Carbon Metabolism

In C3 plants, the metabolite fructose 2,6-bisphosphate (Fru 2,6-P2) has an important role in the regulation of carbon partitioning during photosynthesis. To investigate the impact of Fru 2,6-P2 on carbon metabolism during Crassulacean acid metabolism (CAM), we have developed an Agrobacterium tumefaciens-mediated transformation system in order to alter genetically the obligate CAM plant Kalanchöe daigremontiana. To our knowledge, this is the first report to use genetic manipulation of a CAM species to increase our understanding of this important form of plant metabolism. Transgenic plants were generated containing a modified rat liver 6-phosphofructo-2-kinase gene. In the plants analyzed the activity of 6-phosphofructo-2-kinase ranged from 175% to 198% of that observed in wild-type plants, resulting in Fru 2,6-P2 concentrations that were 228% to 350% of wild-type plants after 2 h of illumination. A range of metabolic measurements were made on these transgenic plants to investigate t...

Growth, net photosynthetic rate (P N ), chlorophyll fluorescence induction kinetics, and stromal fructose-1,6-bisphosphatase (sFBPase) in annual legumes native to the Mediterranean region, two clovers (Trifolium subterraneum L. ssp. oxaloides Nyman cv. Clare and T. michelianum Savi cv. Giorgia) and two Medicago species (M. polymorpha L. cv. Anglona and M. truncatula Gaertn. cv. Paraggio), shifted from 20 to 10 ºC for 1 d or developed at 10 ºC were compared with controls kept at 20 ºC. Cold development produced a larger stimulation of growth in the clover cv. Giorgia and the Medicago cv. Paraggio. Transferring plants to low temperatures affected P N relatively less in clovers than in Medicago plants. Development at 10 ºC relieved the inhibition of photosynthesis in Giorgia and Paraggio, but not in Clare and Anglona, which correlated with increases in the maximum rate of carboxylation by ribulose-1,5-bisphosphate carboxylase/oxygenase, RuBPCO (V cmax ), and the photon-saturated rate of electron transport (J max ). In Medicago, transfer from high to low temperature inhibited photosynthesis in a lesser extent in Anglona than in Paraggio, which showed severe limitations at level of V cmax and J max . Development at 10 ºC in Paraggio produced an efficient photosynthetic cold acclimation, this being associated with a two-fold increase of quantum yield of photosystem 2 electron transport (∆F/F' m ) and with the activity of sFBPase. By contrast, Anglona showed an irreversible inhibition of P N coupled with the reduction of carbon metabolism by impairment of Calvin cycle enzyme activities such as RuBPCO and sFBPase, resulting in a poor cold acclimation of photosynthesis in this cultivar.

A full-length cDNA clone encoding carbonic anhydrase (CA) was isolated from a soybean nodule cDNA library. In situ hybridization and immunolocalization were performed in order to assess the location of CA transcripts and protein in developing soybean nodules. CA transcripts and protein were present at high levels in all cell types of young nodules, whereas in mature nodules they were absent from the central tissue and were concentrated in cortical cells. The results suggested that, in the earlier stages of nodule development, CA might facilitate the recycling of CO2 while at later stages it may facilitate the diffusion of CO2 out of the nodule system. In parallel, sucrose metabolism was investigated by examination of the temporal and spatial transcript accumulation of sucrose synthase (SS) and phosphoenolpyruvate carboxylase (PEPC) genes, with in situ hybridization. In young nodules, high levels of SS gene transcripts were found in the central tissue as well as in the parenchymateou...

In eukaryotic cells tRNA synthesis is negatively regulated by the protein Maf1, conserved from yeast to humans. Maf1 from yeast Saccharomyces cerevisiae mediates repression of trna transcription when cells are transferred from medium with glucose to medium with glycerol, a non-fermentable carbon source. The strain with deleted gene encoding Maf1 (maf1Δ) is viable but accumulates tRNA precursors. In this study tRNA precursors were analysed by RNA-Seq and Northern hybridization in wild type strain and maf1Δ mutant grown in glucose medium or upon shift to repressive conditions. A negative effect of maf1Δ mutant on the addition of the auxiliary CCA nucleotides to the 3' end of pre-tRNAs was observed in cells shifted to unfavourable growth conditions. This effect was reduced by overexpression of the yeast CCA1 gene encoding ATP(CTP):tRNA nucleotidyltransferase. The CCA sequence at the 3' end is important for export of tRNA precursors from the nucleus and essential for tRNA chargi...

Background: Methyltetrahydrofolate, corrinoid iron-sulfur protein methyltransferase (MeTr), catalyzes a key step in the Wood-Ljungdahl pathway of carbon dioxide fixation. It transfers the N 5 -methyl group from methyltetrahydrofolate (CH 3 -H 4 folate) to a cob(I)amide center in another protein, the corrinoid iron-sulfur protein. MeTr is a member of a family of proteins that includes methionine synthase and methanogenic enzymes that activate the methyl group of methyltetra-hydromethano(or -sarcino)pterin. We report the first structure of a protein in this family. We determined the crystal structure of MeTr from Clostridium thermoaceticum at 2.2 Å resolution using multiwavelength anomalous diffraction methods. The overall architecture presents a new functional class of the versatile triose phosphate isomerase (TIM) barrel fold. The MeTr tertiary structure is surprisingly similar to the crystal structures of dihydropteroate synthetases despite sharing less than 20% sequence identity. This homology permitted the methyl-H 4 folate binding site to be modeled. The model suggests extensive conservation of the pterin ring binding residues in the polar active sites of the methyltransferases and dihydropteroate synthetases. The most significant structural difference between these enzymes is in a loop structure above the active site. It is quite open in MeTr, where it can be modeled as the cobalamin binding site. The MeTr structure consists of a TIM barrel that embeds methyl-H 4 folate and cobamide. All related methyltransferases are predicted to fold into a similar TIM barrel pattern and have a similar pterin and cobamide binding site. The observed structure is consistent with either a 'front' (N 5 ) or 'back' (C 8a ) side protonation of CH 3 -H 4 folate, a key step that enhances the electrophilic character of the methyl group, activating it for nucleophilic attack by Co(I).

Background Heliobacterium modesticaldum is a gram-positive nitrogen-fixing phototrophic bacterium that can grow either photoheterotrophically or chemotrophically but not photoautotrophically. Surprisingly, this organism is lacking only one gene for the complete reverse tricarboxylic acid (rTCA) cycle required for autotrophic carbon fixation. Along with the genomic information reported recently, we use multiple experimental approaches in this report to address questions regarding energy metabolic pathways in darkness, CO2 fixation, sugar assimilation and acetate metabolism. Results We present the first experimental evidence that D-ribose, D-fructose and D-glucose can be photoassimilated by H. modesticaldum as sole carbon sources in newly developed defined growth medium. Also, we confirm two non-autotrophic CO2-fixation pathways utilized by H. modesticaldum: reactions catalyzed by pyruvate:ferredoxin oxidoreductase and phosphoenolpyruvate carboxykinase, and report acetate excretion du...

Understanding terrestrial carbon metabolism is critical because terrestrial ecosystems play a major role in the global carbon cycle. Furthermore, humans have severely disrupted the carbon cycle in ways that will alter the climate system and directly affect terrestrial metabolism. Changes in terrestrial metabolism may well be as important an indicator of global change as the changing temperature signal. Improving our understanding of the carbon cycle at various spatial and temporal scales will require the integration of multiple, complementary and independent methods that are used by different research communities. Tools such as air sampling networks, inverse numerical methods, and satellite data (top-down approaches) allow us to study the strength and location of the global-and continental-scale carbon sources and sinks. Bottom-up studies provide estimates of carbon fluxes at finer spatial scales and examine the mechanisms that control fluxes at the ecosystem, landscape, and regional scales. Bottom-up approaches include comparative and process studies (for example, ecosystem manipulative experiments) that provide the necessary mechanistic information to develop and validate terrestrial biospheric models. An iteration and reiteration of top-down and bottom-up approaches will be necessary to help constrain measurements at various scales. We propose a major international effort to coordinate and lead research programs of global scope of the carbon cycle.

Although common bean (Phaseolus vulgaris) is the most important grain legume in the developing world for human consumption, few genomic resources exist for this species. The objectives of this research were to develop expressed sequence tag (EST) resources for common bean and assess nodule gene expression through high-density macroarrays. We sequenced a total of 21,026 ESTs derived from 5 different cDNA libraries, including nitrogen-fixing root nodules, phosphorus-deficient roots, developing pods, and leaves of the Mesoamerican genotype, Negro Jamapa 81. The fifth source of ESTs was a leaf cDNA library derived from the Andean genotype, G19833. Of the total high-quality sequences, 5,703 ESTs were classified as singletons, while 10,078 were assembled into 2,226 contigs producing a nonredundant set of 7,969 different transcripts. Sequences were grouped according to 4 main categories, metabolism (34%), cell cycle and plant development (11%), interaction with the environment (19%), and u...

Root colonisation by arbuscular mycorrhizal fungi (AMF) can improve plant resistance/tolerance to biotic stresses. Although this bioprotection has been amply described in different plant systems, the underlying mechanisms remain largely unknown. Besides mechanisms such as improved plant nutrition and competition, experimental evidence supports the involvement of plant defence mechanisms in the observed protection. During mycorrhiza establishment, modulation of plant defence responses occurs upon recognition of the AMF in order to achieve a functional symbiosis. As a consequence of this modulation, a mild, but effective activation of the plant immune responses may occur, not only locally but also systemically. This activation leads to a primed state of the plant that allows a more efficient activation of defence mechanisms in response to attack by potential enemies.

Effect of different CO 2 concentrations on sucrose metabolizing enzymes and on carbohydrate metabolism was studied for eight blackgram (Vigna mungo L. Hepper) genotypes grown in open top chambers under ambient (380 µmol mol -1 ) vs. elevated CO 2 (550 and 700 µmol mol -1 ) levels. The higher acid invertase activity over neutral invertase indicated the major role of acid invertase in sucrose breakdown. Higher acid invertase activity over Sucrose Synthase (SuSy) suggested the major role of invertase in sucrose breakdown and sucrolysis. Sucrose Phosphate Synthase (SPS) activity did not match with sucrose pool sizes in mature leaves and rather varied among genotypes. Plants exposed to higher CO 2 concentrations showed higher starch and sucrose contents as compared with those exposed to ambient CO 2 . Leaf starch content being found several-folds higher than sucrose throughout the study indicated its major role in regulating assimilate partitioning. Increase in glucose vs. fructose concentrations for genotypes grown under elevated CO 2 conditions ranged from 20 to 90% and from 10 to 140%, respectively. The hexoses/sucrose ratio for elevated CO 2 concentration was approximately 0.8-1.6, however for ambient CO 2 content it approximately amounted to unity. Genotypes IC436720, IC519805, IC343952, and IC282009 with low hexose/sucrose ratio representing high CO 2 assimilation along with high sucrose formation indicated better tolerance to elevated CO 2 for carbon partitioning and carbohydrate metabolism. The up-regulation of leaf carbohydrate metabolizing enzymes of low hexose/sucrose as well as low sucrose/starch ratios for the genotype IC436720 (as compared with other genotypes) improved its photosynthetic capability which coupled with its better efficiency of carbon partitioning (indicative of better acclimation to elevated CO 2 ) could prove beneficial to its growth and productivity in the future change of climatic conditions.

Mycorrhizae are symbiotic associations between soil fungi belonging to diverse taxa and the roots of about 90% of all terrestrial plant species. The mutualistic nature of these symbioses is based on the nutritional exchanges between the partners. However, the benefits to the plant partner are not limited to an improved mineral nutrition because they also include a general increase in stress tolerance and health. Because of these benefits, mycorrhizae are of great interest in sustainable agriculture and forestry. In the past few years, the development of highthroughput molecular tools, in addition to the advancements in microscopy techniques, has allowed us to gain a deeper insight on the molecular mechanisms underlying the establishment and functioning of these symbioses. In this chapter, we focus on the use of proteomic tools to better understand the molecular bases of cell communication and the regulation of developmental and metabolic pathways in mycorrhizal associations.

Perturbations of the source-sink balances were performed in field-grown coffee (Coffea arabica) trees to investigate the possible role of carbohydrates in feedback regulation of photosynthesis. • Four treatments were applied at the whole-plant level: (i) complete defruiting and maintenance of the full leaf area, (ii) the half crop load and full leaf area, (iii) the full crop load and full leaf area and (iv) the full crop load and half leaf area. Sampling and measurements were performed twice during the phase of dry matter accumulation of fruits. Gas exchange, chlorophyll a fluorescence, carbon isotope labelling and steady-state metabolite measurements were assessed in source leaves. • The average rate of net photosynthetic rate (A) and stomatal conductance (g s ) were larger (> 50%), and carbon isotope composition ratio was lower, in trees with a full crop load and half leaf area than in defruited trees, with individuals of the other two treatments showing intermediate values. However, differences in A seem unlikely to have been caused either by photochemical impairments or a direct end-product-mediated feedback down-regulation of photosynthesis. • It is proposed that the decreased A in defruited coffee trees was independent of carbon metabolism and was rather directly related to a lower CO 2 availability coupled to lower g s .

Escherichia coli MG1655 cells expressing Vitreoscilla hemoglobin (VHb), Alcaligenes eutrophus flavohemoprotein (FHP), the N-terminal hemoglobin domain of FHP (FHPg), and a fusion protein which comprises VHb and the A. eutrophus C-terminal reductase domain (VHb-Red) were grown in a microaerobic bioreactor to study the effects of low oxygen concentrations on the central carbon metabolism, using fractional 13 C-labeling of the proteinogenic amino acids and two-dimensional [ 13 C, 1 H]-correlation nuclear magnetic resonance (NMR) spectroscopy. The NMR data revealed differences in the intracellular carbon fluxes between E. coli cells expressing either VHb or VHb-Red and cells expressing A. eutrophus FHP or the truncated heme domain (FHPg). E. coli MG1655 cells expressing either VHb or VHb-Red were found to function with a branched tricarboxylic acid (TCA) cycle. Furthermore, cellular demands for ATP and reduction equivalents in VHb- and VHb-Red-expressing cells were met by an increased f...

CK2 is a highly conserved protein kinase involved in different cellular processes, which shows a higher activity in actively proliferating mammalian cells and in various types of cancer and cancer cell lines. We recently demonstrated that CK2 activity is strongly influenced by growth rate in yeast cells as well. Here, we extend our previous findings and show that, in cells grown in either glucose or ethanol-supplemented media, CK2 presents no alteration in K m for both the ATP and the peptide substrate RRRADDSDDDDD, while a significant increase in V max is observed. In chemostat-grown cells, no difference of CK2 activity was observed in cells grown at the same dilution rate in media supplemented with either ethanol or glucose, excluding the contribution of carbon metabolism on CK2 activity. By using the eIF2b-derived peptide, which can be phosphorylated by the holoenzyme but not by the free catalytic subunits, we show that the holoenzyme activity requires the concurrent presence of both b and b 0 encoding genes. Finally, conditions of nitrogen deprivation leading to a G0-like arrest result in a decrease of total CK2 activity, but have no effect on the activity of the holoenzyme. These findings newly indicate a regulatory role of b and b 0 subunits of CK2 in the nutrient response.

Fhosphofructokinase has been identified and purified from extracts of Rhodotorula glutinlus. Kinetic studies of the enzyme indicated high eooperativity with respect to fructose 6-phosphate. The kinetics for ATP shows no cooperativity as indicated by the hyperbolic behavior of the enzyme. The enzyme is inhibited by ADF. Citrate and phosphate have no effect on the enzyme activity. The role of ATP, fructose 6-phosphate, and ADP is discussed.

Fhosphofructokinase has been identified and purified from extracts of Rhodotorula glutinlus. Kinetic studies of the enzyme indicated high eoopera-tivity with respect to fructose 6-phosphate. The kinetics for ATP shows no cooperativity as indicated by the hyperbolic behavior of the enzyme. The enzyme is inhibited by ADF. Citrate and phosphate have no effect on the en-zyme activity. The role of ATP, fructose 6-phosphate, and ADP is discussed.

Daily and annual courses of leaf transpiration, stomatal conductance and shoot water potential of four Quercus suber individuals were compared in a semi-natural stand in southwest Portugal, from spring 1989 to early summer 1990. The trees investigated showed annual patterns typical of evergreen sclerophyllous species but varied in their range of stomatal operation. This appeared to be related to differences in hydraulic conductivity in the root-to-leaf pathway. Maximum stomatal conductance and transpiration rates occurred from March to June.

Euglena gracilis produces several important health-enhancing metabolites including ascorbate, α-tocopherol and free amino acids (faa). The yield of metabolites is dependent on the strain of E. gracilis and the metabolic growth condition. Here we investigated the effects of photoautotrophic (PT), mixotrophic (MT) and heterotrophic (HT) cultivation on the synthesis of ascorbate, α-tocopherol and faa in E. gracilis var. saccharophila, using label-free shotgun proteomics, and metabolite analysis using colourimetric assay, high-performance and ultra-performance liquid chromatography (HPLC/UPLC). PT cultivation resulted in the production of more antioxidants (up to 4.13 mg g -1 ascorbate and 2.52 mg g -1 α-tocopherol) than the MT and HT growth conditions (up to 0.97 and 0.50 mg g -1 ascorbate, and 1.40 and 0.21 mg g -1 α-tocopherol, respectively). The relative abundance of several faa varied between mid-log and initial stationary growth phases, but the total amount of faa remained about the same, with arginine as the most abundant amino acid. Proteomic analysis revealed a total of 3843 nonredundant proteins in E. gracilis var. saccharophila, of which 1890 were common among all cultivations. Gene ontology annotations suggested derivatisation of metabolic pathways from different organisms, such as lysine biosynthesis from fungi and serine biosynthesis from plants, while a few pathways were unique to Euglena, such as those of ascorbate and arginine. Some enzymes exhibited several isoforms that were influenced by the metabolic growth condition. For example, one of the isozymes of threonine aldolase was expressed in HT/MT cultures only, and one of the isozymes of phosphoglycerate dehydrogenase was expressed in PT cultures only. This is the first proteomic study of E. gracilis var. saccharophila, which provides a mechanistic insight into the biosynthetic pathway dynamics of primary metabolites (antioxidants and faa). This new information can serve as a framework for further development of Euglena as a producer of nutraceuticals.

Total respiration, capacities of cytochrome (CytR) and alternative respiration (AR) were studied in two varieties of barley (Horedum vulgare) and wheat (Triticum aestivum) each and one variety of pea (Pisum sativum) at low (Palampur; 1300 m) and high altitudes (Kibber; 4200 m). Similar studies were carried out in naturally growing Rumex nepalensis and Trifoilum repenses at Palampur, Palchan (2250 m) and Marhi (3250 m). All the plants species exhibited lower CytR but significantly higher AR capacity at high altitude (HA) (72-1117% higher) as compared to those at low altitude (LA). Glycolytic product, pyruvate and tricarboxylic acid cycle intermediate, citrate increased with increase in altitude. While the role of these metabolites in relation to HA biology is discussed, significantly higher AR at HA is proposed to be an adaptive mechanism against the metabolic perturbations wherein it might act to lower reactive oxygen species and also provides metabolic homeostasis to plants under the environment of HA.

Ammonium supplied to Acer cells incubated in Tricine buffer raised net dark incorporation of C-bicarbonate by 1.5 to 3,3 fold; this stimulation was not abolished by prior inhibition of glutamine synthetase by methionine sulphoximine. With cells in phosphate buffer, ammonium gave a smaller 1,15 to 1,3 fold stimulation which was abolished when glutamine synthetase was inhibited. Ammonium had no direct effect on the activities of phosphoenolpyruvate carboxyiase and pyruvate kinase assayed in partially purified extracts or on the enzymes catalysing release of label.

An accurate most-probable-number enumeration method was developed for counting the number of Beggiatoa trichomes from various freshwater sediments. The medium consisted of extracted hay, diluted soil ex tract, 0.05% sodium acetate, and 15-35 units per ml of catalase. The The Enumeration, Isolation, and Characterization of Beggiatoa from freshwater sediments.

In the present work, the response to NaCl applied at the vegetative stage to Medicago truncatula and Lotus japonicus has been evaluated in order to ascertain whether the effect of salt stress on nitrogen fixation is due to a limitation on nodular carbon metabolism. Results show maximum sucrose synthase (SS) and alkaline invertase (AI) activities were obtained at the vegetative stage, when maximum nitrogenase activity was detected in both species. SS activity decreased with the salt treatment, providing evidence of the regulatory role of this enzyme for the carbon supply to the bacteroids. Phosphoenolpyruvate carboxylase (PEPC) and malate dehydrogenase (MDH) activities could account for higher nitrogen fixation efficiency detected in L. japonicus nodules and isocitrate dehydrogenase (ICDH) activity compensated for the carbon limitations that occur under salt stress. These results support that nitrogenase inhibition in nodules experiencing salt stress is doubt to a carbon flux shortage, as result of carbon metabolism enzymes activities down-regulation.

Transgenic tomato (Solanum lycopersicum) plants were generated expressing a fragment of the mitochondrial NAD-dependent isocitrate dehydrogenase gene (SlIDH1) in the antisense orientation. The transgenic plants displayed a mild reduction in the activity of the target enzyme in the leaves but essentially no visible alteration in growth from the wild-type. Fruit size and yield were, however, reduced. These plants were characterized by relatively few changes in photosynthetic parameters, but they displayed a minor decrease in maximum photosynthetic efficiency (Fv/Fm). Furthermore, a clear reduction in flux through the tricarboxylic acid (TCA) cycle was observed in the transformants. Additionally, biochemical analyses revealed that the transgenic lines exhibited considerably altered metabolism, being characterized by slight decreases in the levels of amino acids, intermediates of the TCA cycle, photosynthetic pigments, starch, and NAD(P)H levels, but increased levels of nitrate and protein. Results from these studies show that even small changes in mitochondrial NAD-dependent isocitrate dehydrogenase activity lead to noticeable alterations in nitrate assimilation and suggest the presence of different strategies by which metabolism is reprogrammed to compensate for this deficiency.

Numerous studies have been carried out to investigate the hypergravity effect on plants, where seedlings (4-5 days old) were continuously exposed and grown under hypergravity condition. Here, we have used a novel 'shortterm hypergravity exposure experimental method' where imbibed caryopses (instead of seedlings) were exposed to higher g values ranging from 500 g to 2500 g for a short interval time of 10 minutes and post short-term hypergravity treated caryopses were grown under 1 g conditions for five days. Changing patterns in caryopsis germination and growth, along with various photosynthetic and biochemical parameters were studied. Results revealed the significant inhibition of caryopsis germination and growth in shortterm hypergravity treated seeds over control. Photosynthesis

Background Pichia pastoris is an established eukaryotic host for the production of recombinant proteins. Most often, protein production is under the control of the strong methanol-inducible aox1 promoter. However, detailed information about the physiological alterations in P. pastoris accompanying the shift from growth on glycerol to methanol-induced protein production under industrial relevant conditions is missing. Here, we provide an analysis of the physiological response of P. pastoris GS115 to methanol-induced high-level production of the Hepatitis B virus surface antigen (HBsAg). High product titers and the retention of the protein in the endoplasmic reticulum (ER) are supposedly of major impact on the host physiology. For a more detailed understanding of the cellular response to methanol-induced HBsAg production, the time-dependent changes in the yeast proteome and ultrastructural cell morphology were analyzed during the production process. Results The shift from growth on gl...

The effectiveness of homocysteine-lowering therapy on stroke prevention is still unclear. Although randomized controlled epidemiological trials have yielded mixed findings, a multicenter trial did not show any beneficial effect. Genetic studies are still lacking. Therefore, we report on a female patient with transient ischemic attacks and the thermolabile variant of methylenetetrahydrofolate reductase (TT genotype), who benefited from supplemental therapy for homocysteine lowering.

Understanding and optimizing the CO 2 fixation process would allow human beings to address better current energy and biotechnology issues. We focused on modeling the C 3 photosynthetic Carbon metabolism pathway with the aim of identifying the minimal set of enzymes whose biotechnological alteration could allow a functional re-engineering of the pathway. To achieve this result we merged in a single powerful pipe-line Sensitivity Analysis (SA), Single-(SO) and Multi-Objective Optimization (MO), and Robustness Analysis (RA). By using our recently developed multipurpose optimization algorithms (PAO and PMO2) here we extend our work exploring a large combinatorial solution space and most importantly, here we present an important reduction of the problem search space. From the initial number of 23 enzymes we have identified 11 enzymes whose targeting

Neurofibromatosis (NF1) affects 1/2500 people throughout the world. Children with NF1 require a multidisciplinary service ideally, delivered on a single site. NF1 is a very variable condition with children requiring the expertise of genetics, paediatricians, ophthalmologists, dermatologists, neurologists and other specialities as required. Building such a service concentrates expertise, facilitates coordination of care and fosters ideal opportunities for research. Aims: 1) To develop a service ensuring children had access to a multidisciplinary clinic on an annual basis. 2) Hold monthly clinics offering ophthalmology, medical, developmental and dermatology follow up. 3) To create a registry of patients which captures the incidence and prevalence of NF1 in Ireland. To offer best possible care for the children attending the service by following international consensus guidelines. 4) To liaise with NF1 Association, families and research authorities. Methods: 1) Appointment of a CNS/CNM2 in Neurofibromatosis as funded by the NCH Foundation. 2) Visit to the complex NF1 Clinic in Manchester's Children's Hospital and learn from their service, MDT and guidelines. 3) Establish links with genetics, oncology, radiology and orthopaedic depts. in OLCHC. 4) Create a referral pathway for HCPs to ensure children with NF1 are referred to most appropriate service in a timely fashion. 5) To register the service on Orphanet and gain entry into an ERN as a multi-site service in conjunction with OLCHC. Results/Conclusion: To date, the service has been running for 9 months. The CNM2 provides telephone service and coordinates clinics. The Clinic has been registered in Orphanet and the process has begun to create a patient registry and enter the service in the ERN.

The macroalga Gracilariopsis lemaneiformis undergoes dynamic nitrogen (N) fluctuations during cultivation, driven by climate change and eutrophication. However, our understanding of how G. lemaneiformis responds to different N environments remains limited. This study investigates the physiological and molecular metabolic responses of G. lemaneiformis to various N chemical forms (ammonium, nitrate, and urea). Our results indicate that G. lemaneiformis preferentially utilizes ammonium, consuming it more rapidly than nitrate and urea. Furthermore, the transcript profiles of N transporters exhibited distinct regulatory patterns between inorganic N and urea. Metabolic flux analysis revealed significant changes under different conditions of N. Notably, ammonium treatment upregulated the expression of photosynthesis-related proteins in PS II and led to an accumulation of amino acids with higher N content, such as arginine and citrulline. In contrast, nitrate treatment downregulated carbon (C) fixation while promoting the accumulation of amino acids with higher C/N ratios, particularly leucine and valine. Urea treatment resulted in elevated levels of polyamines and aromatic amino acids, which displayed the highest C/N ratios. As a result, the overall C/N ratio in G. lemaneiformis corresponded with that of the dominant amino acids across the three N treatments. Our findings revealed that ammonium, nitrate, and urea differentially affect C and N allocation in G. lemaneiformis. Furthermore, strategic use of specific forms of N can significantly enhance the production of industrially valuable metabolites, highlighting the potential of this macroalga as a sustainable resource for aquaculture and nutraceutical applications.

Transgenic tomato (Solanum lycopersicum) plants were generated expressing a fragment of the mitochondrial NAD-dependent isocitrate dehydrogenase gene (SlIDH1) in the antisense orientation. The transgenic plants displayed a mild reduction in the activity of the target enzyme in the leaves but essentially no visible alteration in growth from the wild-type. Fruit size and yield were, however, reduced. These plants were characterized by relatively few changes in photosynthetic parameters, but they displayed a minor decrease in maximum photosynthetic efficiency (Fv/Fm). Furthermore, a clear reduction in flux through the tricarboxylic acid (TCA) cycle was observed in the transformants. Additionally, biochemical analyses revealed that the transgenic lines exhibited considerably altered metabolism, being characterized by slight decreases in the levels of amino acids, intermediates of the TCA cycle, photosynthetic pigments, starch, and NAD(P)H levels, but increased levels of nitrate and protein. Results from these studies show that even small changes in mitochondrial NAD-dependent isocitrate dehydrogenase activity lead to noticeable alterations in nitrate assimilation and suggest the presence of different strategies by which metabolism is reprogrammed to compensate for this deficiency.

Thermoanaerobacter sp. strain X514 has great potential in biotechnology due to its capacity to ferment a range of C 5 and C 6 sugars to ethanol and other metabolites under thermophilic conditions. This study investigated the central metabolism of strain X514 via 13 C-labeled tracer experiments using either glucose or pyruvate as both carbon and energy sources. X514 grew on minimal medium and thus contains complete biosynthesis pathways for all macromolecule building blocks. Based on genome annotation and isotopic analysis of amino acids, three observations can be obtained about the central metabolic pathways in X514. First, the oxidative pentose phosphate pathway in X514 is not functional, and the tricarboxylic acid cycle is incomplete under fermentative growth conditions. Second, X514 contains ( Re )-type citrate synthase activity, although no gene homologous to the recently characterized ( Re )-type citrate synthase of Clostridium kluyveri was found. Third, the isoleucine in X514 ...

The role of dietary one-carbon determinants remains largely unexplored for non-Hodgkin's lymphoma (NHL). In a population-based case-control study of non-African-American adult (aged 20-74 years) women and men from four US Surveillance, Epidemiology, and End Results study centers (Detroit, Michigan; Iowa; Los Angeles, California; and Seattle, Washington; 1998, the authors examined folate; vitamins B 2 , B 6 , and B 12 ; methionine; and a one-carbon antagonist, alcohol, in 425 incident NHL cases and 359 controls who completed a detailed food frequency questionnaire. Adjusted odds ratios and 95% confidence intervals were estimated by using unconditional logistic regression. Higher intake of one-carbon determinants from food was associated with a lower risk of NHL, but that for only vitamin B 6 (highest vs. lowest quartile: odds ratio ¼ 0.57, 95% confidence interval: 0.34, 0.95; p trend ¼ 0.01) and methionine (odds ratio ¼ 0.49, 95% confidence interval: 0.31, 0.76; p trend ¼ 0.002) reached statistical significance. Folate from food was inversely associated with diffuse subtype (odds ratio ¼ 0.47, 95% confidence interval: 0.23, 0.94; p trend ¼ 0.03). The authors found no association between total (food plus supplement) vitamins and NHL. Nonusers of alcohol had an elevated NHL risk compared with users, and alcohol did not modify other nutrient-NHL associations. Findings suggest that one-carbon nutrients, particularly vitamin B 6 and methionine, may be protective against NHL.

Pedunculate (Quercus robur L.) and sessile (Q. petraea [Matt.] Liebl.) oaks are known to display different ecological requirements, particularly relative to root hypoxia induced by water-logging. Q. robur is more tolerant to hypoxia than Q. petraea. We designed an experiment aiming at identifying morphological and physiological responses to root hypoxia that might differ between the two species. Potted seedlings were submitted during seven weeks to a water-logging treatment with O 2 concentrations below 3 mg L -1 in the vicinity of roots. The treatment induced growth cessation in both species. Q. petraea displayed a lower tolerance to hypoxia as demonstrated by the higher number of seedlings suffering shoot dieback and leaf chlorosis as compared to Q. robur. This difference should be related to the high number of adventitious roots and hypertrophied lenticels that were formed in Q. robur, compared to Q. petraea. In the fine roots of the two species, the activity of pyruvate decarboxylase (PDC), the key enzyme of the fermentative pathway, was stimulated after 24 h of water-logging. Transcripts of PDC increased after 48 h of water-logging in Q. robur and not in Q. petraea. Interestingly, transcripts of haemoglobin (Hb) (possibly involved in the putative nitric oxide cycle) followed the same pattern of response than those of PDC. Enzymes of the sucrose degradation pathway displayed decreased activities after 3 weeks of water-logging, probably due to decreased carbohydrate availability. Alcohol dehydrogenase (ADH), sucrose synthase (Susy), and pyruvate kinase (PK) activities were higher in Q. robur after 3 weeks of water-logging. This study provided a set of markers characterizing the differences of tolerance to hypoxia between the two species for further studies on intra and inter-specific diversity. water-logging / hypoxia / adventitious root / hypertrophied lenticel / carbon metabolism Résumé -Différences de réponses morphologiques et physiologiques à l'ennoyage entre deux espèces sympatriques de chêne (Quercus petraea [Matt.] Liebl., Quercus robur L.). Les chênes pédonculé (Quercus robur L.) et sessile (Quercus petraea [Matt.] Liebl.) présentent des différences de tolérance à l'hypoxie racinaire induite par ennoyage, Q. robur étant plus tolérant que Q. petraea. Nous avons mené une expérience visant à identifier des différences inter-spécifiques dans les réponses morphologiques et physiologiques à l'hypoxie racinaire. Des semis en pots ont été soumis à un ennoyage de 7 semaines avec une concentration en O 2 maintenue en dessous de 3 mg L -1 au voisinage des racines. Le traitement a provoqué un arrêt de croissance chez les deux espèces. Q. petraea a montré une plus faible tolérance que Q robur, avec un nombre plus élevé de plants présentant un dessèchement de l'appareil aérien ainsi qu'une plus forte chlorose des feuilles. Cette différence pourrait être due au plus grand nombre de racines adventives et de lenticelles hypertrophiées formées au collet de Q. robur. Dans les racines fines des deux espèces, l'activité pyruvate décarboxylase (PDC), enzyme clef de la fermentation alcoolique, a été stimulée après 24 h d'ennoyage. Les transcrits de PDC ont augmenté après 48 h d'ennoyage uniquement chez Q. robur. De façon intéressante, les transcrits d'hémoglobine (Hb) (qui pourrait être impliquée dans la voie de signalisation de l'oxyde nitreux), ont suivi le même profil de réponse que ceux de la PDC. Les enzymes du catabolisme du saccharose ont présenté une diminution d'activité après 3 semaines d'ennoyage, probablement consécutivement à une baisse de la disponibilité en hydrates de carbone. Les activités alcool-déshydrogénase (ADH), saccharose-synthase (Susy), et pyruvate-kinase (PK), ont été plus fortes après 3 semaines d'ennoyage. Cette étude a fourni des marqueurs caractérisant des différences inter-spécifiques de tolérance, qui pourront être utilisés lors d'études ultérieures de diversité intra et inter-spécifique de traits liés la tolérance à l'hypoxie racinaire.

Background E. coli B (BL21), unlike E.coli K-12 (JM109) is insensitive to glucose concentration and, therefore, grows faster and produces less acetate than E. coli K-12, especially when growing to high cell densities at high glucose concentration. By performing genomic analysis, it was demonstrated that the cause of this difference in sensitivity to the glucose concentration is the result of the differences in the central carbon metabolism activity. We hypothesized that the global transcription regulator Cra (FruR) is constitutively expressed in E. coli B and may be responsible for the different behaviour of the two strains. To investigate this possibility and better understand the function of Cra in the two strains, cra - negative E. coli B (BL21) and E. coli K-12 (JM109) were prepared and their growth behaviour and gene expression at high glucose were evaluated using microarray and real-time PCR. Results The deletion of the cra gene in E. coli B (BL21) minimally affected the growt...

Background The effect of high glucose concentration on the transcription levels of the small RNA SgrS and the messenger RNA ptsG, (encoding the glucose transporter IICBGlc), was studied in both E. coli K-12 (MG1655 and JM109) and E. coli B (BL21). It is known that the transcription level of sgrS increases when E. coli K-12 (MG1655 and JM109) is exposed to the non-metabolized glucose alpha methyl glucoside (αMG) or when the bacteria with a defective glycolysis pathway is grown in presence of glucose. The increased level of sRNA SgrS reduces the level of the ptsG mRNA and consequently lowers the level of the glucose transporter IICBGlc. The suggested trigger for this action is the accumulation of the corresponding phospho-sugars. Results In the course of the described work, it was found that E. coli B (BL21) and E. coli K-12 (JM109 and MG1655) responded similarly to αMG: both strains increased SgrS transcription and reduced ptsG transcription. However, the two strains reacted differen...

Transgenic tobacco (Nicotiana tabacum L.) plants expressing the 30-kDa movement protein of tobacco mosaic virus (TMV-MP) were employed to investigate the influence of a localized change in mesophyllbundle sheath plasmodesmal size exclusion limit on photosynthetic performance and on carbon metabolism and allocation. Under conditions of saturating irradiance, tobacco plants expressing the TMV-MP were found to have higher photosynthetic CO2-response curves compared with vector control plants. However, this difference was significant only in the presence of elevated CO2 levels. Photosynthetic measurements made in the greenhouse, under endogenous growth conditions, revealed that there was little difference between TMV-MP-expressing and control tobacco plants. However, analysis of carbon metabolites within source leaves where a TMV-MP-induced increase in plasmodesmal size exclusion limit had recently taken place established that the levels of sucrose, glucose, fructose and starch were considerably elevated above those present in equivalent control leaves. Although expression of the TMV-MP did not alter total plant biomass, it reduced carbon allocation to the lower region of the stem and roots. This difference in biomass distribution was clearly evident in the lower root-to-shoot ratios for the TMV-MP transgenic plants. Microinjection (dye-coupling) studies established that the TMV-MP-associated reduction in photosynthate delivery (allocation) to the roots was not due to a direct effect on root cortical plasmodesmata. Rather, this change appeared to result from an alteration in phloem transport from young source leaves in which the TMV-MP had yet to exert its influence over plasmodesmal size exclusion limits. These results are discussed in terms of the rate-limiting steps involved in sucrose movement into the phloem.

This study aimed to detect catalase [CAT] promoter haplotypes relation to DNA oxidation damage in depression disorder cases, using 8-Oxo-2'-deoxyadenosine [8-oxo-dG], Total Antioxidant [TAO] and Reactive Oxygen Species [ROS] levels, the results showed significant differences in BMI, ROS and TAO, non-sig 8-oxodG between groups. The haplotypes types showed nonsignificant changes [p=0.133]. ROS in patients showed high level in triple haplotype in nonsignificant elevation [p= 0.330]. There were slightly changes in TAO levels between haplotypes in patients [p=0.204].The 8-oxo-dG in patients with double haplotypes was higher in elevation [p=0.582]. Physical activity significant effected in the CAT haplotype [p=0.042]. No association of CAT haplotypes with depression, disease duration and family history.

Data deposition: The atomic coordinates and structure factors reported in this paper have been deposited in the Protein Data Bank, www.wwpdb.org (PDB ID codes 6H7G and 6H7H for CrPRK and AtPRK, respectively). The SEC-SAXS data of CrPRK reported in this paper have been deposited in the Small Angle Scattering Biological Data Bank, (ID code SASDDH9).

Average concentrations of glycolytic intermediates involved in the tryptophan (TRP) synthesis are of major importance in regulating and modulating the TRP synthesis. It is well known that oscillations of glycolytic intermediates often occur in living cells, according to the environment conditions and characteristics of the ATPrecovery system. As the TRP synthesis is connected to glycolysis through the PEP (phosphoenolpyruvate) node, the study of the major role played by the PEP glycolytic intermediate for the tryptophan (TRP) oscillatory metabolic synthesis is of high interest in industrial biosynthesis. Based on a reduced kinetic model for TRP synthesis from literature, this paper performs an in-silico analysis of the way by which the PEP level involved in the oscillatory glycolysis influences the TRP synthesis in E. coli bacteria. The analysis allows further TRP synthesis optimization.

The interaction between Candida albicans and cells of the innate immune system is a key determinant of disease progression. Transcriptional profiling has revealed that C. albicans has a complex response to phagocytosis, much of which is similar to carbon starvation. This suggests that nutrient limitation is a significant stress in vivo, and we have shown that glyoxylate cycle mutants are less virulent in mice. To examine whether other aspects of carbon metabolism are important in vivo during an infection, we have constructed strains lacking FOX2 and FBP1 , which encode key components of fatty acid β-oxidation and gluconeogenesis, respectively. As expected, fox2 Δ mutants failed to utilize several fatty acids as carbon sources. Surprisingly, however, these mutants also failed to grow in the presence of several other carbon sources, whose assimilation is independent of β-oxidation, including ethanol and citric acid. Mutants lacking the glyoxylate enzyme ICL1 also had more severe carbo...

salinity is one of the major environmental constraints on agriculture in many regions of the world (Boyer, 1982; Two cultivars of Cicer arietinum with differential toler- . Chick-pea (Cicer arietinum ance to salinity have been compared by analysing L.) nodules are indeterminate and export growth, photosynthesis, nodulation, nitrogenase activfixed N as amides and ureides (Schubert, 1986). This ity, and carbon metabolism in the nodule cytosol. The legume is considered to be sensitive to salt stress, which aim was to help elucidate the relationships between, affects its growth , on the one hand, sucrose and malate metabolism in nodulation and nitrogenase activity ( Elsheikh and Wood, nodules and, on the other, the inhibition of nitrogen 1990; , and photosynthesis and fixation under salt stress. Chick-pea cultivars Pedrocarbon metabolism in root nodules . Nevertheless, some authors have reported that nitrogen with Mesorhizobium ciceri strain Ch-191 were grown fixation in indeterminate nodules is more tolerant of salt in a controlled environmental chamber and were stress that in determinate ones (Bordeleau and Prevost, treated with salt (0, 50, 75, and 100 mM NaCl) from 1994; Sinclair and Serraj, 1995). sowing to harvest time (28 d). Plant growth and photo- The microsymbiont, i.e. the bacteroid, depends upon synthesis were more affected by salt in Pedrosillano than in ILC1919. Also the effect of salt on nodulation oxygen and photoassimilates supplied by the host plant. and nitrogen fixation was much more pronounced in The photosynthate translocated to the root nodules, Pedrosillano. The increase in nodular mass in ILC1919 mainly sucrose (Gordon et al., 1992), is metabolized to can partially counteract the inhibition of nitrogenase produce dicarboxylic acid, mainly malate, which is the activity. The enzymes of sucrose breakdown were preferred respiratory substrate for bacteroids ( Kim and inhibited by NaCl, but in ILC1919 a rise in alkaline Copeland , 1996). Although photosynthesis is inhibited invertase was observed with salinity, which could comby salt, limitation in the photosynthate supply to the pensate for the lack of the sucrose synthase hydrolytic nodules does not appear to inhibit nitrogenase, given that activity. The activity of PEPC was stimulated by salt in stress promotes an accumulation of soluble sugar in ILC1919. Also, this cultivar showed higher malate connodules ( Fouge `re et al., 1991;. In centrations in root nodules. addition, it has been observed that the reduction of nitrogenase activity under salt stress was associated with Key words: Chick-pea, salinity, nitrogen fixation, carbon the decrease in sucrose synthase (SS ) expression (Gordon metabolism, malate. et al., 1997). Another possible explanation for the inhibition of

The phenomenon of a midday decrease in quantum and all other efficiencies of photosynthesis has been acknowledged. Quantum efficiency of PSII in nettle is known to be lower at midday than in the morning or evening. On other hand, the parameters of induction of Chla fluorescence in stinging nettle (Urtica dioica L.) during the daytime were shown to depend on temperature alone. An increase in temperature also slowed down processes in reaction centres (RCs) and on the acceptor side of PSII, which showed its effect on electron transport in PSII and overall photosynthesis. A similar situation was found for red currant (Ribes spp.). The temperature effect was considered to be transient thermal inhibition of photosynthesis caused by diurnal changes in temperature.Razmatran je fenomen podnevne inhibicije kvantne efikasnosti i drugih aspekata fotosinteze. Parametri kvantne efikasnosti PSII kod koprive su niži u podne nego ujutro ili uveče. Nađeno je, u oglednim uslovima, da parametri indukci...

Na 1200 chlorofylmetingen, 80 als gevolg daarvan gesprongen reageerbuisjes, 2 spijkerbroeken met gaten door zwavelzuur-spetters, 800 fotosynthese-lichtcurves (met soms gekmakend kromme lijnen), een aantal herzen gehoorverhes door het ultrasone tnlapparaat, 357 meter slang, 4532 gebruikte pipettipjes 8 liter aceton, 4 liter methanol, 80 liter demi, 900 liter koffie, 67000 lichtflitsen, 6,78854 10 17 met de coulter-counter getelde deeltjes, 5 kapotte ballonpipetten, 23 overstromingen in de klimaatkamer, 380 koude rillingen in de vriescel, 20 keer verkeerd installeren van alle slangen en draadjes van de PAM-fluorometer, 380 grappen van Jan Peene, 440 fietstochtjes van Goes naar Yerseke, 440 fietstochtjes van Yerseke naar Goes, 10 als gevolg daarvan verloren kilo's, 12 bactenele infecties in de cultures, 35 gesprongen pompslangetjes, 80 vaten met medium en een halve hernia van het sjouwen ervan, 160 zoektochten naar altijd in gebruik zijnde karretjes, 300 dutoclaveerbeurten, 200 discussies met Jacco, 190 foutmeldingen van de spellingscheck vanwege het woord "spectrophotometncally", een twee jaar durende intensieve correspondentie met "Limnology & Oceanography", werkbezoeken aan Southampton,

Endosymbiotic bacteria from different species can live inside cells of the same eukaryotic organism. Metabolic exchanges occur between host and bacteria but also between different endocytobionts. Since a complete genome annotation is available for both, we built the metabolic network of two endosymbiotic bacteria, Sulcia muelleri and Baumannia cicadellinicola, that live inside specific cells of the sharpshooter Homalodisca coagulata and studied the metabolic exchanges involving transfers of carbon atoms between the three. We automatically determined the set of metabolites potentially exogenously acquired (seeds) for both metabolic networks. We show that the number of seeds needed by both bacteria in the carbon metabolism is extremely reduced. Moreover, only three seeds are common to both metabolic networks, indicating that the complementarity of the two metabolisms is not only manifested in the metabolic capabilities of each bacterium, but also by their different use of the same environment. Furthermore, our results show that the carbon metabolism of S. muelleri may be completely independent of the metabolic network of B. cicadellinicola. On the contrary, the carbon metabolism of the latter appears dependent on the metabolism of S. muelleri, at least for two essential amino acids, threonine and lysine. Next, in order to define which subsets of seeds (precursor sets) are sufficient to produce the metabolites involved in a symbiotic function, we used a graph-based method, PITUFO, that we recently developed. Our results highly refine our knowledge about the complementarity between the metabolisms of the two bacteria and their host. We thus indicate seeds that appear obligatory in the synthesis of metabolites are involved in the symbiotic function. Our results suggest both B. cicadellinicola and S. muelleri may be completely independent of the metabolites provided by the co-resident endocytobiont to produce the carbon backbone of the metabolites provided to the symbiotic system (i:e., thr and lys are only exploited by B. cicadellinicola to produce its proteins).

The coordinated expression of the genes involved in respiration in the photosynthetic cyanobacterium Synechocystis sp. PCC 6803 during the early period of glucose (Glc) treatment is poorly understood. When photoautotrophically grown cells were supplemented with 10 mm Glc in the light or after a dark adaptation period of 14 h, significant increases in the respiratory activity, as determined by NAD(P)H turnover, respiratory O2 uptake rate, and cytosolic alkalization, were observed. At the same time, the transcript levels of 18 genes coding for enzymes associated with respiration increased with differential induction kinetics; these genes were classified into three groups based on their half-rising times. Transcript levels of the four genes gpi, zwf, pdhB, and atpB started to increase along with a net increase in NAD(P)H, while the onset of net NAD(P)H consumption coincided with an increase in those of the genes tktA, ppc, pdhD, icd, ndhD2, ndbA, ctaD1, cydA, and atpE. In contrast, the...

Homeostasic balance of one-carbon metabolism is highly dependent upon derivatives of folate and is vital for the prevention of multiple disease states, including vascular disease, birth defects, neurological disorders, and cancer. Homocysteine, a non-protein forming amino acid, accumulates in the absence of 5-methyltetrahydrofolate and represents both a marker for reduced cellular methylation potential and elicitor of pathogenesis . In addition to cofactors like folate, vitamin B 6 , B 12 , and B 2 , various bioactive compounds are believed to influence the same cellular processes either directly or indirectly . The isoflavone genistein exhibits estrogen-like properties and has been implicated in the prevention of various diseases, including those involving hyperhomocysteinemia . Genistein has been shown to stably alter DNA methyltransferase and histone acetyltransferase activities independently of folate . This study was conducted to determine if genistein could modulate one-carbon metabolism during diet-induced folate deficiency. Specifically, we aimed to determine whether genistein is able to prevent hyperhomocysteinemia caused by moderate folate deficiency and classify any changes in relevant enzyme function, including those related to transmethylation, remethylation, and transsulfuration pathways, as well as alterations in genomic DNA methylation patterns. Administration of genistein (300 mg/kg diet) reduced plasma homocysteine concentrations in rats fed a folate-deficient diet in three independent studies without consistent alterations in enzyme activity or expression. Attenuation of plasma homocysteine was similar and occurred independently across each study at 10, 24, and 59 d of supplementation. There were no differences in genomic DNA methylation patterns

Background Saccharomyces cerevisiae and Pichia pastoris are two of the most relevant microbial eukaryotic platforms for the production of recombinant proteins. Their known genome sequences enabled several transcriptomic profiling studies under many different environmental conditions, thus mimicking not only perturbations and adaptations which occur in their natural surroundings, but also in industrial processes. Notably, the majority of such transcriptome analyses were performed using non-engineered strains. In this comparative study, the gene expression profiles of S. cerevisiae and P. pastoris, a Crabtree positive and Crabtree negative yeast, respectively, were analyzed for three different oxygenation conditions (normoxic, oxygen-limited and hypoxic) under recombinant protein producing conditions in chemostat cultivations. Results The major differences in the transcriptomes of S. cerevisiae and P. pastoris were observed between hypoxic and normoxic conditions, where the availabili...