Utilization of gluconate by Escherichia coli. Induction of gluconate kinase and 6-phosphogluconate dehydratase activities

1. A mutant of Escherichia coli, devoid of phosphopyruvate synthetase, glucosephosphate isomerase and 6-phosphogluconate dehydrogenase activities, grew readily on gluconate and inducibly formed an uptake system for gluconate, gluconate kinase and 6-phosphogluconate dehydratase while doing so. 2. This mutant also grew on glucose 6-phosphate and inducibly formed 6-phosphogluconate dehydratase; however, the formation of the gluconate uptake system and gluconate kinase was not induced under these conditions. 3. The use of the Entner–Doudoroff pathway for the dissimilation of 6-phosphogluconate, derived from either gluconate or glucose 6-phosphate, by this mutant was also demonstrated by the accumulation of 2-keto-3-deoxy-6-phosphogluconate (3-deoxy-6-phospho-l-glycero-2-hexulosonate) from both these substrates in a similar mutant that also lacked phospho-2-keto-3-deoxygluconate aldolase activity. 4. Glucose 6-phosphate inhibits the continued utilization of fructose by cultures of the mutants growing on fructose, as it does in wild-type E. coli. 5. The mutants do not use glucose for growth. This is shown to be due to insufficiency of phosphopyruvate, which is required for glucose uptake.     

Sequential utilization of substrates by Pseudomonas putida CSV86: signatures of intermediate metabolites and online measurements

Pseudomonas putida CSV86 preferentially utilizes aromatics over glucose and co-metabolizes them with organic acids. On aromatics plus glucose, CSV86 utilized aromatics first with concomitant appearance of transient metabolites such as salicylate, benzaldehyde and benzoate. Citrate was the main extracellular metabolite observed during glucose uptake. The strain showed simultaneous utilization of organic acids and aromatic compounds. Based on the metabolite analysis and growth profiles, we hypothesize that the repression of glucose utilization could be due to organic acid intermediates generated from aromatic compound metabolism. The online measurements indicate the instantaneous metabolic state of the culture. For example, the CO₂ evolution and agitation speed show peak values during the two growth phases in the diauxic growth while dissolved oxygen values show decrease at the corresponding durations. These measurements correlated well with the offline measurements but provided a better time resolution of the process.     

31P Nuclear magnetic resonance studies of ethanol inhibition in Zymomonas mobilis

Ethanol inhibition of glucose catabolism in Zymomonas mobilis was investigated using ³¹P NMR spectroscopy in vivo and of perchloric acid extracts from cell suspensions incubated with 0, 5 and 10% (w/v) ethanol. In vivo ³¹P NMR experiments revealed slower glucose utilization and decreased levels of nucleoside triphosphates in the presence of 10% ethanol as compared to controls. Using ³¹P NMR spectroscopy of perchloric acid extracts, intracellular accumulation of 3.4 mM 3-phosphoglycerate was found when 10% ethanol was present in the medium. No accumulation of this metabolite occurred in cells incubated with 0 and 5% ethanol. Enzyme assays confirmed that phosphoglycerate-mutase and enolase were inhibited 31 and 40%, respectively, in the presence of 10% ethanol in the test system. Therefore, under the conditions used the decrease in the fermentative activity of Z. mobilis at high ethanol concentrations is due to inhibition of phosphoglycerate-mutase and enolase.Abbreviation KDPG 2-keto-3-deoxy-6-phosphogluconate     

Phenotypic properties of <Emphasis Type="Italic">Sulfobacillus thermotolerans</Emphasis>: Comparative aspects

The phenotypic characteristics of the species Sulfobacillus thermotolerans Kr1^T, as dependent on the cultivation conditions, are described in detail. High growth rates (0.22–0.30 h^?1) and high oxidative activity were recorded under optimum mixotrophic conditions at 40 °C on medium with inorganic (Fe(II), S⁰, or pyrite-arsenopyrite concentrate) and organic (glucose and/or yeast extract) substrates. In cells grown under optimum conditions on medium with iron, hemes a, b, and, most probably, c were present, indicating the presence of the corresponding cytochromes. Peculiar extended structures in the form of cylindrical cords, never observed previously, were revealed; a mucous matrix, likely of polysaccharide nature, occurred around the cells. In the cells of sulfobacilli grown litho-, organo-, and mixotrophically at 40 °C, the enzymes of the three main pathways of carbon utilization and some enzymes of the TCA cycle were revealed. The enzyme activity was maximum under mixotrophic growth conditions. The growth rate in the regions of limiting temperatures (55 °C and 12–14 °C) decreased two-and tenfold, respectively; no activity of 6-phosphogluconate dehydrogenase, one of the key enzymes of the oxidative pentose phosphate pathway, could be revealed; and a decrease in the activity of almost all enzymes of glucose metabolism and of the TCA cycle was observed. The rate of ¹⁴CO₂ fixation by cells under auto-, mixo-, and heterotrophic conditions constituted 31.8, 23.3, and 10.3 nmol/(h mg protein), respectively. The activities of RuBP carboxylase (it peaked during lithotrophic growth) and of carboxylases of heterotrophic carbon dioxide fixation were recorded. The physiological and biochemical peculiarities of the thermotolerant bacillus are compared versus moderately thermophilic sulfobacilli.     

Production of 1,3-Propanediol from Glucose by Recombinant <Emphasis Type="Italic">Escherichia coli</Emphasis> BL21(DE3)

A range of recombinant strains of Escherichia coli were developed to produce 1,3-propanediol (1,3-PDO), an important C3 diol, from glucose. Two modules, the glycerol-producing pathway converting dihydroxyacetone phosphate to glycerol and the 1,3-PDO-producing pathway converting glycerol to 1,3-PDO, were introduced into E. coli. In addition, to avoid oxidative assimilation of the produced glycerol, glycerol oxidative pathway was deleted. Furthermore, to enhance the carbon flow to the Embden- Meyerhof-Parnas pathway, the Entner-Doudoroff pathway was disrupted by deleting 6-phosphogluconate dehydratase and 2-keto-3-deoxy-6-phosphogluconate aldolase. Finally, the acetate production pathway was removed to minimize the production of acetate, a major and toxic by-product. Flask experiments were carried out to examine the performance of the developed recombinant E. coli. The best strain could produce 1,3-PDO with a yield of 0.47 mol/mol glucose. Along with 1,3-PDO, glycerol was produced with a yield of 0.33 mol/mol glucose.     

Flow of 14C-methanol via assimilatory and dissimilatory sequences with yeast in presence of glucose

Experiments were performed to reveal the extent to which individual heterotrophic substrates of a mixture contribute to the overall carbon and energy metabolism. For this reason Hansenula polymorpha MH 20 was chemostatically (C-limited) cultivated at different growth rates on mixtures of methanol and glucose fed at proportions of 3:1 and 1:3 (in weight units), respectively. The distributions of ¹⁴C-carbon from methanol in biomass as well as carbon dioxide (and supernatant) fractions were determined. From these results it followed, firstly, that energy derived from methanol dissimilation was used in part for the incorporation of glucose carbon, resulting in carbon conversion efficiencies for this substrate equivalent to yield coefficients of 0.61–0.69 g/g. Secondly, the growth yield data revealed that the efficiency of methanol conversion had to be increased in order to account for the experimentally determined yield figures. This was further confirmed by theoretical treatment of the growth yield data which showed that these could only be obtained if P/O-quotients for methanol conversion similar to those for glucose, i.e. 2.0–2.5, were considered. The latter property was regarded as the main reason for the observed improvement of growth yield accompanying the simultaneous utilization of methanol and glucose in this yeast.Abbreviations ATP_M,a ATP required for incorporation of assimilated methanol at a given P/O-quotient - ATP_M,d ATP generated from dissimilated methanol at a given P/O-quotient - G and M glucose and methanol; respectively (the indices u, a, d and e mean utilized, assimilated, dissimilated and incorporated by excess energy, respectively) - PGA 3-phosphoglyceric acid - Y _G ^app apparent growth yield on glucose in presence of methanol - Y _G ^P/O theoretical growth yield on glucose at a given P/O-quotient     

Dynamics of carbon assimilation by St Lawrence estuary phytoplankton at the end of summer

This study presents data of in situ measurements of inorganic carbon assimilation by phytoplankton communities of the St Lawrence estuary during the end of summer 1982. We used carboxylase activity measurements (ribulose-1,5-bisphosphate carboxylase, carboxylases) and the ¹³C/¹²C ratio of phytoplankton organic carbon, expressed as ¹³C, to study patterns of assimilation. Upper estuary phytoplankton communities showed a smaller turn-over rate in carbon assimilation than lower estuary phytoplankton communities. Carbon assimilation was limited by light intensity in the upper estuary and by CO₂ availability in the lower estuary. In the St Lawrence estuary, stable carbon isotope ratios of phytoplankton organic carbon seemed to be controlled by inorganic carbon availability rather than by phytoplankton metabolism.     

Nitrogen fixation in Pseudomonas stutzeri

A recently developed oxygen gradient system and a complex medium were used to isolate a microaerobically N₂-fixing heterotrophic bacterium from the rhizosphere of a high fixing Sorghum nutans cultivar. The isolate was identified as nif(+) phenotype of Pseudomonas stutzeri on the basis of cultural, physiological and biochemical characteristics, including DNA/DNA hybridization. N₂ fixation was demonstrated by assimilation of ¹⁵N₂ into cellular protein; the physiology of nitrogen fixation was studied. The isolate contains one 30 MD plasmid and can be cured with associated loss of N₂ fixation capability.Dedicated to Prof. Dr. W. Nultsch on the occasion of his 60th birthday     

Metabolism of [1-13C]glucose in a synaptosomally enriched fraction of rat cerebrum studied by1H/13C magnetic resonance spectroscopy

This study explored the utility of¹H and¹³C magnetic resonance spectroscopy to study a standard synaptosomally enriched fraction (P2 pellet) made from rat cerebrum. The preparations contained high concentrations of N-acetylaspartate and -aminobutyric acid and low concentrations of glutamine, indicating that they were in fact rich in neuronal cytosol. The metabolic competence of the preparation was assessed by quantitative measurements of its ability to convert [1-¹³C]glucose into lactate, glutamate, aspartate, and other metabolites under well oxygenated conditions in 30 minutes. The minimum mean glycolytic rate was 0.8 mM glucose/min and the flow through the tricarboxylic acid cycle was equivalent to 0.2 mM glucose/min.Abbreviations ppm parts per million (chemical shift scale) - NMR nuclear magnetic resonance - GABA -aminobutyric acid - PBS phosphate-buffered normal saline solution - TSP 3-trimethylsilylpropionate During the performance of these studies Dr. A.P. Burlina was on leave from Instituto di Clinica delle Malattie Nervose e Mentali, University of Padua, Padua, Italy.     

Light and dark uptake and loss of 14C: methodological problems with productivity measurements in oceanic waters

Measurements of the uptake and loss of ⁴C in the light and in the dark in the Tasman and Coral Seas have revealed methodological problems with the estimation of productivity in these waters. Rates of productivity estimated without replication, time series incubations and dark controls frequently overestimated the true rates of autotrophic production. The data showed unexpectedly high rates of both uptake and loss in the dark in oligotrophic waters. In oligotrophic oceanic waters, dark incorporation of ¹⁴C sometimes equalled the uptake of ¹⁴C in the light bottle. Rapid uptake of isotope in the dark controls appeared to be the result of rapid bacterial growth and metabolism. This problem was exacerbated by agitation of the sample before or during the incubation. Tropical samples were particularly susceptible to problems arising from the agitation of the samples. Latitudinal gradients of dark uptake and loss were revealed in these incubations. The loss of label during 8–12 hours in the dark (after 12 hr in the light) was as high as 50% in subtropical waters. The loss was frequently unmeasurable (< 10%) in temperate waters. The time course of ¹⁴C uptake indicated active grazing in the bottles and suggested that most of the nighttime losses of label were due to grazing by microheterotrophs. Respiratory losses appeared to be small. Calculated values of the assimilation number (or photosynthetic capacity) which did not correct for dark ¹⁴C uptake were too high to be biochemically realistic. The errors were due to the heterotrophic uptake of label and the lack of dark controls. Rapid release of ¹⁴C in the dark after incubation in the light meant that the estimate of productivity was dependant on the trophic state of the sample and on the period of incubation.     

The Influence of Leaf Windows on the Utilization and Absorption of Radiant Energy in Seven Desert Succulents

Four fluorescence parameters [F_v/F_m = the intrinsic efficiency of energy conversion via photosystem 2 (PS2); F_v/F_m= the efficiency of energy conversion via PS2 in the light; P = fraction of absorbed radiant energy utilized for photosynthesis; and D = fraction of absorbed radiant energy dissipated as heat] were measured on leaves of seven species of succulents having epidermal windows. While the function of leaf windows has reportedly been to increase absorption of radiant energy and, hence, the rate of photosynthesis in these species, recent evidence indicates that this translucent portion of epidermal tissue, lacking chlorophyll, may also result in photoinhibition in these species, especially for those with growth habits aboveground. Species with aboveground and belowground growth habits were compared with their leaf windows covered with reflective tape and with windows unobstructed. Results showed no increase in photoinhibition for these species resulting from the radiant energy penetrating the window tissue. Although the efficiency of the photosynthetic mechanism was not significantly influenced by the additional radiant energy provided by the window for individual species, there were significant differences in the efficiencies of radiant energy capture (F_v/F_m) and utilization (P) between the two growth habits. Species with an aboveground growth habit were less efficient in radiant energy utilization compared with the species having a belowground growth habit.     

Cultivation of Escherichia coli with mixtures of 3-phenylpropionic acid and glucose: Dynamics of growth and substrate consumption

In technical as well as natural ecosystems, pollutants are often mineralised in the presence of easily degradable carbon sources. A laboratory model system consisting of Escherichia coli ML 30 growing with mixtures of 3-phenylpropionic acid (3ppa, pollutant) and glucose (easily degradable substrate) was investigated in batch and carbon-limited continuous culture. Untypically, a linear growth pattern was observed during batch cultivation with 3ppa as the only carbon/energy source. When exposed to mixtures of both substrates in batch culture, E. coli utilised the two compounds sequentially. However, 3ppa and glucose were consumed simultaneously in continous culture. Whereas a pulse of excess glucose to a batch culture growing with 3ppa led to the repression of 3ppa utilisation, an excess of glucose added into continuous culture did not inhibit the utilisation of 3ppa. During continuous cultivation the 3ppa-degrading enzyme system operated close to saturation.     

Heterotrophic Metabolism of the Chemolithotroph Thiobacillus ferrooxidans

Glucose-6-phosphate dehydrogenase and the enzymes of the Entner-Doudoroff pathway, 6-phosphogluconate dehydrase and 2-keto-3-deoxy-6-phosphogluconate aldolase (assayed together), are induced during heterotrophic growth of Thiobacillus ferrooxidans on an iron-glucose-supplemented medium or on glucose alone. By contrast, autotrophic cells (iron-grown) contain low levels of these enzymes. Fructose 1, 6-diphosphate aldolase, an enzyme of the Embden-Meyerhof pathway, is present at low levels irrespective of the growth medium, suggesting that this enzyme is not involved in energy-yielding reactions but merely provides intermediates for biosynthesis. The Entner-Doudoroff and pentose-phosphate pathways are the principle means through which glucose is dissimilated and is presumed to be concerned with energy production. Isotopic studies showed that a high rate of CO(2) formation from specifically labeled glucose came from carbon atoms 1 and 4. An unexpectedly high rate of evolution of CO(2) also came from carbon 6, suggesting that the triose phosphate formed during glucose breakdown and specifically as a result of 2-keto-3-deoxy-6-phosphogluconate aldolase activity, was metabolized via some unorthodox metabolic route. Cells grown in the iron-supplemented and glucose-salts media have a complete tricarboxylic acid cycle, whereas autotrophically grown T. ferrooxidans lacked both alpha-ketoglutarate dehydrogenase and reduced nicotinamide adenine dinucleotide oxidase. Two isocitrate dehydrogenases [nicotinamide adenine dinucleotide (NAD) and NAD phosphate (NADP) specific] were present. NAD-linked enzyme was constitutive, whereas the NADP-linked enzyme was induced upon adaptation of autotrophic cells to heterotrophic growth.     

Mixotrophic and autotrophic growth of Thiobacillus acidophilus on tetrathionate

Thiobacillus acidophilus can grow in batch and chemostat culture as a heterotroph on glucose, a chemolithoautotroph on tetrathionate and CO₂, or as a mixotroph. Mixotrophically it obtains energy from the simultaneous oxidation of tetrathionate and glucose, and carbon from both glucose and CO₂. Mixotrophic cultures contain lower activities of ribulose 1,5-bisphosphate carboxylase and exhibit lower specific rates of tetrathionate oxidation than do autotrophic cultures. Mixotrophic cultures with low concentrations of glucose have growth rates that are intermediate between slow autotrophic growth and fast heterotrophic growth. Slightly more glucose-carbon is assimilated by mixotrophic cultures than by heterotrophic ones provided with the same concentrations of glucose. Mixotrophic yield in the chemostat is also slightly greater than predicted from autotrophic and heterotrophic yields. These observations indicate that there is preferential assimilation of glucose, at the expense of energy from tetrathionate oxidation, during mixotrophy, resulting in an overall energy saving that produces enhanced growth yield. These observations are relevant to understanding the regulatory behaviour of T. acidophilus in its acidic, mineral-leaching habitats.     

Diacetyl production and growth of Lactobacillus rhamnosus on multiple substrates

Lactobacillus rhamnosus is a heterolactic acid bacterium, which can be used to produce flavour compounds like diacetyl and acetoin. Various startegies have been applied to improve the growth rate and diacetyl yield. The use of multiple substrates affected growth as well as the yield of diacetyl. Growth on a medium containing glucose demonstrated a diauxic growth profile, with the second phase of growth being on the product, lactic acid. L. rhamnosus also grew on a medium containing citrate. Growth on medium containing glucose+citrate demonstrated simultaneous utilization of carbon sources. L. rhamnosus did not grow in a medium containing acetate and also did not co-metabolize it with glucose. Maximum specific growth rate ( _max) was found to increase in the case of simultaneous utilization of glucose+citrate (0.38 h^–1) as compared to glucose as the sole carbon source (0.28 h^–1). The yields of diacetyl were also found to increase for glucose + pyruvate and glucose + citrate (0.10 and 0.05 g g^–1 of glucose, respectively) as compared to glucose alone (0.01 g g^–1 of glucose). The productivity of diacetyl on medium containing glucose and citrate was double that of a medium containing only citrate, although the yields were comparable.     

Energy production and growth of Pseudomonas oxalaticus OX1 on oxalate and formate

The efficiency of oxidative phosphorylation in Pseudomonas oxalaticus during growth on oxalate and formate was estimated by two methods. In the first method the amount of ATP required to synthesize cell material of standard composition was calculated during growth of the organism on either of the two substrates. The [Y _ATP ^max ] theor. values thus obtained were 12.5 and 6.5 for oxalate and formate respectively, if the assumption were made that no energy is required for transport of oxalate or carbon dioxide. When active transport of oxalate requiring an energy input equivalent to 1 mole of ATP per mole of oxalate was taken into account, [Y _ATP ^max ]theor. for oxalate was 9.4. True Y _ATP ^max values were derived from these data on the assumption that the energy produced in the catabolism of Pseudomonas oxalaticus is used with approximately the same efficiency as in a range of other chemoorganotrophs. P/O ratios were calculated using the equation P/O=Y _O/Y _ATP. The data for Y _O and m _e required for these calculations were obtained from cultures of Pseudomonas oxalaticus growing on oxalate or formate in carbon-limited continuous cultures. The P/O ratios calculated by this method were, for oxalate, 1.3 (or 1.0 if active transport were ignored), and for formate, 1.7.In the second method the stoicheiometries of the respiration-linked proton translocations with oxalate and formate were measured in washed suspensions of cells grown on the two substrates. The H⁺/O ratios obtained were 4.3 with oxalate and 3.9 with formate. These data indicate the presence of two functional phosphorylation sites in the electron transport chain of Pseudomonas oxalaticus during growth on both substrates. A comparison of the P/O ratio on oxalate obtained with the two methods indicated that the energy requirement for active transport of oxalate has a major effect on the energy budget of the cell; about 50% of the potentially available energy in oxalate is required for its active transport across the cell membrane. Translocation of formate requires approximately 25% of the energy potentially available in the substrate. These results offer an explanation for the fact that molar growth yields of Pseudomonas oxalaticus on oxalate and formate are not very different.Abbreviations PMS phenazinemethosulphate - DCPIP 2,6-dichlorophenolindophenol - TMPD N,N,N,N-tetramethyl-1,4-phenylene-diamine dihydrochloride - SD standard deviation - PEP Phosphoenol-pyruvate     

Physiology of Geobacter metallireducens under excess and limitation of electron donors. Part I. Batch cultivation with excess of carbon sources

For microorganisms that play an important role in bioremediation, the adaptation to swift changes in the availability of various substrates is a key for survival. The iron-reducing bacterium Geobacter metallireducens was hypothesized to repress utilization of less preferred substrates in the presence of high concentrations of easily degradable compounds. In our experiments, acetate and ethanol were preferred over benzoate, but benzoate was co-consumed with toluene and butyrate. To reveal overall physiological changes caused by different single substrates and a mixture of acetate plus benzoate, a nano-liquid chromatography–tandem mass spectrometry-based proteomic approach (nano-LC–MS/MS) was performed using label-free quantification. Significant differential expression during growth on different substrates was observed for 155 out of 1477 proteins. The benzoyl-CoA pathway was found to be subjected to incomplete repression during exponential growth on acetate in the presence of benzoate and on butyrate as a single substrate. Peripheral pathways of toluene, ethanol, and butyrate degradation were highly expressed only during growth on the corresponding substrates. However, low expression of these pathways was detected in all other tested conditions. Therefore, G. metallireducens seems to lack strong carbon catabolite repression under high substrate concentrations, which might be advantageous for survival in habitats rich in fatty acids and aromatic hydrocarbons.     

Simultaneous operation of three catabolic pathways in the metabolism of glucose by Thiobacillus A2

Enzymes essential to the operation of the Embden-Meyerhof glycolytic pathway, the Entner-Doudoroff pathway and oxidative pentose phosphate pathway were present in Thiobacillus A2 grown on glucose and other sugars. Radiorespirometry under various conditions with Thiobacillus A2 oxidising glucose specifically labelled with ¹⁴C in carbon atoms 1, 2, 3, 3+4, 6 or universally labelled demonstrated the simultaneous operation of the Embden-Meyerhof (48%), Entner-Doudoroff (28%), and pentose phosphate (24%) pathways in release of carbon dioxide from glucose. Growth on succinate, or autotrophically on formate or thiosulphate resulted in repression of most enzymes of the pathways, but high aldolase levels were retained indicating its role in gluconeogenesis and the Calvin cycle. Different fructose diphosphatase activities were found in succinate- and thiosulphate-grown organisms. The results indicate that all three major catabolic pathways for glucose function in Thiobacillus A2 grown on sugars. Thiobacillus acidophilus showed a different radiorespirometric pattern and apparently used the Entner-Doudoroff (64.5%) and pentose phosphate (35.5%) pathways, but showed unusually high release of carbon atom 6, as was also found for T. ferrooxidans.Abbreviations EM Embden-Meyerhof - ED Entner-Doudoroff - EDTA ethylene diamine tetra-acetic acid, disodium salt - FDP fructose 1,6-diphosphate - KDPG 2-keto-3-deoxy-6-phosphogluconate - 6-PG 6-phosphogluconate - Pa Pascal (10⁵ Pa=1 bar) - PP pentose phosphate - POPOP 1,4-di[2-(5-phenyloxazolyl)] benzene - PPO 2,5-diphenyloxazole     

Light utilization and competition between Echinacea purpurea, Panicum virgatum and Ratibida pinnata under greenhouse and field conditions

Competitive interactions were compared under field and greenhouse conditions for three representative tallgrass prairie species, Echinacea purpurea, Panicum virgatum and Ratibida pinnata. These were planted in monoculture and in mixtures of two species using a replacement series design with groups of four or eight plants. Competition was determined from shoot dry weight data collected during 120days in the greenhouse and after 415days in the field. Yields declined with increased density of a single species in the greenhouse from 40days onward and in the field. Relative yields were up to 100% higher in mixtures than in monocultures for all species early on in the greenhouse experiment. Later in the experiment and in the field relative yields of E.purpurea decreased in the presence of P.virgatum and R.pinnata, whereas relative yields of these two species increased in the presence of E.purpurea. There were correlations in relative yield between the field and the greenhouse experiment at 80 and 120days. In the greenhouse P.virgatum maintained higher net assimilation rates than the other species. Relative growth rates of all species were higher in monoculture and in mixtures up to 40days, after which they declined, especially for E.purpurea in mixtures. In the field, higher light intensities occurred in pure stands of E.purpurea than in mixed stands with other species. The order of competitive ability that was apparent from these field and greenhouse studies, P.virgatum=R.pinnata>E.purpurea, could be partially explained by photosynthetic rates in relation to canopy light interception.     

Improved growth of the red yeast, Phaffia rhodozyma (Xanthophyllomyces dendrorhous), in the presence of tricarboxylic acid cycle intermediates

Catabolites related to tricarboxylic acid cycle affected growth and carotenogenesis in Phaffia rhodozyma. Glutamate, glutamine, aspartate, asparagine and proline at 75 mM of N increased biomass from 2 g l^–1 to 2.9–4.7 g l^–1 but decreased carotenoid from 420 g g^–1 yeast to 200–260 g g^–1 yeast in strain 67-385. However, simple nitrogen sources did not decrease carotenoid formation. Tricarboxylic acid intermediates repressed carotenogenesis to a less degree than the corresponding amino acids. Carotenoid hyper-producing mutants were impaired in nitrogen utilization. These results indicated that nitrogen assimilation and the concentrations of tricarboxylic acid cycle intermediates are involved in regulation of carotenoid biosynthesis.