Isolation and characterization of a thermophilic sulfate-reducing bacterium Desulfotomaculum thermoacetoxidans sp. nov.

An obligately anaerobic thermophilic sporeforming sulfate-reducing bacterium, named strain CAMZ, was isolated from a benzoate enrichment from a 58°C thermophilic anaerobic bioreactor. The cells of strain CAMZ were 0.7 m by 2–5 m rods with pointed ends, forming single cells or pairs. Spores were central, spherical, and caused swelling of the cells. The Gram stain was negative. Electron donors used included lactate, pyruvate, acetate and other short chain fatty acids, short chain alcohols, alanine, and H₂/CO₂. Lactate and pyruvate were oxidized completely to CO₂ with sulfate as electron acceptor. Sulfate was required for growth on H₂/CO₂, and both acetate and sulfide were produced from H₂/CO₂-sulfate. Sulfate, thiosulfate, or elemental sulfur served as electron acceptors with lactate as the donor while sulfite, nitrate, nitrite, betaine, or a hydrogenotrophic methanogen did not. The optimum temperature for growth of strain CAMZ was 55–60°C and the optimum pH value was 6.5. The specific activities of carbon monoxide dehydrogenase of cells of strain CAMZ grown on lactate, H₂/CO₂, or acetate with sulfate were 7.2, 18.1, and 30.8 mol methyl viologen reduced min^–1 [mg protein]^–1, respectively, indicating the presence of the CO/Acetyl-CoA pathway in this organism. The mol%-G+C of strain CAMZ's DNA was 49.7. The new species name Desulfotomaculum thermoacetoxidans is proposed for strain CAMZ.     

Desulfomonile tiedjei gen. nov. and sp. nov., a novel anaerobic,dehalogenating, sulfate-reducing bacterium

An anaerobic, dehalogenating, sulfate-reducing bacterium, strain DCB-1, is described and nutritionally characterized. The bacterium is a Gram-negative, nonmotile, non-sporeforming large rod with an unusual morphological feature which resembles a collar. The microorganism reductively dehalogenates meta substituted halobenzoates and also reduces sulfate, sulfite and thiosulfate as electron acceptors. The bacterium requires nicotinamide, 1,4-naphthoquinone and thiamine for optimal growth in a defined medium. The microorganism can grow autotrophically on H₂:CO₂ with sulfate or thiosulfate as terminal electron acceptors. It can also grow heterotrophically with pyruvate, several methoxybenzoates, formate plus sulfate or benzoate plus sulfate. It ferments pyruvate to acetate and lactate in the absence of other electron acceptors. The bacterium is inhibited by MoO _inf4 ^sup2- or SeO _inf4 ^sup2- as well as tetracycline, chloramphenicol, kanamycin or streptomycin. Cytochrome c₃ and desulfoviridin have been purified from cells grown in defined medium. 16S rRNA sequence analysis indicates the organism is a new genus of sulfate-reducing bacteria in the delta subdivision of the class Proteobacteria. We propose that the strain be named Desulfomonile tiedjei.Non-standard abbreviations PIPES piperazine-N,N-bis[2-ethanesulfonic acid] - MES 2-[N-morpholino]ethanesulfonic acid - TES N-tris[hydroxymethyl]methyl-2-aminoethanesulfonic acid - HQNO 2-N-heptyl-4-hydroxy-quinoline-N-oxide - CCCP carbonyl-cyanide-m-chlorophenylhydrazine - CM carboxymethyl     

Desulfacinum subterraneum sp.nov.--a new thermophilic sulfate-reducing bacterium isolated from a high temperature oil field

A new thermophilic sulfate-reducing bacterium isolated from the high-temperature White Tiger oil field (Vietnam) is described. Cells of the bacterium are oval (0.4-0.6 by 0.6-1.8 microns), nonmotile, non-spore-forming, and gram-negative. Growth occurs at 45 to 65 degrees C (with an optimum at 60 degrees C) at NaCl concentrations of 0 to 50 g/l. In the course of sulfate reduction, the organism can utilize lactate, pyruvate, malate, fumarate, ethanol, salts of fatty acids (formate, acetate, propionate, butyrate, caproate, palmitate), yeast extract, alanine, serine, cysteine, and H2 + CO2 (autotrophically). In addition to sulfate, the bacterium can use sulfite, thiosulfate, and elemental sulfur as electron acceptors. In the absence of electron acceptors, the bacterium can ferment pyruvate and yeast extract (a yet unrecognized capacity of sulfate reducers) with the formation of acetate and H2. The G + C content of DNA is 60.8 mol %. The level of DNA-DNA hybridization of the isolate (strain 101T) and Desulfacinum infernum (strain B alpha G1T) is as low as 34%. Analysis of the nucleotide sequence of 16S rDNA places strain 101T in the phylogenetic cluster of the Desulfacinum species within the sulfate reducer subdivision of the delta subclass of Proteobacteria. All these results allowed the bacterium studied to be described as a new species, Desulfacinum subterraneum sp. nov., with strain 101 as the type strain.     

Modeling of the pyruvate production with<Emphasis Type="Italic"> Escherichia coli</Emphasis> in a fed-batch bioreactor

A family of 10 competing, unstructured models has been developed to model cell growth, substrate consumption, and product formation of the pyruvate producing strain Escherichia coli YYC202 ldhA::Kan strain used in fed-batch processes. The strain is completely blocked in its ability to convert pyruvate into acetyl-CoA or acetate (using glucose as the carbon source) resulting in an acetate auxotrophy during growth in glucose minimal medium. Parameter estimation was carried out using data from fed-batch fermentation performed at constant glucose feed rates of q_VG=10 mL h^–1. Acetate was fed according to the previously developed feeding strategy. While the model identification was realized by least-square fit, the model discrimination was based on the model selection criterion (MSC). The validation of model parameters was performed applying data from two different fed-batch experiments with glucose feed rate q_VG=20 and 30 mL h^–1, respectively. Consequently, the most suitable model was identified that reflected the pyruvate and biomass curves adequately by considering a pyruvate inhibited growth (Jerusalimsky approach) and pyruvate inhibited product formation (described by modified Luedeking–Piret/Levenspiel term).List of symbols c_A acetate concentration (g L^–1) - c_A,0 acetate concentration in the feed (g L^–1) - c_G glucose concentration (g L^–1) - c_G,0 glucose concentration in the feed (g L^–1) - c_P pyruvate concentration (g L^–1) - c_P,max critical pyruvate concentration above which reaction cannot proceed (g L^–1) - c_X biomass concentration (g L^–1) - K_I inhibition constant for pyruvate production (g L^–1) - K_I^A inhibition constant for biomass growth on acetate (g L^–1) - K_P saturation constant for pyruvate production (g L^–1) - K_P inhibition constant of Jerusalimsky (g L^–1) - K_S^A Monod growth constant for acetate (g L^–1) - K_S^G Monod growth constant for glucose (g L^–1) - m_A maintenance coefficient for growth on acetate (g g^–1 h^–1) - m_G maintenance coefficient for growth on glucose (g g^–1 h^–1) - n constant of extended Monod kinetics (Levenspiel) (–) - q_V volumetric flow rate (L h^–1) - q_VA volumetric flow rate of acetate (L h^–1) - q_VG volumetric flow rate of glucose (L h^–1) - r_A specific rate of acetate consumption (g g^–1 h^–1) - r_G specific rate of glucose consumption (g g^–1 h^–1) - r_P specific rate of pyruvate production (g g^–1 h^–1) - r_P,max maximum specific rate of pyruvate production (g g^–1 h^–1) - t time (h) - V reaction (broth) volume (L) - Y_P/G yield coefficient pyruvate from glucose (g g^–1) - Y_X/A yield coefficient biomass from acetate (g g^–1) - Y_X/A,max maximum yield coefficient biomass from acetate (g g^–1) - Y_X/G yield coefficient biomass from glucose (g g^–1) - Y_X/G,max maximum yield coefficient biomass from glucose (g g^–1) - growth associated product formation coefficient (g g^–1) - non-growth associated product formation coefficient (g g^–1 h^–1) - specific growth rate (h^–1) - _max maximum specific growth rate (h^–1)     

Pyruvate metabolism of the hyperthermophilic archaebacterium Pyrococcus furiosus

The hyperthermophilic anaerobe Pyrococcus furiosus was found to grow on pyruvate as energy and carbon source. Growth was dependent on yeast extract (0.1%). The organism grew with doublings times of about 1 h up to cell densities of 1–2×10⁸ cells/ml. During growth 0.6–0.8 mol acetate and 1.2–1.5 mol CO₂ and 0.8 mol H₂ were formed per mol of pyruvate consumed. The molar growth yield was 10–11 g cells(dry weight)/mol pyruvate. Cell suspensions catalyzed the conversion of 1 mol of pyruvate to 0.6–0.8 mol acetate, 1.2–1.5 mol CO₂, 1.2 mol H₂ and 0.03 mol acetoin. After fermentation of [3-¹⁴C]pyruvate the specific radioactivities of pyruvate, CO₂ and acetate were equal to 1:0.01:1. Cellfree extracts contained the following enzymatic activities: pyruvate: ferredoxin (methyl viologen) oxidoreductase (0.2 U mg^-1, T=60°C, with Clostridium pasteurianum ferredoxin as electron acceptor; 1.4 U mg^-1 at 90°C, with methyl viologen as electron acceptor); acetyl-CoA synthetase (ADP forming) [acetyl-CoA+ADP+Piacetate+ATP+CoA] (0.34 U mg^-1, T=90°C), and hydrogen: methyl viologen oxidoreductase (1.75 U mg^-1). Phosphate acetyl-transferase activity, acetate kinase activity, and carbon monoxide:methyl viologen oxidoreductase activity could not be detected. These findings indicate that the archaebacterium P. furiosus ferments pyruvate to acetate, CO₂ and H₂ involving only three enzymes, a pyruvate:ferredoxin oxidoreductase, a hydrogenase and an acetyl-CoA synthetase (ADP forming).Non-standard abbreviations DTE dithioerythritol - MV methyl viologen - MOPS morpholinopropane sulfonic acid - Tricine N-tris(hydroxymethyl)-methylglycine Part of the work was performed at the Laboratorium für Mikrobiologie, Fachbereich Biologie, Philipps-Universität, Karlvon-Frisch-Strasse, W-3550 Marburg/Lahn, Federal Republic of Germany     

Lactate content and lactate dehydrogenase activity inPalaemon serratus abdominal muscle during temperature changes

Summary Lactate concentration was measured in the abdominal muscle of the shrimpPalaemon serratus. Rapid and seasonal temperature changes result in an increase of the lactate content of approximately 3–4 fold.Lactate dehydrogenase from the abdominal muscle exhibits a temperature dependent pyruvate inhibition with pyruvate as substrate.The kinetic parameters of lactate dehydrogenase fromPalaemon serratus are found to vary during rapid temperature changes: V_max increases with temperature from 0.06 mol min^–1 (mg protein)^–1 at 10°C to 0.28 mol min^–1 (mg protein)^–1 at 30°C with lactate as substrate, and from 5.5 mol min^–1 (mg protein)^–1 at 10°C to 26.2 mol min^–1 (mg protein)^–1 at 30°C, with pyruvate (Table 1). The Hill coefficientn _H, decreases with temperature from 2.2 to 1.2 when the pyruvate reduction is examined, but remains near 1.2 when the activity is measured with lactate as substrate (Table 1). The S_0.5 values for lactate show a tendency to increase below 30 °C (18.9 mM l^–1 at 20 °C) whereas the S_0.5 for pyruvate is found to increase greatly with temperature (0.004 mM l^–1 at 10 °C and 0.06 mM l^–1 at 20 °C).Long term temperature changes involve variations of lactate dehydrogenase activity leading to inverse thermal compensation (Table 2).Activation energy (about 56 kJ both with pyruvate and lactate) does not vary during the year, suggesting that temperature adaptation does not induce important catalytic changes (Table 3).Abbreviation LDH lactate dehydrogenase     

Thermoanaerobium lactoethylicum spec. nov. a new anaerobic bacterium from a hot spring of Kamchatka

A new anaerobic thermophilic Gram-positive, nonsporeforming bacterium strain ZE-1 was isolated from a hot spring of Kamchatka (USSR). The cells are rod-shaped, (0.5–0.8 · 2.0–20 m), non-motile. The bacterium can grow between 42 and 75°C; the optimal temperature is 65°C. The growth is possible between pH values 5.0 and 8.5; optimal pH is 7.0. The cultures grow on the media containing peptone, yeast extract, or casein hydrolysate as nitrogen sources in the presence of glucose or some other sugars, mannitol or starch. The main fermentation products of glucose are ethanol, acetate, lactate, H₂, CO₂; byproducts are propionic, butyric and isovaleric acids. Glucose is metabolized via Embden-Meyerhoff-Parnas pathway. Molecular hydrogen does not inhibit growth. The bacterium does not reduce aceton to isopropanol, but is able to form H₂S from elemental sulfur. The bacterium contains a soluble hydrogenase. This enzyme catalyzes both evolution and uptake of H₂ and is active in the presence of methyl viologen. The DNA-base composition is 34.6 mol%; the genome size 2.08x10⁹ D. The name proposed for the isolated bacterium strain ZE-1 is Thermoanaerobium lactoethylicum spec. nov.     

Uptake of sulfate,sulfite and thiosulfate by proton-anion symport in Desulfovibrio desulfuricans

pH changes and sulfide production upon addition of sulfate, sulfite or thiosulfate to non-buffered H₂-saturated cell suspensions of Desulfovibrio desulfuricans were studied by means of electrodes. The addition of these electron acceptors resulted in a rapid alkalinization of the suspension which was accompanied by sulfide production. At-2° C, alkalinization without immediate sulfide production could be obtained. After addition of ³⁵S-labelled sulfate at-2° C, the label was found to be concentrated 7,500-fold in the cells, while 2 protons per sulfate molecule had disappeared from the outer bulk phase. Alkalinization and sulfide production from micromolar electron acceptor additions depended on the transmembraneous proton gradient ( pH), and were reversibly inhibited in alkaline solution (pH>8.0) or by the protonophore carbonylcyanide m-chlorophenylhydrazone (CCCP). Protonophore-inhibited sulfide production from sulfite or thiosulfate could be restored if the cell membranes were permeabilized by the detergent cetyltrimethylammonium bromide (CTAB), or if downhill transport was made possible by the addition of electron acceptors at millimolar concentrations. Sulfate was not reduced under these conditions, presumably because the cells did not contain ATP for its activation. K⁺-and Na⁺-ionophores such as nigericin, valinomycin or monensin appeared to be of limited efficiency in D. desulfuricans. In most experiments, sulfate reduction was inhibited by the K⁺–H⁺ antiporter nigericin in the presence of K⁺, but not by the thiocyanate anion or the K⁺-transporter valinomycin. The results indicate that sulfate, sulfite and thiosulfate are taken up by proton-anion symport, presumably as undissociated acids with an electroneutral mechanism, driven by the transmembraneous pH gradient ( pH) or by a solute gradient. Kinetics of alkalinization and sulfide production in cells grown with different electron acceptors revealed that D. desulfuricans has different specific uptake systems for sulfate and thiosulfate, and obviously also for sulfite. It is proposed that the electron acceptor transport finally will not consume net energy during growth in buffered medium: The protons taken up during active electron acceptor transport leave the cell with the reduced end-product by simple passive diffusion of H₂S.Abbreviations CCCP carbonyl cyanide m-chlorophenylhydrazone - FCCP carbonyl cyanide p-trifluoromethoxy phenylhydrazone - CTAB cethyltrimethylammonium bromide     

Purification and characterization of hyperthermotolerant leucine aminopeptidase from Geobacillus thermoleovorans 47b

A thermophilic bacterium, which we designated as Geobacillus thermoleovorans 47b was isolated from a hot spring in Beppu, Oita Prefecture, Japan, on the basis of its ability to grow on bitter peptides as a sole carbon and nitrogen source. The cell-free extract from G. thermoleovorans 47b contained leucine aminopeptidase (LAP; EC 3.4.11.10), which was purified 164-fold to homogeneity in seven steps, using ammonium sulfate fractionation followed by the column chromatography using DEAE-Toyopearl, hydroxyapatite, MonoQ and Superdex 200 PC gel filtration, followed again by MonoQ and hydroxyapatite. The enzyme was a single polypeptide with a molecular mass of 42,977.2 Da, as determined by matrix-assisted laser desorption ionization and time-of-flight mass spectrometry, and was found to be thermostable at 90°C for up to 1 h. Its optimal pH and temperature were observed to be 7.6–7.8 and 60°C, respectively, and it had high activity towards the substrates Leu-p-nitroanilide (p-NA)(100%), Arg-p-NA (56.3%) and LeuGlyGly (486%). The K_m and V_max values for Leu-p-NA and LeuGlyGly were 0.658 mM and 25.0 mM and 236.2 mol min^–1 mg^–1 protein and 1,149 mol min^–1 mg^–1 protein, respectively. The turnover rate (k_cat) and catalytic efficiency (k_cat/ K_m) for Leu-p-NA and LeuGlyGly were 10,179 s^–1 and 49,543 s^–1 and 15,470 mM^–1 s^–1 and 1981.7 mM^–1 s^–1, respectively. The enzyme was strongly inhibited by EDTA, 1,10-phenanthroline, dithiothreitol, -mercaptoethanol, iodoacetate and bestatin; and its apoenzyme was found to be reactivated by Co²⁺ .     

<Emphasis Type="Italic">Thialkalivibrio halophilus</Emphasis> sp. nov., a novel obligately chemolithoautotrophic,facultatively alkaliphilic,and extremely salt-tolerant,sulfur-oxidizing bacterium from a hypersaline alkaline lake

A new chemolithoautotrophic, facultatively alkaliphilic, extremely salt-tolerant, sulfur-oxidizing bacterium was isolated from an alkaline hypersaline lake in the Altai Steppe (Siberia, Russia). According to 16S rDNA analysis and DNA–DNA hybridization, strain HL 17^T was identified as a new species of the genus Thialkalivibrio belonging to the subdivision of the Proteobacteria for which the name Thialkalivibrio halophilus is proposed. Strain HL 17^T is an extremely salt-tolerant bacterium growing at sodium concentrations between 0.2 and 5 M, with an optimum of 2 M Na⁺. It grew at high concentrations of NaCl and of Na₂CO₃/NaHCO₃ (soda). Strain HL 17^T is a facultative alkaliphile growing at pH range 7.5–9.8, with a broad optimum between pH 8.0 and 9.0. It used reduced inorganic sulfur compounds (thiosulfate, sulfide, polysulfide, elemental sulfur, and tetrathionate) as energy sources and electron donors. In continuous culture under energy limitation, thiosulfate was stoichiometrically oxidized to sulfate. In sodium carbonate medium under alkaline conditions, the maximum growth rate was similar, while the biomass yield was lower as compared with the NaCl-grown culture. The maximum sulfur-oxidizing capacity measured in washed cells was higher in the soda buffer independent of the growth conditions. The compatible solute content of the biomass was higher in the sodium chloride-grown culture than in the sodium carbonate/bicarbonate-grown culture. The data suggest that the osmotic pressure differences between soda and NaCl solutions might be responsible for the difference observed in compatible solutes production. This may have important implications in overall energetic metabolism of high salt adaptation.     

Life in extreme environments: Investigations on the ecophysiology of a desert bird,the Australian Diamond Dove (Geopelia cuneata Latham)

Summary The Diamond Dove, Geopelia cuneata, is the world's second smallest (ca. 35 g) species of the columbid order. The Diamond Dove is endemic in the arid and semiarid Mulga and Spinifex regions of Central and Western Australia. It regularly encounters ambient temperatures (T _a ) in its habitat above +40° C, especially when foraging for seeds on bare ground cover, and may be found at up to 40 km from water. This entails extreme thermal stress, with evaporative cooling constrained by limited water supply. Energy metabolism (M), respiration, body temperature (T _a ) and water budget were examined with regard to physiological adaptations to these extreme environmental conditions. The zone of thermal neutrality (TNZ) extended from +34° C to at least +45° C. Basal metabolic rate (BMR) was 34.10±4.19 J g^–1h^–1, corresponding to the values predicted for a typical columbid bird. Thermal conductance (C) was higher than predicted. Geopelia cuneata showed the typical breathing pattern of doves, a combination of normal breathing at a stable frequency (ca. 60 min^–1) at low T _a and panting followed by gular flutter (up to 960 min^–1) at high T _a . At T _a > +36° C, T _a increased to considerably higher levels without increasing metabolic rate, i.e. Q₁₀=1. This enabled the doves not only to store heat but also to save the amout of water that would have been required for evaporative cooling if T _a had remained constant. The birds were able to dissipate more than 100% of the metabolic heat by evaporation at T _a +44° C. This was achieved by gular flutter (an extremely effective mechanism for evaporation), and also by a low metabolic rate due to the low Q₁₀ value for metabolism during increased T _b . At lower T _a , Geopelia cuneata predominantly relied on non-evaporative mechanisms during heat stress, to save water. Total evaporative water loss over the whole T _a range was 19–33% lower than expected. In this respect, their small body size proved to be an important advantage for successful survival in hot and arid environments.Abbreviations and units Body Mass W (g) - Ambient Temperature T _a (°C) - Body Temperature T _b (°C) - Thermoneutral Zone (TNZ) - Metabolism M (J g^–1 h^–1) - Thermal Conductance C - wet Thermal Conductance C _wet (J g^–1 h^–1 °C^–1) - Evaporative Water Loss EWL (mg H₂O g^–1 h^–1) - Evaporative Heat Loss EHL (J g^–1 h^–1) - Breathing Frequency F (breaths min^–1) - Tidal Volume V _t (ml breath^–1) - Standard Temperature Pressure Dry STPD - Body Temperature Pressure Saturated BTPS - Respiratory Quotient RQ - n.s. not significant (P>0.05) - n number of experiments     

Isolation and characterization of Dehalospirillum multivorans gen. nov., sp. nov., a tetrachloroethene-utilizing,strictly anaerobic bacterium

A strictly anaerobic bacterium dechlorinating tetrachloroethene (perchloroethylene, PCE) via trichloroethene (TCE) to cis-1,2-dichloroethene (DCE) was isolated from activated sludge with pyruvate plus PCE as energy substrates. The organism, called Dehalospirillum multivorans, is a gram-negative spirillum that does not form spores. The G+C content of the DNA was 41.5 mol%. According to 16S rRNA gene sequence analysis, D. multivorans represents a new genus and a new species belonging to the epsilon subdivision of Proteobacteria. Quinones, cytochromes b and c, and corrinoids were extracted from the cells. D. multivorans grew in defined medium with PCE and H₂ as sole energy sources and acetate as carbon source; the growth yield under these conditions was 1.4g of cell protein per mol chloride released. Alternatively to PCE, fumarate and nitrate could serve as electron acceptors; sulfate could not replace fumarate, nitrate, or PCE in this respect. In addition to H₂, the organism utilized a variety of electron donors for dechlorination (pyruvate, lactate, ethanol, formate, glycerol). Upon growth on pyruvate plus PCE, the main fermentation products formed were acetatc, lactate, DCE, and H₂. At optimal pH (7.3–7.6) and temperature (30°C), and in the presence of pyruvate (20mM) and PCE (160M), a dechlorination rate of about 50 nmol min^-1 (mg cell protein)^-1 and a doubling time of about 2.5h were obtained with growing cultures. The ability to reduce PCE to DCE appears to be constitutive under the experimental conditions applied since cultures growing in the absence of PCE for several generations immediately started dechlorination when transferred to a medium containing PCE. The organism may be useful for bioremediation of environments polluted with tetrachloroethene.Abbreviations PCE Perchloroethylene, tetrachloroethene - TCE Trichloroethene - DCE cis-1,2-Dichloroethene - CHC Chlorinated hydrocarbon     

Superoxide production by thylakoids during chilling and its implication in the susceptibility of plants to chilling-induced photoinhibition

Factors influencing the rate of superoxide (O ₂ ^- ) production by thylakoids were investigated to determine if increased production of the radical was related to injury induced by chilling at a moderate photon flux density (PFD). Plants used were Spinacia oleracea L., Cucumis sativus L. and Nerium oleander L. grown at either 200° C or 45° C. Superoxide production was determined by electron-spin-resonance spectroscopy of the (O ₂ ^- )-dependent rate of oxidation of 2-ethyl-1-hydroxy-2,5,5-trimethyl-3-oxazolidine (OXANOH) to the corresponding oxazolidinoxyl radical, OXANO ·. For all plants, the steady-state rate of O ₂ ^- production by thylakoids, incubated at 25° C and 350 mol photon · m^–2 · s^–1 (moderate PFD) with added ferredoxin and NADP, was between 7.5 and 12.5 mol · (mg chlorophyll)^–1 · h^–1. Incubation at 5° C and a moderate PFD, decreased the rate of O ₂ ^- production 40% and 15% by thylakoids from S. oleracea and 20° C-grown N. oleander, chillinginsensitive plants, but increased the rate by 56% and 5% by thylakoids from C. sativus and 45° C-grown N. oleander, chilling-sensitive plants. For all plants, the addition of either ferredoxin or methyl viologen increased the rate of O ₂ ^- -production at 25° C by 75–100%. With these electron acceptors, lowering the temperature to 5° C caused only a slight decrease in O ₂ ^- production. In the absence of added electron acceptors, thylakoids produced O ₂ ^- at a rate which was about 45% greater than that when ferredoxin and NADP were present. The addition of 3-(3,4-dichlorophenyl)-1,1-dimethylurea reduced O ₂ ^- production under all conditions tested. The results show that the rate of O ₂ ^- production increases in thylakoids when the rate of electron transfer to NADP is reduced. This could explain differences in the susceptibility of thylakoids from chilling-sensitive and chilling-insensitive plants to chilling at a moderate PFD, and is consistent with the proposal that O ₂ ^- production is involved in the injury leading to the inhibition of photosynthesis induced under these conditions.Abbreviations Chl chlorophyll - DCMU 3-(3,4-dichlorophen-yl)-1,1-dimethylurea - Fd ferredoxin - MV methyl viologen - 20°oleander Nerium oleander grown at 20° C - 45°-oleander N. oleander grown at 45° C - OXANOH 2-ethyl-1-hydroxy-2,5,5-tri-methyl-3-oxazolidine - PFD photon flux density (photon fluence rate) - TEMED tetramethyl ethylenediamine We would like to thank R.T. Furbank, R.S.B.S., Australian National University, Canberra, A.C.T., and C.B. Osmond, now of Duke University, Durham, N.C., USA, for the gift of ferredoxin, R.A.J.H. was supported by a Commonwealth Postgraduate Research Award.     

Degradation of polyaromatic hydrocarbons by Burkholderia cepacia 2A-12

A new strain of bacterium degrading polyaromatic hydrocarbons (PAHs), Burkholderia cepacia 2A-12, was isolated from oil-contaminated soil. Of three PAHs, the isolated strain could utilize naphthalene (Nap) and phenanthrene (Phe) as a sole carbon source but not pyrene (Pyr). However, the strain could degrade Pyr when a cosubstrate such as yeast extract (YE) was supplemented. The PAH degradation rate of the strain was enhanced by the addition of other organic materials such as YE, peptone, glucose, and sucrose. YE was a particularly effective additive in stimulating cell growth as well as PAH degradation. When 1 g YE l^–1, an optimum concentration, was supplemented into the basal salt medium (BSM) with 215 mg Phe l^–1, the specific growth rate (0.30 h^–1) and Phe-degrading rate (29.6 mol l^–1 h^–1) were enhanced approximately ten and three times more than those obtained in the BSM with 215 mg Phe l^–1, respectively. Both cell growth and PAH degradation rates were increased with increasing Phe and Pyr concentrations, and B. cepacia 2A-12 had a tolerance against Phe and Pyr toxicity at the high concentration of 730–760 mg l^–1. Through kinetic analysis, the maximum specific growth rate ( _max) and PAH degrading rate ( _max) for Phe were obtained as 0.39 h^–1 and 300 mol l^–1 h^–1, respectively. Also, _max and _max for Pyr were 0.27 h^–1 and 52 mol l^–1 h^–1, respectively. B. cepacia 2A-12 could simultaneously degrade crude oil as well as PAHs, indicating that this bacterium is very useful for the removal of oils and PAHs contaminants.     

Modelling of temperature effects on batch microbial transglutaminase fermentation with Streptoverticillium mobaraense

Batch transglutaminase (MTG) fermentations by Streptovertivillium mobaraense WSH-Z2 at various temperatures ranging from 25 to 35 °C were studied. Dry cell weight and MTG activity could reach their maximal values of 25.1 g/l and 2.94 U/ml, respectively at 30 °C. One typical equation was used to describe the relationship between specific growth rate and culture temperature by comparing several typical equations. Different lag time was observed under various culture temperature. The low lag time was observed under high culture temperature. X = –a ₀(T – T ₀)² + X ₁ + a ₁ (1 – exp(a ₂ (T – T ₁))) and U = –a ₀ (T – T ₀ )² + U ₀ + a ₁ (1 – exp(a ₂ (T – T ₁ ))) could be used to describe the relationship between temperature and the maximal dry cell weight as well as the maximal MTG activity at each temperature.     

Tepidimonas taiwanensis sp. nov., a novel alkaline-protease-producing bacterium isolated from a hot spring

The bacterial strain designated I1-1^T was isolated from a hot spring located in the Pingtung area, southern Taiwan. Cells of this organism were Gram reaction negative rods, motile by a single polar flagellum. Optimum conditions for growth were 55°C and pH 7. Strain I1-1^T grew well in lower nutrient media such as 5–10% Luria–Bertani broth, and its extracellular products expressed alkaline protease activity. The 16S rRNA gene sequence analysis indicates that strain I1-1^T is a member of -Proteobacteria. On the basis of a phylogenetic analysis of 16S rDNA sequences, DNA–DNA similarity data, whole-cell protein analysis, physiological and biochemical characteristics, as well as fatty acid compositions, the organism belonged to the genus Tepidimonas and represented a novel species within this genus. The predominant cellular fatty acids of strain I1-1^T were 16:0 (about 41%), 18:1 7c (about 13%), and summed feature 3 [16:1 7c or 15:0 iso 2OH or both (about 26%)]. Its DNA base ratio was 68.1 mol%. We propose to classify strain I1-1^T (=BCRC 17406^T=LMG 22826^T) as Tepidimonas taiwanensis sp. nov.     

Screening yeast cells for absorption of selenium oxyanions

Certain yeast cells on solid nutrient medium produced colonies surrounded by a light zone of selenite absorption. This screening procedure resulted in the selection of 22 strains out of 200 isolates with different Se⁴⁺-absorbing capacity ranging from 16 to 98.8 g Se⁴⁺ g^–1 l^–1 h^–1. The highest rate of Se⁴⁺ elimination from the Na₂SeO₃ solution was observed with an oval shaped, cream pigmented fermentative yeast, tentatively called Candida sp. strain MS4. This strain was isolated from wastewater and found to accumulate selenium oxyanions. Se⁴⁺ uptake involved both inactive and active phenomena. The amounts of selenium (initial concentration 2 mg Se⁴⁺ l^–1) removed from aqueous solution by inactive and active phenomena were 667 g Se⁴⁺ g^–1 l^–1, and 1580 g Se⁴⁺ g^–1 l^–1, respectively. The strain also removed selenate inactively (135 g Se⁶⁺ g^–1 l^–1).     

Isolation and characterization of Desulfovibrio senezii sp. nov., a halotolerant sulfate reducer from a solar saltern and phylogenetic confirmation of Desulfovibrio fructosovorans as a new species

A new halotolerant Desulfovibrio, strain CVL^T (T = type strain), was isolated from a solar saltern in California. The curved, gram-negative, nonsporeforming cells (0.3 × 1.0–1.3 μm) occurred singly, in pairs, or in chains, were motile by a single polar flagellum and tolerated up to 12.5% NaCl. Strain CVL^T had a generation time of 60 min when grown in lactate-yeast extract medium under optimal conditions (37°C, pH 7.6, 2.5% NaCl). It used lactate, pyruvate, cysteine, or H₂/CO₂ + acetate as electron donors, and sulfate, sulfite, thiosulfate, or fumarate as electron acceptors. Elemental sulfur, nitrate, or oxygen were not used. Sulfite and thiosulfate were disproportionated to sulfate and sulfide. The G+C content of the DNA was 62 mol%. Phylogenetic analysis revealed that Desulfovibrio fructosovorans was the nearest relative. Strain CVL^T is clearly different from other Desulfovibrio species, and is designated Desulfovibrio senezii sp. nov. (DSM 8436). Received: 27 February 1998 / Accepted: 15 June 1998     

Halonatronum saccharophilum gen. nov. sp. nov.: A New Haloalkaliphilic Bacterium of the Order Haloanaerobiales from Lake Magadi

A new alkaliphilic and moderately halophilic chemoorganotrophic anaerobic bacterium (strain Z-7986), which is spore-forming, rod-shaped, and has a gram-negative cell wall pattern, was isolated from the coastal lagoon mud of the highly mineralized Lake Magadi (Kenya). The organism is an obligatorily carbonate- and sodium chloride-dependent motile peritrichously flagellated rod that grows within a 3–17% NaCl concentration range (with an optimum at 7–12% NaCl) and within a pH range of 7.7–10.3 (with an optimum at pH values of 8–8.5). It is a moderate thermophile with a broad temperature optimum at 36–55°C; maximum growth temperature is 60°C. The bacterium catabolizes glucose, fructose, sucrose, maltose, starch, glycogen, N-acetyl-D-glucosamine, and, to a slight degree, peptone and yeast extract. Its anabolism requires yeast extract or casamino acids. Glucose fermentation yields formate, acetate, ethanol, H₂, and CO₂. The bacterium is sulfide-tolerant and capable of the nonspecific reduction of S⁰ to H₂S. The G+C content of the DNA is 34.4 mol %. The analysis of the 16S rRNA sequence revealed that strain Z-7986 belongs to the order Haloanaerobiales and represents a new genus in the family Halobacteroidaceae. We suggest the name Halonatronum saccharophilum gen. nov. sp. nov. The type strain of this species is Z-7986^T (= DSM13868, = Uniqem*211).     

Novel Aerobic Methylotrophic Isolates from the Soda Lakes of the Southern Transbaikal Region

Twenty-one bacterial associations isolated from the soda lakes of the southern Transbaikal region were found to be able to actively grow at pH 9–10 on methanol as the source of carbon and energy. Two alkalitolerant facultatively methylotrophic strains, Bur 3 and Bur 5, were obtained in pure cultures. Both strains represent gram-negative, nonmotile, bean-shaped, encapsulated cells that reproduce by binary fission. The strains are able to grow at temperatures ranging from 6 to 42°C, with an optimum growth temperature of 25–29°C (strain Bur 3) and 35–37°C (strain Bur 5) and at pH between 6.5 and 9.5, with an optimum pH value of 8.0–8.5. At pH 9.0, strain Bur 3 exhibits an increased content of phosphatidylglycerol and a decreased content of phosphatidylethanolamine. Strains Bur 3 and Bur 5 are similar in the G+C content of their DNAs (66.2 and 65.5 mol %, respectively) and in the type of the dominant ubiquinone (Q ₁₀). Unlike Bur 5, strain Bur 3 is able to grow autotrophically in an atmosphere of CO₂+ O₂+ H₂. The strains oxidize, by the respective dehydrogenases, methanol to CO₂, which is assimilated by the ribulose bisphosphate pathway. Ammonium ions are assimilated in the glutamate cycle and by the reductive amination of -ketoglutarate. The strains are highly homologous to each other (92%) and are much less homologous (at a level of 28–35%) to representatives of the genus Ancylobacter, A. aquaticusATCC 25396^Tand A. vacuolatumDSM 1277. Based on the results obtained, both strains are assigned to a new species, Ancylobacter natronumsp. nov.