Experimental supply-demand analysis of anaerobic yeast energy metabolism

Experimental supply-demand analysis of yeast fermentative energy metabolism shows that control of the glycolytic flux is shared between supply and demand. In glucose limited chemostat cultures the supply block was modulated in a dilution rate change and demand block via a benzoic acid titration. Under these conditions the supply block had a flux control of 0.90 and the demand block a flux control of 0.10.     

Carbon monoxide-dependent energy metabolism in anaerobic bacteria and archaea

Despite its toxicity for the majority of living matter on our planet, numerous microorganisms, both aerobic and anaerobic, can use carbon monoxide (CO) as a source of carbon and/or energy for growth. The capacity to employ carboxidotrophic energy metabolism anaerobically is found in phylogenetically diverse members of the Bacteria and the Archaea. The oxidation of CO is coupled to numerous respiratory processes, such as desulfurication, hydrogenogenesis, acetogenesis, and methanogenesis. Although as diverse as the organisms capable of it, any CO-dependent energy metabolism known depends on the presence of carbon monoxide dehydrogenase. This review summarizes recent insights into the CO-dependent physiology of anaerobic microorganisms with a focus on methanogenic archaea. Carboxidotrophic growth of Methanosarcina acetivorans, thought to strictly rely on the process of methanogenesis, also involves formation of methylated thiols, formate, and even acetogenesis, and, thus, exemplifies how the beneficial redox properties of CO can be exploited in unexpected ways by anaerobic microorganisms.     

厌氧甲烷氧化微生物物质代谢与能量代谢研究进展

甲烷既是一种温室气体,也是一种潜在的能源物质,其源与汇的平衡对地球化学循环及工程应用均有重要意义。厌氧甲烷氧化(anaerobic oxidation of methane,AOM)过程是深海、湿地和农田等自然生境中重要的甲烷汇,在缓解温室气体排放方面发挥了巨大作用。AOM微生物的中枢代谢机制及其能量转化途径则是介导厌氧甲烷氧化耦合其他物质还原的关键所在。因此,本文从电子受体多样性的视角,主要分析了硫酸盐型,硝酸盐/亚硝酸盐型,金属还原型厌氧甲烷氧化微生物的生理生化过程及环境分布,并对近些年发现的新型厌氧甲烷氧化进行了梳理;重点总结了厌氧甲烷氧化微生物细胞内电子传递路径以及胞外电子传递方式;根据厌氧甲烷氧化微生物环境分布及反应特征,就其生态学意义及在污染治理与能源回收方面的潜在应用价值进行了展望。本综述以期深化对厌氧甲烷氧化过程的微生物学认知,并为其潜在的工程应用方向提供新的思路。     

Effects of different recovery interventions on anaerobic performances following preseason soccer training

In the preseason soccer training, morning and afternoon training sessions often are scheduled daily. The high frequency of training sessions could place heavy strain on biological systems, and it is necessary to apply proper recovery strategies for improving the players' capability to regain an adequate working state for subsequent training units. However, the effect of recovery interventions following soccer training units is debatable, due to a lack of studies performed in field situations. The aim of this study was to examine, during a 21-day preseason soccer training, the most effective recovery intervention (i.e., passive, dry-aerobic exercises, water-aerobic exercises, electrostimulation) on anaerobic performances (i.e., squat jump, countermovement jump, bounce jumping, and 10-m sprint) and subjective ratings (i.e., perceived exertion and muscle pain), with the conditions before the intervention controlled and standardized. Twelve young (age: 18.1 +/- 1.2 years) elite soccer players participated. Data were collected on 4 occasions 2 days apart and at the same time of the day. Activity and dietary intake were replicated on each occasion. After baseline measurements, participants performed a standardized training during which their heart rates and ratings of perceived exertion were recorded. This was followed by a 20-minute recovery intervention. After a 5-hour rest, athletes' ratings of muscle pain were recorded prior to a second test session. There were no significant differences in exercise intensities and baseline anaerobic performances. Significantly (p < 0.01) better performances were observed in the afternoon. Although no main effect of recovery intervention was observed on anaerobic performances, dry-aerobic exercises (0.6 +/- 0.9) and electrostimulation (0.6 +/- 1.2) were more beneficial (p < 0.01) than water-aerobic exercises (2.1 +/- 1.1) and passive rest (2.1 +/- 1.7) for reducing muscle pain, which could affect the player's working ability.     

Comparative study of the energy metabolism of anaerobic alkaliphiles from soda lakes

We investigated the influence of inhibitors of energy metabolism and ionophores on the growth and formation of metabolic products in alkaliphilic anaerobes characterized by various catabolism types. It was shown that blockage of oxidative phosphorylation by the addition of N,N'-dicyclohexylcarbodiimide (DCCD), an inhibitor of F1F0 ATP synthase, resulted in a complete arrest of the growth of the acetogenic bacterium Tindallia magadiensis with arginine as electron acceptor. In the presence of pyruvate, substrate-level phosphorylation occurred. The methylotrophic methanogenic archaebacterium Methanosalus zhilinae did not grow with DCCD and vanadate, an inhibitor of E1E2 ATPase, suggesting the presence of two ATPase types in this species. In the saccharolytic alkaliphiles Halonatronum, Amphibacillus tropicus, and Spirochaeta alkalica (which are characterized by different pH optima), the contribution of the H+ gradient to the energy metabolism and, presumably, to the maintenance of the intracellular pH level decreased with an increase in the degree of alkaliphily. Based on the data of an inhibitor assay using protonophores, monensin, and amiloride, we suggest that all of the bacteria tested depend on H+- and Na+-gradients. The Na+/H+ antiport appears to be a universal mechanism of regulating the intracellular pH level and the interaction between the Na+ and the H+ cycles in bacterial cells cultivated under alkaline conditions.     

Extractive biofilm membrane bioreactor with energy recovery from excess aeration and new membrane fouling control

Hybrid biofilm membrane bioreactor (BF-MBR) system featuring new mechanisms for recovering the excess energy from air bubbling flow in the biofilm reactor and for controlling membrane biofouling was preliminarily investigated in this study. Alternative design of the biofilm reactor was developed to utilize the bubbling flow from the lower aerobic chamber to generate a mechanical mixing in the upper anoxic chamber in the vertical biofilm reactor. Suspended solid (SS) concentration in the system was hydrodynamically controlled to be lower than 70 mg/L. The ultraviolet (UV) inactivation unit was integrated with the membrane filtration tank to limit biological activities for biofoulant productions and to decelerate the unwanted biofilm formation in the permeate tube. Membrane relaxations at various operating conditions were studied for optimum membrane fouling reductions under low SS environment. Combinations of membrane relaxation and the UV inactivation significantly prolonged sustainable operation periods of the membrane filtration in the BF-MBR process.     

The kinetics of anaerobic metabolism following the initiation of high-intensity exercise.

A mathematical study is made of the kinetics of anaerobic metabolism following the initiation of high-intensity exercise. Power and energy relationships are proposed for oxygen-independent glycolysis, phosphocreatine utilisation and the utilisation of endogenous ATP. The power relations consist of two components, one describing the build-up phase, the other the controlled-utilisation phase. The controlled-utilisation phase of oxygen-independent glycolysis and the build-up of aerobic metabolism are shown to be closely inter-related. The theoretical relations display trends consistent with published experimental results. Some property values are derived, but because of the scatter of the experimental results, the values are, in general, to be regarded as tentative.     

Effects of hypothermic hypoxia on anaerobic energy metabolism in isolated anuran livers

Many lower vertebrates (reptilian and amphibian species) are capable of surviving natural episodes of hypoxia and hypothermia. It is by specific metabolic adaptations that anurans are able to tolerate prolonged exposure to harsh environmental stresses. In this study, it was hypothesized that livers from an aquatic frog would possess an inherent metabolic ability to sustain high levels of ATP in an isolated organ system, providing insight into a metabolic system that is well-adapted for low temperature in vitro organ storage. Frogs of the species, R. pipiens were acclimated at 20 °C and at 5 °C. Livers were preserved using a clinical preservation solution after flushing. Livers from 20 °C-acclimated frogs were stored at 20 °C and 5 °C and livers from 5 °C-acclimated frogs were stored at 5 °C. The results indicated that hepatic adenylate status was maintained for 96 h during 5 °C storage, but not longer than 4–10 h during 20 °C storage. In livers from 5 °C-acclimated animals subjected to 5 °C storage, ATP was maintained at 100% throughout the 96-h period. Warm acclimation (20 °C) and 20 °C storage resulted in poorer maintenance of ATP; energy charge values dropped to 0.50 within 2 h and by 24 h, only 24% of control ATP remained. Lactate levels remained less than 25 μ mol/g dry weight in all 5 °C-stored livers; 20 °C-stored livers exhibited greater accumulation of this anaerobic end-product (lactate reached 45–50 μ mol/g by 10 h). The data imply that hepatic adenylate status is largely dependent on exposure to hypothermic hypoxia and although small amounts of ATP were accounted for by anaerobic glycolysis, there must have been either a substantial reduction in cellular energy-utilization or an efficient use of low oxygen tensions. Accepted: 24 August 1998     

Fluid replacement following dehydration reduces oxidative stress during recovery

To investigate the effects of hydration status on oxidative DNA damage and exercise performance, 10 subjects ran on a treadmill until exhaustion at 80% VO_2max during four different trials [control (C), 3% dehydration (D), 3% dehydration + water (W) or 3% dehydration + sports drink (S)]. Dehydration significantly decreased exercise time to exhaustion (D < C and S). Plasma MDA levels were significantly higher at pre-exercise in D than C. Plasma TAS was significantly lower at pre-exercise in C and S than in D, and was significantly lower in S than D at 60 min of recovery. Dehydration significantly increased oxidative DNA damage during exercise, but fluid replacement with water or sports drink alleviated it equally. These results suggest that (1) dehydration impairs exercise performance and increases DNA damage during exercise to exhaustion; and (2) fluid replacement prolongs exercise endurance and attenuates DNA damage.     

The effect of mechanical stress on cartilage energy metabolism

Cartilage is routinely subjected to varying mechanical stresses which are known to affect matrix turnover by a variety of pathways. Here we show that mechanical loads which suppress sulphate incorporation or protein synthesis by articular chondrocytes, also inhibit rates of oxygen uptake and of lactate production. Although the mechanisms have not been definitively identified, it has been shown that high hydrostatic pressures reduce the activity of the glucose transporter GLUT. Furthermore, fluid expression consequent on static loading changes intracellular pH and ionic strength; intracellular changes which would reduce the activity of glycolytic enzymes. Both pathways would thus lead to a fall in rates of glycolysis and a reduction in intracellular ATP, and - since ATP concentrations directly affect sulphation of proteoglycans - a rapid fall in sulphate incorporation. Our results suggest that load-induced changes in matrix synthesis in cartilage can occur by means other than changes in gene expression.     

Studies on the energy metabolism during anaerobic fermentation of glucose by baker's yeast

Summary As a result of the intimate association of ADP phosphorylation with alcoholic fermentation, resulting in the synthesis of 2 mole ATP per mole glucose fermented, it may be calculated that a minimum of 672 µcal heat development may be expected for every mm³ CO₂ developed during alcoholic fermentation. When all ATP produced would be fully de-phosphorylated to ADP + Pi (e.g. by ATP-ase activity) a maximum heat development of 1200 µcal per mm³ CO₂ could be expected.Using the LKB-Flow-Microcalorimeter for measurement of heat development and at the same time the Warburg technique for measuring CO₂ development during anaerobic glucose fermentation of a baker's yeast suspension, the heat development per mm³ CO₂ produced was calculated over a fermentation period of 90 min.Maintenance of strict anaerobic conditions in the Flow-Microcalorimeter vessel was complicated by diffusion of traces of oxygenvia the Teflon transport lines, resulting in excessive heat development values, not representative for the alcoholic fermentation. This problem could be circumvented by removal of traces of oxygen by means of addition of the enzyme glucose-oxidase.Poisoning the respiratory enzyme system of the yeast by addition of KCN or azide, or using respiratory-deficient mutants of the yeast also resulted in heat development values, inherent with alcoholic fermentation.The values obtained were very close to the minimum of 672 µcal per mm³ CO₂, at least during the initial phases of fermentation, indicating that ADP regeneration from ATP, essential for maintaining the high fermentation rate, is not primarily the result of ATP-ase activity, but must be due to participation of ATP in energy-requiring synthetic reactions.     

The influence of maximal aerobic power on recovery of skeletal muscle following anaerobic exercise

Phosphorus magnetic resonance spectroscopy (³¹P-MRS) was used to investigate the influence of maximal aerobic power (˙VO _2max) on the recovery of human calf muscle from high-intensity exercise. The (˙VOO_2max) of 21 males was measured during treadmill exercise and subjects were assigned to either a low-aerobic-power (LAP) group (n = 10) or a high-aerobic-power (HAP) group (n = 11). Mean (SE) ˙VO _2max of the groups were 46.6 (1.1) and 64.4 (1.4) ml · kg⁻¹ · min⁻¹, respectively. A calf ergometry work capacity test was used to assign the same relative exercise intensity to each subject for the MRS protocol. At least 48 h later, subjects performed the rest (4 min), exercise (2 min) and recovery (10 min) protocol in a 1.5 T MRS scanner. The relative concentration of phosphocreatine (PCr) was measured throughout the protocol and intracellular pH (pH_i) was determined from the chemical shift between inorganic phospate (P_i) and PCr. End-exercise PCr levels were 27 (3.4) and 25 (3.5)% of resting levels for LAP and HAP respectively. Mean resting pH_i was 7.07 for both groups, and following exercise it fell to 6.45 (0.04) for HAP and 6.38 (0.04) for LAP. Analysis of data using non-linear regression models showed no differences in the rate of either PCr or pH_i recovery. The results suggest that ˙VO_2max is a poor predictor of metabolic recovery rate from high-intensity exercise. Differences in recovery rate observed between individuals with similar ˙VO_2max imply that other factors influence recovery. Accepted: 17 December 1996     

Effects of low temperature storage upon subsequent energy metabolism of rabbit embryos

The oxidation of pyruvate by two-cell rabbit embryos occurred at a very low rate at 10 °C, but increased 10–15 times upon rewarming to 37 °C. Pyruvate utilization of these embryos increased with continued development parallel to embryos not cooled to 10 °C, but incubated directly at 37 °C. Storage at 10 °C did not affect the amount of glucose subsequently oxidized at 37 °C, nor the shift from the pentose shunt to glycolysis and the Krebs cycle that normally occurs between the morula and blastocyst stage. Thus, the temporary inhibition of carbohydrate metabolism induced through temperature reduction did not impair the subsequent ability of the embryos to metabolize pyruvate and glucose normally.     

Partial recovery of the endothelial glycocalyx upon rosuvastatin therapy in patients with heterozygous familial hypercholesterolemia

The endothelial glycocalyx has been shown to serve as a protective barrier between the flowing blood and the vessel wall in experimental models. The aim of this study was to evaluate whether hypercholesterolemia is associated with glycocalyx perturbation in humans, and if so, whether statin treatment can restore this. We measured systemic glycocalyx volume (V(G)) in 13 patients with heterozygous familial hypercholesterolemia (FH) after cessation of lipid-lowering therapy for a minimum of 4 weeks and 8 weeks after initiating rosuvastatin therapy. Normocholesterolemic subjects were used as controls. V(G) was estimated by subtracting the intravascular distribution volume of a glycocalyx permeable tracer (dextran 40) from that of a glycocalyx impermeable tracer (labeled erythrocytes). V(G) in untreated FH patients [LDL 225 +/- 57 mg/dl (mean +/- SD)] was significantly reduced compared with controls (LDL 93 +/- 24 mg/dl) (V(G) 0.8 +/- 0.3 vs. 1.7 +/- 0.6, respectively, P < 0.001). After normalization of LDL levels (95 +/- 33 mg/dl) upon 8 weeks of statin treatment, V(G) recovered only partially (V(G) 1.1 +/- 0.4 L, P = 0.04). The endothelial glycocalyx is profoundly reduced in FH patients, which may contribute to increased atherogenic vulnerability. This perturbation is partially restored upon short-term statin therapy.