Growth and physiology of potassium-limited chemostat cultures of Paracoccus denitrificans

In potassium-limited chemostat cultures of Paracoccus denitrificans the maximum specific growth rate (µ_max) was found to depend on the input potassium concentration: At 0.21mM µ_max was 0.10–0.11 h^-1; at 0.44 mM 0.15–0.16 h^-1 and at 0.66 mM 0.20–0.21 h^-1. The plots of the specific rates of oxygen-, succinate-and potassium consumption against gave straight lines. The intracellular potassium concentration was a linear function of and varied from 1% (0.13 M) at a value of 0.034 h^-1 to 2.2% (0.29 M) at =0.26 h^-1; the potassium concentration gradient and the potassium concentration in the culture fluid in the steady state were dependent on the input potassium concentration. The potassium concentration gradient varied from 8,900-1,200. At all values 20–25% of the total energy production was used for potassium transport. 350,100 and 30 ATP molecules were calculated to be required to maintain one potassium ion intracellular during 1 h at values of 0.034, 0.197 and 0.257 h^-1 respectively. It is concluded that the amount of circulation of potassium is dependent on the potassium concentration gradient or on the potassium concentration in the culture in the steady state. The dependency of µ_max on the input potassium concentration was explained by the assumption that at low input potassium concentrations the net uptake of potassium (influx-efflux) is not rapidly enough to maintain the high potassium gradient in the existing cells and to establish it in the newly formed cells. At high values and at high input potassium concentrations µ_max is limited by the specific rate of oxygen consumption, which was found to be 11–12 mmol O₂ g dry weight^-1 h^-1 at µ_max for potassium-, succinate-and sulphate-limited chemostat cultures.     

Invertase and phosphatase of yeast in a phosphate-limited continuous culture

Summary Deficiency of inorganic phosphate caused the hyper production of invertase and the derepression of acid phosphatase in a continuous culture ofSaccharomyces carlsbergensis. The specific invertase activity was 40,000 enzyme units per g dry cell weight at a dilution rate lower than 0.05 h^–1 with a synthetic glucose medium of which the molecular ratio of KH₂PO₄ to glucose was less than 0.006. This activity is eight fold higher than in a batch growth and 1.5 fold as much as the highest enzyme activity observed so far in a glucose-limited continuous culture.For the hyper production of invertase, it is necessary to culture the yeast continuously by keeping the Nyholm's conservative inorganic phosphate concentration at less than 0.2 m mole per g dry weight cell. The derepression of acid phosphatase brought about by phosphate deficiency, was similar in both batch and continuous cultures.Nomenclature D dilution rate of continuous culture (h^–1) - E_i invertase concentration in culture (enzyme unit l^–1) - E_p acid phosphatase concentration in culture (enzyme unit l^–1) - P inorganic phosphate concentration in culture (mM) - S glucose concentration in culture (mM) - X cell concentration in culture (g dry weight cell l^–1) Greek Letter specific rate of growth (h^–1) Suffix f feed - 0 initial value     

Optimization of lipid production in the oleaginous yeastApiotrichum curvatum in wheypermeate

Summary Lipid production of the oleaginous yeastApiotrichum curvatum was studied in wheypermeate to determine optimum operation conditions in this medium. Studies on the influence of the carbon to nitrogen ratio (C/N-ratio) of the growth medium on lipid production in continuous cultures demonstrated that cellular lipid content in wheypermeate remained constant at 22% of the cell dry weight up to a C/N-ratio of about 25. The maximal dilution rate at which all lactose is consumed in wheypermeate with excess nitrogen was found to be 0.073 h^-1. At C/N-ratios higher than 25–30 lipid content gradually increased to nearly 50% at C/N=70 and the maximal obtainable dilution rate decreased to 0.02 h^-1 at C/N=70. From these studies it could be derived that maximal lipid production rates can be obtained at C/N-ratios of 30–35 in wheypermeate. Since the C/N-ratio of wheypermeate normally has a value between 70 and 101, some additional nitrogen is required to optimize the lipid production rate. Lipid production rates ofA. curvatum in wheypermeate were compared in four different culture modes: batch, fed-batch, continuous and partial recycling cultures. Highest lipid production rates were achieved in culture modes with high cell densities. A lipid production rate of nearly 1 g/l/h was reached in a partial recycling culture. It was calculated that by using this cultivation technique lipid production rates of even 2.9 g/l/h may be reached when the supply of oxygen can be optimized.Nomenclature C/N-ratio carbon to nitrogen ratio of the growth medium (g/g) - C/N_crit C/N-ratio at which there is just enough nitrogen to allow all carbon source to be converted to biomass - D dilution rate=volume of incoming medium per unit time/volume of medium in the culture vessel (h^-1) - D_max maximum dilution rate (h^-1) - DW cell dry weight - L lipid yield (g storage lipid/g carbon source) - specific growth rate (h^-1) - _max maximum specific growth rate (h^-1) - Q_L lipid production rate (g/l/h) - Y_i molecular fraction of carbon substrate that is converted to storage carbohydrate (C-mol/C-mol) - Y_ls maximal amount of storage lipid that can be produced per mol carbon source (C-mol/C-mol)     

The effect of carbon dioxide on the growth kinetics of fructose-limited chemostat cultures of Acetobacterium woodii DSM 1030

The growth of the anaerobic acetogenic bacterium Acetobacterium woodii DSM 1030 was investigated in fructose-limited chemostat cultures. A defined medium was developed which contained fructose, mineral salts, cysteine · HCl and Ca pantothenate (1 mg · 1^–1) supplied in a vitamin supplement. Growth at high dilution rates was dependent on the presence of CO₂ in the gas phase. The ^max was found to be 0.16 h^–1 and the fructose maintenance requirement was 0.1 to 0.13 mmol fructose · (g dry wt)^–1 · h^–1. A growth yield of 61 g dry wt · (mol fructose)^–1, corrected for the cell maintenance requirement and for incorporation of fructose carbon into cell biomass, was determined from the fructose consumption. A corresponding growth yield of 69 g dry wt · (mol fructose)^–1 was calculated from the acetate production assuming that fructose fermentation was homoacetogenic. A Y_ATP of 12.2 to 13.8 g dry wt · (mol ATP)^–1 was calculated from these growth yields using a value of 5 mol ATP · (mol fructose)^–1 as an estimate of the amount of ATP synthesised from fructose fermentation. The addition of yeast extract (0.5 g · 1^–1) to the medium did not influence the ^max or cell yield. After prolonged growth under fructose-limited conditions the requirement of the culture for CO₂ in the gas phase was reduced.Abbreviations YE yeast extract - IC inorganic carbon - D fermenter dilution rate : h^–1 - M_X maintenance requirement for X: mmol X · (g dry wt)^–1 · h^–1 - X may be fructose (Fruct), fructose consumed in energy metabolism (Fruct [E]), acetate (Ac) - ATP CO₂, NH _inf4 ^sup+ or Pi - qX specific rate of utilisation or consumption of X: mmol X · (g dry wt)^–1 · h^–1 - V fermenter volume: litre - rC · Cell, fermenter cell carbon production: mmol C · h^–1 - Y_X yield of cells on X: g dry wt · (mol X)^–1 - Y _infx ^supmax the yield corrected for cell maintenance: g dry wt · (mol X)^–1 - S_ATP stoichiometry of ATP synthesis from fructose: mol ATP · (mol frucose)^–1 - x cell concentration: g dry wt · 1^–1 - specific growth rate : h^–1 - ^max maximum specific growth rate: h^–1     

Ferric chelate reduction by suspension culture cells and roots of soybean: A kinetic comparison

The abilities of suspension cultures and intact roots of soybean (Glycine max L. cv. Hawkeye) to reduce ferric chelate were compared. Ferric chelate was supplied as ferric hydroxyethylethylenediaminetriacetic acid (FeHEDTA) and reduction was measured spectrophotometrically using bathophenan-throlinedisulfonic acid (BPDS) as the ferrous scavenger. Ferric chelate reduction by cell suspension cultures showed typical saturation kinetics; however, no difference was observed between cells that had been continuously grown with Fe (+Fe) and those that had been grown for four days without added Fe (–Fe). Values for K_m and V_max, determined from a Lineweaver-Burk plot, were 57 M and nmoles mg^-1 dry weight for the +Fe cells and 50 M and 22 nmoles mg^-1 dry weight for the -Fe cells, respectively. Ferric chelate reduction by Fe-deficient roots also exhibited saturation kinetics, while roots grown with adequate Fe did not reduce ferric chelate. The K_m and V_max values for Fe-deficient roots were 45 M and 20 nmoles mg^-1 dry weight, respectively, and did not differ from values obtained for cells in culture. This study offers strong evidence that the mechanism responsible for the reduction of ferric chelate is the same for cultured cells and roots and that the process is controlled at the cellular level. We propose that suspension cultures can be used as an alternative to intact roots in the study of ferric chelate reduction.     

Inhibition of the acidogenic dissimilation of glucose in anaerobic continuous cultures by free butyric acid

Summary In anaerobic wastewater treatment the separation of fermentative and methanogenic bacteria is aimed at an increased performance of the total digestion process. It is known that the attainable growth rate of the acidogenic population in continuous culture decreases at increasing influent concentrations of glucose. To account for this phenomenon, a new kinetic model was developed that combines substrate and product inhibition. In the present research product inhibition was investigated quantitatively in a continuous culture fermenting 50 mmol/l glucose. Extra acetate and butyrate were added up to 200 mmol/l at different pH values, and it turned out that only free butyric acid inhibited growth. The lower attainable growth rates of cultures producing comparable amounts of butyrate when fed with concentrated influents, strongly indicated substrate inhibition. Evidence is presented that transitions to low-conversion steady states predicted by the kinetic model, play a role and decrease the stability of the culture.Nomenclature D dilution rate, h^-1 - D_att highest D using certain experimental procedure h^-1 - K_i substrate inhibition constant, mol·m^-3 - K_p product inhibition constant mol·m^-3 - K_s substrate saturation constant, mol·m^-3 - P concentration inhibitory product mol·m^-3 - S substrate concentration, mol·m^-3 - S_o influent substrate concentration, mol·m^-3 - S _max ^c substrate concentration at _max ^c , mol·m^-3 - S _max ^h substrate concentration at _max ^h , mol·m^-3 - specific growth rate, h^-1 - experimental realization of at D_att, h^-1 - _max maximum specific growth rate, h^-1 - _max ^c maximum attainable specific growth rate according to combined substrate/product inhibition model, h^-1 - _h ⁰ specific growth rate at S₀ according to Haldane kinetics, h^-1 - _max ^c maximum attainable specific growth rate according to Haldane kinetics, h^-1 - Y_p yield inhibitory product, mol·mol^-1 - Y_x yield biomass, kg dry weight·kg^-1 - bio biomass - EtOH ethanol - gluc glucose - HAc acetate - HBt butyrate - HCap caproate - HFo formate - HPr propionate - HVal valerate - prod produced - lact lactate     

Growth of Thermoanaerobium brockii in batch and continuous culture at supraoptimal temperatures

Summary Thermoanaerobium brockii was grown in batch and continuous culture at supraoptimal temperatures (>65° C). Specific growth rates were lower in batch (_max>1.0 h^-1) than in continuous cultures (_max1.2–1.4 h^-1). Acetone addition to the medium did not increase critical dilution rate significantly. The media used contained significantly less organic material and sulfide than previously reported media; however, yeast extract requirements were shown to be exceptionally high (60% of the glucose concentration used). Organic substrates inhibited growth and product formation in chemostat cultures whereas the slow formation of acetic acid was observed in batch cultures, but also with virtually no growth. The inhibiting concentration was found to be approximately 15 g organic carbon·l^-1. The maintenance requirements of T. brockii were in the same range as expected of aerobic extreme thermophiles (m_s0.5 g·g^-1·h^-1) and could be met only by glucose and not by yeast extract. Maintenance was obviously not independent of specific growth rate. Production of the stereospecific alcohol-aldehyde/ketone oxidore-ductase was strictly growth associated and its formation was not affected by acetone added to medium.     

Transient state characteristics of changes in light conditions for the cyanobacterium Oscillatoria agardhii

The cyanobacterium Oscillatoria agardhii was subjected to changes in irradiance and to changes in light period. During transient states parameters as growth rate, pigment contents, photosynthetic activities and pool sizes of carbohydrate and proteins were followed. The changes in pigments and photosynthesis were similar for irradiance transitions and transitions in light period length. Carbohydrates served for the supply of carbon and energy during adaptation to low light conditions until a basal level of 125 g · mg dry wt^-1 was reached. After transfer to high light conditions excess carbon fixation led to the storage of carbohydrate reserve polymers up to 600 g · mg dry wt^-1. During adaptation to longer light periods cells showed an overcapacity for carbohydrate accumulation even in the presence of a high carbohydrate content at the start of the light period. A model for the feed back repression of photosynthesis related to carbohydrate accumulation was presented. In all cases protein synthesis was directly maximized under the given conditions. Growth rate defined as specific rate of change in carbon showed the fastest response after a shift in light conditions. It was concluded that adaptation of O. agardhii to changes in light conditions was directed to the optimization of growth. The observation that carbohydrate is used to supply carbon and/or energy during adaptation leads to the conclusion that changes on survival in low light depend on carbohydrate level, the efficiency of its conversion in cell material and the maintenance requirements. Such a survival strategy enables cyanobacteria to cope succesfully with light limiting conditions.Abbreviations HL high irradiance Em^-2s^-1 - LL low irradiance Em^-2s^-1 - L/D light-dark cycle h - specific growth rate h^-1 - _e specific maintenance coefficient h^-1 - _max maximal specific growth rate h^-1 - _c specific rate of change of carbon h^-1 - _protein specific rate of change of protein h^-1 - Chl a chlorophyll a - CPC C-phycocyanin - dry wt dry weight mg - light utilization efficiency nmol O₂ · mg dry wt^-1 · min^-1/Em^-1 s^-1 - P _max photosynthetic capacity nmol O₂ · mg dry wt^-1 · min^-1 - PSI photosystem I - PS II photosystem II - RC II reaction center of PS II - PQ plastoquinone     

Energetic aspects of anaerobic growth of Aerobacter aerogenes in complex medium

Molar growth yields for anaerobic growth of Aerobacter aerogenes in complex medium were much higher than for growth in minimal medium. In batch cultures the molar growth yield for glucose varied from 44 to 50 and Y _ATP from 17.1 to 18.8. For glucose-limited chemostat cultures a value of 17.5 g/mole was found for Y _ATP ^max and a value of 2.3 mmoles ATP/g dry weight h for the maintenance coeficient. Growth dependent pH changes were used to control the addition of fresh medium, containing excess of glucose to a continuous culture. The specific growth rate and the population density were dependent on the pH difference between the inflowing medium and the culture. At a value of 1.44 h^-1 the molar growth yield for glucose was about 70 and Y _ATP about 28.5. An-equation is presented, which gives the relation between theoretical and experimental Y _ATP ^max values.     

Effect of endogenous proteins on growth and antibody productivity in hybridoma batch cultures

It has been shown that some B-cell hybridomas secrete autocrine factorsin vitro which can influence cell metabolic processes. Rather than screen specifically for suspected cytokines, that may or may not affect our cell line, we have examined the lumped effects of intracellular and secreted factors on cell proliferation and monoclonal productivity in hybridoma batch cultures. Firstly, supplements of total soluble intracellular proteins combined with other intracellular metabolites were found to both decrease the specific growth rate and increase the antibody production rate at higher concentrations in batch culture. This is an important consideration in high cell density cultures, such as perfusion systems, where a reduction of growth by the presence of intracellular factors may be compensated by an increase in MAb production. In addition, flow cytometry data revealed that the average cell cycle G₁ phase fraction was unaffected by the variation in the maximum specific growth rates during the exponential growth phase, caused by the addition of intracellular factors; this suggests that higher MAb productivity at lower growth rates are not a result of cell arrest in the G₁ phase. Secondly, secreted extracellular proteins larger than 10,000 Daltons, which were concentrated from spent culture supernatant, were shown to have no significant effect on growth and specific MAb productivity when supplemented to batch culture at levels twice that encountered late in normal batch culture. This indicates that endogenous secreted cytokines, if at all present, do not play a major autocrine role for this cell line.Abbreviations FBS fetal bovine serum - MAb monoclonal antibody - MWCO molecular weight cut off - SDS-PAGE sodium dodecyl-sulphate-polyacrylamide gel electrophoresis - k _d exponential phase death rate, h^–1 - q _MAb exponential phase specific monoclonal antibody productivity, pg/(cell·h) - t time, h - X _d dead cell density, cells/mL - X _v viable cell density, cells/mL - specific growth rate, h^–1 - _{max app} apparent maximum specific growth rate, h^–1 - _max maximum specific growth rate, h^–1 _max = _{max app + Kd}      

High cell density fermentation of recombinant Escherichia coli expressing human interferon alpha 1

Summary A defined medium was developed which, by means of a specific fed-batch mode, allows growth of the recombinant Escherichia coli strain TG1 (pBB210) up to a cell density of 60 g dry weight/l. Apart from glucose and aqueous ammonia fed as carbon and nitrogen sources, it was unnecessary to supply other nutrients or O₂-enriched air. Aqueous ammonia also served for pH control. The pO₂ level was kept at 20% saturation via closed-loop controls operating the two output variables of stirrer speed and glucose feeding rate. This fed-batch method prevented significant accumulation of acetate and other metabolic by-products. The recombinant E. coli expressed interferon alpha 1 more efficiently at a lower specific growth rate (_Pr 0.15 h^–1) than at the maximum specific growth rate (_max = 0.45 h^–1). Therefore, fermentation in the batch phase at _max was only allowed to continue up to a medium cell density. In the succeeding fed-batch phase, the specific growth rate was reduced to _Pr by increasing the stirrer speed according to an empirically developed time scale. Offprint requests to: D. Riesenberg     

Growth kinetics of Thermus thermophilus

Summary The nonsporulating extreme thermophile Thermus thermophilus was grown in continuous culture at dilution rates up to 2.65 h^–1 at 75°C and pH 6.9 on complex medium. Concomitantly very low yield (Y=0.12 g cell dry weight g^–1 utilized organic carbon) and incomplete substrate utilization (always less than 45%) were found. In batch cultures T. thermophilus could be grown with _max =h^–1, in shake flasks only with _max =h^–1 with the same low yield and incomplete substrate utilization. Stable steady states at 84C and 45°C were realized at a dilution rate of 0.3 h^–1 whereas at 86°C and 40°C no growth could be detected. Artefacts arising from wall growth (in bioreactors) or improper materials must be ruled out. Inhibition of growth by organic substrates was demonstrated at low concentrations: a decrease in the yield obtained was found when more than 0.7 gl^–1 of meat extract were supplied in the medium. The maintenance requirement for oxygen is potentially very high and was determined to be 10 to 15 mmol g^–1 h^–1.     

Phenol degradation by a psychrotrophic strain of Pseudomonas putida

Summary Cell growth and phenol degradation kinetics were studied at 10°C for a psychrotrophic bacterium, Pseudomonas putida Q5. The batch studies were conducted for initial phenol concentrations, S_o, ranging from 14 to 1000 mg/1. The experimental data for 14<=S_o<=200 mg/1 were fitted by non-linear regression to the integrated Haldane substrate inhibition growth rate model. The values of the kinetic parameters were found to be: _m=0.119 h^–1, K _S=5.27 mg/1 and K _I=377 mg/1. The yield factor of dry biomass from substrate consumed was Y=0.55. Compared to mesophilic pseudomonads previously studied, the psychrotrophic strain grows on and degrades phenol at rates that are ca. 65–80% lower. However, use of the psychrotrophic microorganism may still be economically advantageous for waste-water treatment processes installed in cold climatic regions, and in cases where influent waste-water temperatures exhibit seasonal variation in the range 10–30°C.Nomenclature K _S saturation constant (mg/l) - K _I substrate inhibition constant (mg/l) - specific growth rate (h^–1) - _m maximum specific growth rate without substrate inhibition (h^–1) - _max maximum achievable specific growth rate with substrate inhibition (h^–1) - S substrate (phenol) concentration (mg/l) - S_o initial substrate concentration (mg/l) - S_max substrate concentration corresponding to _max (mg/l) - t time (h) - X cell concentration, dry basis (mg DW/l) - X_f final cell concentration, dry basis (mg DW/l) - X_o initial cell concentration, dry basis (mg DW/l) - Y yield factor (mg DW cell produced/mg substrate consumed)     

Growth and energy production in Bacteroides amylophilus

The molar growth yield (Y _m) of Bacteroides amylophilus strain WP91 on maltose was 68±2 g/mol when determined from batch cultures at the peaks of maximal growth. Continued incubation led to considerable cell lysis. When calculated from batch cultures in exponential phase (specific growth rate, =0.57 h^-1) Y _m was 101 g/mol. The maximum value of Y _m in maltose-limited chemostat cultures at the maximum dilution rate (D) attainable (D==0.39 h^-1) was about 79 g/mol. Ammonia-Fmited chemostat cultures metabolized maltose with a much reduced efficiency and this was associated with a difference in morphology and chemical composition of the cells. The theoretical maximum molar growth yields (Y _m ^max ) were 55 and 114 g/mol for ammonia- and maltose-limited growth respectively. However, if account was taken of extracellular nitrogen-containing material in ammonia-limited cultures, Y _m ^max became 60. The maintenance coefficient (m _s), estimated from the lines relating the specific rate of maltose consumption (q _m) and D (where m _s=q _m at D=0), was 7.4±0.6×10^-4 mol maltose/g x h for both nutrient limitations. A difference in maintenance energy demand, independent of growth-rate, could not account, therefore, for the observed differences in Y _m between ammonia- and maltose-limited growth.     

Feeding in the rotifer Brachionus calyciflorus

Summary The laboratory feeding behavior of Brachionus calyciflorus varies depending upon the type of food cell available in suspension. When feeding on the yeast Rhodotorula glutinis, rotifers show a continuous increase in ingestion with increased cell density between 0.01 and 1000 g dry weight ml^-1. Effective clearance rates drop from ca. 50 l animal^-1 h^-1 to less than 0.5 l animal^-1 h^-1 over this food density range. When feeding on Englena gracilis, B. calyciflorus ingestion rates are constant between 1.0 and 100 g ml^-1 of available food, averaging close to 25 ng animal^-1 h^-1. The decrease in clearance rate is more striking than with R. glutinis, dropping from 45 l animal^-1 h^-1 at 0.1 g ml^-1 to 0.13 l animal^-1 h^-1 at 100 g ml^-1. Differences between the patterns obtained with the two food types indicate fundamental dissimilarities in the feeding behavior of this rotifer species when presented with these different foods.     

Growth inhibition of biomass adapted to the degradation of toluene and xylenes in mixture in a batch reactor with substrates supplied by pulses

A biomass adapted to degrade toluene and xylenes in mixture was grown in a batch reactor with substrates supplied by pulses. The inhibition of biomass growth in the course of substrate degradation was investigated. The maximal biomass concentration of 7 g l^–1 was obtained using 150 l of toluene and 15 l of a mixture of xylenes in one litre of liquid medium, and the maximal biomass productivity and yield were 53 mg l^–1 h^–1 and 0.32 g_DW g _s ^–1 , respectively. Higher quantities of substrate added by pulses, that is 200 l of toluene with 20 l of xylenes and 300 l of toluene with 30 l of xylenes, caused an accumulation of metabolites. These higher quantities of substrates caused inhibition of microbial growth. Among the metabolites produced, 4-methyl catechol was found in large quantities in the culture medium and in the cells.     

Comparison of photomixotrophic and heterotrophic callus and suspension cultures of Pinus elliottii. 1. Photosynthetic properties and ultrastructural evidence for coexistence of starch granules and secondary metabolites

Photomixotrophic callus and suspension cultures of salsh pine (Pinus elliottii var. elliottii Engelm.) have been established. Callus tissues contained up to 2.76 g chlorophyll mg^-1 dry wt and suspensions 2.98 g chlorophyll mg^-1 dry wt. Maximum photosynthetic oxygen evolution was 25–32 mol O₂ h^-1 mg^-1 chlorophyll for callus and 35–39 mol O₂h^-1 mg^-1 chlorophyll for suspension, respectively. Photomixotrophic callus was friable with a high moisture content during early and exponential growth, but evolved into a compact and dense tissue during the latter stage of growth. Compact photomixotrophic callus accumulated and deposited secondary metabolites in the central vacuole and developed large starch granules in the chloroplasts. Secondary metabolites were not observed in photomixotrophic suspensions or in heterotrophic calli and suspensions. Photomixotrophic callus contained numerous mitochondria closely associated with well-developed chloroplasts containing 2–6 thylakoids per granum. Heterotrophic callus was characterized by a poorly developed cytoplasm and cup-shaped mitochondria.     

Kinetics of malate transport and decomposition in acid soils and isolated bacterial populations: The effect of microorganisms on root exudation of malate under Al stress

The kinetics and characteristics of malate degradation were studied in four acid soils ranging in both pH (4.30 to 5.00) and vegetation type. The breakdown of malate was rapid in all soils with a half life of approximately 1.7 h, K_m of 1.7 mM and V_max of 70 nmol g^–1 soil h^–1. No relationship was observed between malate decomposition rate and pH. Co-metabolism studies with other C and N substrates (glucose, glycine, glutamate, citrate and succinate) indicated that the microorganisms were not N limited and competitive inhibition of malate breakdown was only observed in the presence of succinate. Studies with isolated mixed bacterial cultures indicated that the bacterial malate uptake was mediated by an energy dependent, dicarboxylate transporter which can be inhibited by succinate and is independent of pH between pH 5.0 and 7.0. The K_m and V_max parameters ranged from 279–955 M and 0.1–17 mol mg^–1 protein h^–1 for the mixed bacterial cultures depending on the bacteria's previous C source. The results indicate that in acid topsoils where microbial populations are high, the microbes may provide a considerable sink for organic acids. If organic acids are being released by roots in response to an environmental stress (e.g. Al toxicity, P deficiency) it can be expected that the efficiency of these root mediated metal resistance mechanisms will be markedly reduced by rapid microbial degradation.     

Effect of molybdenum on the efficiency of tetrathionate utilisation by chemostat cultures ofSulfolobus BC

Summary Continuous cultures ofSulfolobus BC oxidising tetrathionate were found not to be tetrathionate limited, but molybdenum limited. Incorporation of only 1.3M Na₂MoO₄ into the medium of autotrophically grown, continuous cultures ofSulfolobus BC oxidising 15mM tetrathionate at a dilution rate of 0.029h^–1 increased the steady state biomass from absorbance (660nm) 0.160 to 0.315. The biomass yield gave a corresponding increase from 5.93±0.84 to 11.52±0.68 g dry weight mol^–1 tetrathionate oxidised.     

Phosphate uptake capacity of summer phytoplankton of the Loosdrecht Lakes in relation to phosphorus loading and irradiance

Summer populations of the phytoplankton of the Loosdrecht Lakes were enclosed in laboratory scale enclosures (LSE), supplied with 7.5 g P.l^–1.d^–1 and 105 g P.l^–1.d^–1, respectively. The maximum initial phosphate uptake rate (V_m) was related to irradiance and primary production. At phosphate uptake saturating light-irradiance V_m values up to 4 times the V_m values in the dark were measured.The phosphate uptake capacity per unit dry weight remained more or less constant throughout the experiments in the LSE receiving the lower amount of phosphorus, and declined in the LSE receiving the higher amount of phosphorus. Within the range of V_m values measured (<10 g P.mg DW^–1.h^–1 or 1,3 g P. g chla ^–1.h^–1), the growth rate of the phytoplankton was not influenced by alterations in phosphorus availability.