in the superovulated rat ovary. It is suggested that the stimulatory effect of glucose on steroidogenesis from acetate results from an increased rate of NADPH formation through one or both dehydrogenases, brought about by the increases in the concentrations of malate, isocitrate or both. Possible pathways involving the two enzymes are discussed. 相似文献
19.
Lipid metabolism by rat lung in vitro. Effect of starvation and re-feeding on utilization of [U-14C]glucose by lung slices 总被引:4,自引:2,他引:2
1. The incorporation of [U-(14)C]glucose into several lipid components of lung and liver slices, and the activities of glucose 6-phosphate dehydrogenase (EC 1.1.1.49), 6-phosphogluconate dehydrogenase (EC 1.1.1.44), ;malic' enzyme (EC 1.1.1.40) and NADP-isocitrate dehydrogenase (EC 1.1.1.42) of the cell cytosol were examined in normal, starved and re-fed rats. 2. Lipogenesis and the activities of these enzymes in liver were decreased markedly in rats starved for 72h. Re-feeding starved rats on a fat-free diet for 72h resulted in the well documented hyperlipogenic response in liver, particularly in its ability to convert glucose into neutral lipid, and increased activities of glucose 6-phosphate dehydrogenase, ;malic' enzyme and 6-phosphogluconate dehydrogenase to values approx. 700, 470 and 250% of controls respectively. 3. Approx. 70% of the total label in lung lipids was present in the phospholipid fraction. Hydrolysis of lung phospholipids revealed that lipogenesis from glucose was considerable, with approx. 40% of the total phospholipid radioactivity present in the fatty acid fraction. 4. Incorporation of glucose into total lung lipids was decreased by approx. 40% in lung slices of starved rats and was returned to control values on re-feeding. Although phospholipid synthesis from glucose was decreased in lung slices of starved rats, the decrease proportionally was greater for the fatty acid fraction (approx. 50%) as compared with the glycerol fraction (approx. 25%). 5. The activities of lung glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase and NADP-isocitrate dehydrogenase were not affected by the dietary alterations. ;Malic' enzyme activity was not detected in lung cytosol preparations. 6. The results are discussed in relation to the surface-active lining layer (surfactant) of the lung. 相似文献
20.
Energy transduction by electron transfer via a pyrrolo-quinoline quinone-dependent glucose dehydrogenase in Escherichia coli, Pseudomonas aeruginosa, and Acinetobacter calcoaceticus (var. lwoffi). 总被引:13,自引:9,他引:4
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B J van Schie K J Hellingwerf J P van Dijken M G Elferink J M van Dijl J G Kuenen W N Konings 《Journal of bacteriology》1985,163(2):493-499
The coupling of membrane-bound glucose dehydrogenase (EC 1.1.99.17) to the respiratory chain has been studied in whole cells, cell-free extracts, and membrane vesicles of gram-negative bacteria. Several Escherichia coli strains synthesized glucose dehydrogenase apoenzyme which could be activated by the prosthetic group pyrrolo-quinoline quinone. The synthesis of the glucose dehydrogenase apoenzyme was independent of the presence of glucose in the growth medium. Membrane vesicles of E. coli, grown on glucose or succinate, oxidized glucose to gluconate in the presence of pyrrolo-quinoline quinone. This oxidation led to the generation of a proton motive force which supplied the driving force for uptake of lactose, alanine, and glutamate. Reconstitution of glucose dehydrogenase with limiting amounts of pyrrolo-quinoline quinone allowed manipulation of the rate of electron transfer in membrane vesicles and whole cells. At saturating levels of pyrrolo-quinoline quinone, glucose was the most effective electron donor in E. coli, and glucose oxidation supported secondary transport at even higher rates than oxidation of reduced phenazine methosulfate. Apoenzyme of pyrrolo-quinoline quinone-dependent glucose dehydrogenases with similar properties as the E. coli enzyme were found in Acinetobacter calcoaceticus (var. lwoffi) grown aerobically on acetate and in Pseudomonas aeruginosa grown anaerobically on glucose and nitrate. 相似文献