Recording the oxygen production of a single Acetabularia cell for a prolonged period

A method for recording the O₂ evolution of an individual Acetabularia cell or cell fragment over a period of weeks is described. The method is based on the polarographic O₂ determination by means of a platinum electrode in a flow-through system. The mean O₂ evolution of a full-grown cell under constant conditions (2 5001m/m², 20 °C) was 3–6 μ1 O₂ per cell perh. Under these conditions the O₂ evolution exhibited a pronounced circadian rhythm with an average period of about 23 h and an amplitude of about 2.3 μ1 O₂ per cell per h. No significant differences were found between nucleate and anucleate cells.     

Effects of aeration and of corn steep concentration on L-asparaginase production in Escherichia coli

Summary The production of L-asparaginase was investigated in Escherichia coli, growing under different conditions of aeration in a medium containing 2% or 6% corn steep. At both concentrations, excessive aeration decreased enzyme production. In the medium with 2% corn steep, L-asparaginase activity began to decline as soon as the oxygen absorption exceeded 0.22 mmol O₂ l^–1 min^–1, and when the oxygen absorption rate was 1.26 mmol O₂ l^–1 min^–1, enzyme activity reached only about 5% of maximum. In the medium with 6% corn steep, a decline of L-asperaginase activity did not appear until the oxygen absorption rate value exceeded 0.54 mmol O₂ l^–1 min^–1, at the oxygen absorption rate of 1.26 mmol O₂ l^–1 min^–1, the enzyme activity still reached about 50% of maximum.     

Promotion of indole alkaloid production in Catharanthus roseus cell cultures by rare earth elements

Production of the indole alkaloids, ajmalicine or catharanthine, in cell suspension cultures of Catharanthus roseus was enhanced by cerium (CeO₂ and CeCl₃), yttrium (Y₂O₃) and neodymium (NdCl₃). The yield of ajmalicine in these treated-cultures reached 51 mg l^–1 (CeO₂), 40 mg l^–1 (CeCl₃), 41 mg l^–1 (Y₂O₃) and 49 mg l^–1 (NdCl₃) while catharanthine production reached to 36 mg l^–1 (CeO₂) and 31 mg l^–1 (CeCl₃). A major portion of increased alkaloids was released into medium in these treatments. But Sm₂O₃, SmCl₃, La₂O₃, LaCl₃, complex of chromium (III)-titanium (IV) and NaSeO₄ treatments had little effect on alkaloid production of C. roseus cell cultures.     

Oxygen availability for bacterial growth with a flow microcalorimeter

Summary The enthalpy change associated with aerobic growth of E. coli K₁₂ on minimal media with succinic acid as sole carbon and energy source, determined by flow microcalorimetry (with aerobic mixing cell) was 733.01±15.32 kJ·mol^–1. Molar growth yield was 39.6±1.2 g·mol^–1. When the microcalorimetric growth was limited by oxygen supply, the power-time curve was altered and the total heat evolved was less than the enthalpy change. The maximum thermal output corresponding to a fully aerobic growth in the calorimetric cell was 1.89×10^–3 W·ml^–1. Thus, the oxygen uptake rate was about 0.39 ml O₂·h^–1·ml^–1.     

Nitrite uptake by Chlamydomonas reinhardtii cells immobilized in calcium alginate

When an initial cell loading of about 30–40 µg chlorophyll (Chl)·g^–1 gel and alginate suspension of 3% (w/v) were used for immobilization of Chlamydomonas reinhardtii, the resulting cell beads showed optimum nitrite uptake rate, at 30° C and pH 7.5, of 14 µmol NO _inf2 ^sup– ·mg^–1 Chl·h^–1, the photosynthetic and respiratory activities being about 120 µmol O₂ produced·mg^–1 Chl·h^–1, and 40 µmol O₂ consumed ·mg^–1 Chl·h^–1, respectively. The nitrite uptake activity required CO₂ in the culture and persisted after 8 days of cells immobilization, or in the presence of 0.2 mm ammonium in the medium. Our data indicate that alginate-entrapped C, reinhardtii cells may provide a stable and functional system for removing nitrogenous contaminants from waste-waters.Correspondence to: C. Vílchez     

Selective production of bikaverin in a fluidized bioreactor with immobilized Gibberella fujikuroi

The best culture medium composition for the production of bikaverin by Gibberella fujikuroi in shake-flasks, i.e. 100 g glucose l^–1; 1 g NH₄Cl l^–1; 2 g rice flour l^–1; 5 g KH₂PO₄ l^–1 and 2.5 g MgSO₄ l^–1, was obtained through a fractional factorial design and then scaled-up to a fluidized bioreactor. The effects of carbon and nitrogen concentrations, inoculum size, aeration, flow rate and bead sizes on batch bikaverin production using immobilized G. fujikuroi in a fluidized bioreactor were determined by an orthogonal experimental design. Concentrations of up to 6.83 g bikaverin l^–1 were obtained when the medium contained 100 g glucose l^–1 and 1 g NH₄Cl l^–1 with an inoculum ratio of 10% v/v, an aeration rate of 3 volumes of air per volume of medium min^–1, and a bead size of 3 mm. Based on dry weight, the bikaverin production was 30–100 times larger than found in submerged culture and approximately three times larger than reported for solid substrate fermentation.     

Removal of bisphenol A by polymerization and precipitation method using Coprinus cinereus peroxidase

Bisphenol A was efficiently removed by the polymerization and precipitation method using Coprinus cinereus peroxidase. The removal efficiency was optimal between pH 9–10 and at 40 °C with a molar ratio of H₂O₂ to bisphenol A of about 2. To remove 100 mg bisphenol A l^–1, peroxidase was required 5 U ml^–1 at pH 7 and 25 °C and 3 U ml^–1 at pH 10 and 40 °C.     

Induction of astaxanthin formation by reactive oxygen species in mixotrophic culture of Chlorococcum sp.

The induction of astaxanthin formation by reactive oxygen species in mixotrophic culture of Chlorococcum sp. was investigated. H₂O₂ (0.1 mM) enhanced the total astaxanthin formation from 5.8 to 6.5 mg g^–1 cell dry wt. Fe²⁺ (0.5 mM) added to the medium with H₂O₂ (0.1 mM) further promoted astaxanthin formation to 7.1 mg g^–1 cell dry wt. Similarly, Fe²⁺ (0.5 mM) together with methyl viologen (0.01 mM) promoted astaxanthin formation to 6.3 mg g^–1 cell dry wt. In contrast, an addition of KI (1 mM), a specific scavenger for hydroxyl radicals (OH), together with H₂O₂ (0.1 mM) and Fe²⁺ (0.5 mM), to the medium decreased astaxanthin formation to 1.8 mg g^–1 cell dry wt. KI (1 mM) also inhibited the enhancement of carotenogenesis by superoxide anion radicals (O₂ ^–), with a decrease of astaxanthin formation to 1.7 mg g^–1 cell dry wt. This suggested that O₂ ^– might be transformed to OH before promoting carotenogenesis in Chlorococcum sp.     

Fluorescence and Phosphorescence Study of Tet Repressor–Operator Interaction

Fluorescence and phosphorescence measurements have been carried out on single-p tryptophan (Trp 43 or Trp 75)-containing mutants of Tet repressor (Tet R). Tet R containing Trp 43, the residue localized in the DNA recognition helix of the repressor, has been used to observe the binding of Tet R to two 20-bp DNA sequences of tet O₁ and tet O₂ operators. Binding of Tet R to tet O₁ operator leads to a 78% decrease of the repressor fluorescence intensity, with an accompanying 20-nm blue shift of its fluorescence emission maximum to 330 nm. Upon binding of Tet R to tet O₂ operator, the Trp 43 fluorescence intensity is quenched by 60%, and a 10-nm shift of its emission maximum to 340 nm occurs. Solute fluorescence quenching studies, using acrylamide, performed at low ionic strength indicate that in both the complex of Tet R with the O₁ and that with the O₂ operator, Trp 43 is moderately buried, as indicated by a bimolecular rate quenching constant of about 1.8 × 10⁹ M^–1 sec^–1. In contrast to the Tet R–tet O₂ complex, the Stern–Volmer acrylamide quenching constant K _sv of the complex with tet O₁ operator changes from 7.5 M^–1 at 5 mM NaCl to 22 M^–1 at 200 mM NaCl, indicating different exposures of Trp 43 in the two complexes in solutions of higher ionic strength. Phosphorescence studies showed a 0–0 vibronic transition at 408 and 403 nm for Trp 43 and Trp 75, respectively. Upon binding of Tet R to the tet operators, we observed red shifts of 0–0 vibronic bands of Trp 43 to 413 and 412 nm for tet O₁ and tet O₂ operator, respectively, and the phosphorescence triplet lifetime of Trp 43 at 75 K was quenched from 6.0–5.5 to 3.5–3.3 sec. The thermal phosphorescence quenching profile ranged from –200°C to –20°C, and differed drastically for the two complexes, suggesting different dynamics of the microenvironment of the Trp 43 residue. The luminescence data for Trp 43 of Tet R suggest that the recognition helix of Tet R interacts in different fashions with the tet O₁ and tet O₂ operators.     

Energy metabolism of Chironomus anthracinus (Diptera: Chironomidae) from the profundal zone of Lake Esrom,Denmark, as a function of body size,temperature and oxygen concentration

The profundal zone of Lake Esrom, Denmark has a dense population of Chironomus anthracinus, which survives 2–4 months of oxygen depletion each summer during stratification. The metabolism of 3rd and 4th instar larvae was examined in regard to variation in biomass and temperature. Respiration at air saturation was described by a curvilinear multiple regression relating oxygen consumption to individual AFDW and temperature. At 10 °C and varying oxygen regimes the O₂ consumption and CO₂ production of 4th instar larvae were almost unaltered from saturation to about 3 mg O₂ l^–1, but decreased steeply below this level. The respiratory quotient increased from 0.82 at saturation to about 3.4 at oxygen concentrations near 0.5 mg O₂ l^–1. This implied a shift from aerobic to partially anaerobic metabolism. At 0.5 mg O₂ l^–1 the total energy production equalled 20% of the rate at saturation of which more than one third was accounted for by anaerobic degradation of glycogen. This corresponded to a daily loss of 12 µg mg AFDW^–1 or approximately 5% of the body reserves. At unchanged metabolic rate the glycogen store would last three weeks, but long term oxygen deficiency causes a further suppression of the energy metabolism in C. anthracinus.     

Comparative effects of TiO2-immobilized photocatalytic supporters on the bactericidal efficiency of a novel photoreactor

A novel and highly effective UV-TiO₂ photocatalytic reactor was developed for killing microorganisms, including Escherichia coli. Among tested four types of TiO₂-immobilized photocatalytic supporters (glass bead, muscovite bead, alginate bead, and TiO₂ thin film coated quartz tube), the muscovite bead had a 99.9% percent bactericidal activity within 5 min along with permanent longevity. Adding air bubbles or H₂O₂ (<50 mg l^–1) to the sample solution significantly enhanced the killing activity in that 100% percent of bacterial cells were killed within 3 min.     

Plankton and primary productivity of Lake Manzala,Egypt

Primary production and distribution and abundance of phyto- and zooplankton of lake Manzala were investigated from June 1985 to June 1986.Primary production varied from 4.1 to 28.7 g O₂ m^–2 d^–1 with the highest values 24.2 and 28.7 g O₂ m^–2 d^–1 recorded in the eastern and southern sectors and the lowest value 4.1 g O₂ m^–2 d^–1 recorded in the northern sector. The seasonal mean production for the lake was estimated at 13.3 g O₂ m^–2 d^–1.Mean phytoplankton abundance ranged from 32.7 10⁷ to 76.1 10⁷ cells m^–3 with a mean value of 48.10⁷ cells m^–3. Diatoms were the dominant phytoplankton group comprising 52 to 90 % by number. The greatest relative abundance (87 to 90%) was recorded in the southern sector.Mean zooplankton abundance ranged from 30.1 10³ to 44.4 10³ organisms m^–3 in the eastern sector to 5.5.10³ in the northern sector. In response of eutrophication, the species composition changed significantly over the last 20 years. Cladocerans represented less than 1% of zooplankton during 1959/60, but 75% in 1985/86. Rotifers constituted 40% in 1959/60, and only 1% in 1985/86. Cirriped larvae declined from 21% to 1%.     

Cell death unlikely contributes to taxol production in fungal elicitor-induced cell suspension cultures of Taxus chinensis

Cell suspension cultures ofTaxus chinensis, with 20, 40 and 100 mg fungal elicitor l^–1 from Aspergillus niger, underwent rapid cell death after 24 h, which was about 2, 3.7 and 5-fold of that of the control. At the same time, Taxol production was increased, respectively, to about 5, 8 and 3-fold of that of the control. Inhibition of phenolics biosynthesis resulted in a 150% increase in cell death but a 54% decrease in Taxol production compared with 40 mg elicitor l^–1 alone. O₂-free N₂ inhibited cell death but had little effect on Taxol production as induced by 40 mg fungal elicitor l^–1.     

Chemolithoautotrophy in the marine,magnetotactic bacterial strains MV-1 and MV-2

Magnetite-producing magnetotactic bacteria collected from the oxic–anoxic transition zone of chemically stratified marine environments characterized by O₂/H₂S inverse double gradients, contained internal S-rich inclusions resembling elemental S globules, suggesting they oxidize reduced S compounds that could support autotrophy. Two strains of marine magnetotactic bacteria, MV-1 and MV-2, isolated from such sites grew in O₂-gradient media with H₂S or thiosulfate (S₂O₃^2–) as electron sources and O₂ as electron acceptor or anaerobically with S₂O₃^2– and N₂O as electron acceptor, with bicarbonate (HCO₃^–)/CO₂ as sole C source. Cells grown with H₂S contained S-rich inclusions. Cells oxidized S₂O₃^2– to sulfate (SO₄^2–). Both strains grew microaerobically with formate. Neither grew microaerobically with tetrathionate (S₄O₆^2–), methanol, or Fe²⁺ as FeS, or siderite (FeCO₃). Growth with S₂O₃^2– and radiolabeled ¹⁴C-HCO₃^– showed that cell C was derived from HCO₃^–/CO₂. Cell-free extracts showed ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO) activity. Southern blot analyses indicated the presence of a form II RubisCO (cbbM) but no form I (cbbL) in both strains. cbbM and cbbQ, a putative post-translational activator of RubisCO, were identified in MV-1. MV-1 and MV-2 are thus chemolithoautotrophs that use the Calvin–Benson–Bassham pathway. cbbM was also identified in Magnetospirillum magnetotacticum. Thus, magnetotactic bacteria at the oxic–anoxic transition zone of chemically stratified aquatic environments are important in C cycling and primary productivity.     

Effects of specific versus cross-training on running performance

The cross-training (XT) hypothesis suggests that despite the principle of specificity of training, athletes may improve performance in one mode of exercise by training using another mode. To test this hypothesis we studied 30 well-trained individuals (10 men, 20 women) in a randomized longitudinal trail. Subjects were evaluated before and after 8 weeks of enhanced training (+10%/week), accomplished by adding either running (R) or swimming (XT) to baseline running, versus continued baseline running (C). Both R ( – 26.4s) and XT (– 13.2s) improved time trial (3.2 km) performance, whereas C did not (– 5.4s). There were no significant changes during treadmill running in maximum oxygen uptake (O_2peak; – 0.2, – 6.0, and + 2.7%), steady state submaximal O₂ at 2.68 m · s^–1 ( – 1.2, – 3.3 and + 0.2 ml · kg^–1 · min^–1), velocity at O_2peak (+0.05, +0.25 and +0.09 m · s^–1) or accumulated O₂ deficit (+ 11.2, – 6.1 and + 9.4%) in the R, XT or C groups, respectively. There was a significant increase in velocity associated with a blood lactate concentration of 4 mmol · l^–1 in R but not in XT or C ( + 0.32, + 0.07 and + 0.08 m · s^–1). There were significant changes in arm crank O_2peak ( + 5%) and arm crank O₂ at 4 mmol · l^–1 ( + 6.4%) in XT. There was no significant changes in arm crank O_2peak ( + 1.3 and – 7.7%) or arm crank O₂ at 4 mmol · l^–1 ( + 0.8 and + 0.4%) in R or C, respectively. The data suggest that muscularly non-similar XT may contribute to improved running performance but not to the same degree as increased specific tranining.     

Growth ofCatharanthus roseus cell suspensions in bioreactors: On-line analysis of oxygen and carbon dioxide levels in inlet and outlet gas streams

Summary Catharanthus roseus cells (C87N) grown in a 30 litre airlift vessel achieved a growth rate of 0.366 day^–1. The maximum biomass yield (9.13 gl^–1) was recorded after 168 hours (7 days). On-line analysis of the composition of inlet and outlet gas streams during the growth cycle allowed calculation of the metabolic activity of the cultures. Oxygen uptake on a dry weight basis reached a maximum of 4.5×10^–4 Moles O₂ g dry weight^–1 h^–1 after 96 hours (during the mid-logarithmic phase of growth) and a maximum of 2.7×10^–3 Moles O₂ l^–1 h^–1 on a volume basis (towards the end of the logarithmic phase). Carbon dioxide production ran in parallel with oxygen use with maxima at 4.2×10^–4 Moles CO₂ g dry weight^–1 h^–1 and 3.4×10^–3 Moles g l^–1 h^–1 respectively.     

Energy cost and energy sources in karate

Energy costs and energy sources in karate (wado style) were studied in eight male practitioners (age 23.8 years, mass. 72.3 kg, maximal oxygen consumption (VO_2max) 36.8 ml · min^–1 · kg^–1) performing six katas (formal, organized movement sequences) of increasing duration (from approximately. 10 s to approximately 80 s). Oxygen consumption (VO₂) was determined during pre-exercise rest, the exercise period and the first 270 s of recovery in five consecutive expired gas collections. A blood sample for lactate (la^–) analysis was taken 5 min after the end of exercise. The overall amount of O₂ consumed during the exercise and in the following recovery increased linearly with the duration of exercise (t) from approximately 1.51 (for t equal to 10.5 s (SD 1.6)) to approximately 5.81, for t equal to 81.5 s (SD 1.0). The energy release from la^– production (VO_21a ^–) calculated assuming that an increase of 1 mmol · l^–1 la^– corresponded to a VO₂ of 3 mlO₂ · kg^–1 was negligible for t equal to or less than 20 s and increased to 17.3 ml · kg^–1 (la^– = 5.8 mmol · l^–1 above resting values) for t equal approximately to 80 s. The overall energy requirement (VO_2eq) as given by the sum of VO₂ and VO_2la ^– was described by VO_2eq = 0.87 + 0.071 · t (n = 64; r ² = 0.91), where VO_2eq is in litres and t in seconds. This equation shows that the metabolic power (VO_2eq · t ^–1) for this karate style is very high: from approximately 9.51 · min^–1 for t equal to 10 s to approximately 4.91 · min^–1 for t equal to 80 s, i.e. from 3.5 to 1.8 times the subjects' VO_2max. The fraction of VO_2eq derived from the amount of O₂ consumed during the exercise increased from 11% for t equal to 10 s to 41 % for t equal to 80 s whereas VO_21a ^– was negligible far t equal to or less than 20 s and increased to 13 % o for t equal to 80 s. The remaining fraction (from 90% for t equal to 10 s to 46% for t equal to 80 s), corresponding to the amount of O₂ consumed in the recovery after exercise, is derived from anaerobic alactic sources, i.e. from net splitting of high energy phosphates during the exercise.     

On-line electrocatalyzed luminol chemiluminescence determination of d-glucose and dissolved oxygen in simulated mammalian cell bioreactor perfusion fluid using solid phase reagent modules

On-line instrumentation and methods for the chemiluminescence based real-time monitoring of d-glucose and O₂ levels in mammalian cell bioreactor perfusion fluid are described. The unit processes required for the analysis include: pH adjustment using solid phase flow-through modules, immobilized enzyme catalyzed oxidation of glucose by molecular oxygen to produce hydrogen peroxide, controlled release of luminol using a solid phase flow-through module, electrocatalyzed luminescence using gold electrodes, and photodetection of chemiluminescent emissions. Calibration curves for d-glucose and dissolved O₂ in simulated bioreactor perfusion fluid have been generated using fully integrated reagentless test systems from 0–800 mg l^–1 and 0–10 mg l^–1, respectively.     

Nitrification rates in a saline medium at different dissolved oxygen concentrations

A kinetic study was carried out in a saline medium to assess the effect of O₂ on the two-step nitrification process: for the first nitritation step, 2–26 mg dissolved O₂ (DO) l^–1 was used and for the second nitratation step, 0.5–24 mg DO l^–1 was used. Nitritation rate was measured in the presence of sodium azide so as to inhibit nitratation. Ammonia-oxidizing (AOB) and nitrite-oxidizing (NOB) bacterial in the mixed culture were determined by 16 S rRNA hybridization. The affinity constants for oxygen O₂ of the AOB and the NOB were 1.66 mg O₂ l^–1 and 3 mg O₂ l^–1 respectively. The larger than the previously reported values of these constants might be due to the high salt content in the medium. High O₂ concentrations did inhibit the nitrification rate.     

The roles of malate and aspartate in C4 photosynthetic metabolism of Flaveria bidentis (L.)

In C₄ grasses belonging to the NADP-malic enzyme-type subgroup, malate is considered to be the predominant C₄ acid metabolized during C₄ photosynthesis, and the bundle sheath cell chloroplasts contain very little photosystem-II (PSII) activity. The present studies showed that Flaveria bidentis (L.), an NADP-malic enzyme-type C₄ dicotyledon, had substantial PSII activity in bundle sheath cells and that malate and aspartate apparently contributed about equally to the transfer of CO₂ to bundle sheath cells. Preparations of bundle sheath cells and chloroplasts isolated from these cells evolved O₂ at rates between 1.5 and 2 mol · min^–1 · mg^–1 chlorophyll (Chl) in the light in response to adding either 3-phosphoglycerate plus HCO ₃ ^– or aspartate plus 2-oxoglutarate. Rates of more than 2 mol O₂ · min^–1 · mg^–1 Chl were recorded for cells provided with both sets of these substrates. With bundle sheath cell preparations the maximum rates of light-dependent CO₂ fixation and malate decarboxylation to pyruvate recorded were about 1.7 mol · min^–1 · mg^–1 Chl. Compared with NADP-malic enzyme-type grass species, F. bidentis bundle sheath cells contained much higher activities of NADP-malate dehydrogenase and of aspartate and alanine aminotransferases. Time-course and pulse-chase studies following the kinetics of radiolabelling of the C-4 carboxyl of C₄ acids from ¹⁴CO₂ indicated that the photosynthetically active pool of malate was about twice the size of the aspartate pool. However, there was strong evidence for a rapid flux of carbon through both these pools. Possible routes of aspartate metabolism and the relationship between this metabolism and PSII activity in bundle sheath cells are considered.Abbreviations DHAP dihydroxyacetone phosphate - NADP-ME(-type) NADP-malic enzyme (type) - NADP-MDH NADP-malate dehydrogenase - OAA oxaloacetic acid - 2-OG 2-oxoglutarate - PEP phosphoenolpyruvate - PGA 3-phosphoglycerate - Pi orthophosphate - Ru5P ribulose 5-phosphate