Carotenoids and Reaction Center II-D1 Protein in Light Regulation of the Photosynthetic Apparatus in Aphanocapsa*

A new cyanobacterial isolate, morphologically closely resembling Aphanocapsa, was characterized for its growth requirements, as well as pigmentation, photosynthetic activity and dynamics of the D1 protein in the reaction center (RC) of photosystem II (PSII). It was shown to be able to grow on glucose in the dark in the presence of DCMU. The cyanobacterium turned light yellow at high light intensity in the absence, and dark emerald green in high light in the presence of sublethal concentrations of the DCMU-type inhibitor atrazine. While total carotenoids per cell slightly decreased with increasing light intensity during growth, the cells still turned pale yellow due to decreased levels of chlorophyll and phycocyanin. In contrast to β-carotene, zeaxanthin and echinenone which decreased with increasing light intensity during growth, the carotenoid glycoside, myxoxanthophyll, continuously increased in concentration. Extremely high rates of light-saturated O₂ evolution were recorded for the high light cultures after a 0.5 h recovery period in the dark. The recovery measured after 2.5 h was shown to be less effective in darkness than in dim light and was prohibited by chloramphenicol. The degree of recovery was dependent on the light intensity during growth. A fast light intensity-dependent RC II-D1 protein turnover was found for the bleached yellow cells rich in myxoxanthophyll. The half-life of the RC II-D1 protein, plotted against the light intensity during growth and experimentation, yielded a curve the slope of which was considerably steeper for Aphanocapsa than for Anacystis. Apparently, the isolated strain of Aphanocapsa reacts more vigorously to changes in the environment than other strains tested and may, therefore, turn out to be a suitable organism in the attempt to elucidate the molecular mechanism of light intensity adaptation.     

Metabolism of glucose by unicellular blue-green algae

Summary A facultative photo- and chemoheterotroph, the unicellular bluegreen alga Aphanocapsa 6714, dissimilates glucose with formation of CO₂ as the only major product. A substantial fraction of the glucose consumed is assimilated and stored as polyglucose (probably glycogen). The oxidation of glucose proceeds through the pentose phosphate pathway. The first enzyme of this pathway, glucose-6-phosphate dehydrogenase, is partly inducible. In addition, the rate of glucose oxidation is controlled, at the level of glucose-6-phosphate dehydrogenase function, by the intracellular level of an intermediate of the Calvin cycle, ribulose-1,5-diphosphate, which is a specific allosteric inhibitor of this enzyme. As a consequence, the rate of glucose oxidation is greatly reduced by illumination, an effect reversed by the presence of DCMU, an inhibitor of photosystem II.Two obligate photoautotrophs, Synechococcus 6301 and Aphanocapsa 6308, produce CO₂ from glucose at extremely low rates, although their levels of pentose pathway enzymes and of hexokinase are similar to those in Aphanocapsa 6714. Failure to grow with glucose appears to reflect the absence of an effective glucose permease. A general hypothesis concerning the primary pathways of carbon metabolism in blue-green algae is presented.Abbreviations A (U)DPG ADP-glucose or UDP-glucose - G-1-P glucose-1-phosphate - G-6-P glucose-6-phosphate - G_(int.) intracellular glucose - F-6-P fructose-6-phosphate - 6-PG 6-phosphogluconate - Ru-5-P ribulose-5-phosphate - RUDP ribulose-1,5-diphosphate - PGA 3-phosphoglycerate - GAP glyceraldehyde-3-phosphate     

Truepera radiovictrix gen. nov., sp. nov., a new radiation resistant species and the proposal of Trueperaceae fam. nov

Two isolates, belonging to a new species of a novel genus of the Phylum “Deinococcus/Thermus ”, were recovered from hot spring runoffs on the Island of São Miguel in the Azores. Strains RQ-24^T and TU-8 are the first cultured representatives of a distinct phylogenetic lineage within this phylum. These strains form orange/red colonies, spherical-shaped cells, have an optimum growth temperature of about 50 °C, an optimum pH for growth between about 7.5 and 9.5, and do not grow at pH below 6.5 or above pH 11.2. These organisms grow in complex media without added NaCl, but have a maximum growth rate in media with 1.0% NaCl and grow in media containing up to 6.0% NaCl. The organisms are extremely ionizing radiation resistant; 60% of the cells survive 5.0 kGy. These strains are chemoorganotrophic and aerobic; do not grow in Thermus medium under anaerobic conditions with or without nitrate as electron acceptor and glucose as a source of carbon and energy, but ferment glucose to d-lactate without formation of gas. The organisms assimilate a large variety of sugars, organic acids and amino acids. Fatty acids are predominantly iso- and anteiso-branched; long chain 1,2 diols were also found in low relative proportions; menaquinone 8 (MK-8) is the primary respiratory quinone. Peptidoglycan was not detected. Based on 16S rRNA gene sequence analysis, physiological, biochemical and chemical analysis we describe a new species of one novel genus represented by strain RQ-24^T (CIP 108686^T = LMG 22925^T = DSM 17093^T) for which we propose the name Truepera radiovictrix. We also propose the family Trueperaceae fam. nov. to accommodate this new genus.     

Physiological and biochemical contributions to the taxonomy of the genus Prototheca

Prototheca zopfii (12 strains) is able to use glucose, fructose, propanol, glycerol, and acetate as sources of carbon for growth. One of the strains is biochemically (utilization also of galactose and mannose), and two strains are morphologically slightly different.Two strains can be identified as P. wikerhamii. They exhibit good growth with glucose, fructose, galactose, trehalose, propanol, glycerol, acetate, and glutamate as sources of carbon. P. spec. 263-2 grows only with glucose and acatate. P. zopfii and P. wickerhamii are able to use urea, glycine, and glutamate as sources of nitrogen. P. spec. 263-2, on the other hand, cannot utilize these organic nitrogen compounds for growth.Four strains of Chlorella protothecoides are able to use glucose, fructose, galactose, and acetate as sources of carbon for growth in the dark. Three of them utilize also mannose, trehalose, and glutamate. Two strains can grow with glycerol, and one is able to use lactose. — Urea and glycine can serve as sources of nitrogen for the four strains of C. protothecoides. Glutamate supports growth of three strains, and one strain is able to use nicotinamide.     

Pelotomaculum terephthalicum sp. nov. and Pelotomaculum isophthalicum sp. nov.: two anaerobic bacteria that degrade phthalate isomers in syntrophic association with hydrogenotrophic methanogens

An anaerobic phthalate isomer-degrading strain (JT^T) that we previously isolated was characterized. In addition, a strictly anaerobic, mesophilic, syntrophic phthalate isomer-degrading bacterium, designated strain JI^T, was isolated and characterized in this study. Both were non-motile rods that formed spores. In both strains, the optimal growth was observed at temperatures around 37°C and neutral pH. In syntrophic co-culture with the hydrogenotrophic methanogen Methanospirillum hungatei, both strains could utilize two or three phthalate isomers for growth, and produce acetate and methane as end products. Strain JT^T was able to grow on isophthalate, terephthalate, and a number of low-molecular weight aromatic compounds, such as benzoate, hydroquinone, 2-hydroxybenzoate, 3-hydroxybenzoate, 2,5-dihydroxybenzoate, 3-phenylpropionate in co-culture with M. hungatei. It could also grow on crotonate, hydroquinone and 2,5-dihydroxybenzoate in pure culture. Strain JI^T utilized all of the three phthalate isomers as well as benzoate and 3-hydroxybenzoate for growth in co-culture with M. hungatei. No substrates were, however, found to support the axenic growth of strain JI^T. Neither strain JT^T nor strain JI^T could utilize sulfate, sulfite, thiosulfate, nitrate, fumarate, Fe (III) or 4-hydroxybenzoate as electron acceptor. Phylogenetically, strains JT^T and JI^T were relatively close to the members of the genera Pelotomaculum and Cryptanaerobacter in ‘Desulfotomaculum lineage I’. Physiological and chemotaxonomic characteristics indicated that the two isolates should be classified into the genus Pelotomaculum, creating two novel species for them. Here, we propose Pelotomaculum terephthalicum sp. nov. and Pelotomaculum isophthalicum sp. nov. for strain JT^T and strain JI^T, respectively. The type strains are strains JT^T (= DSM 16121^T = JCM 11824^T = NBRC 100523^T) and JI^T (= JCM 12282^T = BAA-1053^T) for P. terephthalicum and P. isophthalicum, respectively.Nucleotide sequence accession number: The GenBank/EMBL/DDBJ accession numbers of the 16S rRNA gene sequences of strains JT^T and JI^T are AB091323 and AB232785, respectively     

31P NMR and 13C NMR studies of recombinant Saccharomyces cerevisiae with altered glucose phosphorylation activities

Manipulation of cellular metabolism to maximize the yield and rate of formation of desired products may be achieved through genetic modification. Batch fermentations utilizing glucose as a carbon source were performed for three recombinant strains of Saccharomyces cerevisiae in which the glucose phosphorylation step was altered by mutation and genetic engineering. The host strain (hxk1 hxk2 glk) is unable to grow on glucose or fructose; the three plasmids investigated expressed hexokinase PI, hexokinase PII, or glucokinase, respectively, enabling more rapid glucose and fructose phosphorylation in vivo than that provided by wild-type yeast.Intracellular metabolic state variables were determined by ³¹P NMR measurements of in vivo fermentations under nongrowth conditions for high cell density suspensions. Glucose consumption, ethanol and glycerol production, and polysaccharide formation were determined by ¹³C NMR measurements under the same experimental conditions as used in the ³¹P NMR measurements. The trends observed in ethanol yields for the strains under growth conditions were mimicked in the nongrowth NMR conditions.Only the strain with hexokinase PI had higher rates of glucose consumption and ethanol production in comparison to healthy diploid strains in the literature. The hexokinase PII strain drastically underutilized its glucose-phosphorylating capacity. A regulation difference in the use of magnesium-free ATP for this strain could be a possible explanation. Differences in ATP levels and cytoplasmic pH values among the strains were observed that could not have been foreseen. However, cytoplasmic pH values do not account for the differences observed among in vivo and in vitro glucose phosphorylation activities of the three recombinant strains.     

Screening <Emphasis Type="BoldItalic">Arthrospira</Emphasis> (Spirulina) strains for heterotrophy

Thirty-five clonal, axenic Arthrospira strains were screened for their ability to grow heterotrophically on six carbon sources (20 mM). Glucose (34 strains) and fructose (24 strains) were the only substrates permitting growth in the dark. In some assays, however, not every replicate grew and, in at least one strain (D867), repeat assays over 2 years on material maintained in the light indicated that the ability to use fructose in the dark had become lost. Four further strains from other culture collections were compared, because they are presumed duplicates of three of the main set of strains; in at least three cases the duplicates of a pair differed in their ability to use fructose in the dark. In another comparison, where straight and helical morphotypes of the same strain could be compared, two of the four straight morphotypes (including one duplicate strain) grew with glucose in the dark, whereas none of the helical morphotypes did so. It is suggested that genetic drift has led to losses in the ability to grow heterotrophically in some strains.Ten of the main set of strains were tested for their ability to grow photoheterotrophically with four of the carbon sources (glucose, maltose, fructose, sucrose). All grew with glucose and maltose, but none with fructose or sucrose, in spite of the fact that eight (of this subset) grew with fructose in the dark. Sucrose led to most of a culture lysing, but often with short fragments of trichome surviving and subsequently giving rise to a normal culture. All the surviving cells in the transitory stage showed the presence of large intrathylakoidal granules, which had disappeared by the time that the culture had returned to a healthy state. Bleaching and recovery were more rapid at 70 than 10 μmol photon m⁻² s⁻¹. The presence of DCMU prevented this recovery. There was no bleaching when an inoculum of the recovered material was subcultured to medium with sucrose.     

Arginine catabolism in Aphanocapsa 6308

The catabolic products of arginine metabolism were observed in Aphanocapsa 6308, a unicellular cyanobacterium, by thin layer chromatography of growth media, by limiting growth conditions, and by enzymatic analysis. Of the organic, nitrogenous compounds examined, only arginine supported growth in CO₂-free media. The excretion of ornithine at a concentration level greater than citrulline suggested the existence in Aphanocapsa 6308 of the arginine dihydrolase pathway which produced ornithine, CO₂, NH₄, + adenosine 5-triphosphate. Its existence was confirmed by enzymatic analysis. Although cells could not grow on urea as a sole carbon source a very active urease and subsequently an arginase were also demonstrated, indicating that Aphanocapsa can metabolize arginine via the arginase pathway. The level of enzymes for both pathways indicates a lack of genetic control. It is suggested that the arginase pathway provides only nitrogen for the cells whereas the arginine dihydrolase pathway provides not only nitrogen, but also CO₂ and adenosine 5-triphosphate.Nonstandard Abbreviations CCCP carbonylcyanide mchlorophenyl hydrazone - DCMU 3-(3,4-dichlorophenyl)-1,1-dimethyl urea - CGP cyanophycin granule protein - PS II photosystem II - PSI photosystem I - TLC thin layer chromatography - TCA trichloroacetic acid - DPM disintegrations per min     

Anaerobic,alkaliphilic, saccharolytic bacterium <Emphasis Type="Italic">Alkalibacter saccharofermentans</Emphasis> gen. nov., sp. nov. from a soda lake in the Transbaikal region of Russia

Three strains of new obligately anaerobic alkaliphilic bacteria have been isolated as a saccharolytic component from the cellulolytic community of alkaline Lake Nizhnee Beloe (Transbaikal region, Russia), a lake with low salt concentration. DNA analysis of these strains showed an interspecies level of DNA similarity of 96–100%. Strain Z-79820 was selected for further investigations. Cells were Gram-positive, asporogenous, nonmotile short rods with pointed ends. The strain was a true alkaliphile: growth occurred from pH 7.2 to 10.2 with the optimum at pH 9.0. Strain Z-79820 was halotolerant and could grow in medium with up to 10% (w/v) NaCl, with the optimum between 0 and 4% NaCl. The new isolate obligately depended on Na⁺ ions in the form of carbonates or chlorides. Total Na⁺ content needed for optimal growth was 0.46 M Na⁺, with a wide range from 0.023–0.9 M Na⁺ at which growth also occurred. The isolate was a mesophile and grew at temperatures from 6 to 50°C (slow growth at 6 and 15°C) with an optimum at 35°C. The organotrophic organism fermented ribose, xylose, glucose, mannose, fructose, sucrose, mannitol, and peptone. The products of glucose fermentation were acetate, ethanol, formate, H₂, and CO₂. Yeast extract was required for some anabolic needs. The DNA G+C content of the type strain Z-79820 was 42.1 mol%. The new bacterium fell into the 16S rRNA gene cluster XV of the Gram-positive bacteria with low G+C content, where it formed an individual branch. Based on its growth characteristics and genotype traits, we propose the new genus and species named Alkalibacter saccharofermentans with the type strain Z-79820 (=DSM14828), Uniqem-218 (Institute Microbiology, RAS; ).     

Energetics of glucose uptake in Salmonella typhimurium

We have studied the energetics of glucose uptake in Salmonella typhimurium. Strain PP418 transprots glucose via the phosphoenolpyruvate: glucose phosphotransferase system, while strain PP1705 lacks this system and can only use the galactose permease for glucose uptake. These two strains were cultured anaerobically in glucose-limited chemostats. Both strains produced ethanol and acetate in equimolar amounts but a significant difference was observed in the molar growth yield on glucose (Y _Glc). It is suggested that this difference is due to a difference in the energetics of the glucose uptake systems in the two strains.Assuming an equal Y _ATP for both strains, we could calculate that uptake of 1 mole of glucose via the galactose permease consumes the equivalent of 0.5 mole of ATP. With the additional assumption that one proton is transported in symport with one glucose molecule, these results imply a stoichiometry of two protons per ATP hydrolysed.Abbreviations PTS Phosphoenolpyruvate: carbohydrate phosphotransferase system - D dilution rate (h^-1 - DW dry weight - GalP galactose permease - EtOH ethanol - HAc acetate - Lact lactate - Suc succinate - HFo formate - Glc Glucose - Y _Glc, Y _ATP yield of cells per glucose or ATP - q specific production rate     

Sulfobacillus sibiricus sp. nov., a New Moderately Thermophilic Bacterium

In the course of pilot industrial testing of a biohydrometallurgical technology for processing gold-arsenic concentrate obtained from the Nezhdaninskoe ore deposit (East Siberia, Sakha (Yakutiya)), a new gram-positive rod-shaped spore-forming moderately thermophilic bacterium (designated as strain N1) oxidizing Fe²⁺, S⁰, and sulfide minerals in the presence of yeast extract (0.02%) was isolated from a dense pulp. Physiologically, strain N1 differs from previously described species of the genus Sulfobacillus in having a somewhat higher optimal growth temperature (55°C). Unlike the type strain of S. thermosulfidooxidans, strain N1 could grow on a medium with 1 mM thiosulfate or sodium tetrathionate as a source of energy only within several passages and failed to grow in the absence of an inorganic energy source on media with sucrose, fructose, glucose, reduced glutathione, alanine, cysteine, sorbitol, sodium acetate, or pyruvate. The G+C content of the DNA of strain N1 was 48.2 mol %. The strain showed 42% homology after DNA–DNA hybridization with the type strain of S. thermosulfidooxidans and 10% homology with the type strain of S. acidophilus. The isolate differed from previously studied strains of S. thermosulfidooxidans in the structure of its chromosomal DNA (determined by the method of pulsed-field gel electrophoresis), which remained stable as growth conditions were changed. According to the results of the 16S rRNA gene analysis, the new strain forms a single cluster with the bacteria of the species Sulfobacillus thermosulfidooxidans (sequence similarity of 97.9–98.6%). Based on these genetic and physiological features, strain N1 is described as a new species Sulfobacillus sibiricus sp. nov.     

Cytochrome c peroxidase from a methylotrophic yeast: physiological role and isolation

Mutant strains of the methylotrophic yeast Hansenula polymorpha defective in catalase (cat) and in glucose repression of alcohol oxidase synthesis (gcr1) have been isolated following multiple UV mutagenesis steps. One representative gcr1 cat mutant C-105 grows during batch cultivation in a glucose/methanol medium. However, growth is preceded by a prolonged lag period. C-105 and other gcr1 cat mutants do not grow on methanol medium without an alternative carbon source. A large collection of second-site suppressor catalase-defective (scd) revertants were isolated with restored ability for methylotrophic growth (Mth⁺) in the absence of catalase activity. These Mth⁺ gcr1 cat scd strains utilize methanol as a sole source of carbon and energy, although biomass yields are reduced relative to the wild-type strain. In contrast to the parental C-105 strain, H₂O₂ does not accumulate in the methanol medium of the revertants. We show that restoration of methylotrophic growth in the suppressor strains is strongly correlated with increased levels of the alternative H₂O₂-destroying enzyme, cytochrome c peroxidase. Cytochrome c peroxidase from cell-free extracts of one of the scd revertants has been purified to homogeneity and crystallized. Received: 9 December 1996 / Received revision: 5 May 1997 / Accepted: 25 May 1997     

Physiological and biochemical contributions to the taxonomy of the genus prototheca

Five physiological and biochemical characters, which had proved to be valuable for the taxonomy of the genus Chlorella, were studied in the genus Prototheca. There is no hydrogenase activity and no liquefaction of gelatin. Most strains are very acidtolerant (limit of growth at pH 2.0 or 2.5) and very salt-tolerant (limit of growth at 4 or 5% NaCl). Two strains grow well at 38°C. The 16 strains, which were previously assigned to seven taxa, fall into four different groups. Our results tend to support the assumption that Prototheca might be related to Chlorella protothecoides.     

Mineralization of 2-aminobenzenesulfonate by a bacterial consortium

A variety of environmental inocula were tested for the development of 2-aminobenzenesulfonate (2-ABS, Orthanilic acid) degrading bacterial enrichment. A bacterial consortium (BC), which could utilize 2-ABS as the sole carbon and energy source, could only be developed from the sludge derived from a wastewater treatment unit of a large chemical industry manufacturing nitro and aminoaromatics. BC consisted of two bacterial strains. Based on 16S rDNA sequence analysis, these strains were identified to be belonging to the genus, Acinetobacter and Flavobacterium. The consortium could degrade 1,000 mg l⁻¹ 2-ABS within 40 h. Evidence for the extensive mineralization of 2-ABS, during the growth of BC, was derived from U.V-spectral and total organic carbon analysis. BC was highly specific for 2-ABS, as other benzene sulfonates tested in this study, including other ABS isomers, were not utilized as growth substrates. 2-ABS removal pattern in the presence of glucose was significantly influenced by acclimation characteristics of the culture. Consortium adapted to 2-ABS/glucose demonstrated the concomitant removal of both substrates, whereas glucose exerted catabolic repression on 2-ABS removal with glucose adapted culture. Presence of chloramphenicol inhibited 2-ABS degradation by cells, pregrown on succinate, indicating that the 2-ABS degrading enzymes are inducible in nature. Thus the presence of 2-ABS is essential for maintaining the high degradation potential. This enrichment culture can find an application in the treatment of 2-ABS containing wastewaters.     

Thermus sediminis sp. nov., a thiosulfate-oxidizing and arsenate-reducing organism isolated from Little Hot Creek in the Long Valley Caldera,California

Thermus species are widespread in natural and artificial thermal environments. Two new yellow-pigmented strains, L198^T and L423, isolated from Little Hot Creek, a geothermal spring in eastern California, were identified as novel organisms belonging to the genus Thermus. Cells are Gram-negative, rod-shaped, and non-motile. Growth was observed at temperatures from 45 to 75 °C and at salinities of 0–2.0% added NaCl. Both strains grow heterotrophically or chemolithotrophically by oxidation of thiosulfate to sulfate. L198^T and L423 grow by aerobic respiration or anaerobic respiration with arsenate as the terminal electron acceptor. Values for 16S rRNA gene identity (≤ 97.01%), digital DNA–DNA hybridization (≤ 32.7%), OrthoANI (≤ 87.5%), and genome-to-genome distance (0.13) values to all Thermus genomes were less than established criteria for microbial species. The predominant respiratory quinone was menaquinone-8 and the major cellular fatty acids were iso-C_15:0, iso-C_17:0 and anteiso-C_15:0. One unidentified phospholipid (PL1) and one unidentified glycolipid (GL1) dominated the polar lipid pattern. The new strains could be differentiated from related taxa by β-galactosidase and β-glucosidase activity and the presence of hydroxy fatty acids. Based on phylogenetic, genomic, phenotypic, and chemotaxonomic evidence, the novel species Thermus sediminis sp. nov. is proposed, with the type strain L198^T (= CGMCC 1.13590^T = KCTC XXX).

     

Strain-related differences in bacterivory and demography of Diaphanosoma mongolianum (Cladocera) in relation to diet and previous exposure to cyanobacteria in nature

We compared the demographic variables and bacterivory of two strains of Diaphanosoma mongolianum from two water bodies in Spain, one without Microcystis (Maidevera in Zaragoza) and the other with dense Microcystis (La Albufera of Valencia). We hypothesized that the strain rarely exposed to Microcystis would be unable to grow on this cyanobacterial diet. We fed both strains Monoraphidium caribeum and Microcystis aeruginosa, together and separately, and compared their demographic variables. Monoraphidium caribeum was cultured in the laboratory on a defined medium, while the cyanobacteria were collected from La Albufera and sonicated before feeding the cladocerans (at 0.5?×?10⁶ cells ml^?1). We also tested the growth of D. mongolianum on bacterial diets by using seston (0–15 µm), bacterioplankton (0–3 µm) and mixed fractions (3–15 µm), from sieving Lake Albufera. We conducted population growth and life table demography experiments at 25 °C, using the two strains of D. mongolianum. Both strains had r (population growth rate) ranging from 0.05 to 0.3 d^?1, on all diets. The r was higher (0.18 d^?1) on the 0–15 µm seston compared to the mixed fraction (0.12 d^?1) although D. mongolianum also grew well on bacterioplankton (0.16 d^?1) alone. The response of the strains collected from two different water bodies was different to the test diets. We found that both strains of D. mongolianum could effectively utilize Microcystis for survival and growth, regardless of previous exposure to the cyanobacteria. The tested cladocerans could also grow well on small sized food particles (0–3 µm and 0–15 µm). Our results explain why D. mongolianum is common in eutrophic water bodies

     

Characterization of a new metal-mobilizing Thiobacillus isolate

A new acidophilic, mineral sulphide oreoxidizing bacterium was isolated from a uranium mine near Salamanca, Spain. Cells were rod-shaped, motile and gram-negative. They were aerobes, could grow on pyrite and use sulphur or thiosulphate as sole energy source, suggesting this new isolate belongs to the genus Thiobacillus. It could grow neither with glucose nor with yeast extract as sole substrates. It could not grow on ferrous sulphate as the only energy source, although it grew in the same medium supplemented with glucose, yeast extract or thiosulphate. It was a mesophilic and extremely acidophilic Thiobacillus, with an optimal pH of 1.5 2. The G+C content of the DNA was 58%. The new isolate could grow in cultures on pyrite where electrophoretic pattern was clearly different from those of other thiobacilli, such as T. ferrooxidans.Abbreviations G+C Guanine + Cytosine     

<Emphasis Type="Italic">Singulispaera mucilagenosa</Emphasis> sp. nov., a novel acid-tolerant representative of the order <Emphasis Type="Italic">Planctomycetales</Emphasis>

Two novel strains of budding bacteria, Z-0071^T and Z-0072, were isolated from dystrophic humified waters formed by xylotrophic fungi in the course of spruce wood degradation. The cells of both strains are coccoid (0.95–1.80 μm), nonmotile, single or arranged in pairs. The cells have a complex system of intracellular membranes and are covered with fimbriae and surrounded by a mucous capsule up to 0.3 μm thick. Both strains are aerobic organoheterotrophic, mesophilic, and acid-tolerant microorganisms that are able to grow under microaerobic conditions. They utilize N-acetyl-glucosamine, carbohydrates, and lactate as growth substrates. The strains grow in a pH range of 4.0–7.5 with an optimum at 6.0–6.5. The temperature range for growth is 4–30°C, with an optimum at 25–28°C. Strains Z-0071^T and Z-0072, inhabitants of dystrophic low-mineral waters, are NaCl-sensitive: the NaCl content in the media above 0.5 g/l inhibited growth. The main fatty acids of strains Z-0071^T and Z-0072 are C_16:0, C_18:1ω9c, and C_{18:2ω9c, 12c}. The DNA G + C base content is 51.2–51.7 mol %. The sequences of the 16S rRNA gene fragments (1310 bp) of strains Z-0071^T and Z-0072 were found to be identical. The obtained sequences showed a 94.3% similarity with the sequences of the type strain of the most closely related species Singulisphaera acidiphila MOB10≅T. The phenotypic and phylogenetic properties of strains Z-0071^T and Z-0072 support classification of these strains within the genus Singulisphaera as a new species Singulisphaera mucilagenosa sp. nov., with the type strain Z-0071^T (VKM B-2626).     

Screening and Identification of Endomannanase-Producing Microfungi from Hypersaline Environments

A culture-dependent enrichment technique was used to isolate endo-1,4-β-mannanase–producing fungi from a hypersaline environment. Galactomannan was used as carbon source and resulted in isolation of strains of Scopulariopsis brevicaulis, S. candida, and Verticillium dahliae. The Scopulariopsis isolates were found to be more dominant and could be isolated from consecutive evaporation ponds, whereas Verticillium was only isolated from one pond. The Scopulariopsis strains exhibited only endomannanase activity, whereas Verticillium displayed broad-activity spectrum by secreting endoxylanases and cellulases in addition to endomannanases. S. candida LMK004 and LMK008 produced 7420 and 14750 nkat g⁻¹ biomass, respectively. Endomannanase production in these strains increased with an increase in NaCl concentration up to 10% (w/v), after which both growth and enzyme production was decreased. V. dahliae LMK006 grew and produced up to 5000 nkat g⁻¹ biomass endomannanase in the absence of NaCl. Increased NaCl concentration had a negative effect on this strain. S. brevicaulis LMK002 showed poor endomannanase production but a similar growth trend as the other Scopulariopsis strains. In general, the Scopulariopsis strains exhibited better halotolerance than V. dahliae and could grow in the presence of 20% NaCl on solid medium.     

Experimental determination of control by the H+-ATPase inEscherichia coli

Strains carrying deletions in theatp genes, encoding the H⁺-ATPase, were unable to grow on nonfermentable substrates such as succinate, whereas with glucose as the substrate the growth rate of anatp deletion mutant was surprisingly high (some 75–80% of wild-type growth rate). The rate of glucose and oxygen consumption of these mutants was increased compared to the wild-type rates. In order to analyze the importance of the H⁺-ATPase at its physiological level, the cellular concentration of H⁺-ATPase was modulated around the wild-type level, using genetically manipulated strains. The control coefficient by the H⁺-ATPase with respect to growth rate and catabolic fluxes was measured. Control on growth rate was absent at the wild-type concentration of H⁺-ATPase, independent of whether the substrate for growth was glucose or succinate. Control by the H⁺-ATPase on the catabolic fluxes, including respiration, was negative at the wild-type H⁺-ATPase level. Moreover, the turnover number of the individual H⁺-ATPase enzymes increased as the H⁺-ATPase concentration was lowered. The negative control by the H⁺-ATPase on catabolism may thus be involved in a homeostatic control of ATP synthesis and, to some extent, explain the zero control by the H⁺-ATPase onE. coli growth rate.