共查询到20条相似文献,搜索用时 31 毫秒
1.
The developmental profile of the activities of some enzymes involved in malate metabolism, namely phosphoenolpyruvate carboxylase (PEPC; EC 4. 1. 1. 31), NAD+ -linked (EC 1. 1. 1. 37) and NADP+ -linked (EC 1. 1. 1. 82) malate dehydrosenase (MDH), NAD+ linked (EC 1. 1. 1. 39) and NADP+ -linked (EC 1. 1. 1. 40) malic enzyme (ME), has been determined in leaves of peach [ Prunus persica (L.) Batsch cv. Maycrest], a woody C3 species. In order to study the role of these enzymes, their activities were related to developmental changes of photosynthesis, respiration, and capacity for N assimilation. Activities of PEPC, NAD(P)+ -MDH and NADP+ -ME were high in young expanding leaves and decreased 2- to 3-fold in mature ones, suggesting that such enzymes play some role during the early stages of leaf expansion. In leaves of peach, such a role did not seem to be linked to C3 photosynthesis or nitrate assimilation, in that photosynthetic O2 evolution and activities of nitrate reductase (EC 1. 6. 6. 1) and glutamine synthetase (EC 6. 3. 1. 2) increased during leaf development. In contrast, leaf respiration strongly decreased with increasing leaf age. We suggest that in expanding leaves of this woody species the enzymes associated with malate metabolism have anaplerotic functions, and that PEPC may also contribute to the recapture of respiratory CO2 . 相似文献
2.
Protein content and activities of the enzymes glutamine synthetase (EC 6.3.1.2), NADH-glutamate synthase (EC 1.4.1.14), NADH-glutamate dehydrogenase (reductive amination (EC 1.4.1.2) and NAD+ -glutamate dehydrogenase (oxidative deamination) (EC 1.4.1.2) from the plant fraction of root nodules of alfalfa ( Medicago sativa L. cv. Aragon) were determined under water stress. Only NADH-glutamate synthase activity was inhibited during drought. The results indicate that the glutamine synthetase/NADH-glutamate synthase cycle was fully operational in alfalfa nodules of control or even mildly stressed plants when N2 -fixation was not inhibited, but that the coupling between glutamine synthetase and NADH-glutamate synthase was lost as drought progressed. Patterns of glutamine synthetase and NADH-/NAD+ -gluta-mate dehydrogenase activities reflect changes in ammonia content of nodules and/or availability of carbon substrates, and indicate that nodules maintain sufficient enzyme activity for ammonia assimilation throughout water stress. 相似文献
3.
P. B. Kavi Kishor 《Physiologia plantarum》1989,75(3):395-398
The activity during root and shoot initiation of key enzymes involved in aromatic amino acid metabolism was examined in rice ( Oryza sativa L. cv. Bala) callus cultures. Increased activities of the enzymes quinate:NAD+ oxidoreductase (EC 1.1.1.24), shikimate kinase (EC 2.7.1.71), chorismate mutase (EC 5.4.99.5), anthranilate synthase (EC 4.1.3.27) and tryptophan synthetase (EC 4.2.1.20) were noticed in organ-forming callus compared to proliferating callus of rice, especially prior to the visible manifestation of form. These results suggest a correlation between organogenesis and the aromatic amino acid pathway. 相似文献
4.
It was possible to extract simultaneously several active enzymes involved in the carbohydrate or the amino acid metabolism from spruce needles [ Picea abies (L.) Karst.] when a) a 100 m M Na-Pi buffer of pH 7.5 containing 5% PVPP and 0.5% Triton X-100 was used and when b) the resulting crude extracts were freed from lowmolecular-weight compounds by gel-chromatography using the separation medium Fractogel TSK HW-40. Besides Triton X-100, Triton X-305, Myrij-52 and Brij-35 were tested, but 0.5% Triton X-100 brought about the most active enzyme extracts. In crude extracts prepared from spruce needles during the early summer a high increase in absorbance at 334 nm was observed when the co-substrate NADP+ was added, thus making reliable spectrophotometric assays impossible. The interfering low-molecular-weight substances could be eliminated by gel chromatography. As separation media Bio-Gel P-6 DG, Sephadex G-25 m, Trisacryl GF 05 and Fractogel TSK HW-40 (F) were tested, with Fractogel yielding the highest activities.
With the methods described in this paper the activities of the following enzymes were determined: glucose-6-phosphate dehydrogenase (EC 1.1.1.49), 6-phosphogluconate dehydrogenase (EC 1.1.1.44), glucose-6-phosphate isomerase (EC 5.3.1.9), shikimate dehydrogenase (EC 1.1.1.25), NAD+ -malate dehydrogenase (EC 1.1.1.37), glutamate dehydrogenase (EC 1.4.1.2), aspartate aminotransferase (EC 2.6.1.1) and alanine aminotransferase (EC 2.6.1.2). The activities estimated for NAD+ -malate dehydrogenase and 6-phosphogluconate dehydrogenase are in the range of those published for the needle enzymes of white spruce and Scots pine, respectively. 相似文献
With the methods described in this paper the activities of the following enzymes were determined: glucose-6-phosphate dehydrogenase (EC 1.1.1.49), 6-phosphogluconate dehydrogenase (EC 1.1.1.44), glucose-6-phosphate isomerase (EC 5.3.1.9), shikimate dehydrogenase (EC 1.1.1.25), NAD
5.
Urszula Maciejewska 《Physiologia plantarum》1990,80(1):133-135
Low temperature (2°C) caused an increase in the activity of NAD+ kinase (EC 2.7.1.23) in leaves of winter rape plants ( Brassica napus L. var. oleifera L. cv. Górezański). The enzyme activity markedly increased between day 4 and 11 of plant exposure to cold, then tended to decrease. Changes in activity of NAD+ kinase coincided with the previously observed changes in the levels of pyridine nucleotides, NADP(H) (U. Maciejewska and A. Kacperska, Physiol. Plant. 69: 687–691, 1987). As a result of cold treatment, Ca2+ –calmodulin–dependent and Ca2+ –calmodulin–independent NAD+ kinase activities increased to almost the same extent. It seems therefore, that the cold–induced activation of NAD+ kinase does not depend on the Ca2+ –calmodulin complex. 相似文献
6.
The influence of nicotianamine (NA) and iron on the activities of 4 iron-containing and two iron-free enzymes in leaves and roots of the NA-free tomato mutant chloronerva and its NA-containing wild-type ( Lycopersicon esculentum Mill. cv. Bonner Beste) was investigated. Aconitase (EC 4.2.1.3) activity in both leaves and roots was much higher in the mutant under normal iron supply (10 μ M FeEDTA) and in wild-type under iron deficiency than in wild-type supplied with 10 μ M FeEDTA. Application of NA to chloronerva leaves led to a decrease of aconitase activity in leaves and roots. NA had no effect on the enzyme activity when added to the assay medium.
Similar results were obtained for the iron-containing enzymes catalase (EC 1.11.1.6), ascorbate-dependent peroxidase (EC 1.11.1.11) and guaiacol-dependent peroxidase (EC 1.11.1.7) in roots. NA treatment of the mutant leaves decreased enzyme activities in roots down to wild-type values. In vivo NA application had no effect on enzyme activities in leaf extracts.
The activities of the iron-free enzymes NAD+ -malate dehydrogenase (EC 1.1.1.37) and phosphofructokinase (EC 2.7.1.11) in root and leaf extracts were not influenced by the iron supply to the plants. 相似文献
Similar results were obtained for the iron-containing enzymes catalase (EC 1.11.1.6), ascorbate-dependent peroxidase (EC 1.11.1.11) and guaiacol-dependent peroxidase (EC 1.11.1.7) in roots. NA treatment of the mutant leaves decreased enzyme activities in roots down to wild-type values. In vivo NA application had no effect on enzyme activities in leaf extracts.
The activities of the iron-free enzymes NAD
7.
The seven NAD(H)-glutamate dehydrogenase isoenzymes exhibit similar anabolic and catabolic activities 总被引:4,自引:0,他引:4
The specific activities of aminating NADH- and deaminating NAD+ -glutamate dehydrogenase (GDH, EC 1.4.1.2) varied considerably in crude extracts of grapevine ( Vitis vinifera L. cv. Sultanina) callus and were dependent on the nitrogen source of the culture medium. However, dialysis of the enzyme preparations resulted in a significant decrease in the deaminating GDH specific activity while the aminating activity was not affected. The presence of malate in the crude extract resulted in erroneous overestimation of the NAD+ -GDH activity through the malate dehydrogenase reaction. Thus, in dialysed extracts, the ratio of the NADH-GDH/NAD+ -GDH specific activities remained relatively constant irrespective of the nitrogen source. In view of this evidence, we now have modified methods for staining both the NADH-GDH and NAD+ -GDH activities on gels in order to compare the aminating and deaminating activities of each of the 7 GDH isoenzymes. The results from the staining of NADH-GDH and NAD+ -GDH activity of enzyme preparations from calluses revealed the same isoenzyme profile. Furthermore, separated leaf isoenzymes showed similar activity ratios and kinetic properties. These results may suggest that each one of the 7 isoenzymes have similar in vitro anabolic and catabolic activities. 相似文献
8.
Seeds of Phaseolus vulgaris L. cv. White Kidney were germinated and grown either in a nitrogen-free or in an ammonia-supplied medium. The changes in the soluble protein concentration and in the levels of glutamine synthetase (GS, EC 6.3.1.2), NADH–glutamate synthase (NADH-GOGAT, EC 1.4.1.14), ferredoxin-glutamate synthase (Fd-GOGAT, EC 1.4.7.1) and glutamate dehydrogenase (GDH, EC 1.4.1.2), both NADH- and NAD+ -dependent, were examined in cotyledons and roots during the first 10 days after sowing. Soluble protein declined rapidly in the cotyledons and increased slightly in the roots. GS activity was initially high both in cotyledons and roots but subsequently decreased during seedling growth. Exogenous ammonia hardly affected GS activity. High levels of NADH-GOGAT were present both in cotyledons and roots during the first days of germination. The activity then gradually declined in both organs. In contrast, Fd-GOGAT in cotyledons was initially low and progressively increased with seedling development. In roots, the levels of Fd-GOGAT were higher in young than in old seedlings. Supply of ammonia to the seedlings increased the levels of NADH-GOGAT and Fd-GOGAT both in cotyledons and roots. NADH-GDH (aminating) activity gradually increased during germination. In contrast, the levels of NAD+ -GDH (deaminating) activity were highest during the first days of germination. Exogenous ammonia did not significantly affect the activities of GDH. 相似文献
9.
Frankia vesicle clusters were prepared from Alnus incana (L.) Moench root nodules containing a local source of Frankia by an improved homogenization-filtration procedure. The capacity of the vesicle clusters to metabolize hexoses was investigated by respirometric and enzymological studies. The vesicle clusters could utilize glucose, glucose-6-phosphate and 6-phosphogluconate provided that appropriate cofactors were added to the preparations. The enzymes hexokinase (EC 2.7.1.1), NADP+ : glucose-6-phosphate dehydrogenase (EC 1.1.1.49) and NAD+ ;6-phosphogluconate dehydrogenase (EC 1.1.1.44) were found in cell-free extracts of the vesicle clusters and kinetic constants for the enzymes were determined. Hexokinase had a lower Km for glucose than for fructose. Extracts from both symbiotic and propionate grown Frankia AvcII also showed activity of these hexose-degrading enzymes, indicating that their presence is not necessarily dependent on sugars as carbon source. The NAD+ - dependent 6-phosphogluconate dehydrogenase was only present in Frankia cells and not in alder root cells, which makes this enzyme a useful Frankia -specific marker in these symbiotic systems. 相似文献
10.
NADH is central to the functioning of mitochondrial respiration. It is produced by enzymes in, or associated with, the tricarboxylic acid cycle in the matrix, and it is oxidized by two respiratory chain enzymes in the inner membrane, the rotenone-sensitive complex I and the rotenone-insensitive internal NADH dehydrogenase (NDin ). A simplified kinetic model for NADH turnover in the matrix of plant mitochondria is presented. Only the two main NADH-producing enzymes, NAD-malate dehydrogenase [EC 1.1.1.37] (MDH) and NAD-malic enzyme [EC 1.1.1.39] (ME), are considered. This model reproduces the complex behaviour of malate oxidation by isolated mitochondria in response to additions of ADP (state 3/state 4), NAD+ and/or rotenone, as well as to changes in pH. It is found that MDH always operates at or close to equilibrium. Changes in the activity of complex I, NDin , or ME are predicted to cause clear changes in the pattern of malate oxidation. In general, the model predicts high sensitivity to changes in the ME activity. In contrast, MDH activity can be reduced 100-fold without detectable changes in malate oxidation. It is demonstrated that it is not the high activity, but the equilibrium properties of MDH that are important for the redox-buffering function of MDH in the mitochondrial matrix. Binding of NAD+ and NADH in the matrix reduces the concentrations of free NAD+ and NADH, depending on the concentration of binding sites and the binding strength. On the basis of the modelling results it is estimated that a significant proportion of the mitochondrial NAD is bound. 相似文献
11.
Andrés P. Sanz Conrado Moreno-Vivián José M. Maldonado Agustín González-Fontes 《Physiologia plantarum》1995,95(1):39-44
The effect of the nitrogen source on carbohydrate and protein contents and on several enzymatic activities involved in the carbon and nitrogen metabolism was studied in Anabaena variabilis ATCC 29413 cells grown under a constant supply of either N, NO− 3 or NH+ 4 at different concentrations. An enhancement of protein content accompanied by a parallel decrease of carbohydrates was observed with increasing NO− 3 or NH+ 4 concentrations in the medium. In cultures containing 0.1 m M NO− 3 or 0.1 m M NH+ 4 nitrogenase (EC 1.18.6.1) activity was 74 and 66%, respectively, of that found in N2 -grown cells. This activity was still present with 1 m M NO− 3 or 1 m M NH+ 4 in the medium and even with 10 m M NO− 3 , but it was completely inhibited by 5 m M NH+ 4 . Ferredoxin-nitrate reductase (EC 1.7.7.2) activity was detected only in NO− 3 grown cells and simultaneously with nitrogenase activity. Increasing concentrations of combined nitrogen in the medium, especially NH+ 4 , promoted a concomitant decline of glutamine synthetase (EC 6.3.1.2), NADP+ -isocitrate dehydrogenase (EC 1.1.1.42), and NAD+ -malate dehydrogenase (EC 1.1.1.37) activities, suggesting that these enzymes play an important role in the regulation of carbon-nitrogen metabolism in cyanobacteria. 相似文献
12.
Coordination of photosynthetic and respiratory metabolism in Chlorella vulgaris UAM 101 in the light
In Chlorella vulgaris UAM 101, the presence of glucose altered the photosynthetic and respiratory metabolism in the light. When glucose was added to the growth medium, an increase in the cellular level of enzymes involved in glucose oxidation, namely glucose-6-P dehydrogenase (EC 1.1.1.49) and NAD+ -glyceraldehyde-3-P dehydrogenase (EC 1.2.1.12), was observed. Glucose also enhanced respiratory O2 consumption. In addition, CO2 released by glucose oxidation was refixed in photosynthesis. The presence of glucose also affected photosynthesis. Phosphoribulokinase (EC 2.7.1.19) and NADP+ -dependent glyceraldehyde-3-P dehydrogenase (EC 1.2.1.13), two regulatory enzymes of the reductive pentose phosphate cycle, were increased by glucose. However, Rubisco (EC 4.1.1.39) activity of these cells was lower than that of autotrophic cells. Despite these alterations, the photosynthetic O2 evolution was not significantly inhibited by glucose. On the other hand, an increase in the cytosolic NADP+ -glyceraldehyde-3-P dehydrogenase (EC 1.2.1.9) that is involved in obtaining reducing power for anabolic processes was observed. The CO2 levels in the growth medium did not significantly affect the cellular level of enzymes measured in this work, except those involved in biosynthetic pathways. These data suggest that the effect of glucose on photosynthesis and respiration can be explained by alteration of the cellular level of photosynthetic enzymes and respiratory substrates, respectively. 相似文献
13.
The subcellular distribution of pyruvate-degrading enzymes has been determined in Chlamydomonas reinhardtii (Dangeard) by protoplast induction with autolysine, dig-itonin lysis and further fractionation by differential centrifugation using a Percoll cushion. Mitochondrial and plastidic fractions contained intact and physiologically competent organelles - RC 1.7, ADP/O 2.7 and rate of malate oxidation 76 nmol O, (mg protein)-1 min-1 for mitochondria, CO2; fixation 46.8 μmol (mg Chi)-1 h-1 for chloroplasts.
Results from protoplast fractionation were further confirmed by the determination of enzyme activities within trypsin-treated organelles. Mitochondria (formate fermentation) and chloroplasts (chlorofermentation) were shown to possess the capacity for anaerobic pyruvate degradation. Pyruvate dehydrogenase (NAD+ , EC 1.2.4.1), pyruvate formate-lyase (EC 2.3.1.54) and lactate dehydrogenase (NADH, EC 1.1.1.27) showed equal distribution between mitochondria and chloroplasts, whereas activities of phosphotransacetylase (EC 2.3.1.8) and acetate kinase (EC 2.7.2.1) were only detectable in the mitochondrial fraction. NADH- and NADPH-dependent activities of both alcohol dehydrogenase (EC 1.1.1.1) and aldehyde dehydrogenase (acylating, EC 1.2.1.10) were localized in the mitochondrial and cytoplasmic or the plastidic and cytoplasmic fractions, respectively, whereas pyruvate decarboxylase (EC 4.1.1.1) was only detected in the cytoplasmic fraction. 相似文献
Results from protoplast fractionation were further confirmed by the determination of enzyme activities within trypsin-treated organelles. Mitochondria (formate fermentation) and chloroplasts (chlorofermentation) were shown to possess the capacity for anaerobic pyruvate degradation. Pyruvate dehydrogenase (NAD
14.
In tobacco callus, the induction of nicotine synthesis, which stimulates enzyme activities of the ornithine-methylpyrroline route (see the preceding paper), also leads to marked changes in the enzyme activities of the pyridine-nucleotide cycle. This cycle provides the metabolite (probably nicotinic acid) for condensation with methylpyrroline to produce nicotine. The activities of eight enzymes of the pyridine-nucleotide cycle and of quinolinic-acid phosphoribosyltransferase, the anaplerotic enzyme, were determined by high-performance liquid chromatography assays. The distinct changes of their activities upon induction of nicotine synthesis lead to the following conclusions: i) nicotinic acid is the relevant metabolite which is provided by the pyridine-nucleotide cycle and consumed for nicotine synthesis. ii) The enhancement of the nicotinic-acid pool arises in two ways, by synthesis of NAD and degradation via nicotinamide mononucleotide and by a direct route from nicotinic-acid mononucleotide (NaMN) which is degraded by a glycohydrolase with a rather high K
m value. Such a K
m value prevents the complete depletion of the NaMN pool.Abbreviations HPLC
high-performance liquid chromatography
- NAD-PPase
NAD-pyrophosphatase
- NaMN-ATase
nicotinic-acid mononucleotide (NaMN) adenylyltransferase
- NaMN-GHase
NaMN-glycohydrolase
- Na-PRTase
nicotinic-acid phosphoribosyltransferase
- NMN-ATase
nicotinamide mononucleotide (NMN) adenylyltransferase
- NMN-Ghase
NMN-glycohydrolase
- PMT
putrescine methyltransferase
- Qa-PRTase
quinolinic acid phosphoribosyltransferase 相似文献
15.
Marc Bonzon Patrice Simon Robert Degli Agosti Hubert Greppin Edgar Wagner 《Physiologia plantarum》1987,70(4):577-582
The activity of NAD+ and NADP+ -dependent glyceraldehyde-3-phosphate dehydrogenase isozymes (EC 1.2.1.12 and EC 1.2.1.13, respectively) were measured in spinach ( Spinacia oleracea L. cv. Nobel) leaves grown under different photoperiodic treatments in order to discriminate between the early events of floral induction and processes related to acclimation. Glycolysis-linked isozyme activities were increased not only during floral induction and acclimation, but also during acclimation alone, suggesting that the changes in cytosolic activities were most probably associated with acclimation. In contrast, the chloroplast-linked isozyme activities only increased during flower induction and appeared to be specifically associated with the initiation of the flowering process. The relative activity changes in the chloroplast and cytosol compartments may thus be supposed to be among the first signs of translation of the photoperiodic signal into cytosolic and cellular metabolic adaptation, whereby the leaf moves rapidly into a new metabolic state. 相似文献
16.
A survey for the enzyme L-myo-inositol-1-phosphate synthase (EC 5.5.1.4) has been conducted among various members of the lower plant groups, mainly algac, bryophytes and fungi; some properties of the partially purified enzyme from Euglena gracilis Z . are presented. The enzyme was detected in Chloropycean algae, Marchantiales and the Basidiomycetous fungi. The enzyme from Euglena had a pH optimum at 7.5. The Km for glucose-6-P was 2.1 m M and for NAD+ 80 μ M . When assayed in the absence of added NAD+ , the enzyme showed a basal activity suggesting the presence of bund NAD+ in the system. NH4 Cl increased the enzyme activity two-fold, altough the enzyme was inactivated by (NH4 )2 SO4 . 相似文献
17.
The nature of the aggregated form of chloroplast glyceraldehyde-3-phosphate dehydrogenase isozymes (GPD, EC 1.2.1.13) from Chenopodium rubrum leaves was investigated. After disaggregation of the isozymes in NADP + buffer, and resuspension of the disaggregated isozymes in NAD+ buffer, complete reaggregation could only be achieved by remixing the enzyme with a high molecular weight fraction, from which the isozymes had dissociated during the NADP+ filtration. After separation of the isozymes by inverse ammonium sulphate gradient solubilization, spontaneous extensive reaggregation of each isozyme was observed in NAD+ buffer. The high molecular weight material consisted of ribonucleoprotein, and RNase treatment impaired its ability to promote reaggregation of chloroplast GPD. It is proposed that pyridine nucleotide-controlled aggregation and binding to ribonucleoprotein in vitro are artifacts which reflect an in situ binding to cellular components. Since uncontrolled NAD+ -linked activities of the bifunctional isozymes in the chloroplast would lead to an equalization of the NAD + and NADP + redox couples, it is suggested that the reversible binding of the isozymes forms the basis of a regulatory system in vivo. 相似文献
18.
Hatsumi Taniguchi Hironori Aramaki Yoshihiko Nikaido yasuo Mizuguchi Masahiro Nakamura Toshihiko Koga Shin-ichi Yoshida 《FEMS microbiology letters》1996,144(1):103-108
Abstract The bradyzoite and tachyzoite forms of Toxoplasma gondii , purified from infected animals, were analysed for their activities of phosphofructokinase, pyruvate kinase, lactate dehydrogenase, NAD+ - and NADH-linked isocitrate dehydrogenases, and succinic dehydrogenase. Both developmental stages contained high activities of phosphofructokinase (specific for pyrophosphate rather than ATP), pyruvate kinase and lactate dehydrogenase, suggesting that energy metabolism in both forms may centre around a high glycolytic flux linked to lactate production. The markedly higher activity of the latter two enzymes in bradyzoites suggests that lactate production is particularly important in this developmental form. NAD+ -specific isocitrate dehydrogenase was not detectable in either stage of the parasite (and proved useful as a measure of the purity of the bradyzoite preparation), whereas both parasite forms contained low activities of NADP+ -linked isocitrate dehydrogenase. The results are consistent with the bradyzoites lacking a functional TCA cycle and respiratory chain and are suggestive of a lack of susceptibility of this developmental stage to atovaquone. 相似文献
19.
Murine Glial Cells Regenerate NAD, After Peroxide-Induced Depletion, Using Either Nicotinic Acid, Nicotinamide, or Quinolinic Acid as Substrates 总被引:2,自引:1,他引:1
Abstract: The potential for regeneration of intracellular pyridine nucleotide levels from different precursors, after peroxide-induced NAD depletion, in cultured glial cells was investigated. Cultured murine glial cells showed a decrease in intracellular NAD levels of >40% after treatment with H2 O2 (100 µ M ). Removal of the H2 O2 followed by a 2-h incubation did not result in NAD recovery in the absence of precursors. However, NAD levels increased significantly in these cells after the following substrate additions, at minimum effective concentrations of 1 m M for quinolinic acid (QUIN), 500 µ M for nicotinamide, and 2 µ M for nicotinic acid. The regeneration of significant amounts of NAD from nicotinic acid at doses 250 and 500 times lower than either nicotinamide or QUIN indicates a preferred route for NAD biosynthesis in glial cells in vitro, probably via nicotinic acid phosphoribosylation. 相似文献
20.
Pyridine nucleotide cycle of Salmonella typhimurium: in vitro demonstration of nicotinamide adenine dinucleotide glycohydrolase, nicotinamide mononucleotide glycohydrolase, and nicotinamide adenine dinucleotide pyrophosphatase activities. 总被引:6,自引:4,他引:2
下载免费PDF全文
![点击此处可从《Journal of bacteriology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
J W Foster 《Journal of bacteriology》1981,145(2):1002-1009
Extracts of Salmonella typhimurium were chromatographed by using Sephadex G-150 to separate the various enzymes involved with pyridine nucleotide cycle metabolism. This procedure revealed a previously unsuspected nicotinamide adenine dinucleotide (NAD) glycohydrolase (EC 3.2.2.5) activity, which was not observed in crude extracts. In contrast to NAd glycohydrolase, NAD pyrophosphatase (EC 3.6.1.22) was readily measured in crude extracts. This enzyme possessed a native molecular weight of 120,000. Other enzymes examined included nicotinamide mononucleotide (NMN) deamidase (EC 3.5.1.00), molecular weight of 43,000; NMN glycohydrolase (EC 3.2.2.14), molecular weight of 67,000; nicotinic acid phosphoribosyl transferase (EC 2.4.2.11), molecular weight of 47,000; and nicotinamide deamidase (EC 3.5.1.19), molecular weight of 35,000. NMN deamidase and NMN glycohydrolase activities were both examined for end product repression by measuring their activities in crude extracts prepared from cells grown with and without 10(-5) M nicotinic acid. No repression was observed with either activity. Both activities were also examined for feedback inhibition by NAD, reduced NAD, and NADP. NMN deamidase was unaffected by any of the compounds tested. NMN glycohydrolase was greatly inhibited by NAD and reduced NAD, whereas NADP was much less effective. Inhibition of NMN glycohydrolase was found to level off at an NAD concentration of ca. 1 mN, the approximate intracellular concentration of NAD. 相似文献