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Archives of Microbiology (v.176, #4)
No Title
by Pascale Servant; Philippe Mazodier (pp. 237-242).
All organisms respond to a sudden increase in temperature by inducing the synthesis of a set of proteins called heat shock proteins (HSPs). Although the induction of HSPs is a universal response, a diversity of mechanisms control HSP synthesis in different organisms. In Streptomyces, the synthesis of major HSPs, such as the widespread molecular chaperones DnaK, ClpB, GroEL and HSP18, is negatively controlled at the transcriptional level by at least three different repressors. The control of groE gene expression involves an inverted repeat (called the CIRCE element) that is highly conserved among eubacteria, and the HrcA repressor. The dnaK operon and clpB belong to the HspR /HAIR regulon. The HspR repressor-HAIR operator system is used in some bacteria but is not widespread. In particular, it has not been found in gram-positive bacteria with low G+C content. Transcription of hsp18, which encodes a small HSP, is regulated by the RheA repressor. This repressor, which has intrinsic thermosensor activity, has to date been identified only in Streptomyces.
Keywords: Heat regulation Streptomyces Repressor HrcA HspR RheA CtsR
No Title
by Johannes F. Imhoff (pp. 243-254).
Anoxygenic phototrophic bacteria are widely distributed in marine sediments and shallow waters of the coastal zone, where they often form intensely colored mass developments. The phototrophic bacteria have adapted to the whole spectrum of salt concentrations, from freshwater to saturated brines, and it is apparent that individual species have adapted well to particular habitats and mineral salts compositions, both qualitatively and quantitatively. This adaptation is reflected not only in the demand for defined ranges of salt concentrations, but also in the phylogenetic relationships of these bacteria, as established by 16S rDNA sequences. Major phylogenetic branches of purple sulfur bacteria are represented by: (1) marine and extremely halophilic Ectothiorhodospiraceae, (2) truly marine and halophilic Chromatiaceae and (3) freshwater Chromatiaceae, some of which are tolerant to low salt concentrations and are successful competitors in brackish and marine habitats. Quite similarly, salt-dependent green sulfur bacteria form distinct phylogenetic lines. In addition, also among the phototrophic α-Proteobacteria (purple nonsulfur bacteria), distinct phylogenetic lines of salt-dependent species are recognized. Available data give rise to the assumption that salt concentrations of natural habitats are an important selective factor that determines the development of a selected range of phototrophic bacteria in an exclusive way. As a consequence, the salt responses of these bacteria are reflected in their phylogenetic relationships.
Keywords: Anoxygenic phototrophic bacteria Marine phototrophic bacteria Phylogeny Salt adaptation
No Title
by Shiping Deng; James G. Elkins; Linda H. Da; Lina M. Botero; Timothy R. McDermott (pp. 255-263).
Sinorhizobium meliloti has two nonspecific periplasmic acid phosphatases. The NapD enzyme has been previously described, and a second acid phosphatase, NapE, is described in this report. NapE was partially purified from an S. meliloti napD mutant and characterized with respect to molecular mass and substrate range. As predicted from SDS-PAGE analysis, the subunit molecular mass of NapE is approximately 35.8 kDa and gel filtration experiments estimated the native molecular mass to be approximately 70 kDa, indicating that the active enzyme is a homodimer. NapE demonstrated significant activity with p-nitrophenyl phosphate, phenyl phosphate, and α-naphthyl-phosphate. The pH optimum was between 4.5 and 5.0. The gene encoding NapE was also sequenced and the inferred amino acid sequence from the predicted ORF was found to be 60% identical and 75% similar to that encoded by napD. An S. meliloti napE mutant was constructed and assessed for symbiotic competence. This mutant did not differ from the wild-type parent strain in nodulation and symbiotic efficiency.
Keywords: Phosphorus Sinorhizobium Alfalfa Phosphatase
No Title
by Yoh Takahata; Toshihiro Hoaki; Tadashi Maruyama (pp. 264-270).
The starvation survivability of seven Thermococcus strains isolated from four Japanese oil reservoirs was compared with that of Thermococcus strains from marine hydrothermal fields. 16S rDNA analyses showed the isolates to be closely related to Thermococcus litoralis. Growth of the isolates was dependent on amino acids, which were present at low concentrations in the oil reservoirs. At 80 °C in the formation water, strain CKU-1 from the oil reservoir showed a higher starvation survivability than strain KS-1 from the marine hydrothermal field. Crude oil did not affect the starvation survivability of strain CKU-1, but it reduced that of strain KS-1. These results indicate that strain CKU-1 could survive longer than stain KS-1 under the conditions of an oil reservoir. At 90 °C in artificial seawater without organic nutrients, the half-lives of the isolates were between 7.7 and 25.1 days. However, those of the strains from marine hydrothermal fields, except Thermococcus litoralis and Thermococcus chitonophagus, were less than 1.0 day. The higher starvation survivability is probably important for the hyperthermophiles to continue to exist in a hot subterranean oil reservoir where the supply of nutrients seems to be limited.
Keywords: Hyperthermophilic archaea Thermococcus Starvation survivability Oil reservoir Hydrothermal field Half-life Amino acid
No Title
by Song-Gun Kim; Hee-Sung Bae; Sung-Taik Lee (pp. 271-277).
The aerobic and anaerobic degradation of trimethylamine by a newly isolated denitrifying bacterium from an enrichment culture with trimethylamine inoculated with activated sludge was studied. Based on 16S rDNA analysis, this strain was identified as a Paracoccus sp. The isolate, strain T231, aerobically degraded trimethylamine, dimethylamine and methylamine and released a stoichiometric amount of ammonium ion into the culture fluid as a metabolic product, indicating that these methylated amines were completely degraded to formaldehyde and ammonia. The strain degraded trimethylamine also under denitrifying conditions and consumed a stoichiometric amount of nitrate, demonstrating that complete degradation of trimethylamine was coupled with nitrate reduction. Cell-free extract prepared from cells grown aerobically on trimethylamine exhibited activities of trimethylamine mono-oxygenase, trimethylamine N-oxide demethylase, dimethylamine mono-oxygenase, and methylamine mono-oxygenase. Cell-free extract from cells grown anaerobically on trimethylamine and nitrate exhibited activities of trimethylamine dehydrogenase and dimethylamine dehydrogenase. These results indicate that strain T231 had two different pathways for aerobic and anaerobic degradation of trimethylamine. This is a new feature for trimethylamine metabolism in denitrifying bacteria.
Keywords: Trimethylamine Denitrification Anaerobic degradation Aerobic degradation Trimethylamine metabolism
No Title
by Remy Guyoneaud; Carles M. Borrego; Asunción Martínez-Planells; Erik T. Buitenhuis; Jesús L. Garcia-Gil (pp. 278-284).
The morphology (mainly prosthecae length), ultrastructure, and antenna pigment composition of the green sulfur bacterium Prosthecochloris aestuarii changed when grown under different light intensities. At light intensities of 0.5 and 5 µmol quanta m–2 s–1, the cells had a star-like morphology. Prosthecae, the characteristic appendages of the genus Prosthecochloris, were 232 nm and 194 nm long, respectively. In contrast, when grown at 100 µmol quanta m–2 s–1, these appendages were shorter (98 nm) and the cells appeared more rod-shaped. Transmission electron microscopy revealed a significant decrease in the cell perimeter to area ratio and in the number of chlorosomes per linear µm of membrane as light intensity increased. In addition to these morphological and ultrastructural responses, Prosthecochloris aestuarii exhibited changes in its pigment composition as a function of light regime. Lower specific pigment content and synthesis rates were found in cultures grown at light intensities above 5 µmol quanta m–2 s–1. A blue shift in the bacteriochlorophyll (BChl) c Qy absorption maximum of up to 17.5 nm was observed under saturating light conditions (100 µmol quanta m–2 s–1). This displacement was accompanied by changes in the composition of BChl c homologs and by a very low carotenoid content. The morphological, ultrastructural and functional changes exhibited by Prosthecochloris aestuarii revealed the strong light-response capacity of this bacterium to both high and low photon-flux densities.
Keywords: Prosthecochloris Light response Prosthecae Chlorosomes Antenna pigments
No Title
by Michael Hoppert; Carsten Gentzsch; Kurt Schörgendorfer (pp. 285-293).
The electron microscopic image of native cyclosporin synthetase molecules showed large globular complexes of 25 nm in diameter, built up by smaller interconnected units. Compartmentation of cyclosporin synthetase and the functionally interconnected D-alanine racemase was revealed after sucrose density gradient centrifugation of subcellular fractions and immunoelectron microscopy. A considerable proportion of cyclosporin synthetase and D-alanine racemase was detected at the vacuolar membrane. The product cyclosporin was localized in the fungal vacuole.
Keywords: Cyclosporin A Cyclosporin synthetase Tolypocladium inflatum Compartmentation Immunoelectron microscopy
No Title
by Antti Nyyssölä; Matti Leisola (pp. 294-300).
The extremely halophilic actinomycete Actinopolyspora halophila is a rare example of a heterotrophic eubacterium producing betaine from simple carbon sources. A. halophila synthesized remarkably high intracellular concentrations of betaine. The highest betaine concentration, determined at 24% (w/v) NaCl, was 33% of the cellular dry weight. Trehalose was synthesized as a compatible solute, accounting for up to 9.7% of the cellular dry weight. The betaine concentration was shown to increase with increasing NaCl concentration, whereas the trehalose concentration was highest at the lowest NaCl concentration used (15% w/v). A. halophila was capable of accumulating betaine from the medium, while at the same time betaine was also excreted back into the medium by the cells. Along with the de novo synthesis of betaine, A. halophila was able to take up choline from the medium and oxidize it to betaine. Some basic characteristics of the choline oxidation system are described. Choline was oxidized to betaine aldehyde in a reaction in which H2O2 generation and oxygen consumption were coupled. Betaine aldehyde was also oxidized, but with lesser efficiency. In addition, betaine aldehyde was oxidized further to betaine in a reaction in which NAD(P)+ was reduced.
Keywords: Betaine Osmoregulation Extreme halophiles Actinopolyspora halophila Choline oxidation Compatible solutes
No Title
by Olga Sánchez; Isabel Ferrera; Christiane Dahl; Jordi Mas (pp. 301-305).
Adenosine-5′-phosphosulfate (APS) reductase participates in the oxidation of sulfite to APS in Allochromatium vinosum. Oxidation of sulfite via the APS pathway yields ATP through substrate-level phosphorylation. An alternative enzyme for the oxidation of sulfite to sulfate, sulfite:acceptor oxidoreductase, has also been reported in Ach. vinosum. Oxidation of sulfite through this enzyme does not yield ATP. APS reductase is expressed constitutively in Ach. vinosum, suggesting that it performs an important role in this organism. However, studies carried out with batch cultures of an APS reductase mutant showed little or no differences in growth or in the rates of substrate oxidation when compared to the wild-type, therefore questioning the role of this enzyme. In an attempt to establish whether the ATP gain derived from APS-reductase-mediated oxidation of sulfite is relevant for energy-limited cultures, we compared growth of the wild-type SM50 and the APS-reductase-deficient mutant D3 when grown in continuous culture under different degrees of illumination. Little differences in the specific growth rates of the two strains were observed at light-limiting irradiances, suggesting that the ATP gained during sulfite oxidation through the APS reductase pathway does not constitute a significant energy input. However, at saturating irradiances, wild-type Ach. vinosum grew considerably faster than the mutant. Increasing the irradiance even further resulted in inhibition of the wild-type strain down to the level of the APS reductase mutant. The implications of these results are discussed.
Keywords: Allochromatium vinosum Adenosine-5′-phosphosulfate reductase Sulfite oxidation Sulfite:acceptor oxidoreductase
No Title
by Antje Baumgarten; Insa Redenius; Jan Kranczoch; Heribert Cypionka (pp. 306-309).
Washed cells of Desulfovibrio vulgaris strain Marburg (DSM 2119) reduced oxygen to water with H2 as electron donor at a mean rate of 253 nmol O2 min–1 (mg protein)–1. After separating the periplasm from the cells, more than 60% of the cytochrome c activity and 90% of the oxygen-reducing activity were found in the periplasmic fraction. Oxygen reduction and the reduction of cytochrome c with H2 were inhibited by CuCl2. After further separation of the periplasm by ultrafiltration (exclusion sizes 30, 50, and 100 kDa), oxygen reduction with H2 occurred with the retentates only. Ascorbate plus tetramethyl-p-phenylenediamine (TMPD), however, were also oxidized by the filtrates. The stoichiometry of 1 mol O2 reduced per 2 mol ascorbate oxidized indicated the formation of water. Our experiments present evidence that in D. vulgaris periplasmic hydrogenase and cytochrome c play a major role in oxygen reduction. Preliminary studies with other Desulfovibrio species indicated a similar function of periplasmic c-type cytochromes in D. desulfuricans CSN and D. termitidis KH1.
Keywords: Sulfate-reducing bacteria Aerobic respiration Terminal oxidase Cytochrome c3 Periplasm Hydrogen TMPD Ascorbate Desulfovibrio vulgaris strain Marburg Desulfovibrio desulfuricans strain CSN Desulfovibrio termitidis strain KH1
No Title
by Roland Gross; Jörg Simon; Achim Kröger (pp. 310-313).
The cell homogenate and the soluble cell fraction of Wolinella succinogenes grown with formate and fumarate catalyzed the oxidation of benzyl viologen radical by methacrylate [apparent K m=0.23 mM, V max=1.0 U (mg cell protein) –1] or acrylate [apparent K m=0.50 mM, V max=0.77 U (mg cell protein) –1]. Crotonate did not serve as an oxidant. A mutant of W. succinogenes lacking the fccABC operon was unable to catalyze methacrylate or acrylate reduction. In contrast, the inactivation of fccC alone had no effect on these activities. Methacrylate reduction by benzyl viologen radical was not catalyzed by fumarate reductase isolated from the membrane of W. succinogenes. Cells grown with formate and fumarate did not catalyze methacrylate reduction by formate, and W. succinogenes did not grow with formate and methacrylate as catabolic substrates. The results suggest that the reduction of methacrylate or acrylate by benzyl viologen radical is most likely catalyzed either by the periplasmic flavoprotein FccA or by a complex consisting of FccA and the predicted c-type cytochrome FccB. The metabolic function of the fccABC operon remains unknown.
Keywords: Methacrylate reduction Acrylate reduction Fumarate reduction Anaerobic respiration Flavoprotein Wolinella succinogenes
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