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1.
Various electron donors were found to stimulate C2H2 reduction (N2 fixation) by isolated heterocysts from Anabaena variabilis and Anabaena cylindrica. Intermediates of glycolysis and the tricarboxylic acid cycle as well as unphosphorylated sugars like glucose, fructose and erythrose were among these electron donors. The transfer of electrons from donors like H2, NADH, glyoxylate and glycollate was strictly light-dependent, whereas others like NADPH or pyruvate plus coenzyme A supported C2H2 reduction also in the dark. In all cases, the overall activity was enhanced by light. The stimulation by light was more distinct with heterocysts from A. variabilis than with heterocysts from A. cylindrica.The present communication establishes that pyruvate supports C2H2 reduction by heterocysts from either A. variabilis or A. cylindrica with rates comparable to those with other electron donors. Pyruvate could, however, support C2H2 reduction only in the presence of coenzyme A, and the concentrations of both coenzyme A and pyruvate were crucial. A pyruvate-dependent reduction of ferredoxin by extracts from heterocysts was recorded spectrophotometrically. Glyoxylate, which is an inhibitor of thiamine pyrophosphate-dependent decarboxylations, inhibited pyruvate-dependent C2H2 reduction. This result supports the conclusion that pyruvate is metabolised by pyruvate: ferredoxin oxidoreductase in heterocysts. High concentrations of pyruvate and other electron donors inhibited C2H2 reduction which suggests that nitrogenase activity in heterocysts may be controlled by the availability of electron donors.Dedicated to Professor Norbert Pfennig, Konstanz, on the occasion of his 60th birthday 相似文献
2.
The cyanobacterium Anabaena variabilis showed a pH dependent uptake of ethylenediamine. No uptake of ethylenediamine was detected at pH 7.0. At higher pH values (e.g. pH 8.0 and pH 9.0) accumulation did occur and was attributed to diffusion of uncharged ethylenediamine in response to a pH gradient. A biphasic pattern of uptake was observed at these higher pH values. Treatment with l-methionine-d,l-sulphoximine (MSX) to inactivate glutamine synthetase (GS) inhibited the second slower phase of uptake without any significant alteration of the initial uptake. Therefore for sustained uptake, metabolism of ethylenediamine via GS was required. NH
4
+
did not alter the uptake of ethylenediamine. Ethylenediamine was converted in the second phase of uptake to an analogue of glutamine which could not be detected in uptake experiments at pH 7.0 or in uptake experiments at pH 9.0 following pretreatment of cells with MSX. Ethylenediamine treatment inhibited nitrogenase activity and this inhibition was greatest at high pH values.Abbreviations EDA
1,2-diaminoethane (ethylenediamine)
- GS
glutamine synthetase
- HEPES
4-(2-hydroxyethyl)-1 piperazine ethanesulphonic acid
- MSX
l-methionine-dl-sulphoximine
-
membrane potential
- Tricine
N-tris(hydroxymethyl) methylglycine 相似文献
3.
Herbert Böhme 《Archives of microbiology》1986,146(1):99-103
Addition of nitrite to rapidly growing, nitrogen-fixing filaments ofAnabaena variabilis caused an immediate drop in nitrogenase activity. This was followed by a transient induction of nitrite reductase, recovery of nitrogen fixation and cyanobacterial growth. The experiments with isolated heterocysts and a partially purified nitrogenase preparation from heterocysts showed that nitrite primarily exerted its inhibitory effect by inactivating nitrogenase irreversibly, rather than interfering with photosynthetic energy conservation.Abbreviations ATCC
American type culture collection
- Chl
chlorophyll
- FCCP
carbonyl cyanide p-trifluoromethoxy phenylhydrazone
- Tes
2-{[2 hydroxy-1,1-bis(hydroxymethyl)ethyl] amino} ethane sulfonic acid 相似文献
4.
5.
Pawan Sharma 《Archives of microbiology》1984,139(2-3):196-201
Neo-peptone B119 (Difco) was found to have a significant effect on differentiation of heterocysts and akinetes in Anabaena cylindrica. On adding neopeptone (0.4 g/l) to exponential phase culture of A. cylindrica, the following effects were observed (i) increased heterocyst frequency with altered heterocyst spacing and presence of double and multiple heterocysts after 24 h in cultures grown on N-free medium, (ii) induction of regular pattern of heterocysts after 48 h, in culture grown on medium supplemented with NH4Cl, (iii) induction of pro-akinetes after 48 h in both N-free and ammonium-grown cultures. The higher concentrations of neo-peptone were lytic to A. cylindrica, and, its lytic and inductive effects could be decreased by acid hydrolysis or supplementation of NH4Cl. Gel-filtration of neo-peptone showed that the inductive as well as the lytic effect was associated with some active factor(s) with molecular weight between 10,000–20,000. The retention of the inductive effect on autoclavation but its loss on trypsin digestion suggested that active factor(s) may be heat stable polypeptide(s). The heterocyst induction by active factor(s) decreased and akinete induction increased with increasing culture age. The pro-akinetes induced during exponential phase divided before maturation, while those induced during late exponential phase, could achieve full maturity. Growth and nitrogenase activity was unaffected while there was an increase in mean cell length on treatment of A. cylindrica with active factor(s) from neo-peptone, indicating that the effect may be mediated through cell division process(es).Abbreviations used N
Nitrogen
- chl
chlorophyll 相似文献
6.
Simon V. Avery Marvin E. Miller Geoffrey M. Gadd Geoffrey A. Codd Joseph J. Cooney 《FEMS microbiology letters》1991,84(2):205-210
Abstract Inhibition of photosynthesis by a range of organotin compounds in Plectonema boryanum was concentration-dependent and decreased in the order tributyltin (Bu3 SnCl) > tripropyltin (Pr3 SnCl) ≥ dibutyltin (Bu2 SnCl2 ) ≥ triphenyltin (Ph3 SnCl) > triethyltin (Et3 SnCl) > trimethyltin (Me3 SnCl) > monobutyltin (BuSnCl3 ). IC50 values were determined for the most toxic organotin species and varied from approximately 1.2 μM for Bu3 SnCl to approximately 13 μM for Ph3 SnCl. A similar order of inhibition of photosynthesis was observed in Anabaena cylindrica , although here IC50 values were slightly lower (e.g. approximately 1 μM for Bu3 SnCl and 5 μM for Ph3 SnCl).Nitrogenase activity was generally more sensitive to inhibition by organotin compounds than photosynthesis in A. cylindrica and this was particularlyy evident for Bu2 SnCl2 ; approximate IC50 values for Bu2 SnCl2 were 3 and 9 μM, as estimated by nitrogenase activity and photosynthesis, respectively. These results indicate that organotin compounds have the potential to inhibit cyanobacterial metabolism in aquatic systems. 相似文献
7.
A comparative study has been made on the pigment composition and nitrogenase activity of whole filaments and isolated beterocysts from a mutant strain of Anabaena CA. The whole cell absorption spectra of intact filaments and isolated heterocysts showed close resemblance especially between 550–700 nm region. On a quantitative basis the chlorophyll a content was found almost equal between the vegetative cell and heterocyst but the c-phycocyanin content in the heterocyst was about 1/2 that of the vegetative cell. The purification of the phycobiliprotein on DEAE-cellulose showed the presence of c-phycocyanin (max 615 nm) and allophycocyanin (max 645 nm, shoulder 620 nm). Isolated heterocysts under H2 showed acetylene reduction rates of 57 nmol C2H4/mg dry wt·min (342 mol C2H4/mg chl a·h), whereas intact filaments reduced at the rate of 18 nmol C2H4/mg dry wt·min (108 mol C2H4/mg chl a·h). This rate accounts for 30% recovery of nitrogenase activity in isolated heterocysts compared to whole filaments. The activity was strictly light dependent and was linear under H2 for more than 3 h. Addition of as little as 5% H2 under argon stimulated the C2H2 reductionseveral fold. The acetylene reduction (nitrogenase activity) also showed tolerance to 5% added O2 either under H2 or argon. The results suggest that the heterocyst of Anabaena CA-V is different in some characteristics (viz., higher endogenous C2H2 reduction rate, prolonged activity and higher levels of phycobiliproteins) than those reported in other Anabaena species. 相似文献
8.
Transport of the ammonium analogue [(14)C]methylammonium was similar in non-growing, fully differentiated heterocysts as compared to vegetative, multiplying cells of the filamentous cyanobacterium Anabaena variabilis. NH(4)(+) inhibited uptake into the cells and released accumulated methylammonium from the cells. These observations suggest that the main function of ammonium transport in heterocysts may not be NH(4)(+) acquisition but cyclic retention of ammonia produced by nitrogenase. 相似文献
9.
Mutants ofAnabaena variabilis Kütz. that have a decreased ability to grow photoautotrophically have been isolated by a modification of the techniques used to isolate auxotrophic mutants of that filamentous cyanobacterium, and have been stably propagated. Three mutants have a reduced content of phycocyanin and, as determined by in situ assays of partial reaction sequences of photosynthesis, an impairment in photosystem II. Three other strains, all of which appear to have a normal complement of carotenoids when grown heterotrophically, are sensitive to light.Abbreviations Used TES
N-tris(hydroxymethyl)-methyl-2-aminoethanesulfonic acid, sodium salt
- HEPES
N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid, sodium salt
- MV
methylviologen
- DBMIB
2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone
- DAB
3,3-diaminobenzidine
- P-BQ
p-benzoquinone
- DCMU
3-(3,4-dichlorophenyl)-1,1-dimethylurea
- Fecy
K-ferricyanide
- NTG
N-methyl-N-nitro-N-nitrosoguanidine 相似文献
10.
A method is described for the isolation of heterocysts that are virtually free of contaminating cell debris after sonication of aerobically grown Anabaena 7120. Isolated heterocysts reduced acetylene in a light-dependent process in the absence of exogenously provided ATP; heterocysts supplied with ATP and Na2S2O4 reduced acetylene slowly in the dark but still showed a marked light activation. Nitrogenase activity was greatest in fractions containing intact heterocysts. Up to 13% of the activity of the intact filaments was accounted for in the isolated heterocyst preparation.Isolated heterocysts took up O2 in a light-independent process; O2 uptake with added NADP+ was enhanced by pyruvate, isocitrate and intermediates of the oxidative pentose pathway. 相似文献
11.
Glutathione reductase activity was detected and characterized in heterocysts and vegetative cells of the cyanobacterium Nostoc muscorum. The activity of the enzyme varied between 50 and 150 nmol reduced glutathione· min-1·mg protein-1, and the apparent Km for NADPH was 0.125 and 0.200 mM for heterocysts and vegetative cells, respectively. The enzyme was found to be sensitive to Zn+2 ions, however, preincubation with oxidized glutathione rendered its resistance to Zn+2 inhibition. Nostoc muscorum filaments were found to contain 0.6–0.7mM glutathione, and it is suggested that glutathione reductase can regenerate reduced glutathione in both cell types. The combined activity of glutathione reductase and isocitrate dehydrogenase in heterocysts was as high as 18 nmol reduced glutathione·min-1·mg protein-1. A relatively high superoxide dismutase activity was found in the two cell types; 34.2 and 64.3 enzyme units·min-1·mg protein-1 in heterocysts and vegetative cells, respectively.We suggest that glutathione reductase plays a role in the protection mechanism which removes oxygen radicals in the N2-fixing cyanobacterium Nostoc muscorum.Abbreviations DTNB
5-5-dithiobis-(2-nitrobenzoic acid)
- EDTA
ethylenediaminetetra-acetic acid
- GR
glutathione reductase (EC1.6.4.2)
- GSH
reduced glutathione
- GSSG
oxidized glutathione
- OPT
O-phtaldialdehyde
- SOD
superoxide dismutase (EC 1.15.1.1) 相似文献
12.
Summary
Anabaena variabilis ATCC 29413 contains two cryptic plasmids. Clones of the smaller (41 kb) plasmid, designated pRDS1, in cosmid vectors were
used to construct a physical map. A clone bank of pRDS1 constructed by ligating fragments from aXhoII digest of a pRDS1 cosmid clone into a mobilizable plasmid was used to locate an origin of replication of pRDS1. Because we
were unable to cureA. variabilis of pRDS1, the clone bank was transferred by conjugation to another strain ofAnabaena sp., strain M-131. A 5.3 kb fragment of pRDS1 contained all of the sequences necessary for replication inAnabaena sp. strain M-131 as judged by the ability to rescue the hybrid vector from exconjugants in unchanged form after many generations.
Hybrid plasmids derived from pRDS1, one bearing genes for luciferase, were also transferred by conjugation toA. variabilis, where they appeared to recombine with pRDS1. 相似文献
13.
Bent Borg Jensen 《Archives of microbiology》1990,154(1):92-98
We have investigated the utilization of [14C]-fructose by whole filaments and isolated heterocysts of Anabaena variabilis ATCC 29413, a strain which is capable of fructose-dependent heterotrophic growth. The experimental conditions were chosen such that both transport and subsequent metabolism were studied. The apparent Km for fructose was 60 mM, close to the results of previous studies. Rates of fructose utilization were the same in light and darkness. When photosynthetic CO2 fixation was possible, almost all the label appeared as cell-carbon. In darkness or in the presence of DCMU appreciable amounts of label were released as CO2. Isolated heterocysts with high rates of endogenous metabolism were not capable of utilizing added fructose at significant rates. The effects of oxygen concentration on the metabolism of added fructose in darkness showed that uptake was saturated at low pO2 values. Increasing the pO2 values lead to an increase in the ratio between the lable released as CO2 and that recovred as cell-carbon. These results suggest that fructose is taken up only by the vegetative cells but carbon derived from added fructose can be released as CO2 as a result of respiration in the heterocysts. Fructose utilization was inhibited by uncouplers. The greatest inhibition was found when both (delta) (psi) and (delta) pH were abolished. High concentrations of erythrose inhibited fructose utilization. None of the other potential analogs tested had any effect. 相似文献
14.
The photophobic responses in the Cyanobacterium Anabaena variabilis which belongs to the Nostocaceae have been studied with aid of a population method as well as by single trichome observations. In white light experiments both step-up and step-down photophobic responses were observed. The wavelength dependence was examined at a constant fluence rate. The photophobically active light is absorbed by the photosynthetic pigments, mainly by the phycobiliproteins and chlorohyll a. Above 690 nm only negative reactions were observed, i.e. the trichomes left the light trap. In white light experiments DCMU strongly inhibited the photophobic responses, whereas photokinesis was not affected to the same extent indicating that the reaction is coupled with the non cyclic photosynthetic electron transport. DBMIB impaired the photophobic behaviour only slightly. It seems that the photophobic responses of A. variabilis are controlled by a similar mechanism as in Phormidium uncinatum (Oscillatoriaceae) although the two families and, hence, the two species differ in their movement mechanism as well as in their photoactic behaviour. 相似文献
15.
The presence and localization of a reversible hydrogenase in non-N2-fixing cells of the filamentous cyanobacterium Anabaena variabilis were investigated by in vitro activity measurements, native-PAGE/activity stain, SDS-PAGE/Western immunoblots, and immunogold localization. Reversible hydrogenase activity was induced approximately 100-fold by sparging the cell suspensions with a mixture of 99% argon and 1% CO2 for 20–26 h. Native-PAGE/activity stain demonstrated the presence of an in vitro functional enzyme with an apparent molecular mass of 118 kDa. Native-PAGE/Western immunoblots, using polyclonal antisera directed against purified hydrogenase from the purple sulphur bacterium Thiocapsa roseopersicina, detected two native proteins with molecular masses of 118 and 133 kDa, respectively. SDS-PAGE/Western immunoblots confirmed the presence of a single polypeptide with a molecular mass of approximately 40 kDa in both induced and non-induced cells. Immunocytolocalization experiments using ultrathin sections again demonstrated the presence of hydrogenase in both induced and non-induced cells. A higher specific labeling was associated with the thylakoid regions, which, using an image analyzer, was calculated to be approximately 4 x higher per cell area compared to in the centroplasm. It is suggested that anaerobic incubation induces higher reversible hydrogenase activity, regulated mainly at the level of activating (pre)existing form(s) of inactive enzyme(s)/protein(s), maybe in combination with synthesis of additional subunit(s). 相似文献
16.
Isolated heterocysts from Anabaena variabilis show high rates of light- and hydrogen-dependent acetylene reduction. This heterocyst preparation also shows light-induced redox reactions of cytochromes f-556 and b-564. Both cytochromes are reversibly oxidized by light or by oxygen (in the dark). Oxygen-oxidized cytochromes assume their initially reduced state by addition of dithionite or flushing the reaction vessel with hydrogen or argon. All three agents remove oxygen from solutions more or less efficiently, thereby creating reducing conditions; hydrogen-induced reduction is slow and generally completed after about 10 min. No evidence has been obtained for a direct electron donation of hydrogen to either of the cytochromes measured. Photoreduction of nitrogen in the presence of hydrogen is explained by a combined operation of properly poised cyclic photophosphorylation providing ATP, and a light-independent hydrogenase reaction providing reductant. 相似文献
17.
The free-living cyanobacterium Anabaena variabilis showed a biphasic pattern of 14CH3NH
3
+
uptake. Initial accumulation (up to 60 s) was independent of CH3NH
3
+
metabolism, but long-term uptake was dependent on its metabolism via glutamine synthetase (GS). The CH3NH
3
+
was converted into methylglutamine which was not further metabolised. The addition of l-methionine-dl-sulphoximine (MSX), to inhibit GS, inhibited CH3NH
3
+
metabolism, but did not affect the CH3NH
3
+
transport system.NH
4
+
, when added after the addition of 14CH3NH
3
+
, caused the efflux of free CH3NH
3
+
; when added before 14CH3NH
3
+
, NH
4
+
inhibited its uptake indicating that both NH
4
+
and CH3NH
3
+
share a common transport system. Carbonylcyanide m-chlorophenylhydrazone and triphenyl-methylphosphonium both inhibited CH3NH
3
+
accumulation indicating that the transport system was -dependent. At pH 7 and at an external CH3NH
3
+
concentration of 30 mol dm-3, A. variabilis showed a 40-fold intracellular accumulation of CH3NH
3
+
(internal concentration 1.4 mmol dm-3). Packets of the symbiotic cyanobacterium Anabaena azollae, directly isolated from the water fern Azolla caroliniana, also showed a -dependent NH
4
+
transport system suggesting that the reduced inhibitory effect of NH
4
+
on nitrogenase cannot be attributed to the absence of an NH
4
+
transport system but is probably related to the reduced GS activity of the cyanobiont.Abbreviations CCCP
carbonylcyanide m-chlorophenylhydrazone
- GS
glutamine synthetase
- HEPES
4-(2-hydroxyethyl)-1-piperazine ethanesulphonic acid
- MSX
l-methionine-dl-sulphoximine
-
membrane potential
- pH
transmembrane pH difference
- TPMP+
triphenylmethylphosphonium 相似文献
18.
Five strains of heterocystous blue-green algae capable of high rates of growth and nitrogenase activity were isolated from shallow coastal environments. Growth of the organisms was characterized with respect to temperature, NaCl concentration in the medium, and nitrogen source. The temperature optima ranged from 35–42°C, and all but one of the strains displayed a requirement for added NaCl. The generation times under N2-fixing conditions were 5.1–5.9 h, and were as low as 3.4 h for growth on NH4Cl. Nitrogenase activity (C2H2 reduction) was high throughout the logarithmic growth phase of each strain. The maximum value observed for one strain was 65.5 nmoles C2H4 produced/mg protein x min, and the average values for the five strains ranged from 24.5–46.7 nmoles C2H4/mg protein x min. The organisms all belong to the genusAnabaena. The growth and nitrogenase activity of these strains are much higher than those of the heterocystous blue-green algae commonly used for investigation of nitrogen metabolism, and they thus should prove to be useful physiological tools. Their prevalence, as judged by the ease of their enrichment and isolation, in bay and estuarine environments suggests that they are important contributors of combined nitrogen. 相似文献
19.
Ellen B. Braun-Howland Peter Lindblad Sandra A. Nierzwicki-Bauer Birgitta Bergman 《Planta》1988,176(3):319-322
Transmission electron microscopy and immunocytological labeling were used to study the distribution and ontological occurrence of dinitrogenase reductase (Fe-protein) of nitrogenase in cyanobacterial symbionts within young leaves of the water-ferns Azolla filiculoides Lamarck, A. caroliniana Willdenow, and A. pinnata R. Brown. Rabbit anti-dinitrogenase reductase antisera and goat anti-rabbit-immunoglobulin G antibody conjugated to colloidal gold were used as probes. Western blot analyses showed that a polypeptide of approx. 36 kDa (kdalton) was recognized in the symbionts of all three Azolla species and that the polyclonal sera used were monospecific. In all symbionts, nitrogenase was immunologically recognizable within heterocysts. It was absent from vegetative cells, and also from the akinetes of the A. caroliniana and A. pinnata symbionts. The differentiation of vegetative cells into heterocysts in all three symbionts was initiated by formation of additional external cell-wall layers and narrowing of the neck followed by loss of glycogen, mild vesiculation of thylakoid membranes, and the appearance of polar nodules. No nitrogenase was detected at these early stages, but it appeared in the intermediate proheterocyst stage concomitantly with the formation of contorted membranes, and reached the strongest labeling in mature heterocysts, containing extensive tightly packed membranes. Nitrogenase was evenly distributed throughout heterocysts except at the polar regions, which contained honey-comb configurations and large polar nodules. With increased age of the A. caroliniana and A. pinnata symbionts, heterocysts became highly vesiculated, with a concomitant decrease in the amount of nitrogenase detected.Abbreviations IgG
Immunoglobulin G
- PAGE
polyacrylamide gel electrophoresis
- SDS
sodium dodecyl sulfate
- TEM
transmission electron micrograph 相似文献
20.
Isolated heterocysts of Anabaena 7120 evolve H2 in an ATP-dependent nitrogenase-catalyzed process that is inhibited by N2 and C2H2. Heterocysts have an active uptake hydrogenase that only requires an electron acceptor of positive redox potential, e.g., methylene blue, dichlorophenolindophenol or potassium ferricyanide. O2 supplied at low partial pressures is a very effective physiological oxidant for H2 uptake. High concentrations of O2 are inhibitory to H2 uptake. The oxyhydrogen reaction in heterocysts appears to be mediated by a cytochrome-cytochrome oxidase system, and it supports ATP synthesis via oxidative phosphorylation. Attempts to demonstrate acetylene reduction in isolated heterocysts employing H2 as an electron donor were unsuccessful. It is suggested that the uptake hydrogenase functions to conserve reductant that otherwise would be dissipated via nitrogenase-catalyzed H2 evolution. 相似文献