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91.
The decrease in the electron flow of the aerobic respiratory chain of the bacterium Paracoccus denitrificans, owing to either the drop in the saturation of terminal oxidases by oxygen or to the inhibition of the rate of respiration by azide or nitrite, resulted in the synthesis of dissimilatory nitrate reductase and nitrite reductase. The dependence of the resulting activities of the two enzymes (after a three-hour adaptation) on the initial value of the parameter Vmax/kLa (oxidase activity of the volume unit of the culture divided by the volumetric oxygen transfer coefficient) or on the concentrations of the inhibitors had a similar form, characterized by the appearance of a maximum. The increasing parts of the obtained curves reflect the synthesis of enzymes, probably initiated by the increase in the intracellular degree of reduction, the subsequent drop being evidently in connection with the lack of metabolic energy for biosynthesis. The possible mechanisms of the effect of nitrogenous terminal acceptors (NO-3 and NO-2) on the formation of the denitrification pathway are discussed. 相似文献
92.
I. Martha Schlichtherle David S. Roos Judith L. Van Houten 《Molecular & general genetics : MGG》1996,250(6):665-673
We have cloned the first bifunctional gene dihydrofolate reductase-thymidylate synthase (DHFR-TS) from a free-living, ciliated protozoan,Paramecium tetraurelia, and determined its macronuclear sequence using a modified ligation-mediated polymerase chain reaction (PCR) that can be of general use in cloning strategies, especially where cDNA libraries are limiting. While bifunctional enzyme sequences are known from parasitic protozoa, none had previously been found in free-living protozoa. The AT-rich (68%) coding region spanning 1386 bp appears to lack introns. DHFR-TS localizes to a 500 kb macronuclear chromosome and is transcribed as an mRNA of 1.66 kb, predicted to encode a 53 kDa protein of 462 residues. The N-terminal one-third of the protein is encoded by DHFR, which is joined by a short junctional peptide of 12 amino acids to the highly conserved C-terminal TS domain. Among known DHFR-TS sequences, theP. tetraurelia gene is most similar to that fromToxoplasma gondii, based on primary sequence and parsimony analyses. The predicted secondary protein structure is similar to those of previously crystallized monofunctional sequences. 相似文献
93.
超结瘤大豆(Glycine m ax (L.) Merr.) nts 382 和不结瘤大豆Nod 49 的叶和根组织水提取物经Sephadex G25 过滤、洗脱,再根据洗脱物对硝酸还原酶(NR)活性的影响可划分为4 个组分(fraction)样品,即nts 382(Nod 49) F1、nts 382(Nod 49) F2、nts 382(Nod 49) F3 和nts 382(Nod 49) F4。其中, nts382 F2 和F4 抑制NR 活性作用在接种USDA110 后明显下降, 但接种的nts 382 F2 却能提高大豆Bragg 的结瘤数达一倍, 而接种的nts 382 F3 和F4 的作用不明显。NR 活性抑制因子不是刺激结瘤的因子, 刺激结瘤的因子主要分布在接种的nts382 F2 部分中。与这一现象相反, Nod 49 F2 和F4 抑制NR活性的作用在接种后更强, 且也抑制大豆nts 382 的结瘤, 其中Nod 49 F4 抑制结瘤的作用基本不能逆转。抑制结瘤因子主要分布在接过种的Nod 49 F4 部分中 相似文献
94.
Cathrine Lillo Lucy H. Smith Hugh G. Nimmo Malcolm B. Wilkins 《Physiologia plantarum》1996,98(1):140-146
Nitrate reductase (NR, EC 1.6.6.1) was tested in crude extracts of leaves from Bryophyllum fedtschenkoi plants growing under alternating light/darkness as well as in excised leaves kept in continuous light or darkness. In most extracts NR activity was inhibited 20–80% by 5 m M Mg2+ A light or darkness shift (30 min darkness) during the first part of the photoperiod gave an increase in the Mg2+ inhibition and a decrease in NR activity. Magnesium ion inhibition of NR also showed diurnal variations. Strongest inhibition was found in extracts made during the latter part of the photoperiod and start of the dark period. Pre-incubation of crude extracts with ATP increased Mg2+ inhibition, indicating that phosphorylation of NR is involved in regulation of NR in Crassulacean acid metabolism (CAM) plants. In continuous light an increase in Mg2+ inhibition occurred after 20 h and 40 h, indicating a rhythm in the phosphorylation of NR. A delay in the production of nitrite in the assay (hysteresis) was generally seen in extracts susceptible to Mg2+ inhibition. The rhythms related to NR activity showed the same period length (20 h) as the rhythm in CO2 exchange. However, in contrast to the rhythm in CO2 exchange, NR rhythms were strongly damped in continuous light. In constant darkness the rhythms were even more damped. The results show that post-translational modification of CAM NR is influenced by light/darkness and by an endogenous rhythm. 相似文献
95.
Christophe Bailly Abdelilah Benamar Françoise Corbineau Daniel Come 《Physiologia plantarum》1996,97(1):104-110
Sunflower ( Helianthus annuus L.) seeds progressively lost their ability to germinate at 25°C, the optimal temperature for germination, after accelerated aging was carried out at 45°C (a temperature too high to permit germination) in water or at 76 or 100% relative humidity (RH). The deleterious effects of the high-temperature treatment increased with increasing seed moisture content. Incubation of seeds at 45°C in water resulted in electrolyte leakage, which indicated a loss of membrane integrity. A relationship between leakage and loss of seed viability could not be assumed, since no increase in electrolyte efflux occurred after aging al 100% RH. Accelerated aging induced accumulation of malondialdehyde, suggesting that seed deterioration was associated with lipid peroxidation. However, there was no direct relationship between lipid peroxidation and deterioration in membrane integrity. Loss of seed viability was also associated with a decrease in superoxide dismutase, catalase and glutathione reductase activities. Finally, the results obtained suggest that sunflower seed deterioration during accelerated aging is closely related to a decrease in the activities of detoxifying enzymes and to lipid peroxidation. 相似文献
96.
Barley leaf protoplasts were incubated in light or darkness in the presence of various inhibitors, metabolites or weak acids/bases. Nitrate reductase (NR) and phosphoenolpyruvate carboxylase (PEPCase) were rapidly extracted from the protoplasts and assayed under sub-optimal conditions, i.e. in the presence of Mg2+ and malate, respectively. Under these conditions changes in activities are thought to reflect changes in the phosphorylation states of the enzymes. The NR was activated by illumination to 90% of its maximal activity within 10 min. Photosynthetic electron transport appeared necessary for light activation of NR since activation was inhibited by the photosynthetic electron-transport inhibitor 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU), and, additionally, an electron acceptor (HCO
3
-
) was required. The PEPCase was also activated by light. However, this activation was not prevented by DCMU or lack of HCO
3
-
. Loading of protoplasts in the dark with a weak acid resulted in activation of both NR and PEPCase. For NR, full activation was completed within 5 min, whereas for PEPCase a slower, modest activation continued for at least 40 min. Incubation of protoplasts with a weak base also gave activation of PEPCase, but not of NR. On the contrary, base loading counteracted light activation of NR. Since several treatments tested resulted in the modulation of either NR or PEPCase activity, but not both, signal transduction cascades leading to changes in activities appear to be very different for the two enzymes.Abbreviations DCMU
3-(3,4-dichlorophenyl)-1,1-dimethylurea (diuron)
- DMO
5,5-dimethyl-2,4 oxazolidinedione
- NR
nitrate reductase
- PEPCase
Phosphoenolpyruvate carboxylase
This work was supported by the Norwegian Research Council by a Grant to C.L: L.H.S. was supported by the Biotechnology and Biological Sciences Research Council. 相似文献
97.
Anaerobic metabolism of 2-hydroxybenzoic acid (salicylic acid) by a denitrifying bacterium 总被引:6,自引:0,他引:6
The anaerobic metabolism of 2-hydroxybenzoic acid (salicylic acid) was studied in a denitrifying bacterium. Cells grown with
2-hydroxybenzoate were simultaneously adapted to degrade benzoate. Extract of these cells formed benzoate or benzoyl-CoA when
incubated under reducing conditions with salicylate, MgATP, and coenzyme A, suggesting a degradation of 2-hydroxybenzoate
via benzoate or benzoyl-CoA. This suggestion was supported by enzyme activity measurements. In extracts of 2-hydroxybenzoate-grown
cells, the following enzyme activities were detected: two CoA ligases, one specific for 2-hydroxybenzoate, the other for benzoate,
and two different enzyme activities catalyzing the reductive transformation of 2-hydroxybenzoyl-CoA. These findings suggest
a degradation of salicylic acid by two new enzymes, 2-hydroxybenzoate-CoA ligase (AMP-forming) and 2-hydroxybenzoyl-CoA reductase
(dehydroxylating), catalyzing (1) 2-hydroxybenzoate + MgATP + CoASH → 2-hydroxybenzoyl-CoA + MgAMP + PPi (2) 2-hydroxybenzoyl-CoA + 2[H] → benzoyl-CoA + H2O Benzoyl-CoA was dearomatized by reduction of the ring. This represents another case in which benzoyl-CoA is a central intermediate
in anaerobic aromatic metabolism.
Received: 1 February 1996 / Accepted: 24 February 1996 相似文献
98.
Kanika Misra Arun B. Banerjee Subhankar Ray Manju Ray 《Molecular and cellular biochemistry》1996,156(2):117-124
Two enzymes, one NADPH-dependent and another NADH-dependent which catalyze the reduction of methylglyoxal to acetol have been isolated and substantially purified from crude extracts of Escherichia coli K12 cells. Substrate specificity and formation of acetol as the reaction product by both the enzymes, reversibility of NADH-dependent enzyme with alcohols as substrates and inhibitor study with NADPH-dependent enzyme indicate that NADPH-dependent and NADH-dependent enzymes are identical with an aldehyde reductase (EC 1.1.1.2) and alcohol dehydrogenase (EC 1.1.1.1) respectively. The Km for methylglyoxal have been determined to be 0.77 mM for NADPH-dependent and 3.8 mM for NADH-dependent enzyme. Stoichiometrically equimolar amount of acetol is formed from methylglyoxal by both NADPH- and NADH-dependent enzymes. In phosphate buffer, both the enzymes are active in the pH range of 5.8–6.6 with no sharp pH optimum. Molecular weight of both the enzymes were found to be 100,000 ± 3,000 by gel filtration on a Sephacryl S-200 column. Both NADPH- and NADH-dependent enzymes are sensitive to sulfhydryl group reagents. 相似文献
99.
M. Popović S. KevreŠan J. Kandrač J. Nikolić N. Petrović R. Kastori 《Biologia Plantarum》1996,38(2):281-287
In young sugar beet plants cadmium suppressed the activity of nitrate reductase, glutamine synthetase and glutamate dehydrogenase,
whereas sulphur exhibited a protective role towards activity of these enzymes, except of glutamine synthetase. Protein synthesis
was suppressed in the absence of S in nutrient medium; the lowest level was at 10-3 M Cd2+. Chloroplast pigment contents were increased by S while Cd2+, even in the lowest concentration, (10−5 M) showed a repressive effect. The highest concentrations of Cd2+ (10−3 M) caused a decrease in dry mass, whereas S induced its increase. Nitrate content was increased in the presence of
Cd2+ and decreased by increased concentration of S.
Acknowledgement: The authors acknowledge financial support of the Ministry for Science and Technology of Serbia.
The paper was presented at 9th Congress of the Federation of European Societies of Plant Physiology, Brno, Czech Republic,
3–8 July 1994. 相似文献
100.
Ferredoxin-sulfite reductases (Fd-SiRs) [hydrogen-sulfide: ferredoxin oxidoreductase, EC 1.8.7.1] from leek leaves have been
purified to homogeneity. The enzymes (SiR 1, SiR 2 and SiR 3) were separated by Mono Q chromatography. The collective molecular
mass of the enzymes was estimated to be 65 kDa by gel filtration. In all three cases, subunit analysis by SDS-PAGE yielded
a single protein band corresponding to a molecular mass of 64 kDa, indicating that the enzymes each exist as a monomer. In
the oxidized forms, SiR 1, SiR 2 and SiR 3 all exhibited nearly identical absorption maxima at 279∼280, 389∼390, 588 and 714
nm, indicating that siroheme is involved in the catalysis of sulfite reduction. On enzymatic properties, SiR 1, SiR 2 and
SiR 3 could only react with the physiological electron donor, feriedoxin. The enzymes exhibited different heat stabilities.
The pH active curve obtained from SiR 2 was different from the others. Moreover, SiR 1 exhibited a lower Km value for ferredoxin
than SiR 2. Although the N-terminal sequence was the same, the results of some enzymatic properties, amino acid analysis,
and peptide mapping suggested the presence of the Fd-SiR isozymes in leek leaves. 相似文献