The protonmotive force and phosphorylation potential developed by whole cells of the methylotrophic bacteriumMethylophilus methylotrophus

The and the Gp have been measured in whole cells ofMethylophilus methylotrophus during the oxidation of various respiratory chain substrates. The magnitude of the depended on the external pH and the composition of the assay medium, and varied from-109 to-165 mV. The relative contributions of the and the pH to the were found to vary with the external pH such that the internal pH remained constant; depending on the composition of the assay medium, this value was between 6.6 and 7.0. A Gp of approximately-46 kJ/mol was generated during the oxidation of methanol, and either the or pH alone was fully competent to drive ATP synthesis. Respiration and ATP synthesis were found to be poised far from equilibrium under the conditions of these experiments, and the value of the Gp was thus controlled kinetically. Comparison of the with the Gp yielded an H⁺/ATP quotient >2.6 g-ion H⁺/mol ATP.Abbreviations TMPD N,N,N,N-tetramethyl-p-phenylenediamine - FCCP carbonylcyanidep-trifluoromethoxyphenylhydrazone - DMO 5,5-dimethyloxazolidine-2,4-dione - TPMP⁺ triphenylmethylphosphonium (iodide salt); Tween 20, polyoxyethylenesorbitan monolaurate - TPP⁺ tetraphenylphosphonium (bromide salt) - bulk phase transmembrane electrochemical potential difference of protons ( ) - pH bulk phase transmembrane pH difference (pHin-pHout) - bulk phase transmembrane electrical potential difference (in-out) - p true protonmotive force (incorporating both bulk phase and localised protons; )     

Molar growth yields,respiration and cytochrome profiles of Beneckea natriegens when grown under carbon limitation in a chemostat

The effect of growth rate on the physiology of Beneckea natriegens was studied in chemostat culture. The molar growth yields (Y) from glucose and oxygen, the specific rates of oxygen (q _{O 2}) and glucose (q _glc) consumption and the specific rate of CO₂ production (q _{CO 2}) were linearly dependent on the growth rate over the dilution rate 0.17 h^-1 to 0.60 h^-1. Further increase in the dilution rate resulted in a decrease in growth yield and respiration rate and these changes were coincident with increases in the specific rate of glucose utilisation and of acetate production. The affinity of Beneckea natriegens for glucose was similar when measured either directly in chemostat culture or in a closed oxygen electrode system using harvested bacteria. The total content of cytochromes decreased with increasing growth rate. However, the quantity of CO-binding cytochromes remained independent of growth rate and correlated with the potential respiration rate.     

Simultaneous uptake and utilisation of glucose and xylose by Clostridium thermohydrosulfuricum

Abstract Growth studies of Clostridium thermohydrosulfuricum Rt8.B1 demonstrated that glucose and xylose were used simultaneously when supplied together at nonlimiting concentrations in pH-controlled batch culture. Under conditions of hyperbolic growth, both catabolite repression and inducer exclusion were absent. Glucose did not repress xylose metabolism (i.e. xylose permease and xylose isomerase genes were expressed in the presence of glucose and were not subject to catabolite inhibition when glucose was added to cultures growing on high concentrations of xylose). The kinetics of glucose and xylose utilisation indicated that separate systems were present for the uptake of these substrates when supplied together. Glucose utilisation was biphasic, indicating high- and low-affinity systems for glucose uptake. Xylose utilisation was directly proportional to the xylose concentration, suggesting a facilitated diffusion mechanism was operative for uptake.     

Energy conservation in whole cells of the methylotrophic bacterium Methylophilus methylotrophus

Respiratory chain phosphorylation has been investigated in the methylotrophic bacterium Methylophilus methylotrophus following the addition of oxidisable substrates to aerobic, whole cell suspensions. Initial-rate experiments showed that ATP synthesis occurred at the overall expense of AMP and inorganic phosphate via the sequential action of the ATP phosphohydrolase and adenylate kinase; some of the nascent ATP was rapidly used to synthesis nonadenine nucleoside triphosphates. After being corrected for ATP turnover, Pi/O quotients of 0.46 to 0.54, 0.77 and 1.37 nmol/ng-atom O were obtained for the oxidation of methanol dehydrogenase-linked substrates (methanol, ethanol and acetaldehyde), duroquinol and formate (NAD⁺-linked) respectively. These values were proportional to the H⁺/O and/or K⁺/O quotients exhibited by these substrates, and yielded an average H⁺/ATP (H⁺/Pi) quotient of 4.2 ng-ion H⁺/nmol. Steady-state experiments showed that the extent of cellular energisation varied with the respiration rate but was always in the order methanol > duroquinol > acetaldehyde, thus indicating that under these longer-term conditions methanol was completely oxidised to yield PQQH₂ and 2NAD(P)H. These results are discussed in terms of the various reactions which lead to the generation or utilisation of the protonmotive force in this organism.Abbreviations FCCP carbonylcyanide p-trifluoromethyxyphenyl-hydrazone - bulk phase, transmembrane electrochemical potential difference of protons ( ) - pH bulk phase, transmembrane pH difference (pH_in–pH_out) - bulk phase, transmembrane electrical potential difference (_in - _out) - [P] concentration of anhydride phosphate bonds in adenine nucleotides (2[ATP]+[ADP]) - FPLC fast protein liquid chromatography - PQQ pyrroloquinoline quinone - Gp phosphorylation potential     

Comment on E. Broda's recent publications

Summary E. Broda's recent argument against our concept that nitrate respiration antedated oxygen respiration is criticized.     

Is maximization of molar yield in metabolic networks favoured by evolution?

Stoichiometric analysis of metabolic networks allows the calculation of possible metabolic flux distributions in the absence of kinetic data. In order to predict which of the possible fluxes are present under certain conditions, additional constraints and optimization principles can be applied. One approach of calculating unknown fluxes (frequently called flux balance analysis) is based on the optimality principle of maximizing the molar yield of biotransformations. Here, the relevance and applicability of that approach are examined, and it is compared with the principle of maximizing pathway flux. We discuss diverse experimental evidence showing that, often, those biochemical pathways are operative that allow fast but low-yield synthesis of important products, such as fermentation in Saccharomyces cerevisiae and several other yeast species. Together with arguments based on evolutionary game theory, this leads us to the conclusion that maximization of molar yield is by no means a universal principle.     

Regulation of enzyme synthesis in Hyphomicrobium X: Growth on mixtures of methylamine and ethanol in continuous cultures

Abstract Hyphomicrobium X was grown on a range of mixtures of methylamine and ethanol at various dilution rates in continuous cultures. It was shown that both substrates were utilized simultaneously at all dilution rates below μ_max ethanol, the biomass being the sum of the biomass obtained during growth on the single substrates alone. No evidence was observed for a redistribution of the carbon-flow nor the ability to utilize the 'poorer' growth substrate-ethanol, at dilution rates greater than μ_max ethanol. Work is presented which suggests that the assimilatory pathway enzymes for either the C₁- or the C₂-compound are regulated coordinately, but separately from the dissimilatory pathway enzymes associated with that compound.     

增温对宁夏引黄灌区春小麦生产的影响

近40a来,宁夏引黄灌区年平均增温幅度高于全国平均值,对春小麦生产已经产生了重大影响。基于哥本哈根联合国气候变化大会控制未来50 a全球升温幅度的范围值,设计增温幅度为0.5-2.5 ℃,采用红外线辐射器大田增温模拟实验,研究增温对宁夏引黄灌区春小麦生长发育和产量的影响。结果表明:增温0.5-2.5 ℃,宁夏引黄灌区春小麦全生育期(播种-收获)缩短1-22 d,减产0.5%-18.5%;增温2.0-2.5 ℃,春小麦全生育期缩短18-22 d,减产16.5%-18.5%。增温引起春小麦三叶期和孕穗期光合速率下降,穗粒数减少,千粒重下降,最终导致减产。     

皱纹盘鲍的个体能量收支

对皱纹盘鲍的呼吸、摄食、生长及能量收支等实验研究表明, 鲍的耗氧率与壳长、体重、温度及昼夜变化有关, 耗氧率与壳长、体重均呈幂函数关系, 一天中16~4时(夜间)耗氧率高于4~16时(白天)且在18~20时达峰值.同温度下鲍日摄食率与体重呈幂指数关系, 日(相对)摄食率随温度升高而增加, 而日相对摄食率、日相对生长率均随壳长、体重增加呈下降趋势.鲍在14~20℃内对海带的总转化效率为53%.鲍软体部、海带及鲍粪便干品的比能值分别为19.2、8.57和7.23kJ·g^-1.14~20℃皱纹盘鲍摄入能量的34.6~48.6%为粪能, 22.0~38.2%的能量用于自身代谢, 5.6~28.2%用于贝体软体部的生长.     

微种植体支抗压低上颌磨牙的研究进展

在口腔科诊疗工作中经常会见到因下颌磨牙长期缺失导致对颌磨牙伸长的情况,这将直接导致对颌磨牙修复空间不足。这种病例需要通过压低伸长的上颌磨牙来获取足够的空间,以利于下颌磨牙的修复。另外,高角和开牙合病例要想获得良好的治疗效果也需要通过压低磨牙来进行垂直向控制,因此压低上颌磨牙成为临床工作的重要内容。压低上颌磨牙的方法除了牙合垫、高位牵引头帽、手术等之外,还有微种植体支抗,它又称为\"绝对支抗\"由钛或合金制成,分为自攻型和助攻型。由于创伤小,植入部位灵活,手术简单,压低磨牙效果显著,所以自临床应用以来得到大力推广。因此本文就微种植体支抗压低上颌磨牙的临床应用、压低效果评价、副作用及防治措施和生物力学研究等方面作一综述。     

Extracellular amylase activities ofRhizomucor pusillus andHumicola lanuginosa at initial stages of growth

Among thermophilic fungi,Rhizomucor Pusillus andHumicola lanuginosa have been reported to be among the most prolific producers of amylase, an apparently heat stable enzyme vital to the incorporation of carbon from macromolecular sources such as starch. Yet the highest levels of extracellular amylase in starch-yeast cultures of these fungi were measured after most of the growth had occurred; pre-growth levels appeared to be very small. Since these low levels are the significant ones for growth, a procedure was devised to measure them: 1.162×10^–2 units (mg maltose/ml/min) were measured after two days of growth ofR. pusillus and 6.230×10^–3 units measured after four days of the slower-growingH. lanuginosa. Re-assays of these after dialysis to remove most of the reducing sugars gave 1.689 × 10^–2 units and 1.234 × 10^–2 units, respectively, with all correlation coefficients 0.96 or better.     

Meclizine Inhibits Mitochondrial Respiration through Direct Targeting of Cytosolic Phosphoethanolamine Metabolism

We recently identified meclizine, an over-the-counter drug, as an inhibitor of mitochondrial respiration. Curiously, meclizine blunted respiration in intact cells but not in isolated mitochondria, suggesting an unorthodox mechanism. Using a metabolic profiling approach, we now show that treatment with meclizine leads to a sharp elevation of cellular phosphoethanolamine, an intermediate in the ethanolamine branch of the Kennedy pathway of phosphatidylethanolamine biosynthesis. Metabolic labeling and in vitro enzyme assays confirmed direct inhibition of the cytosolic enzyme CTP:phosphoethanolamine cytidylyltransferase (PCYT2). Inhibition of PCYT2 by meclizine led to rapid accumulation of its substrate, phosphoethanolamine, which is itself an inhibitor of mitochondrial respiration. Our work identifies the first pharmacologic inhibitor of the Kennedy pathway, demonstrates that its biosynthetic intermediate is an endogenous inhibitor of respiration, and provides key mechanistic insights that may facilitate repurposing meclizine for disorders of energy metabolism.     

皱纹盘鲍的个体能量收支

对皱纹盘鲍的呼吸、摄食、生长及能量收支等实验研究表明, 鲍的耗氧率与壳长、体重、温度及昼夜变化有关, 耗氧率与壳长、体重均呈幂函数关系, 一天中16~4时(夜间)耗氧率高于4~16时(白天)且在18~20时达峰值.同温度下鲍日摄食率与体重呈幂指数关系, 日(相对)摄食率随温度升高而增加, 而日相对摄食率、日相对生长率均随壳长、体重增加呈下降趋势.鲍在14~20℃内对海带的总转化效率为53%.鲍软体部、海带及鲍粪便干品的比能值分别为19.2、8.57和7.23kJ·g^-1.14~20℃皱纹盘鲍摄入能量的34.6~48.6%为粪能, 22.0~38.2%的能量用于自身代谢, 5.6~28.2%用于贝体软体部的生长.