Stimulation of Chloride In→Out Permeation Across the Deiters' Neuron Membrane by Pentobarbital on the Cytoplasmic Side: Additional Evidence of GABAA Receptors Acting as Chloride Extrusion Pumps

Pentobarbital stimulates ³⁶Cl^– permeation across single Deiters' membranes in a microchamber system, acting on classical, extracellularly facing, GABA_A receptors. However, when applied on the membrane cytoplasmic side it activates per se labeled chloride inout permeation. No effect was found on chloride outin permeation. Similarly, at lower concentrations it facilitates the increase of ³⁶Cl^– inout permeation by application of GABA on the membrane inside, again via asymmetric chloride channels allowing inout but not outin passage. These data confirm that on the Deiters' membrane cytoplasmic side there are structures behaving pharmacologically as GABA_A receptors whose function is that of a Cl^– extrusion pump. This mechanism involves a cycle of activation-phosphorylation/desensitization-reactivation of the receptor complexes     

Effect of tree distance on N2O and CH4-fluxes from soils in temperate forest ecosystems

Complete annual cycles of N₂O and CH₄ flux in forest soils at a beech and at a spruce site at the Höglwald Forest were followed in 1997 by use of fully automatic measuring systems. In order to test if on a microsite scale differences in the magnitude of trace gas exchange between e.g. areas in direct vicinity of stems and areas in the interstem region at both sites exist, tree chambers and gradient chambers were installed in addition to the already existing interstem chambers at our sites. N₂O fluxes were in a range of –4.6–473.3 g N₂O-N m^–2 h^–1 at the beech site and in a range of –3.7–167.2 g N₂O-N m^–2 h^–1 at the spruce site, respectively. Highest N₂O emissions were observed during and at the end of a prolonged frost period, thereby further supporting previous findings that frost periods are of crucial importance for controlling annual N₂O losses from temperate forests. Fluxes of CH₄ were in a range of +10.4––194.0 g CH₄ m^–2 h^–1 at the beech site and in a range of –4.4––83.5 g CH₄ m^–2 h^–1 at the spruce site. In general, both N₂O-fluxes as well as CH₄-fluxes were higher at the beech site. On a microsite scale, N₂O and CH₄ fluxes at the beech site were highest within the stem area (annual mean: 49.6±3.3 g N₂O-N m^–2 h^–1; –77.2±3.1 g CH₄ m^–2 h^–1), and significantly lower within interstem areas (18.5±1.4 g N₂O-N m^–2 h^–1; –60.2±1.8 g CH₄ m^–2 h^–1). Significantly higher values of total N, C and pH in the organic layer, as well as increased soil moisture, especially in spring, in the stem areas, are likely to contribute to the higher N₂O fluxes within the stem area of the beech. Also for the spruce site, such differences in trace gas fluxes could be demonstrated to exist (mean annual N₂O emission within (a) stem areas: 9.7±0.9 g N₂O-N m^–2 h^–1 and (b) interstem areas: 6.2±0.6 g N₂O-N m^–2 h^–1; mean annual CH₄ uptake within (a) stem areas: –26.1±0.6 g CH₄ m^–2 h^–1 and (b) interstem areas: –38.4±0.8 g CH₄ m^–2 h^–1), though they were not as pronounced as at the beech site.     

The Effects of Hypoxia on Three Sympatric Shark Species: Physiological and Behavioral Responses

Behavioral and physiological responses to hypoxia were examined in three sympatric species of sharks: bonnethead shark Sphyrna tiburo, blacknose shark, Carcharhinus acronotus, and Florida smoothhound shark, Mustelus norrisi, using closed system respirometry. Sharks were exposed to normoxic and three levels of hypoxic conditions. Under normoxic conditions (5.5–6.4mg l^–1), shark routine swimming speed averaged 25.5 and 31.0cm s^–1 for obligate ram-ventilating S. tiburo and C. acronotus respectively, and 25.0cm s^–1 for buccal-ventilating M. norrisi. Routine oxygen consumption averaged about 234.6 mg O₂kg^–1h^–1 for S. tiburo, 437.2mg O₂kg^–1h^–1 for C. acronotus, and 161.4mg O₂ kg^–1 h^–1 for M. norrisi. For ram-ventilating sharks, mouth gape averaged 1.0cm whereas M. norrisi gillbeats averaged 56.0 beats min^–1. Swimming speeds, mouth gape, and oxygen consumption rate of S. tiburo and C. acronotus increased to a maximum of 37–39cm s^–1, 2.5–3.0cm and 496 and 599mg O₂ kg^–1 h^–1 under hypoxic conditions (2.5–3.4mg l^–1), respectively. M. norrisi decreased swimming speeds to 16cm s^–1 and oxygen consumption rate remained similar. Results support the hypothesis that obligate ram-ventilating sharks respond to hypoxia by increasing swimming speed and mouth gape while buccal-ventilating smoothhound sharks reduce activity.     

Spatial and temporal variation in soil CO2 efflux in an old-growth neotropical rain forest, La Selva, Costa Rica

Our objectives were to quantify and compare soil CO₂ efflux of two dominant soil types in an old-growth neotropical rain forest in the Atlantic zone of Costa Rica, and to evaluate the control of environmental factors on CO₂ release. We measured soil CO₂ efflux from eight permanent soil chambers on six Oxisol sites. Three sites were developed on old river terraces (old alluvium) and the other three were developed on old lava flows (residual). At the same time we measured soil CO₂ concentrations, soil water content and soil temperature at various depths in 6 soil shafts (3 m deep). Between old alluvium sites, the two-year average CO₂ flux rates ranged from 117.3 to 128.9 mg C m^–2 h^–1. Significantly higher soil CO₂ flux occurred on the residual sites (141.1 to 184.2 mg C m^–2 h^–1). Spatial differences in CO₂ efflux were related to fine root biomass, soil carbon and phosphorus concentration but also to soil water content. Spatial variability in CO₂ storage was high and the amount of CO₂ stored in the upper and lower soil profile was different between old alluvial and residual sites. The major factor identified for explaining temporal variations in soil CO₂ efflux was soil water content. During periods of high soil water content CO₂ emission decreased, probably due to lower diffusion and CO₂ production rates. During the 2-year study period inter-annual variation in soil CO₂ efflux was not detected.     

Dual Role of Superoxide Radicals in the Chilling-Induced photoinhibition in Maize Seedlings

Maize (Zea mays) seedlings were exposed for 6 h to strong irradiance (1 000 mol m^–1 s^–1 of PPFD) at 5, 12, 17, or 25 °C, followed by an exposure to the darkness for 6 h at 22 °C. Leaf chlorophyll fluorescence, net photosynthetic rate (P _N), and the amount of superoxide radicals (O₂ ^–) in relation to chilling-induced photoinhibition were investigated. During the photophase, a good correlation (r=–0.879) was observed between _PS2 (relative quantum efficiency of PS2 electron transport) and the amount of O₂ ^–. Treatment with exogenous O₂ ^– reduced the P _N and _PS2 as the chilling stress did, that was inhibited by specific scavenger of O₂ ^–. Hence chilling-induced photoinhibition might be due to the production of O₂ ^–. In contrast, in the dark period, P _N and _PS2 of the seedlings treated with the exogenous O₂ ^– were enhanced, but they were inhibited by the specific scavenger of O₂ ^–, showing the photoprotective role of O₂ ^– in the recovery phase. Furthermore, in terms of the effect of exogenous O₂ ^– on the xanthophyll cycle, the O₂ ^– production suggested a promotion effect for the de-epoxidation of violaxanthin during the photophase, the epoxidation of zeaxanthin at the dark stage, and the increase of the xanthophyll pool both in the photophase and dark phase, resulting in an enhancement of the ability of non-photochemical quenching to avoid or alleviate the damage to photosynthetic apparatus.     

Inhibitory ca2+-control of movement of beads coated withphysarum myosin along actin-cables inChara internodal cells

Summary The actin-activated ATPase activityPhysarum myosin was shown to be inhibited of M levels of Ca²⁺. To determine if Ca²⁺ regulates ATP-dependent movement ofPhysarum myosin on actin, latex beads coated withPhysarum myosin were introduced intoChara cells by intracellular perfusion. In perfusion solution containing EGTA, the beads moved along the parallel arrays ofChara actin filaments at a rate of 1.0–1.8 m/sec; however, in perfusion solution containing Ca²⁺, the rate reduced to 0.0–0.7 m/sec. The movement of beads coated with scallop myosin, whose actin-activated ATPase activity is activated by Ca²⁺, was observed only in the perfusion solution containing Ca²⁺, indicating that myosin is responsible for the inhibitory effect of Ca²⁺ onPhysarum myosin movement. The involvement of this myosin-linked regulation in the inhibitory effect of Ca²⁺ on the cytoplasmic streaming observed inChara internodal cell andPhysarum plasmodium was discussed.Abbreviations ATP adenosine 5-triphosphate - DTT dithiothreitol - EDTA ethylenediaminetetraacetic acid - EGTA ethyleneglycolbis(-aminoethylether) N,N,N,N-tetraacetic acid - PIPES piperazine-N,N-bis(2-ethanesulfonic acid)     

Ethanol acetylation by mycelium-bound carboxylesterase of Aspergillus oryzae: estimation of thermodynamic parameters and integral productivity

The results collected at different temperatures for ethanol acetylation by cell-bound carboxylesterase from lyophilized cells of Aspergillus oryzae have been used to investigate the kinetics and thermodynamics of this esterification in n-heptane. The occurrence of reversible unfolding followed by irreversible denaturation of the enzyme has been proposed to explain the increase in the starting rate of ethyl acetate formation with temperature observed up to 55 °C and the consequent fall beyond this threshold. The Arrhenius model has been used to estimate the apparent activation enthalpies of both the acetylation reaction (H = 29–33 kJ mol^–1) and reversible enzyme unfolding (H _u = 56–63 kJ mol^–1). The results of residual activity tests performed with cells previously exposed at different temperatures for variable times enabled us also to estimate the first-order rate constant of irreversible denaturation (2.40 × 10^–3 h^–1 < k _d < 8.11 × 10^–3 h^–1) as well as the related thermodynamic parameters (H _d = 22 kJ mol^–1; S _d = –0.29 kJ mol^–1 K^–1). This last phenomenon proved particularly slow for the system under consideration, probably because the biocatalyst link to the mycelium was able to improve its thermostability. In view of future continuous application, the effects of operating time, starting substrate concentration and temperature on the theoretical integral productivity of a fixed-bed column filled with this biocatalyst have been investigated.     

Exopolysaccharide and extracellular metabolite production by Lactobacillus delbrueckii subsp. bulgaricus, grown on lactose in continuous culture

Lactobacillus delbrueckii subsp. bulgaricus NCFB 2483, when grown on lactose in continuous culture, showed increasing specific yields and volumetric productivities of exopolysaccharide (EPS) with increasing dilution rate. Specific and volumetric productivities of lactate and galactose, as extracellular metabolites, increased in response to the incremental changes in the dilution rate up to 0.4 h^–1. Elevated Y_p/s values determined for EPS (0.025 g EPSg lactose^–1) at the dilution rates of 0.3 h^–1–0.4 h^–1, relative to those determined at lower dilution rates, suggest a diversion of carbon flux towards EPS being associated with the higher rates of growth.     

δ15N as an indicator of N2-fixation by cyanobacterial mats in tropical marshes

Cyanobacterial mats (CBM) are important components of wetland ecosystems in limestone-based regions of the Caribbean. During two sampling periods (July 1999 and January 2000) we measured N₂-fixation in samples from 23 different marshes simultaneously with measurements of relevant environmental factors. Samples were evaluated for abundance of five groups of cyanobacteria: (1) Leptolyngbya, (2) Oscillatoria, (3) Chroococcales, (4) Nostoc-& Stigonematales, and (5) dead sheaths. Differences in nitrogen fixation, expressed as nitrogenase activity in nmol C₂H₄ cm^–2 h^–1, were best explained by the proportion of heterocyst-forming cyanobacteria. The samples were analyzed for the natural abundance of ¹⁵N. ¹⁵N values ranged from –1.99 to 11.44 and were strongly negatively correlated with N₂-fixation. With all data included, ¹⁵N was also strongly correlated with nitrates in water. With the samples from Little Belize (high nitrate content marshes) excluded, the effect of nitrate became insignificant. N₂-fixation predicted from ¹⁵N measured on an independent data set from September 2000 was moderately accurate (r² = 0.68, 0.52 and 0.54 for predictions based on July 1999, January 2000 and combined data sets, respectively). When individual sample sets were divided into two groups with ¹⁵N < 2 and ¹⁵N > 2, the two groups were always highly significantly different in terms of their N₂-fixation. The presented evidence suggests that ¹⁵N can be used as a reliable indicator of N₂-fixation by CBM.     

Optimization of cell density and dilution rate in <Emphasis Type="Italic">Pichia pastoris</Emphasis> continuous fermentations for production of recombinant proteins

This paper provides an approach for optimizing the cell density (X_c) and dilution rate (D) in a chemostat for a Pichia pastoris continuous fermentation for the extracellular production of a recombinant protein, interferon (INF-). The objective was to maximize the volumetric productivity (Q, mg INF- l^–1 h^–1), which was accomplished using response surface methodology (RSM) to model the response of Q as a function of X_c and D within the ranges 150 X_c 450 g cells (wet weight) l^–1 and 0.1 _mD0.9 _m (_m=0.0678 h^–1, the maximum specific growth rate obtained from a fed-batch phase controlled with a methanol sensor). The methanol and medium feed rates that resulted in the desired X_c and D were determined based on the mass balance. From the RSM model, the optimal X_c and D were 328.9 g l^–1 and 0.0333 h^–1 for a maximum Q of 2.73 mg l^–1 h^–1. The model of specific production rate (, mg INF- g^–1 cells h^–1) was also established and showed the optimal X_c=287.7 g l^–1 and D=0.0361 h^–1 for the maximum (predicted to be 8.92×10^–3 mg^–1 g^–1 h^–1). The methanol specific consumption rate (, g methanol g^–1 cells h^–1) was calculated and shown to be independent of the cell density. The relationship between and (specific growth rate) was the same as that discovered from fed-batch fermentations of the same strain. The approach developed in this study is expected to be applicable to the optimization of continuous fermentations by other microorganisms.     

Radial transport of water across cortical sleeves of excised roots ofZea mays L.

The stationary radial volume flows across maize (Zea mays L.) root segments without steles (sleeves) were measured under isobaric conditions. The driving force of the volume flow is an osmotic difference between the internal and external compartment of the root preparations. It is generated by differences in the concentrations of sucrose, raffinose or polyethylene glycol. The flows are linear functions of the corresponding osmotic differences ( ) up to osmotic values which cause plasmolysis. The straight lines obtained pass through the origin. No asymmetry of the osmotic barrier could be detected within the range of driving forces applied ( =±0.5 MPa), corresponding to volume-flow densities of j_{v, s}=±7·10^–8 m·s^–1. Using the literature values for the reflection coefficients of sucrose and polyethylene glycol in intact roots (E. Steudle et al. (1987) Plant Physiol.84, 1220–1234), values for the sleeve hydraulic conductivity of about 1·10^–7 m·s^–1 MPa^–1 were calculated. They are of the same order of magnitude as those reported in the literature for the hydraulic conductivity of intact root segments when hydrostatic pressure is applied.Abbreviations and symbols a _s outer surface of sleeve segment - c concentration of osmotically active solute - j _{v, s} radial volume flow density across sleeve segment - Lp_s hydraulic conductivity of sleeves - Lp_r hydraulic conductivity of intact roots - _N thickness of Nernst diffusion layer - reflection coefficient of root for solute - osmotic value of bulk phase - osmotic coefficient     

Protonmotive force in freshwater sulfate-reducing bacteria,and its role in sulfate accumulation in Desulfobulbus propionicus

The protonmotive force in several sulfate-reducing bacteria has been determined by means of radiolabelled membrane-permeant probes (tetraphenyl-phosphonium cation, TPP⁺, for , and benzoate for pH). In six of ten freshwater strains tested only the pH gradient could be determine, while the membrane potential was not accessible due to nonspecific binding of TPP⁺. The protonmotive force of the other four strains was between –110 and –155 mV, composed of a membrane potential of –80 to –140 mV and a pH gradient between 0.25 and 0.8 (inside alkaline) at pH_out=7. In Desulfobulbus propionicus the pH gradient decreased with rising external pH values. This decrease, however, was compensated by an increasing membrane potential. Sulfate, which can be highly accumulated by the cells, did not affect the protonmotive force, if added in concentrations of up to 4 mM. The highest sulfate accumulation observed (2500-fold), which occurred at external sulfate concentrations below 5 M, could be explained by a symport of three protons per sulfate, if equilibrium with the protonmotive force was assumed. At higher sulfate concentrations the accumulation decreased and suggested an electroneutral symport of two protons per sulfate. At sulfate concentrations above 500 M, the cells stopped sulfate uptake before reaching an equilibrium with the protonmotive force.Abbreviations CCCP carbonyl cyanide m-chlorophenylhydrazone - MOPS morpholinopropanesulfonic acid - TPP⁺ tetraphenylphosphonium cation - EDTA ethylenediaminetetraacetic acid - pH transmembrane pH gradient (pH_in-pH_out) - transmembrane electrical potential difference     

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.     

Isolation and characterization of Saccharomyces cerevisiae mutants supersensitive to G1 arrest by the mating hormone a-factor

Summary Nine independent mutants which are supersensitive (ssl ^–) to G1 arrest by the mating hormone a-factor were isolated by screening mutagenized Saccharomyces cerevisiae MAT cells on solid medium for increased growth inhibition with a-factor. These mutants carried lesions in two complementation groups, ssl1 and ssl2. Mutations at the ssl1 locus were mating type specific: MAT ssl1 ^– cells were supersensitive to -factor but MAT ssl1 ^– were not supersensitive to -factor. In contrast, mutations at the ssl2. locus conferred supersensitivity to the mating hormone of the opposite mating type on both MAT, and MATa cells. The -cell specific capacity to inactivate externally added a-factor was shown to be lacking in MAT ssl1 ^– mutants whereas MAT ssl2. cells were able to inactivate a-factor. Complementation analysis showed that ssl2 and sst2, a mutation originally isolated as conferring supersensitivity to -factor to MATa cells, are lesions in the same gene. The ssl1 gene was mapped 30.5 centi-Morgans distal to ilv5 on chromosome XII.     

Interactions of growth inhibitory factor with hydroxyl and superoxide radicals

To show the effects of growth inhibitory factor (Cu₄Zn₃MT-III) involved in the scavenging of reactive oxygen species (ROS), a pulse radiolytic study was employed using N₂O-saturated Cu₄Zn₃MT-III aqueous solutions. It was demonstrated that the oxidizing OH radical efficiently reacted with Cu₄Zn₃MT-III by forming a thiyl radical RS with a second-order constant of 1.46×10¹¹ mol l^–1s^–1, which was determined by competition kinetics against KSCN. The thiyl radical RS reacted rapidly and reversibly with a thiolate in Cu₄Zn₃MT-III to form radical anion RSSR ^– with a constant of 1.65×10⁹ mol lL^–1s^–1 per thiolate, while the constant of the decay of this radical anion was 2.72×10⁵ s^–1, and the equilibrium constant of the formation for RSSR ^– was 6.08×10³ mol^–1 l. These values were close to those of Cd₅Zn₂MT-II. The SOD activity of Cu₄Zn₃MT-III to quench O₂ ^–was assayed by the riboflavine-methionine-nitrobluetetrazolium (NBT) method which catalyzed the dismutation of superoxide (O₂ ^–) at pH 7.8 with an IC₅₀ value of 1.50×10^–6 M for Cu₄Zn₃MT-III and 1.62×10^–6 M for Cd₅Zn₂MT-II. Additionally, the down-regulation of GIF may be a main factor in the decrease of the scavenging ability for the free OH and O₂ ^–radicals, which is possibly associated with the pathogenesis of neurodegenerative disease.     

Hormone studies inMyxine glutinosa: effects of the eicosanoids arachidonic acid,prostaglandin E1, E2, A2, F2α, thromboxane B2 and of indomethacin on plasma cortisol,blood pressure,urine flow and electrolyte balance

Summary Hagfish,Myxine glutinosa, were used in an investigation of the possible effects of various eicosanoids and the prostaglandin synthetase inhibitor indomethacin, on cortisol production, blood pressure control, urine flow and electrolyte balance.Cortisol levels in plasma of untreated control animals and plasma from animals 1 h following injection of 50 g kg^–1 prostaglandin E₁, E₂, A₂, F₂ TXB₂ and indomethacin were not detectable. However, plasma cortisol levels rose to between 10 and 26 pg ml^–1 1 h following injection of either 50 g kg^–1 arachidonic acid or prostaglandin E₂. This rise was similar in magnitude to that produced 1 h following administration of 50 g kg^–1 porcine ACTH.The resting dorsal aortic blood pressure of between 3.50 and 3.75 mmHg was reduced on average by 50% for 12–15 min when animals received 10 g kg^–1 arachidonic acid, prostaglandin E₁, E₂, A₂, and TXB₂ and was effectively reduced to zero for 20 min or more following 50 g kg^–1 of these eicosanoids. Similar doses of prostaglandin F₂, however, evoked an increase in blood pressure (19–33%) whilst indomethacin was without effect.Control measurements of urine flow inMyxine were estimated to be between 540 and 660 l h^–1 kg^–1. There was a marked reduction in urine output following the arterial vasodepression induced by arachidonic acid, prostaglandin E₁, E₂, A₂ and TXB₂ in doses of 10 g kg^–1, an effect which became even more pronouced following injection of 50 g kg^–1 quantities, leading in some cases to complete anuria. There was no significant change in urine volume following either the vasopressor action of prostaglandin F₂ or following indomethacin.None of the compounds tested in this study significantly influenced the plasma or urine electrolyte status ofMyxine.     

A novel eye morphology induced by a P element in somatic tissue of Drosophila melanogaster.

Summary We found a specific eye morphology designated as Square, which is induced when some Drosophila melanogaster strains harboring P elements are crossed with the 2–3 strain carrying a modified P element, P[ry ⁺, 2–3], which produces transposase in somatic tissue. This phenotype was dominant and also induced in the reciprocal crosses. Square was induced when the 2–3 strain was crossed with Q and M strains such as the sn^w (M) strain carrying three small P elements but not with P strains. Inheritance of Square was also tested and its phenotype was not transmitted to the next generation. These results suggest that Square is caused by the transposition of P elements in somatic cells.     

The Use of the [13C]/[12C] Ratio for the Assay of the Microbial Oxidation of Hydrocarbons

The study deals with a comparative analysis of the relative abundances of the carbon isotopes ¹²C and ¹³C in the metabolites and biomass of the Burkholderia sp. BS3702 and Pseudomonas putida BS202-p strains capable of utilizing aliphatic (n-hexadecane) and aromatic (naphthalene) hydrocarbons as sources of carbon and energy. The isotope compositions of the carbon dioxide, biomass, and exometabolites produced during the growth of Burkholderia sp. BS3702 on n-hexadecane (¹³C = –44.6 ± 0.2) were characterized by the values of ¹³C_{CO 2} = –50.2 ± 0.4, ¹³C_biom = –46.6 ± 0.4, and ¹³C_exo = –41.5 ± 0.4, respectively. The isotope compositions of the carbon dioxide, biomass, and exometabolites produced during the growth of the same bacterial strain on naphthalene (¹³C = –21 ± 0.4) were characterized by the isotope effects ¹³C_{CO 2} = –24.1 ± 0.4, ¹³C_biom = –19.2 ± 0.4, and ¹³C_exo = –19.1 ± 0.4, respectively. The possibility of using the isotope composition of metabolic carbon dioxide for the rapid monitoring of the microbial degradation of petroleum hydrocarbons in the environment is discussed.     

Photosynthetic electrogenic events in native membranes ofChloroflexus aurantiacus. Flash-induced charge displacements within the reaction center-cytochromec 554 complex

The thermophilic phototrophChloroflexus aurantiacus possesses a photosynthetic reaction center (RC) containing a pair of menaquinones as primary (Q_A) and secondary (Q_B) electron acceptors and a bacteriochlorophyll dimer (P) as a primary donor. A tetraheme cytochromec ₅₅₄ with two high(H)- and two low(L)-potential hemes operates as an immediate electron donor for P. The following equilibrium E_m,7 values were determined by ESR for the hemes in whole membrane preparations: 280 mV (H1), 150 mV (H2), 95 mV (L1) and 0 mV (L2) (Van Vliet et al. (1991) Eur. J. Biochem. 199: 317–323). Partial electrogenic reactions induced by a laser flash inChl. aurantiacus chromatophores adsorbed to a phospholipid-impregnated collodion film were studied electrometrically at pH 8.3. The photoelectric response included a fast phase of generation ( < 10 ns, phase A). It was ascribed to the charge separation between P⁺ and Q_A ^– as its amplitude decreased both at high and low E_h values (E_m,high=360±10 mV, estimated E_m,low\s-160 mV) in good agreement with E_m values for P/P⁺ and Q_A/Q_A ^– redox couples. A slower kinetic component appeared upon reduction of the cytochromec ₅₅₄ hemes (phase C). With H1 reduced before the flash the amplitude of phase C was equal to 15–20% of that of phase A and its rise time was 1.2–1.3 s: we attribute this phase to the electrogenic electron transfer from H1 to P⁺. Pre-reduction of H2 decreased the value to about 700–800 ns and increased the amplitude of phase C to 30–35% of that of phase A. Pre-reduction of L1 further accelerated phase C (up to of 500 ns) and induced a reverse electrogenic phase with of 12 s and amplitude equal to 10% of phase A. Upon pre-reduction of L2 the rise time of phase C was decreased to about 300 ns and its amplitude decreased by 30%. The acceleration in the onset of phase C is explained by the acceleration of the rate-limiting H1 P electrogenic reaction after reduction of the other hemes due to their electrostatic influence; a P-H1-(L1-L2)-H2 alignment of redox centers with an approximately rhombic arrangement of the cytochromec ₅₅₄ hemes is proposed. The observed reverse phase is ascribed to the post-flash charge redistribution between the hemes. Redox titration of the amplitude of phase C yielded the E_m,8.3 values of H1, H2 and L2 hemes: 340±10 mV for H1, 160±20 mV for H2 and –40±40 mV for L2.     

Modelling continuous culture of Rhodospirillum rubrum in photobioreactor under light limited conditions

Rhodospirillum rubrum was grown continuously and photoheterotrophically under light limitation using a cylindrical photobioreactor in which the steady state biomass concentration was varied between 0.4 to 4 kg m^–3 at a constant radiant incident flux of 100 W m^–2. Kinetic and stoichiometric models for the growth are proposed. The biomass productivities, acetate consumption rate and the CO₂ production rate can be quantitatively predicted to a high level of accuracy by the proposed model calculations. Nomenclature: C _X, biomass concentration (kg m^–3) D, dilution rate (h^–1) Ea, mean mass absorption coefficient (m² kg^–1) I , total available radiant light energy (W m^–2) K, half saturation constant for light (W m^–2) R _W, boundary radius defining the working illuminated volume (m) r _X, local biomass volumetric rate (kg m^–3 h^–1) <r _X>, mean volumetric growth rate (kg m^–3 h^–1) V _W, illuminated working volume in the PBR (m^–3). Greek letters: , working illuminated fraction (–) _M, maximum quantum yield (–) bar, mean energetic yield (kg J^–1).