Energy Demand For Maintenance Research Articles

Single nucleotide polymorphisms linked to mitochondrial uncoupling protein genes UCP2 and UCP3 affect mitochondrial metabolism and healthy aging in female nonagenarians.

Energy expenditure decreases with age, but in the oldest-old, energy demand for maintenance of body functions increases with declining health. Uncoupling proteins have profound impact on mitochondrial metabolic processes; therefore, we focused attention on mitochondrial uncoupling protein genes. Alongside resting metabolic rate (RMR), two SNPs in the promoter region of UCP2 were associated with healthy aging. These SNPs mark potential binding sites for several transcription factors; thus, they may affect expression of the gene. A third SNP in the 3'-UTR of UCP3 interacted with RMR. This UCP3 SNP is known to impact UCP3 expression in tissue culture cells, and it has been associated with body weight and mitochondrial energy metabolism. The significant main effects of the UCP2 SNPs and the interaction effect of the UCP3 SNP were also observed after controlling for fat-free mass (FFM) and physical-activity related energy consumption. The association of UCP2/3 with healthy aging was not found in males. Thus, our study provides evidence that the genetic risk factors for healthy aging differ in males and females, as expected from the differences in the phenotypes associated with healthy aging between the two sexes. It also has implications for how mitochondrial function changes during aging.

Metabolic network capacity of Escherichia coli for Krebs cycle-dependent proline hydroxylation.

BackgroundUnderstanding the metabolism of the microbial host is essential for the development and optimization of whole-cell based biocatalytic processes, as it dictates production efficiency. This is especially true for redox biocatalysis where metabolically active cells are employed because of the cofactor/cosubstrate regenerative capacity endogenous in the host. Recombinant Escherichia coli was used for overproducing proline-4-hydroxylase (P4H), a dioxygenase catalyzing the hydroxylation of free l-proline into trans-4-hydroxy-l-proline with a-ketoglutarate (a-KG) as cosubstrate. In this whole-cell biocatalyst, central carbon metabolism provides the required cosubstrate a-KG, coupling P4H biocatalytic performance directly to carbon metabolism and metabolic activity. By applying both experimental and computational biology tools, such as metabolic engineering and 13C-metabolic flux analysis (13C-MFA), we investigated and quantitatively described the physiological, metabolic, and bioenergetic response of the whole-cell biocatalyst to the targeted bioconversion and identified possible metabolic bottlenecks for further rational pathway engineering.ResultsA proline degradation-deficient E. coli strain was constructed by deleting the putA gene encoding proline dehydrogenase. Whole-cell biotransformations with this mutant strain led not only to quantitative proline hydroxylation but also to a doubling of the specific trans-4-l-hydroxyproline (hyp) formation rate, compared to the wild type. Analysis of carbon flux through central metabolism of the mutant strain revealed that the increased a-KG demand for P4H activity did not enhance the a-KG generating flux, indicating a tightly regulated TCA cycle operation under the conditions studied. In the wild type strain, P4H synthesis and catalysis caused a reduction in biomass yield. Interestingly, the ΔputA strain additionally compensated the associated ATP and NADH loss by reducing maintenance energy demands at comparably low glucose uptake rates, instead of increasing the TCA activity.ConclusionsThe putA knockout in recombinant E. coli BL21(DE3)(pLysS) was found to be promising for productive P4H catalysis not only in terms of biotransformation yield, but also regarding the rates for biotransformation and proline uptake and the yield of hyp on the energy source. The results indicate that, upon a putA knockout, the coupling of the TCA-cycle to proline hydroxylation via the cosubstrate a-KG becomes a key factor constraining and a target to further improve the efficiency of a-KG-dependent biotransformations.Electronic supplementary materialThe online version of this article (doi:10.1186/s12934-015-0298-1) contains supplementary material, which is available to authorized users.

Iron reduction by the deep-sea bacterium Shewanella profunda LT13a under subsurface pressure and temperature conditions.

Microorganisms influence biogeochemical cycles from the surface down to the depths of the continental rocks and oceanic basaltic crust. Due to the poor recovery of microbial isolates from the deep subsurface, the influence of physical environmental parameters, such as pressure and temperature, on the physiology and metabolic potential of subsurface inhabitants is not well constrained. We evaluated Fe(III) reduction rates (FeRRs) and viability, measured as colony-forming ability, of the deep-sea piezophilic bacterium Shewanella profunda LT13a over a range of pressures (0–125 MPa) and temperatures (4–37∘C) that included the in situ habitat of the bacterium isolated from deep-sea sediments at 4500 m depth below sea level. S. profunda LT13a was active at all temperatures investigated and at pressures up to 120 MPa at 30∘C, suggesting that it is well adapted to deep-sea and deep sedimentary environments. Average initial cellular FeRRs only slightly decreased with increasing pressure until activity stopped, suggesting that the respiratory chain was not immediately affected upon the application of pressure. We hypothesize that, as pressure increases, the increased energy demand for cell maintenance is not fulfilled, thus leading to a decrease in viability. This study opens up perspectives about energy requirements of cells in the deep subsurface.

Metabolic modeling of spatial heterogeneity of biofilms in microbial fuel cells reveals substrate limitations in electrical current generation

Microbial fuel cells (MFCs) have been proposed as an alternative energy resource for the conversion of organic compounds to electricity. In an MFC, microorganisms such as Geobacter sulfurreducens form an anode-associated biofilm that can completely oxidize organic matter (electron donor) to carbon dioxide with direct electron transfer to the anode (electron acceptor). Mathematical models are useful in analyzing biofilm processes; however, existing models rely on Nernst-Monod type expressions, and evaluate extracellular processes separated from the intracellular metabolism of the microorganism. Thus, models that combine both extracellular and intracellular components, while addressing spatial heterogeneity, are essential for improved representation of biofilm processes. The goal of this work is to develop a model that integrates genome-scale metabolic models with the model of biofilm environment. This integrated model shows the variations of electrical current production and biofilm thickness under the presence/absence of NH4 in the bulk solution, and under varying maintenance energy demands. Further, sensitivity analysis suggested that conductivity is not limiting electrical current generation and that increasing cell density can lead to enhanced current generation. In addition, the modeling results also highlight instances such as the transformation into respiring cells, where the mechanism of electrical current generation during biofilm development is not yet clearly understood.

The miRNAome of the postpartum dairy cow liver in negative energy balance.

BackgroundNegative energy balance (NEB) is an altered metabolic state in high yielding cows that occurs during the first few weeks postpartum when energy demands for lactation and maintenance exceed the energy supply from dietary intake. NEB can, in turn, lead to metabolic disorders and to reduced fertility. Alterations in the expression of more than 700 hepatic genes have previously been reported in a study of NEB in postpartum dairy cows. miRNAs (microRNA) are known to mediate many alterations in gene expression post transcriptionally. To study the hepatic miRNA content of postpartum dairy cows, including their overall abundance and differential expression, in mild NEB (MNEB) and severe NEB (SNEB), short read RNA sequencing was carried out. To identify putative targets of differentially expressed miRNAs among differentially expressed hepatic genes reported previously in dairy cows in SNEB computational target identification was employed.ResultsOur results indicate that the dairy cow liver expresses 53 miRNAs at a lower threshold of 10 reads per million. Of these, 10 miRNAs accounted for greater than 95% of the miRNAome (miRNA content). Of the highly expressed miRNAs, miR-122 constitutes 75% followed by miR-192 and miR-3596. Five out of thirteen let-7 miRNA family members are also among the highly expressed miRNAs. miR-143, down-regulated in SNEB, was found to have 4 putative up-regulated gene targets associated with SNEB including LRP2 (low density lipoprotein receptor-related protein 2), involved in lipid metabolism and up-regulated in SNEB.ConclusionsThis is the first liver miRNA-seq profiling study of moderate yielding dairy cows in the early postpartum period. Tissue specific miR-122 and liver enriched miR-192 are two of the most abundant miRNAs in the postpartum dairy cow liver. miR-143 is significantly down-regulated in SNEB and putative targets of miRNA-143 which are up-regulated in SNEB, include a gene involved in lipid metabolism.

Towards habitat-oriented systems biology of “Aromatoleum aromaticum” EbN1

The denitrifying betaproteobacterium "Aromatoleum aromaticum" EbN1 is a well-studied model organism for anaerobic degradation of aromatic compounds. Following publication of its genome in 2005, comprehensive physiological-proteomic studies were conducted to deduce functional understanding from the genomic blueprint. A catabolic network (85 predicted, 65 identified proteins) for anaerobic degradation of 24 aromatic growth substrates (including 11 newly recognized) was established. Newly elucidated pathways include those for 4-ethylphenol and plant-derived 3-phenylpropanoids, involving functional assignment of several paralogous genes. The substrate-specific regulation of individual peripheral degradation pathways is probably initiated by highly specific chemical sensing via dedicated sensory/regulatory proteins, e.g. three different σ⁵⁴-dependent one-component sensory/regulatory proteins are predicted to discriminate between three phenolic substrates (phenol, p-cresol and 4-ethylphenol) and two different two-component systems are assumed to differentiate between two alkylbenzenes (toluene, ethylbenzene). Investigations under in situ relevant growth conditions revealed (a) preferred utilization of benzoate from a mixture with succinate results from repressed synthesis of a C₄-dicarboxylate TRAP transporter; (b) response to alkylbenzene-induced solvent stress comprises metabolic re-routing of acetyl-CoA and reducing equivalents to poly(3-hydroxybutyrate) synthesis, alteration of cellular membrane composition and formation of putative solvent efflux systems; and (c) multifaceted adaptation to slow growth includes adjustment of energy demand for maintenance and preparedness for future nutritional opportunities, i.e. provision of uptake systems and catabolic enzymes for multiple aromatic substrates despite their absence. This broad knowledge base taken together with the recent development of a genetic system will facilitate future functional, biotechnological (stereospecific dehydrogenases) and habitat re-enacting ("eco-"systems biology) studies with "A. aromaticum" EbN1.

Self‐Replenishing Dual Structured Superhydrophobic Coatings Prepared by Drop‐Casting of an All‐In‐One Dispersion

Robust dual structured superhydrophobic coatings which replenish spontaneously their surface chemical composition on new multi‐scale structured surfaces, recreated upon damage, are described. The surface repair occurs at room temperature, via intrinsic elements of the coatings, all covalently bonded. These coatings can be prepared from all‐in‐one dispersions by a simple drop‐cast method, with different thicknesses and on various substrates. The critical factors to optimize the self‐replenishment are described and three main design principles are postulated. The superhydrofobicity of the coatings is maintained even after 500 abrasion cycles. The principles reported can be extended towards self‐healing other surface‐dependent functionalities, that is, anti‐bacteria, anti‐fouling, or drag‐reduction, which will maintain high performance levels all through their life‐cycle with low cost and energy demand for maintenance and surface repair.

Thermal adaptation of decomposer communities in warming soils

Temperature regulates the rate of biogeochemical cycles. One way it does so is through control of microbial metabolism. Warming effects on metabolism change with time as physiology adjusts to the new temperature. I here propose that such thermal adaptation is observed in soil microbial respiration and growth, as the result of universal evolutionary trade-offs between the structure and function of both enzymes and membranes. I review the basis for these trade-offs and show that they, like substrate depletion, are plausible mechanisms explaining soil respiration responses to warming. I argue that controversies over whether soil microbes adapt to warming stem from disregarding the evolutionary physiology of cellular metabolism, and confusion arising from the term thermal acclimation to represent phenomena at the organism- and ecosystem-levels with different underlying mechanisms. Measurable physiological adjustments of the soil microbial biomass reflect shifts from colder- to warmer-adapted taxa. Hypothesized declines in the growth efficiency of soil microbial biomass under warming are controversial given limited data and a weak theoretical basis. I suggest that energy spilling (aka waste metabolism) is a more plausible mechanism for efficiency declines than the commonly invoked increase in maintenance-energy demands. Energy spilling has many fitness benefits for microbes and its response to climate warming is uncertain. Modeled responses of soil carbon to warming are sensitive to microbial growth efficiency, but declines in efficiency mitigate warming-induced carbon losses in microbial models and exacerbate them in conventional models. Both modeling structures assume that microbes regulate soil carbon turnover, highlighting the need for a third structure where microbes are not regulators. I conclude that microbial physiology must be considered if we are to have confidence in projected feedbacks between soil carbon stocks, atmospheric CO2, and climate change.

Different O3 response of sensitive and resistant snap bean genotypes (Phaseolus vulgaris L.): The key role of growth stage, stomatal conductance, and PSI activity

The ozone (O3) biomonitoring system based on sensitive (S156) and resistant (R123) genotypes of snap bean (Phaseolus vulgaris L.), was tested in a fumigation experiment, with the aim of: (i) assessing the performance of the S156/R123 system under simulated climatic conditions and O3 levels often found in Mediterranean sites; (ii) contributing to identify possible ecophysiological mechanisms determining the different O3 sensitivity of S156 and R123, along different plant growth stages. In contrast with what reported by other authors, differences in stomatal O3 uptake were evident between sensitive and resistant plants. In particular, R123 plants showed an O3-induced stomatal closure (−38.1% than the control) during flowering and the onset of visible injury, a behaviour that can be regarded as an O3 avoidance mechanism. At the end of the fumigation period (AOT40=4.66ppmh), despite the reduction of assimilation and the photoprotective down regulation of PSII photochemistry, the capacity to reduce the final electron acceptors beyond PSI was up-regulated, particularly in S156, while the PSI activity remained constant in both genotypes, an unusual response to O3 stress. Increased energy demand for maintenance and repair processes also determined increased dark respiration rates (Rd) in all fumigated plants; nocturnal stomatal conductance (gsN) was also enhanced, differently in the two genotypes, with possible implication for higher nocturnal stomatal uptake of S156 in field conditions. No clear genotype×O3 effect was instead detected on pod biomass, although a significant O3-induced reduction of yield was evident in both genotypes.

Ethanol reduces mitochondrial membrane integrity and thereby impacts carbon metabolism of Saccharomyces cerevisiae

Saccharomyces cerevisiae is an excellent ethanol producer, but is rather sensitive to high concentration of ethanol. Here, influences of ethanol on cellular membrane integrity and carbon metabolism of S.cerevisiae were investigated to rationalize mechanism involved in ethanol toxicity. Addition of 5% (v/v) ethanol did neither significantly change the permeability of the cytoplasmic membrane of the reference strain S.cerevisiae BY4741 nor of the ethanol-tolerant strain iETS3. However, the addition of ethanol resulted in a marked decrease in the mitochondrial membrane potential and in increased concentrations of intracellular reactive oxygen species (ROS). The carbon flux was redistributed under these conditions from mainly ethanol production to the TCA cycle. This redistribution was possibly a result of increased energy demand for cell maintenance that increased from about zero to 20-40mmolATP(g(CDW) h)(-1) . This increase in maintenance energy might be explained by the ethanol-induced reduction of the proton motive force and the required removal of ROS. Thus, the stability of the mitochondrial membrane and subsequently the capacity to keep ROS levels low could be important factors to improve tolerance of S.cerevisiae against ethanol.

Physiological and Proteomic Adaptation of “Aromatoleum aromaticum” EbN1 to Low Growth Rates in Benzoate-Limited, Anoxic Chemostats

"Aromatoleum aromaticum" EbN1 was cultivated at different growth rates in benzoate-limited chemostats under nitrate-reducing conditions. Physiological characteristics, proteome dynamics, phospholipid-linked fatty acid (PLFA) composition, and poly(3-hydroxybutyrate) (PHB) content were analyzed in steady-state cells at low (μ(low)) (0.036 h(-1)), medium (μ(med)) (0.108 h(-1)), and high (μ(high)) (0.180 h(-1)) growth rates. A positive correlation to growth rate was observed for cellular parameters (cell size, and DNA and protein contents). The free energy consumed for biomass formation steadily increased with growth rate. In contrast, the energy demand for maintenance increased only from μ(low) to μ(med) and then remained constant until μ(high). The most comprehensive proteomic changes were observed at μ(low) compared to μ(high). Uniformly decreased abundances of protein components of the anaerobic benzoyl coenzyme A (benzoyl-CoA) pathway, central carbon metabolism, and information processing agree with a general deceleration of benzoate metabolism and cellular processes in response to slow growth. In contrast, increased abundances were observed at μ(low) for diverse catabolic proteins and components of uptake systems in the absence of the respective substrate (aromatic or aliphatic compounds) and for proteins involved in stress responses. This potential catabolic versatility and stress defense during slow growth may be interpreted as preparation for future needs.

Adapting bioenergetic factorial modelling to understand the implications of heat stress on barramundi (Lates calcarifer) growth, feed utilisation and optimal protein and energy requirements - potential strategies for dealing with climate change?

The implications of temperature on bioenergetics for barramundi (Lates calcarifer) were defined in an improved factorial model that encompassed revised parameters accounting for effects over the temperature range of 16–39 °C and size range of 10–3000 g. A revised growth function describing weight gain by barramundi as a function of fish weight and temperature was derived from farm and laboratory data and included a term for a shift in optimal temperature with fish size: Gain (g fish−1 day−1) = (K + xT + yT 2 + zT3) * (weight)ax+b. Maintenance energy and protein demand functions were also derived on a similar form, and all three functions combined to form the basis of a factorial model for energy and protein demand. Using this model, optimal iterative feed specifications were defined for a range of fish sizes at temperatures of 25, 30 and 35 °C. A feed demand model was also developed based on the demand for digestible energy (DE) at each of these temperatures. The model shows that at high temperatures (35 °C), there is an increase in digestible protein (DP) to DE demand, and that with increasing size, there is a decrease in the DP to DE demand.

Seasonal and nutrients intake regulation of lipoprotein lipase (LPL) activity in grazing yak ( Bos grunniens) in the Alpine Regions around Qinghai Lake

Lipoprotein lipase (LPL) is considered as a key enzyme in the lipid deposition and metabolism in tissues. To better understand fat cycling in grazing yak to adapt to the harsh environment on the Qinghai-Tibetan Plateau, and we have therefore explored seasonal and nutritional regulation of LPL in adipose tissue, liver and skeletal muscle. Sixty 3 year old growing yaks (BW. 120.3 ± 15.28 kg) were subdivided into six groups, each used to determine effects on chemical body composition and LPL activity in different tissues. The alpine pastures had the highest Crude protein (CP, 11.06%) contents and the gross energy (GE, 11.49 MJ/kg) in summer. Growth rates and body fat content were responsive to CP and GE intake regimen. Late spring up-regulation of LPL activity in the subcutaneous adipose tissue was consistent with a pronounced increase of body weight (BW) and whole body fat content. The highest LPL activity in the skeletal muscle was found in September (774 ± 64.1 mU/g tissue), which may serve to cover the increased energy demands for compensatory growth and maintenance of adiposity in the coming cold season. Furthermore, the seasonal regulation of LPL involves some factors in addition to insulin and triglycerides (TG). These results suggest that yaks could rely, in part, on LPL activity to adapt to the harsh forage environment. During the growing season, an enhanced synthesis of LPL production in the adipose tissues along with mechanisms for the recycling of fat contributes toward the rapid recovery of body weight.

Development of a nutritional model to define the energy and protein requirements of tilapia, Oreochromis niloticus

This study assessed the protein and energy requirements of tilapia ( Oreochromis niloticus) using a bioenergetic factorial modelling approach. Several model parameters were examined using a series of inter-related studies to define key factorial parameters. Studies included both controlled laboratory experiments and field-data collection from commercial farms in the HaiDuong province of northern Vietnam. The model includes parameters for maintenance energy (25.9 kJ/kg 0.80/d − at 28 °C) and protein (0.45 g/kg − 0.80 /d at 28 °C) demands, the effect of animal live-weight on growth and maintenance energy demands, the efficiencies of energy and protein utilisation at various energy and protein intake levels and the variability in whole body composition with varying live-weights. The energy (0.440 • [Energy intake] − 15.45) utilisation efficiencies were substantially lower than that observed from carnivorous fish species, but similar to other omnivores, but the protein utilisation efficiencies were similar to that seen in most other fish species (0.520 • [Protein intake] − 0.284). Using this modelling approach it was possible to iteratively derive optimal dietary protein and energy specifications for this species.

A factorial approach to defining the energy and protein requirements of Tra Catfish, Pangasianodon hypothalamus

This study assessed the protein and energy requirements of Tra catfish (Pangasianodon hypothalamus) using a bio-energetic factorial approach. To examine these parameters, a series of inter-related studies were undertaken to define the various factorial parameters. Studies included controlled laboratory experiments and field-data collection from commercial farms in the Mekong Delta region of Vietnam. Key components of the model include parameters for maintenance energy (39.7 kJ kg−0.844 day−1 at 32 °C) and protein (0.467 g kg−0.833 day−1 at 32 °C) demands, the effect of animal live-weight and temperature on growth and maintenance energy demands, the efficiencies of energy and protein utilization at various energy and protein intake levels and the variability in whole body composition with varying live-weight. Interestingly, both the energy [0.510· (Energy intake) − 11.143] and protein [0.322·(Protein intake) + 0.665] utilization efficiencies at near satiety feed intake levels were substantially lower than that observed from other fish species. Modelling using these equations was used to iteratively derive optimal diet designs and feeding rations for growing Pangasius catfish.

Application of physiologically based modelling and transcriptomics to probe the systems toxicology of aldicarb for Caenorhabditis elegans (Maupas 1900)

The toxicity of aldicarb on movement, life cycle, population growth rate and resource allocation, and the gene expression changes underpinning these effects, were investigated for Caenorhabditis elegans. A clear effect of aldicarb on nematode movement was found suggesting that this pesticide acts as a neurotoxicant. Aldicarb also had an effect on life cycle traits including low concentration life-span extension; high concentration brood size reduction and a high concentration extension of time to first egg. All life-cycle and growth data were integrated into a biology-based model (DEBtox) to characterise aldicarb effects on life-history traits, resource allocation and population growth rate within a single modelling framework. The DEBtox fits described concentration dependent effects on individual traits and population growth rate and indicated that the most probable mechanism of action of the pesticide was an increase in energy demands for somatic and reproductive tissue maintenance. Transcriptomic profiling indicated that aldicarb was associated with changes in amino acid metabolism, DNA structure, fatty acid metabolism and cytochrome P450 mediated xenobiotic metabolism. The changes in the amino acid and fatty acid pathways suggest an effect of aldicarb on protein integrity; while effects on DNA suggests that aldicarb influence DNA morphology or replication. Both these effects have the potential to incur increased costs for structural maintenance of macromolecules. These effects, coupled to the effect on biotransformation enzymes also seen, represent the materialisation of the maintenance costs indicated by DEBtox modelling.Electronic supplementary materialThe online version of this article (doi:10.1007/s10646-010-0591-z) contains supplementary material, which is available to authorized users.

Bioenergetic function in cardiovascular cells: The importance of the reserve capacity and its biological regulation

The ability of the cell to generate sufficient energy through oxidative phosphorylation and to maintain healthy pools of mitochondria are critical for survival and maintenance of normal biological function, especially during periods of increased oxidative stress. Mitochondria in most cardiovascular cells function at a basal level that only draws upon a small fraction of the total bioenergetic capability of the organelle; the apparent respiratory state of mitochondria in these cells is often close to state 4. The difference between the basal and maximal activity, equivalent to state 3, of the respiratory chain is called the reserve capacity. We hypothesize that the reserve capacity serves the increased energy demands for maintenance of organ function and cellular repair. However, the factors that determine the volume of the reserve capacity and its relevance to biology are not well understood. In this study, we first examined whether responses to 4-hydroxynonenal (HNE), a lipid peroxidation product found in atherosclerotic lesions and the diseased heart, differ between vascular smooth muscle cells, adult mouse cardiomyocytes, and rat neonatal cardiomyocytes. In both types of cardiomyocytes, oxygen consumption increased after HNE treatment, while oxygen consumption in smooth muscle cells decreased. The increase in oxygen consumption in cardiomyocytes decreased the reserve capacity and shifted the apparent respiratory state closer to state 3. Neonatal rat cardiomyocytes respiring on pyruvate alone had a fourfold higher reserve capacity than cells with glucose as the sole substrate, and these cells were more resistant to mitochondrial dysfunction induced by 4-HNE. The integration of the concepts of reserve capacity and state-apparent are discussed along with the proposal of two potential models by which mitochondria respond to stress.

Performance of the auxotrophic Saccharomyces cerevisiae BY4741 as host for the production of IL-1β in aerated fed-batch reactor: role of ACA supplementation, strain viability, and maintenance energy

BackgroundSaccharomyces cerevisiae BY4741 is an auxotrophic commonly used strain. In this work it has been used as host for the expression and secretion of human interleukin-1β (IL1β), using the cell wall protein Pir4 as fusion partner. To achieve high cell density and, consequently, high product yield, BY4741 [PIR4-IL1β] was cultured in an aerated fed-batch reactor, using a defined mineral medium supplemented with casamino acids as ACA (auxotrophy-complementing amino acid) source. Also the S. cerevisiae mutant BY4741 Δyca1 [PIR4-IL1β], carrying the deletion of the YCA1 gene coding for a caspase-like protein involved in the apoptotic response, was cultured in aerated fed-batch reactor and compared to the parental strain, to test the effect of this mutation on strain robustness. Viability of the producer strains was examined during the runs and a mathematical model, which took into consideration the viable biomass present in the reactor and the glucose consumption for both growth and maintenance, was developed to describe and explain the time-course evolution of the process for both, the BY4741 parental and the BY4741 Δyca1 mutant strain.ResultsOur results show that the concentrations of ACA in the feeding solution, corresponding to those routinely used in the literature, are limiting for the growth of S. cerevisiae BY4741 [PIR4-IL1β] in fed-batch reactor. Even in the presence of a proper ACA supplementation, S. cerevisiae BY4741 [PIR4-IL1β] did not achieve a high cell density. The Δyca1 deletion did not have a beneficial effect on the overall performance of the strain, but it had a clear effect on its viability, which was not impaired during fed-batch operations, as shown by the kd value (0.0045 h-1), negligible if compared to that of the parental strain (0.028 h-1). However, independently of their robustness, both the parental and the Δyca1 mutant ceased to grow early during fed-batch runs, both strains using most of the available carbon source for maintenance, rather than for further proliferation. The mathematical model used evidenced that the energy demand for maintenance was even higher in the case of the Δyca1 mutant, accounting for the growth arrest observed despite the fact that cell viability remained comparatively high.ConclusionsThe paper points out the relevance of a proper ACA formulation for the outcome of a fed-batch reactor growth carried out with S. cerevisiae BY4741 [PIR4-IL1β] strain and shows the sensitivity of this commonly used auxotrophic strain to aerated fed-batch operations. A Δyca1 disruption was able to reduce the loss of viability, but not to improve the overall performance of the process. A mathematical model has been developed that is able to describe the behaviour of both the parental and mutant producer strain during fed-batch runs, and evidence the role played by the energy demand for maintenance in the outcome of the process.

Effect of stocking density and feeding level on energy expenditure and stress responsiveness in European sea bass Dicentrarchus labrax

European sea bass (initial weight 72 ± 4 g) were stocked in 200-L tanks at two densities: a low density (LD) ∼ 5.5 kg m − 3 and a high density (HD) ∼ 36 kg m − 3 . The tanks were part of a recirculation system and were equipped to carry out frequent oxygen measurements. At each density the fish were fed at increasing levels from around maintenance requirement up to apparent satiation. The experiment was carried out for 10 weeks at 22 °C, after which the density in the LD treatment had increased to ∼ 10 kg m − 3 and to ∼ 60 kg m − 3 in the HD treatment. At the end of the trial blood samples were taken from several fish to determine the basal levels of cortisol and glucose. Furthermore, to assess the responsiveness to an acute stressor, additional fish were subjected to individual confinement in submerged nets, blood was sampled and cortisol and glucose analysed. At the end of the trial there was no significant difference in growth performance and voluntary feed intake between the groups raised at different densities. The partitioning of energy demand for maintenance and growth highlighted a slightly higher energy maintenance requirement in the LD fish (50.9 kJ (kg) 0.80 day − 1 ) compared to the HD groups (43.15 kJ (kg) 0.80 day − 1 ). In accordance with this, an increased oxygen demand for sea bass kept at the low density was detected through weekly measurements. Analyses of the blood parameters showed, that higher stocking density resulted in higher cortisol levels in both control and stressed groups (after netting), but the effect of stocking density on the acute stress response was less pronounced at the higher feeding level.

Plastic and adaptive responses of plant respiration to changes in atmospheric CO2 concentration

The concentration of atmospheric CO2 has increased from below 200 microl l(-1) during last glacial maximum in the late Pleistocene to near 280 microl l(-1) at the beginning of the Holocene and has continuously increased since the onset of the industrial revolution. Most responses of plants to increasing atmospheric CO2 levels result in increases in photosynthesis, water use efficiency and biomass. Less known is the role that respiration may play during adaptive responses of plants to changes in atmospheric CO2. Although plant respiration does not increase proportionally with CO2-enhanced photosynthesis or growth rates, a reduction in respiratory costs in plants grown at subambient CO2 can aid in maintaining a positive plant C-balance (i.e. enhancing the photosynthesis-to-respiration ratio). The understanding of plant respiration is further complicated by the presence of the alternative pathway that consumes photosynthate without producing chemical energy [adenosine triphosphate (ATP)] as effectively as respiration through the normal cytochrome pathway. Here, we present the respiratory responses of Arabidopsis thaliana plants selected at Pleistocene (200 microl l(-1)), current Holocene (370 microl l(-1)), and elevated (700 microl l(-1)) concentrations of CO2 and grown at current CO2 levels. We found that respiration rates were lower in Pleistocene-adapted plants when compared with Holocene ones, and that a substantial reduction in respiration was because of reduced activity of the alternative pathway. In a survey of the literature, we found that changes in respiration across plant growth forms and CO2 levels can be explained in part by differences in the respiratory energy demand for maintenance of biomass. This trend was substantiated in the Arabidopsis experiment in which Pleistocene-adapted plants exhibited decreases in respiration without concurrent reductions in tissue N content. Interestingly, N-based respiration rates of plants adapted to elevated CO2 also decreased. As a result, ATP yields per unit of N increased in Pleistocene-adapted plants compared with current CO2 adapted ones. Our results suggest that mitochondrial energy coupling and alternative pathway-mediated responses of respiration to changes in atmospheric CO2 may enhance survival of plants at low CO2 levels to help overcome a low carbon balance. Therefore, increases in the basal activity of the alternative pathway are not necessarily associated to metabolic plant stress in all cases.