Aerobic Production Research Articles

The impact of MIG1 and/or MIG2 disruption on aerobic metabolism of succinate dehydrogenase negative Saccharomyces cerevisiae

The zinc finger proteins Mig1 and Mig2 play important roles in glucose repression of Saccharomyces cerevisiae. To investigate whether the alleviation of glucose effect would result in an increase in aerobic succinate production, MIG1 and/or MIG2 were disrupted in a succinate dehydrogenase (SDH)-negative S. cerevisiae strain. Moreover, their impacts on physiology of the SDH-negative S. cerevisiae strain were studied under fully aerobic conditions when glucose was the sole carbon source. Our results showed that the succinate production for the SDH-negative S. cerevisiae was very low even under fully aerobic conditions. Furthermore, deletion of MIG1 and/or MIG2 did not result in an increase in succinate production in the SDH-negative S. cerevisiae strain. However, the synthesis of acetate was significantly affected by MIG1 deletion or in combination with MIG2 deletion. The acetate production for the mig1/mig2 double mutant BS2M was reduced by 69.72% compared to the parent strain B2S. In addition, the amount of ethanol produced by BS2M was slightly decreased. With the mig2 mutant BSM2, the concentrations of pyruvate and glycerol were increased by 26.23% and 15.28%, respectively, compared to the parent strain B2S.

The sunny side of greenhouse gas emissions – quantifying the contribution of aerobic methane production to global methane budgets

The sunny side of greenhouse gas emissions – quantifying the contribution of aerobic methane production to global methane budgets

Flow of energy in the outer retina in darkness and in light

Structural features of neurons create challenges for effective production and distribution of essential metabolic energy. We investigated how metabolic energy is distributed between cellular compartments in photoreceptors. In avascular retinas, aerobic production of energy occurs only in mitochondria that are located centrally within the photoreceptor. Our findings indicate that metabolic energy flows from these central mitochondria as phosphocreatine toward the photoreceptor's synaptic terminal in darkness. In light, it flows in the opposite direction as ATP toward the outer segment. Consistent with this model, inhibition of creatine kinase in avascular retinas blocks synaptic transmission without influencing outer segment activity. Our findings also reveal how vascularization of neuronal tissue can influence the strategies neurons use for energy management. In vascularized retinas, mitochondria in the synaptic terminals of photoreceptors make neurotransmission less dependent on creatine kinase. Thus, vasculature of the tissue and the intracellular distribution of mitochondria can play key roles in setting the strategy for energy distribution in neurons.

The Role of Oxygen in Cardiac Arrest Resuscitation

The heart is incapable of storing significant oxygen or substrates and thus is entirely dependent on a continuous delivery of flow in order to support its high metabolic state. Following cardiac arrest, myocardial tissue oxygen tension falls rapidly and aerobic production of ATP ceases. Without re-oxygenation of the ischemic myocardium, return of spontaneous circula-tion (ROSC) cannot be achieved. The oxygen paradox which has been described regarding other ischemia-reperfusion conditions seems to have application in cardiac arrest. It is clear that some level of oxygenation is necessary to achieve ROSC, however post ROSC there appears to be increased toxicity associated with hyperoxia. The optimal conditions for re-oxygenation in the setting of cardiac arrest remain ill defined at present.

Catalase Overexpression Reduces Lactic Acid-Induced Oxidative Stress in Saccharomyces cerevisiae

Industrial production of lactic acid with the current pyruvate decarboxylase-negative Saccharomyces cerevisiae strains requires aeration to allow for respiratory generation of ATP to facilitate growth and, even under nongrowing conditions, cellular maintenance. In the current study, we observed an inhibition of aerobic growth in the presence of lactic acid. Unexpectedly, the cyb2Delta reference strain, used to avoid aerobic consumption of lactic acid, had a specific growth rate of 0.25 h(-1) in anaerobic batch cultures containing lactic acid but only 0.16 h(-1) in identical aerobic cultures. Measurements of aerobic cultures of S. cerevisiae showed that the addition of lactic acid to the growth medium resulted in elevated levels of reactive oxygen species (ROS). To reduce the accumulation of lactic acid-induced ROS, cytosolic catalase (CTT1) was overexpressed by replacing the native promoter with the strong constitutive TPI1 promoter. Increased activity of catalase was confirmed and later correlated with decreased levels of ROS and increased specific growth rates in the presence of high lactic acid concentrations. The increased fitness of this genetically modified strain demonstrates the successful attenuation of additional stress that is derived from aerobic metabolism and may provide the basis for enhanced (micro)aerobic production of organic acids in S. cerevisiae.

The role of ultraviolet radiation, photosensitizers, reactive oxygen species and ester groups in mechanisms of methane formation from pectin

Ultraviolet (UV) radiation has recently been demonstrated to drive an aerobic production of methane (CH(4)) from plant tissues and pectins, as do agents that generate reactive oxygen species (ROS) in vivo independently of UV. As the major building-blocks of pectin do not absorb solar UV found at the earth's surface (i.e. >280 nm), we explored the hypothesis that UV radiation affects pectin indirectly via generation of ROS which themselves release CH(4) from pectin. Decreasing the UV absorbance of commercial pectin by ethanol washing diminished UV-dependent CH(4) production, and this was restored by the addition of the UV photosensitizer tryptophan. Certain ROS scavengers [mannitol, a hydroxyl radical ((*)OH) scavenger; 1,4-diazabicyclo[2.2.2] octane; and iodide] strongly inhibited UV-induced CH(4) production from dry pectin. Furthermore, pectin solutions emitted CH(4) in darkness upon the addition of (*)OH, but not superoxide or H(2)O(2). Model carbohydrates reacted similarly if they possessed -CH(3) groups [e.g. methyl esters or (more weakly) acetyl esters but not rhamnose]. We conclude that UV evokes CH(4) production from pectic methyl groups by interacting with UV photosensitizers to generate (*)OH. We suggest that diverse processes generating (*)OH could contribute to CH(4) emissions independently of UV irradiation, and that environmental stresses and constitutive physiological processes generating ROS require careful evaluation in studies of CH(4) formation from foliage.

Aerobic bioethenol production by Rhizomucor pusillus

Aerobic bioethenol production by Rhizomucor pusillus

Kinetic and metabolic aspects of Defluviicoccus vanus-related organisms as competitors in EBPR systems

A reactor was successfully enriched (90% as shown by Fluorescence in situ Hybridization) in Defluviicoccus vanus-related organisms presenting a Glycogen Accumulating Organisms (GAO) phenotype. Initial batch tests were performed using anaerobic/aerobic conditions to assess the capacity of different carbon sources utilization frequently abundant in wastewater: acetate, propionate, butyrate, valerate and glucose. Acetate and propionate were totally consumed in the anaerobic phase as well as butyrate and valerate, though these last ones with a very low consumption rate. All substrates were converted to polyhydroxyalkanoates (PHA). Glucose had a very slight anaerobic consumption but failed to disclose a typical GAO phenotype. In aerobic conditions, again all carbon sources were readily consumed except for glucose, with acetate and propionate having the higher consumption rates. Therefore, glucose seems not be used by this type of organisms. Acetate and propionate consumption rates indicated that these GAOs could reveal good competition advantages in EBPR systems where these carbon sources are available, especially propionate. Volatile Fatty Acid (VFA) uptake in aerobic phase and consequential PHA production indicate these organisms as possible candidates for PHA production.

Effect of vitamin concentration on the synthesis of lactate, ethanol, pyruvate, and ethyl acetate in cells of the yeast Dipodascus magnusii

In the yeast Dipodascus magnusii, which is auxotrophic for thiamine and biotin, during cultivation on glucose with excessive thiamine concentration, pyruvate metabolism was shown to result in the synthesis of fermentation products, namely, ethanol and, to a lesser extent, lactate. Substantial synthesis of ethyl acetate was also observed under these conditions. Introduction of nicotinic acid (NA) into the medium resulted in time separation of ethanol and lactate production. It was shown that cultivation of the yeast under biotin deficiency resulted in nearly complete suppression of aerobic production of ethanol and cessation of ethyl acetate synthesis, whereas lactate synthesis was activated as early as in the first hours of cultivation. Upon introduction of NA under these conditions, lactate concentration sharply increased. These results show that the combination of thiamine and biotin with other vitamins can stimulate utilization of the pyruvate pool in yeasts towards formation of considerable amounts of lactate, which is typical only of cells of higher eukaryotes and bacteria.

Effects of chloramphenicol on brain energy metabolism using 31P spectroscopy: influences on sleep‐wake states in rat

Effects of chloramphenicol (antibiotic inhibiting complex-1 of respiratory chain) and thioamphenicol (TAP, a structural analog of CAP inactive on complex-1) were examined on cerebral energy metabolites and sleep-wake cycle architecture in rat. In the first group, animals were chronically equipped with a cranial surface resonator and (31)P spectroscopic measurements were performed using a 2 T magnetic resonance spectrometer (operating frequency 34.46 MHz). CAP administration (400 mg/kg, tail vein, light period) induced deficits in phosphocreatine (-30%, p < 0.01) and ATP (-40%, p < 0.01), whereas TAP (400 mg/kg) had no effect. In the second group, animals were chronically implanted with polygraphic electrodes for EEG and electromyogram recordings. CAP administered intraperitoneally at light-onset reduced rapid-eye movement (REM) sleep (-60% in the first 6 h of light period, p < 0.01), increased waking state (+65% in the first 6 h of light period, p < 0.01), and slightly affected slow-wave sleep (SWS). During waking state, theta and sigma power bands of the EEG were, respectively, increased and decreased (p < 0.05). During SWS, delta power band was reinforced (p < 0.05), while theta, alpha, and sigma bands were decreased (p < 0.05). No changes occurred during REM sleep. TAP had no effect on sleep-wake states and spectral components of the EEG. Overall, these data indicate that REM sleep occurrence is linked to an aerobic production of ATP.

Aerobic production of methane in the sea

Methane is a potent greenhouse gas that has contributed approximately 20% to the Earth’s warming since pre-industrial times. The world’s oceans are an important source of methane, comprising 1–4% of annual global emissions. But despite its global significance, oceanic methane production is poorly understood. In particular, methane concentrations in the surface waters of most of the world’s oceans are supersaturated with respect to atmospheric concentrations, but the origin of this methane, which has been thought to be produced exclusively in anaerobic environments, is not known. Here, we measure methane production in seawater samples amended with methylphosphonate, an organic, phosphorus-containing compound. We show that methane is produced aerobically as a by-product of methylphosphonate decomposition in phosphate-stressed waters. Methylphosphonate decomposition, and thus methane production, may be enhanced by the activity of nitrogen-fixing microorganisms. We suggest that aerobic marine methane production will be sensitive to the changes in water-column stratification and nutrient limitation that are likely to result from greenhouse-gas-induced ocean warming. Surface waters of most of the world’s oceans are supersaturated with respect to atmospheric methane. Measurements in seawater samples suggest that an aerobic methane production pathway, which involves the decomposition of phosphorus-containing organic compounds, may be responsible.

Oral Amino Acid Supplements Improve Exercise Capacities in Elderly Patients with Chronic Heart Failure

We investigated whether 30 days of oral supplementation with a special mixture of amino acids (AAs), together with conventional therapy, could improve exercise capacity in elderly outpatients with chronic heart failure (CHF). A group of 95 outpatients (12 women and 83 men; New York Heart Association class II-III) aged 65-74 years were studied. This was a randomized, double-blind, placebo-controlled study. The patients performed a basal exercise test and were then randomly assigned to a special oral nutritional mixture of AAs 4 g twice daily (n = 43) or placebo (n = 42). After 30 days we repeated the exercise test. In both tests we measured the following: oxygen consumption (VO2), CO2 production (VCO2), minute ventilation (VE), oxygen cost of ventilation (VO2/VE), CO2 elimination per liter of ventilation (VCO2/VE), respiratory exchange ratio (RER; calculated as VCO2/VO2), oxygen pulse (VO2/heart rate [HR]) and anaerobic metabolism during exercise (ANA-VO2). At day 30, exercise capacity in the AA group had improved (+11 +/- 8 W, p <0.01; +67.5 +/- 44 seconds, p <0.02). This improvement was associated with both reduced circulatory dysfunction and increased peripheral oxygen availability. Indeed, peak VO2 increased by 1.2 +/- 1.1 mL/kg per min (+12.7% +/- 13%; p<0.02) and VO2/HR improved by 1.5 +/- 1.4 mL O2 per heartbeat (p <0.05). ANA-VO2 was reduced by >50% in patients on AAs (from 20.2 +/- 10 mL/kg at day 0 to 10.9 +/- 5 mL/kg at day 30; p <0.02). These variables did not significantly change for patients who received placebo. In conclusion, the study showed that oral AA supplementation, in conjunction with standard pharmacologic therapy, appears to increase exercise capacity by improving circulatory function, muscle oxygen consumption, and aerobic production of energy in elderly outpatients with CHF.

Mitochondrial biogenesis: is an old dog still teaching us new tricks?

Figure 1. The FASEB Summer Research Conference on the Assembly of the Mitochondrial Respiratory Chain took place between 5 and 10 August 2007, in Tucson, Arizona, USA, and was organized by D. Winge and E. Shoubridge. Participants of this meeting were deeply saddened to learn of the passing of Ron Butow (Dallas, TX, USA), who died on 8 August 2007. Originally scheduled to present at the meeting, Butow cancelled his participation just shortly beforehand. Butow heroically battled cancer for more than 10 years and had remained an extremely active member of the mitochondrial community during this time. Butow was a giant in this field and throughout his career had used elegant genetic and biochemical approaches to study many areas of mitochondrial function, including protein import and, more recently, the organization and inheritance of the mitochondrial genome. His contributions to this field were enormous and he will be deeply missed by all in the mitochondrial and yeast genetic communities. A FASEB summer research conference held in Tuscon, Arizona, USA in August 2007 focused on the broad field of mitochondrial respiratory chain complexes, their assembly and regulation. Although commonly referred to as the ‘powerhouse of the cell’, mitochondria and their functions are not limited to the aerobic production of ATP. Rather, mitochondrial metabolism interfaces with many cellular metabolic processes, such as lipid metabolism—both catabolic and anabolic—FeS cluster metabolism, haem biosynthesis, programmed cell death and metal metabolism, to name but a few. As mitochondrial function and oxidative phosphorylation (OXPHOS) capacity are crucial to all eukaryotic cells, perturbations in the function of this organelle underlie many cellular metabolic disorders, and therefore give rise to various clinical disorders, from cardiac and muscle myopathies to neuromuscular defects (Shoubridge, 2001; Zeviani & Spinazzola, 2003; Fig 1). This FASEB conference saw the gathering of many scientists active in …

Oxygen-controlled Biosurfactant Production in a Bench Scale Bioreactor

Rhamnolipids have been pointed out as promising biosurfactants. The most studied microorganisms for the aerobic production of these molecules are the bacteria of the genus Pseudomonas. The aim of this work was to produce a rhamnolipid-type biosurfactant in a bench-scale bioreactor by one strain of Pseudomonas aeruginosa isolated from oil environments. To study the microorganism growth and production dependency on oxygen, a nondispersive oxygenation device was developed, and a programmable logic controller (PLC) was used to set the dissolved oxygen (DO) concentration. Using the data stored in a computer and the predetermined characteristics of the oxygenation device, it was possible to evaluate the oxygen uptake rate (OUR) and the specific OUR (SOUR) of this microorganism. These rates, obtained for some different DO concentrations, were then compared to the bacterial growth, to the carbon source consumption, and to the rhamnolipid and other virulence factors production. The SOUR presented an initial value of about 60.0 mgO(2)/g(DW) h. Then, when the exponential growth phase begins, there is a rise in this rate. After that, the SOUR reduces to about 20.0 mgO(2)/g(DW) h. The carbon source consumption is linear during the whole process.

Aerobic production of isoamyl acetate by overexpression of the yeast alcohol acetyl-transferases AFT1 and AFT2 in Escherichia coli and using low-cost fermentation ingredients

Isoamyl acetate, produced via fermentation, is a natural flavor chemical with applications in the food industry. Two alcohol acetyltransferases from Saccharomyces cerevisiae (ATF1 and ATF2) can catalyze the esterification of isoamyl alcohol with acetyl coenzyme A. The respective genes were cloned and expressed in an appropriate ack-pta(-) strain of Escherichia coli. The engineered strains produce isoamyl acetate when isoamyl alcohol is added to the culture medium. Aerobic shake flask experiments examined isoamyl acetate production over various growth times, temperatures, and initial optical densities. The strain carrying the pBAD-ATF1 plasmid exhibited a high molar ester yield from glucose (1.13) after 48 h of aerobic growth at 25 degrees C. Low-cost media components, such as fusel oil, sorghum glucose and corn steep liquor, were found to give a high yield of isoamyl acetate. High-cell-density gave an increased isoamyl acetate yield of 0.18 g/g of glucose consumed.

Individualized Aerobic-Power Training in an Underperforming Youth Elite Association Football Player

A 7-week, 10-session individual training program was implemented with a youth elite football (soccer) player who had been underperforming because of poor aerobic fitness. The intervention focused on developing aerobic power and high lactate production and contributed to a 32.3% improvement in a football-specific performance test. The player was able to return to play and exceed expected performance levels during competitive match play.

Aerobic production of activated sludge polyhydroxyalkanoates from nutrient deficient wastewaters

It was found that aerobic strategies combined with multiple nutrient limitations produced greater quantities of polyhydroxyalkanoates (PHAs) than strategies relying on oxygen limitation (either microaerophilic or anaerobic/aerobic). This was true both for a synthetic wastewater composed of acetic and propionic acid, and also for a nutrient deficient industrial wastewater. PHA/substrate yields were shown to be comparable to axenic systems for many operating strategies analyzed, and it was found that PHA composition could be affected by process operational conditions. The molecular weight and melting point of the PHA produced were found to be in a desirable range with respect to material properties, which have not been well studied in the previous literature for mixed cultures (Salehizadeh and Van Loodsrecht, 2004). The effects of process staging, multiple treatment cycles, and inocula source were also addressed.

Microarray gene expression analysis in atrophying rainbow trout muscle: a unique nonmammalian muscle degradation model

Muscle atrophy is a physiological response to diverse physiological and pathological conditions that trigger muscle deterioration through specific cellular mechanisms. Despite different signals, the biochemical changes in atrophying muscle share many common cascades. Muscle deterioration as a physiological response to the energetic demands of fish vitellogenesis represents a unique model for studying the mechanisms of muscle degradation in non-mammalian animals. A salmonid microarray, containing 16,006 cDNAs, was used to study the transcriptome response to atrophy of fast-switch muscles from gravid rainbow trout compared with sterile fish. Eighty-two unique transcripts were upregulated and 120 transcripts were downregulated in atrophying muscles. Transcripts having gene ontology identifiers were grouped according to their functions. Muscle deterioration was associated with elevated expression of genes involved in the catheptic and collagenase proteolytic pathways; the aerobic production, buffering, and utilization of ATP; and growth arrest; whereas atrophying muscle showed downregulation of genes encoding a serine proteinase inhibitor, enzymes of anaerobic respiration, muscle proteins as well as genes required for RNA and protein biosynthesis/processing. Therefore, gene transcription of the trout muscle atrophy changed in a manner similar to mammalian muscle atrophy. These changes result in an arrest of normal cell growth, protein degradation, and decreased glycolytic cellular respiration that is characteristic of the fast-switch muscle. For the first time, other changes/mechanisms unique to fish were discussed including genes associated with muscle atrophy.

Assembly of mitochondrial cytochrome c-oxidase, a complicated and highly regulated cellular process

Cytochrome c-oxidase (COX), the terminal enzyme of the mitochondrial respiratory chain, plays a key role in the regulation of aerobic production of energy. Biogenesis of eukaryotic COX involves the coordinated action of two genomes. Three mitochondrial DNA-encoded subunits form the catalytic core of the enzyme, which contains metal prosthetic groups. Another 10 subunits encoded in the nuclear DNA act as a protective shield surrounding the core. COX biogenesis requires the assistance of >20 additional nuclear-encoded factors acting at all levels of the process. Expression of the mitochondrial-encoded subunits, expression and import of the nuclear-encoded subunits, insertion of the structural subunits into the mitochondrial inner membrane, addition of prosthetic groups, assembly of the holoenzyme, further maturation to form a dimer, and additional assembly into supercomplexes are all tightly regulated processes in a nuclear-mitochondrial-coordinated fashion. Such regulation ensures the building of a highly efficient machine able to catalyze the safe transfer of electrons from cytochrome c to molecular oxygen and ultimately facilitate the aerobic production of ATP. In this review, we will focus on describing and analyzing the present knowledge about the different regulatory checkpoints in COX assembly and the dynamic relationships between the different factors involved in the process. We have used information mostly obtained from the suitable yeast model, but also from bacterial and animal systems, by means of large-scale genetic, molecular biology, and physiological approaches and by integrating information concerning individual elements into a cellular system network.

DISRUPTION OF MITOCHONDRIAL FUNCTION IN INTERPOPULATION HYBRIDS OF TIGRIOPUS CALIFORNICUS

Electron transport system (ETS) function in mitochondria is essential for the aerobic production of energy. Because ETS function requires extensive interactions between mitochondrial and nuclear gene products, coadaptation between mitochondrial and nuclear genomes may evolve within populations. Hybridization between allopatric populations may then expose functional incompatibilities between genomes that have not coevolved. The intertidal copepod Tigriopus californicus has high levels of nucleotide divergence among populations at mitochondrial loci and suffers F2 hybrid breakdown in interpopulation hybrids. We hypothesize that hybridization results in incompatibilities among subunits in ETS enzyme complexes and that these incompatibilities result in diminished mitochondrial function and fitness. To test this hypothesis, we measured fitness, mitochondrial function, and ETS enzyme activity in inbred recombinant hybrid lines of Tigriopus californicus. We found that (1) both fitness and mitochondrial function are reduced in hybrid lines, (2) only those ETS enzymes with both nuclear and mitochondrial subunits show a loss of activity in hybrid lines, and (3) positive relationships exist between ETS enzyme activity and mitochondrial function and between mitochondrial function and fitness. We also present evidence that hybrid lines harboring mitochondrial DNA (mtDNA) and mitochondrial RNA polymerase (mtRPOL) from the same parental source population have higher fitness than those with mtDNA and mtRPOL from different populations, suggesting that mitochondrial gene regulation may play a role in disruption of mitochondrial performance and fitness of hybrids. These results suggest that disruption of coadaptation between nuclear and mitochondrial genes contributes to the phenomenon of hybrid breakdown.